Substituted oxazolone compounds

Abstract

Equation (I): Compounds represented by TIFF2026521393000060.tif42140, or their stereoisomers, tautomers, or salts are disclosed. Also disclosed are methods for using such compounds to reduce the level of IKZF1-4 protein, and pharmaceutical compositions comprising such compounds. These compounds are useful in treating viral infections and proliferation disorders, such as cancer.

Description

[Technical Field]

[0001] (cross reference) This application claims the interests of U.S. Provisional Patent Application No. 63 / 505,181, filed on 31 May 2023, which is incorporated herein by reference in its entirety.

[0002] (Field of Invention) This application generally relates to substituted oxazolone compounds that reduce the levels of Ikaros, Helios, Aiolos, and Eos proteins. This specification provides substituted oxazolone compounds, compositions containing the compounds, and methods of use. The present invention further relates to pharmaceutical compositions containing the compounds according to the present invention that are useful for treating proliferative disorders such as cancer and viral infections. [Background technology]

[0003] Transcription factors (TFs) of the IKZF family (IKZF) play crucial roles in lymphocyte development and function (Heizmann et al., 2018, Curr Opin Immunol. 51:14-23). ​​In mammals, the following five members of this family—IKZF1 (encoded), Helios (IKZF2), Aeolus (IKZF3), Eos (IKZF4), and Pegasus (IKZF5)—are expressed in immune cells. The amino acid sequences of these proteins show high homology, with IKZF and Aeolus, and Helios and Eos being the most homologous pairs, while Pegasus is the least related IKZF member. These TFs serve both overlapping and unique functions in lymphocytes (Read et al., 2020, Immunological Reviews, 300:1). Decreased levels of IKZF TF protein may boost the antitumor T cell response.

[0004] IKZF1 encodes Ikaros, which is broadly and abundantly expressed in human and mouse B cell, NK cell, and T lymphocyte populations, and moderately expressed in other immune cell types, including myeloid cells. In T cells, loss of Ikaros protein or expression of the dominant-negative protein releases the repression of a locus associated with differentiation into effector T cell state, resulting in increased expression of effector cytokines, including IFN-γ, TNF-α, and GM-CSF (Lyon de Ana et al., 2019, Journal of Immunology 202:1112-1123; Heller et al., 2014, Journal of Immunology, 193:3934-3946; Wang et al., 2020, Cell Transplantation, 29).

[0005] IKZF2 encodes helios, exhibiting a more restricted expression profile limited to human and mouse restriction T (Treg) cells, some CD8+ T cells and MAIT cells, and NK cells (Akimova et al., 2011, PLoS One, 6:e24226; Dias et al., 2017, Proceedings of the National Academy of Sciences USA, 114:E5434-E5443; Thornton and Shevach, 2019, Immunology, 158:161-170).

[0006] IKZF3 encodes Aiolos, which is widely and abundantly expressed in human and mouse B lymphocytes and is widely expressed at low levels in T and NK cells. In T cells, the repression targets of the Aiolos gene show a large overlap with those of the Ikaros target genes (Powell et al., 2019, Frontiers in Immunology, 10:1299). Compared with Ikaros, Aiolos may have a stronger impact on follicular helper T cell and type 17 T helper responses (Quintana et al., 2012, Nature Immunology, 13:770-777; Read et al., 2017, Journal of Immunology, 7:2377-2387), where it is associated with tissue immune responses and, in some cases, anti-tumor immunity.

[0007] IKZF4 encodes Eos, which is abundantly expressed in Treg cells and is also widely expressed at low levels among B, NK, and T lymphocytes. In Treg cells, loss of Eos expression in FoxP3+ Treg cells drives the improvement of anti-tumor responses in preclinical syngeneic tumor models (Gokhale et al., 2019, Journal of Autoimmunity, 105:102300). In addition, Eos expression levels can increase after activation of T cells in previous CD4+ and CD8+ T cells, which can limit effector T cell responses (Rieder et al., 2015, Journal of Immunology, 195:553-563).

[0008] A common function shared among IKZF TFs is the repression of cellular gene expression at specific loci. Each IKZF TF can bind to genomic loci as either a homodimer or heterodimer, such as IKZF:IKZF:IKZF or IKZF:Helios. These dimeric TFs together bind to DNA and interact with nucleosomes and complexes that regulate histone acetylation, which in turn leads to the regulation of gene expression. As for the mechanism of action, IKZF:IKZF:IKZF, Helios, and Aeolus have been shown to repress gene expression by interacting with nucleosome remodeling and deacetylase (NuRD) and Sin3 histone deacetylase (HDAC) complexes, respectively (Zhang et al., 2011, Nature Immunology, 13:86-94; Georgopoulos et al., 2017, Genes and Development, 31:439-450). Similarly, Ikaros, Helios, and Aeolus all associate with centromere heterochromatin and may contribute to the expression of genes located at centromere loci (Brown et al., 1997, Cell, 91:845-854; Thompson et al., 2007, Immunity, 26:335-344). Eos does not cooperate with Aeolus but cooperates with Ikaros and interacts with the C-terminal binding protein 1 (CtBP1) of transcriptional repressors in lymphocytes (Koipally et al., 2002, Journal of Biological Chemistry, 277:27697-27705; Pan et al., 2009, Science, 325:1142-1146). Together, the overlapping functions of IKZF TFs may partially compensate for the loss or degradation of one or more TFs. Therefore, in cells expressing multiple IKZF members, broadly therapeutically degrading TFs of this family is thought to result in greater phenotypic changes compared to selective degradation of one or two IKZF TFs.

[0009] In T cells and Treg cells, the shared role of IKZF TFs in regulating gene loci crucial for anti-tumor immune responses is exemplified by the regulation of the gene encoding interleukin-2 (IL-2). Ikaros can directly bind to the IL-2 gene locus in CD4+ T cells and recruit the HDAC complex; loss of Ikaros results in increased IL-2 production by CD4+ and CD8+ T cells (Bandyopadhyay et al., 2007, Blood, 109:2671-2672; Thomas et al., 2007, Journal of Immunology, 179:7305-7315; O'Brien et al., 2014, Journal of Immunology, 192:5118-5129). Helios directly binds to the IL-2 locus in Treg cells, recruiting the HDAC complex and enhancing IL-2 gene silencing (Blaine et al., 2013, Journal of Immunology, 190:1008-1016). Eos also suppresses IL-2 expression in Treg cells and may act through a mechanism involving interaction with TF FoxP3 (Pan et al., 2009, Science, 325:1142-1146; Sharma et al., 2013, Immunity, 38:998-1012). The role of Aeolus' direct binding on IL-2 loss is unclear, but it has been reported that siRNA knockdown of Aeolus in human Treg cells increases IL-2 production (Gandhi et al., 2010, Nature Immunology, 11:846-853). In summary, IKZF TFs are abundantly expressed in multiple lymphocyte subtypes, particularly all four of these IKZF TFs, and act to regulate IL-2 production by Treg cells, which is normally negligible for IL-2 production.

[0010] Treg cells, marked by the expression of the transcription factor FoxP3, are a subset of immunosuppressive lymphocytes that use multiple mechanisms of action to maintain immune homeostasis (Sakaguchi et al., 2020, Annual Review of Immunology, 38:541-566; Whibley et al., 2019, Nature Immunology, 20:386-396). Patients with deleterious mutations in the gene encoding FoxP3 lack functional Treg cells and exhibit immune dysregulation, polyendocrinopathy, enteropathy, X-linked (IPEX) syndrome, and multi-organ autoimmunity. In the tumor microenvironment (TME), the activity of Treg cells is selected to promote and maintain an immunosuppressive state (Plitas and Rudensky, 2020, Annual Review of Cancer Biology, 4:459-477). By secreting inhibitory molecules, blocking cytokines (e.g., IL-2), and directly inhibiting the activation of T cells and antigen-presenting cells, Treg cells can promote resistance to TME-mediated immunotherapy by regulating multiple axes in the cancer immune cycle (Chen and Mellman, 2013, Immunity, 39:1-10). In preclinical models, the removal of Treg cells results in the regression of established tumors with high malignancy (Bos et al., 2013, Journal of Experimental Medicine, 210:2435-2466).

[0011] When activated by specific antigens, Treg cells can suppress responder T cells in an antigen-nonspecific and passive manner in vitro (Takahashi et al., 1998, Int Immunol. 10:1969-80; Thornton et al., 1998, J Exp. Med. 188:287-96). FoxP3+CD25+CD4+Treg cells have the ability to suppress a wide range of antitumor immune responses involving CD4+ helper T cells, CD8+ T cells, natural killer cells, and natural killer T cells (Tanaka et al., 2017, Cell Research 27:109-118). In preclinical models, depletion of CD25+CD4+Treg cells within tumors induced established tumor regression along with changes in the cytokine environment at the tumor site (Yu et al., 2005, J Exp Med. 201:779-91). In addition, transplantation of CD4+ T cells depleted of Treg cells significantly enhanced the antitumor immune response compared to transplantation of CD4+ T cells with sufficient Treg cells (Antony et al., 2005, J Immunol 174:2591-601). Tumor-infiltrating Treg cells activated by either tumor-induced autoantigens or tumor-associated antigens can similarly suppress the specific antitumor immune response.

[0012] Clinically, increased Treg cell frequency in TMEs correlates with poor outcomes in multiple solid tumor manifestations (Shang et al., 2015, Scientific Reports, 5:15179). Furthermore, the correlation between PD-L1+ Treg cell frequency and response to anti-PD-1 therapy in non-small cell lung cancer (NSCLC) patients (Wu et al., 2018, Journal of Thoracic Oncology, 13:521-532) highlights the therapeutic potential of targeting Treg cells in TMEs. Modulating the activity of key factors that control Treg cell differentiation and / or functional suppressor status may reveal potential therapeutic strategies for treating certain diseases, including cancer and viral infections.

[0013] Furthermore, the removal of FoxP3+ Treg cells has also been reported to enhance vaccine-induced antitumor T cell responses (Nishikawa et al., 2010, Int. J. Cancer 127:759-767), suggesting that a decrease in helios levels may be beneficial in enhancing the effectiveness of cancer vaccines. In addition to antitumor immunotherapy, during viral infections, Treg cells can limit the immunopathology resulting from excessive inflammation, further inhibiting more effective antiviral T cell responses and potentially promoting viral persistence (Schmitz et al., 2013, PLOS Pathogens 9:e1003362). Chronic infection (not acute infection) with lymphocytic vascular meningitis virus in mice results in a significant proliferation of FoxP3+ Treg cells, suggesting a possible mechanism by which certain infectious agents may evade the host immune response by activating and proliferating Treg cells (Punkosdy et al., 2011, PNAS 108:3677-3682). In situations associated with chronic viral infections, therapeutic effects may be achieved by reducing helios levels in activated Treg cells.

[0014] Solutions targeting neoplastic Treg cells include antibody-mediated depletion and / or functional modification (Tanaka and Sakaguchi, 2019, European Journal of Immunology, 49:1140-1146), as well as small molecule-mediated "reprogramming" of the immunosuppressive phenotype of Treg cells by altering gene expression in these cells (Kim et al., 2015, Science, 350:334-339; Sebastian et al., 2016, Journal of Immunology, 196:144-155). Mice with Treg cells genetically engineered to lack helios do not develop the IPEX-like immunopathology characteristic of FoxP3 deficiency or complete removal of Treg cells, but instead possess Treg cells that exhibit a more T-effector-like transcriptional program (Fu et al., 2012, Nature Immunology, 13:972-980; Yates et al., 2018, Proceedings of the National Academy of Sciences USA, 115:2162-2167). Importantly, helios controls the activity of Treg cells that are crucial in TME, as evidenced by the improved control of B16F10 and MC38 tumors in mice with helios-deficient Treg cells (Nakagawa et al., 2016, Proceedings of the National Academy of Sciences USA, 113:6248-6253). Therefore, therapeutic modulation of helios may reprogram tumor Treg cells toward a more effector-like phenotype, potentially driving anti-tumor immunity. Notably, mice lacking Eos expression in FoxP3 Treg cells more effectively controlled allograft tumors compared to controls, suggesting that Eos also drives immunosuppressive Treg cell activity in TMEs in preclinical tumor models (Gokhale et al., 2019, Journal of Autoimmunity, 105:102300).Similarly, individuals with germline loss-of-function IKZF2 mutations do not exhibit IPEX-like symptoms (including diabetes, dermatitis, hepatitis, and generalized lymphadenopathy), but instead express an immunophenotype associated with enhanced T cell activation and increased inflammatory cytokine production (Hetemaki et al., 2021, Science Immunology, 6:eabe3454; Shahin et al., 2021, Science Immunology, 6:eabe3981). These data suggest that reduced levels of helios and eos proteins in Treg cells lead to a less severe suppression of the antitumor T cell response in patients with solid tumors.

[0015] Small molecules that degrade Ikaros and Aeolus in Treg cells can also reduce the inhibitory function of these cells in vitro (Galustian et al., 2008, Cancer Immunology, Immunotherapy, 58:1033-1045). In genetically engineered mouse models, lenalidomide, a degrader of Ikaros and Aeolus, can slightly enhance the antitumor immune response against highly immunogenic allografted tumors (Geng et al., 2022, Cell Chemical Biology, 29:1260-1272). Degraders targeting Ikaros and Aeolus have also been tested clinically in patients with solid tumors, sometimes resulting in a stable, modest disease response. These studies include those on abadomide (CC-122) (Rasco et al., 2019, Clin Cancer Research, 25:90-98), lenalidomide (Semeraro et al., 2013, OncoImmunology, 2:11), and pomalidomide (Cooney et al., 2012, Cancer Chemotherapy and Pharmacology, 70, 755) in advanced malignancies. In addition, lenalidomide has been shown to enhance T and NK cell function in preclinical and clinical trials (Hideshima et al., Leukemia, 2021; D'Souza et al., Frontiers in Immunology, 2021).

[0016] In summary, the IKZF TFs Ikaros, Helios, Aeolus, and Eos are abundantly expressed in Treg cells. Combining and reducing the levels of individual proteins of these four TFs in Treg cells would more effectively reverse immunosuppressive programs, including repression of IL-2 transcription and other T cell effector genes, compared to solutions that selectively target a single IKZF TF or a pair of TFs, i.e., Ikaros and Aeolus, or Helios and Eos. Beyond Treg cells, pan-IKZF1-4 degraders are expected to enhance conventional CD4+ and CD8+ T cell effector function, boost NK cell activity, and drive a robust antitumor response in patients.

[0017] There remains a demand for therapies that can reduce the levels of the four IKZF1-4 proteins: Ikaros, Helios, Aeolus, and Eos. The present invention satisfies the above requirements by providing compounds that are useful in reducing the levels of four types of IKZF1-4 proteins: Ikaros, Helios, Aeolus, and Eos. [Overview of the project]

[0018] The present invention provides substituted oxazolone compounds of formula (I) (including their stereoisomers, tautomers, salts, and prodrugs) that are useful for reducing the levels of four proteins: Ikaros, Helios, Aeolus, and Eos. The present invention also provides a pharmaceutical composition comprising a compound of formula (I), its stereoisomers, tautomers, pharmaceutically acceptable salts, or prodrugs; and a pharmaceutically acceptable carrier.

[0019] The present invention also provides a method for treating a disease or disorder by reducing the levels of four IKZF1-4 proteins, Ikaros, Helios, Aeolus, and Eos, comprising administering to a patient a compound of formula (I), its stereoisomers, tautomers, pharmaceutically acceptable salts, or prodrugs. The present invention also provides methods and intermediates for producing compounds of formula (I), stereoisomers, tautomers, or salts thereof.

[0020] The present invention also provides the use of a compound of formula (I), or its stereoisomers, tautomers, pharmaceutically acceptable salts, or prodrugs, for producing pharmaceuticals that reduce the levels of Icarus, Helios, Aeolus, and Eos proteins, for treating certain diseases, including cancer and viral infections.

[0021] Compounds of formula (I) and compositions containing compounds of formula (I) may be used to treat, prevent or cure various proliferative disorders such as cancer. Pharmaceutical compositions containing the compounds of the present invention are useful for treating, preventing or delaying the progression of diseases or disorders in various therapeutic areas such as cancer.

[0022] Compounds of formula (I) and compositions containing compounds of formula (I) may be used to treat, prevent or cure viral infections. Pharmaceutical compositions containing the compound are useful for treating, preventing, or slowing the progression of diseases or disorders such as viral infections. These and other features of the present invention will be described in expanded forms as the disclosure progresses. [Modes for carrying out the invention]

[0023] The inventors have discovered a substituted oxazolone compound that reduces the levels of Ikaros, Helios, Aeolus, and Eos proteins. This substituted oxazolone compound is thought to promote the interaction between the Ikaros, Helios, Aeolus, and Eos proteins and the corresponding E3 ubiquitin ligase complex (Cullin4-Cereblon, CUL4-CRBN), while simultaneously degrading the Ikaros, Helios, Aeolus, and Eos proteins. The compound reduces the levels of Ikaros, Helios, Aeolus, and Eos proteins. This compound is useful for the treatment of certain diseases, including cancer and viral infections. The compound is provided as a useful pharmaceutical product possessing the desired stability, bioavailability, therapeutic index, and toxicity values, which are important for its efficacy.

[0024] A first aspect of the present invention is formula (I): [ka] The objective is to provide the compound shown, or its stereoisomer or salt.

[0025] One embodiment provides a compound of formula (I), or its stereoisomers, tautomers, or pharmaceutically acceptable salts. One embodiment provides a compound of formula (I), or its stereoisomers or tautomers. One embodiment provides a salt of the compound of formula (I), or a stereoisomer or tautomer thereof. One embodiment provides a pharmaceutically acceptable salt of a compound of formula (I), or a stereoisomer or tautomer thereof.

[0026] One embodiment provides a compound of formula (I) or a tautomer or salt thereof, wherein the compound is (S)-3-(5-(4-(6-amino-5-(difluoromethoxy)-4-methylpyridine-2-yl)-2-fluorophenyl)-2-oxoxazole-3(2H)-yl)piperidine-2,6-dione. In addition, this embodiment also includes one or more pharmaceutically acceptable salts. One embodiment provides a compound of formula (I), or a tautomer or salt thereof, wherein the compound is (R)-3-(5-(4-(6-amino-5-(difluoromethoxy)-4-methylpyridine-2-yl)-2-fluorophenyl)-2-oxoxazole-3(2H)-yl)piperidine-2,6-dione. In addition, this embodiment also includes one or more pharmaceutically acceptable salts.

[0027] One embodiment has the following structural formula: [ka] The present invention provides a compound of formula (I), represented by (I), or its stereoisomers, tautomers, or salts.

[0028] One embodiment has the following structural formula: [ka] The present invention provides a compound of formula (I), or a tautomer or salt thereof, as shown in (I). One embodiment has the following structural formula: [ka] The present invention provides a compound of formula (I), or a tautomer or salt thereof, as shown in (I).

[0029] Compounds of formula (I), or their stereoisomers, tautomers, or salts, are useful in reducing the levels of the four IKZF1-4 proteins, Ikaros, Helios, Aeolus, and Eos.

[0030] As used herein, “reducing the level of” one of the IKZF1-4 proteins means reducing the level of the protein by degradation and / or inactivation and / or inhibition and / or a decrease in its expression level, or a combination thereof, compared to the initial level of the protein before contact with the compound of formula (I), or its stereoisomers, tautomers, or salts, or before treatment with them.

[0031] Various methods can be used to measure the decrease in protein levels of IKZF1-4 proteins, including the following assays described below: (i) IKZF1: Human CD8+ T cell reprogramming assay; (ii) IKZF2: Jurkat cell degradation assay; (iii) IKZF3: Human CD8+ T cell reprogramming assay; and (iv) IKZF4: Human regulatory T cell reprogramming assay.

[0032] The present invention may be implemented in other specific forms without departing from its spirit or essential characteristics. The present invention encompasses any combination of the aspects and / or embodiments of the present invention described herein. It is understood that any embodiment of the present invention may be construed in combination with any other embodiment to illustrate further embodiments. It is also understood that each individual component of an embodiment may be combined with any other component from any embodiment to illustrate further embodiments.

[0033] The features and advantages of the present invention will be more readily understood by those skilled in the art by reading the following detailed description. For clarity, it should be recognized that certain features of the present invention described in the context of the separate embodiments above and below may also be combined to form a single embodiment. Conversely, for brevity, various features of the present invention described in the context of a single embodiment may also be combined to form its subcombinations. Embodiments specified herein as examples or preferred are descriptive and not limiting.

[0034] Unless otherwise specified herein, singular references may also include plural forms. For example, “a” and “an” may refer to one, or one or more. As used herein, the term “compound and / or salt thereof” means a compound, at least one salt of such compound, or a combination thereof. For example, compounds and / or salts of formula (I) include the compound of formula (I); a salt of the compound of formula (I); a salt of the compound of formula (I) and one or more compounds of formula (I); and two or more salts of the compound of formula (I).

[0035] Unless otherwise specified, any atom whose valence is not met is assumed to have enough hydrogen atoms to satisfy that valence. The definitions set forth herein supersede any definitions set forth in any patent, patent application, and / or published patent application incorporated herein by attribution. The following lists the definitions of various terms used to describe the present invention. These definitions apply individually or as part of a larger framework to terms in which their definitions are used throughout the specification (unless such terms are particularly limited in specific cases).

[0036] Throughout the specification, the groups and their substituents can be selected by those skilled in the art to provide stable moieties and compounds. In accordance with the conventions used in that field, [ka] In the structural formulas described herein, this is used to indicate the bonding points where a part or substituent bonds to the core or skeletal structure. The word "amino" refers to the base: -NH2. The word "okiso" refers to the base: = O.

[0037] The compounds of the present invention encompass all isotopes of the atoms present in the compound. Isotopes include atoms with the same atomic number but different mass numbers. Generally speaking, isotopes of hydrogen include deuterium (D) and tritium (T). Isotopes of carbon include: 13 C and 14 C is an example. Compounds of the present invention labeled with isotopes can generally be prepared by means of the prior art known to those skilled in the art, or by means of means similar to those described herein, using the isotope-labeled reagent in place of the otherwise unlabeled reagent.

[0038] As used herein, the term "tautomer" refers to each of two or more isomers of a compound that exist together in equilibrium and are readily interchangeable by the movement of atoms or groups within the molecule. For example, as those skilled in the art will know, 1,2,3-triazole is as follows: [ka] It will be easy to understand that it exists in the two tautomer forms shown.

[0039] Thus, this disclosure covers all possible tautomers, even if only one structure is shown. For example, the compound of formula (I) has the following tautomer forms: [ka] It may exist in [location].

[0040] As another example of tautomer morphology: [ka] These are some examples.

[0041] The term "medically acceptable" is used herein to mean, within the bounds of sound medical judgment, that their compounds, materials, compositions, and / or dosage forms are suitable for use in contact with human and animal tissues, without excessive toxicity, irritation, allergic reactions, or other problems or complications, and that the proportion of benefits / risks is reasonable.

[0042] The compounds of formula (I) may form salts, which are also within the scope of the present invention. Unless otherwise specified, references to the compounds of the present invention are understood to include references to one or more of their salts. The term "salt" means an acidic acid formed from inorganic and / or organic acids. Medicinally acceptable (i.e., non-toxic and physiologically acceptable) salts are preferred. However, other salts may also be useful, for example, for the isolation or purification of salts that can be used in production, and are therefore considered to be within the scope of the present invention. Salts of the compounds of formula (I) may be formed, for example, by reacting the compound of formula (I) with a fixed amount of acid, such as one equivalent, in a solvent in which the salt precipitates, or by reacting it in an aqueous medium and then freeze-drying it.

[0043] Examples of acid addition salts include acetates (salts formed with acetic acid or trihaloacetic acid, e.g., trifluoroacetic acid), adipines, alginates, ascorbic acid, aspartates, benzoates, benzenesulfons, bisulfates, borates, butyrates, citrates, camphorates, camphor sulfons, cyclopentanepropionates, digluconates, dodecyl sulfates, ethanesulfons, fumarates, glucoheptanoates, glycerophosphates, hemisulfates, heptanoates, hexanoates, hydrochlorides (formed with hydrochloric acid), hydrobroms (formed with hydrogen bromide), and aqueous iodide. Examples include nitrates, maleates (formed with maleic acid), 2-hydroxyethanesulfonates, lactates, methanesulfonates (formed with methanesulfonic acid), 2-naphthalenesulfonates, nicotinates, nitrates, oxalates, pectins, persulfates, 3-phenylpropionates, phosphates, picrinates, pivaphosphates, propions, salicylates, succinates, sulfates (such as salts formed with sulfuric acid), sulfonates (such as salts described herein), tartrates, thiocyans, tosylates, and other toluenesulfonates, as well as undecanoates.

[0044] The compound of formula (I) can be provided as an amorphous or crystalline solid. By freeze-drying, the compound of formula (I) can be obtained as a solid. Furthermore, solvates (e.g., hydrates) of the compounds of formula (I) should also be understood to be within the scope of the present invention. The term “solvate” means a physical bond between the compound of formula (I) and one or more solvent molecules, whether organic or inorganic. This physical bond includes hydrogen bonds. In certain cases, the solvate will have isolation ability, for example, when one or more solvent molecules are incorporated into the crystal lattice of a crystalline solid. “Solvates” include both liquid-phase solvates and isolationable solvates. Exemplary solvates include hydrates, ethanolates, methanolates, isopropanolates, acetonitrile solvates, and complex salt ethyl solvates.

[0045] Various forms of prodrugs are well known in this field and are described in Rautio, J. et al., Nature Review Drug Discovery, 17, 559-587 (2018). In addition, after its preparation, the compound of formula (I) can be isolated and purified to obtain a composition containing 99% by weight or more ("substantially pure") of the compound of formula (I), which is then used or formulated as described herein. Such "substantially pure" compound of formula (I) is also considered herein as part of the present invention.

[0046] "Stable compound" and "stable structure" refer to compounds that are robust enough to survive when isolated from a reaction mixture to a useful purity and formulated into an effective therapeutic agent. This invention embodies a stable compound.

[0047] The terms "IKZF1 degrading agent" and "IKZF1 degrading agent" refer to drugs that have the ability to reduce the level of the IKZF1 protein by degrading and / or inactivating and / or inhibiting the IKZF1 protein, and / or by reducing its expression level, or by a combination thereof. The terms "IKZF2 degrading agent" and "Helios degrading agent" refer to drugs that have the ability to lower the level of the IKZF2 protein by degrading and / or inactivating and / or inhibiting the IKZF2 protein, and / or reducing its expression level, or by a combination thereof.

[0048] The terms "IKZF3 degrading agent" and "Aeolus degrading agent" refer to drugs that have the ability to reduce the level of IKZF3 protein by degrading and / or inactivating and / or inhibiting the IKZF3 protein, and / or reducing its expression level, or by a combination thereof. The terms "IKZF4 degrading agent" and "EOS degrading agent" refer to drugs that have the ability to lower the level of IKZF4 protein by degrading and / or inactivating and / or inhibiting the IKZF4 protein, and / or reducing its expression level, or by a combination thereof.

[0049] The term "IKZF1-4 protein" refers to the IKZF1, Helios, Aeolus, and Eos proteins. The term "pan-IKZF1-4 degrader" refers to a drug that has the ability to reduce the protein levels of four types of IKZF1-4 proteins: Ikaros, Helios, Aeolus, and Eos. As used herein, the “IKAROS” protein is encoded by the IKZF1 gene. IKAROS is also known as IKAROS family zinc finger 1, ZNFN1Al, 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. The “IKAROS” protein encompasses isoforms encoded by the following human isoforms listed below.

[0050] Isoform 1 (UniProt Q13422-1) [Table 1]

[0051] Isoform 2 (UniProt Q13422-2) [Table 2]

[0052] Isoform 3 (UniProt Q13422-3) [Table 3]

[0053] Isoform 4 (UniProt Q13422-4) [Table 4]

[0054] Isoform 7 (UniProt Q13422-7) [Table 5]

[0055] Isoform 8 (UniProt Q13422-8) [Table 6]

[0056] The "Ikaros" protein isoforms 1, 2, 3, 4, 7, and 8 listed above contain the degron FQCNQCGASFTQKGNLLRHIKLH (Sequence ID: 22), which is the same as the degron of the "Aeolus" protein. The Ikaros protein also contains isoforms encoded by amino acid sequences Q13422-5 and Q13422-6.

[0057] As used herein, the “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 protein, subfamily 1A,2, and IKAROS family zinc finger protein 2. As used herein, the Helios protein encompasses various isoforms, including those listed below.

[0058] Isoform 1 (UniProt Q9UKS7-1) [Table 7]

[0059] Isoform 2 (UniProt Q9UKS7-2) [Table 8]

[0060] Isoform 4 (UniProt Q9UKS7-4) [Table 9]

[0061] Isoform 6 (UniProt Q9UKS7-6) [Table 10]

[0062] Isoform 7 (UniProt Q9UKS7-7) [Table 11]

[0063] The “Helios” isoforms 1, 2, 4, 6, and 7 listed above contain a degron, which is FHCNQCGASFTQKGNLLRHIKLH (SEQ ID NO: 23). Degron is a part of the protein that plays a role in regulating the rate of protein degradation. The Helios protein also contains isoforms encoded by amino acid sequences Q9UKS7-3, Q9UKS7-5, and Q9UKS7-8.

[0064] The “Aeolus” protein as used herein is encoded by the IKZF3 gene. Aeolus is also known as IKAROS family zinc finger 3, ZNFN1A3, zinc finger protein, subfamily 1A,3, IKAROS family zinc finger protein 3, and AIO. The Aeolus protein as used herein includes the following human isoforms listed below:

[0065] Isoform 1 (UniProt Q9UKT9-1) [Table 12]

[0066] Isoform 3 (UniProt Q9UKT9-3) [Table 13]

[0067] Isoform 4 (UniProt Q9UKT9-4) [Table 14]

[0068] Isoform 6 (UniProt Q9UKT9-6) [Table 15]

[0069] Isoform 7 (UniProt Q9UKT9-7) [Table 16]

[0070] Isoform 8 (UniProt Q9UKT9-8) [Table 17]

[0071] Isoform 9 (UniProt Q9UKT9-9) [Table 18]

[0072] Isoform 14 (UniProt Q9UKT9-14) [Table 19]

[0073] The "Aeolus" protein isoforms 1, 3, 4, 6, 7, 8, 9, and 14 listed above contain the degron FQCNQCGASFTQKGNLLRHIKLH (Sequence ID: 24), which is the same as the degron of the "Ikaros" protein. The Aeolus protein also contains isoforms encoded by the amino acid sequences Q9UKT9-2, Q9UKT9-5, Q9UKT9-10, Q9UKT9-11, Q9UKT9-12, Q9UKT9-13, Q9UKT9-15, and Q9UKT9-16.

[0074] The "Eos" protein as used herein is encoded by the IKZF4 gene and is also known as IKAROS family zinc finger 4, ZNFNIA4, zinc finger protein, subfamily 1A,4, IKAROS family zinc finger protein 4, and KIAAI782. The "Eos" protein includes isoforms encoded by the following two human isoforms: 1 (Q9H2S9-1) and 2 (Q9H2S9-2):

[0075] Isoform 1 (UniProt Q9H2S9-1) [Table 20]

[0076] Isoform 2 (UniProt Q9H2S9-2) [Table 21]

[0077] The "Eos" protein isoforms 1 and 2 listed above contain a degron named FHCNQCGASFTQKGNLLRHIKLH (SEQ ID NO: 25), which is the same as the degron of the "Helios" protein. The "Pegasus" protein as used herein 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.

[0078] As used herein, the term “contact” means bringing together the specified portions in vitro or in vivo. For example, contacting the IKZF1-4 protein with the compound of formula (I) includes administering the compound of the present invention to an individual or patient, such as a human, having the Ikaros protein, Helios protein, Aeolus protein, and Eos protein, as well as introducing, for example, the compound of formula (I) into a sample containing a cell preparation or purified preparation containing the Ikaros protein, Helios protein, Aeolus protein, and Eos protein.

[0079] As used herein, the terms “to treat,” “to treat,” and “treatment” refer to any type of intervention or process performed on or involving the administration of an activator to a subject with the aim of reversing, alleviating, improving, inhibiting, delaying, or preventing the progression, onset, worsening, or recurrence of signs, complications, symptoms, or biochemical manifestations associated with a disease. In contrast, “prevention” or “prevention” refers to administering an agent to a subject that is not suffering from the disease in order to prevent the onset of the disease. “To treat,” “to treat,” and “treatment” do not encompass prevention or prevention.

[0080] "Therapeutically effective amount" includes an amount of the compound of the present invention that is effective in reducing the level of IKZF1-4 protein in cells alone, or an amount that is effective in treating or preventing viral infections and proliferative disorders such as cancer, or an amount of the compound of the present invention in combination with other active ingredients that is effective in reducing such levels, or in treating or preventing such disorders.

[0081] As used herein, the term “cell” means in vitro, ex vivo, or in vivo cells. In some embodiments, ex vivo cells may be a portion of a tissue sample extracted from an organism such as a mammal. In some embodiments, in vitro cells may be cells in a cell culture medium. In some embodiments, in vivo cells may be cells living in an organism such as a mammal. The term "patient" encompasses human subjects.

[0082] As used herein, the term “pharmaceutically acceptable carrier” means a pharmaceutically acceptable material, composition, or vehicle, such as a liquid or solid filler, diluent, excipient, manufacturing aid (e.g., lubricant, talc, magnesium stearate, calcium or zinc, or stearic acid), or solvent encapsulating material, that is involved in transporting or delivering the compound of interest from one organ or part of the body to another organ or part of the body. Each carrier must be compatible with the other components of the formulation (i.e., adjuvants, excipients, or vehicles such as diluents, preservatives, fillers, flow modifiers, disintegrants, wetting agents, emulsifiers, antiseptics, flavoring agents, fragrances, antimicrobial agents, antifungal agents, lubricants, and dispersants) depending on the method of administration and the characteristics of the dosage form; and must be “acceptable” in the sense that it is not harmful to the patient.

[0083] The term "pharmaceutical composition" means a composition comprising the compound of the present invention in combination with at least one further pharmaceutically acceptable carrier.

[0084] usefulness The compound of formula (I) is useful in the treatment of cancer. The compound of formula (I) is useful in treating viral infections. One embodiment provides a method for treating cancer in a patient, comprising administering to the patient a therapeutically effective amount of a compound relating to formula (I), or a stereoisomer, tautomer, or pharmaceutically acceptable salt thereof. One embodiment provides a method for treating a viral infection in a patient, comprising administering to the patient a therapeutically effective amount of a compound relating to formula (I), or a stereoisomer, tautomer, or pharmaceutically acceptable salt thereof.

[0085] In one embodiment, a method for treating cancer in a patient, wherein a therapeutically effective amount of a structural formula is given to the patient: [ka] A method is provided comprising administering a compound represented by, or a stereoisomer, tautomer, or pharmaceutically acceptable salt thereof.

[0086] One embodiment provides a method for treating a disease or disorder by reducing the levels of four IKZF1-4 proteins, iKZF1-4, iKZF1-4, iKZF1-4, iKZF1-4, iKZF1-4, iKZF1-4, the method comprising administering to a patient a therapeutically effective amount of the agent to reduce the levels of iKZF1-4,

[0087] One embodiment provides a method for treating a disease or disorder in a patient, comprising administering to the patient a therapeutically effective amount of a drug to reduce the levels of Ikaros, Helios, Aeolus, and Eos proteins, wherein: a) the Ikaros protein is the amino acid sequence encoded by SEQ ID NO: 1, 2, 3, 4, 5, or 6; b) the Helios protein is the amino acid sequence encoded by SEQ ID NO: 7, 8, 9, 10, or 11; c) the Aeolus protein is the amino acid sequence encoded by SEQ ID NO: 12, 13, 14, 15, 16, 17, 18, or 19; and d) the Eos protein is the amino acid sequence encoded by SEQ ID NO: 20 or 21.

[0088] Embodiment 1 provides a method for treating a disease or disorder in a patient, comprising administering to the patient a therapeutically effective amount of an agent to reduce the levels of IKZF1, Helios, Aeolus, and Eos proteins, wherein: (i) the level of the IKZF1 protein is reduced by at least 30%; (ii) the level of the Helios (IKZF2) protein is reduced by at least 50%; (iii) the level of the Aeolus (IKZF3) protein is reduced by at least 30%; and (iv) the level of the Eos (IKZF4) protein is reduced by at least 50%. This embodiment includes a method when the disease or disorder is cancer. This embodiment also includes a method when the disease or disorder is a viral infection. In addition, this embodiment includes a method when the agent is a compound of formula (I), its stereoisomers, tautomers, or pharmaceutically acceptable salts.

[0089] Embodiment 2 provides a method for treating a disease or disorder in a patient, comprising administering to the patient a therapeutically effective amount of an agent to reduce the levels of IKZF1, Helios, Aeolus, and Eos proteins, wherein: (i) the level of the IKZF1 protein is reduced by at least 40%; (ii) the level of the Helios (IKZF2) protein is reduced by at least 50%; (iii) the level of the Aeolus (IKZF3) protein is reduced by at least 40%; and (iv) the level of the Eos (IKZF4) protein is reduced by at least 50%. This embodiment includes a method when the disease or disorder is cancer. This embodiment also includes a method when the disease or disorder is a viral infection. In addition, this embodiment includes a method when the agent is a compound of formula (I), its stereoisomers, tautomers, or pharmaceutically acceptable salts.

[0090] Embodiment 3 provides a method for treating a disease or disorder in a patient, comprising administering to the patient a therapeutically effective amount of an agent to reduce the levels of IKZF1, Helios, Aeolus, and Eos proteins, wherein: (i) the level of the IKZF1 protein is reduced by at least 50%; (ii) the level of the Helios (IKZF2) protein is reduced by at least 50%; (iii) the level of the Aeolus (IKZF3) protein is reduced by at least 50%; and (iv) the level of the Eos (IKZF4) protein is reduced by at least 50%. This embodiment includes a method when the disease or disorder is cancer. This embodiment also includes a method when the disease or disorder is a viral infection. In addition, this embodiment includes a method when the agent is a compound of formula (I), its stereoisomers, tautomers, or pharmaceutically acceptable salts.

[0091] Embodiment 4 provides a method for treating a disease or disorder in a patient, comprising administering to the patient a therapeutically effective amount of an agent to reduce the levels of IKZF1, Helios, Aeolus, and Eos proteins, wherein: (i) the level of the IKZF1 protein is reduced by at least 60%; (ii) the level of the Helios (IKZF2) protein is reduced by at least 50%; (iii) the level of the Aeolus (IKZF3) protein is reduced by at least 60%; and (iv) the level of the Eos (IKZF4) protein is reduced by at least 50%. This embodiment includes a method when the disease or disorder is cancer. This embodiment also includes a method when the disease or disorder is a viral infection. In addition, this embodiment includes a method when the agent is a compound of formula (I), its stereoisomers, tautomers, or pharmaceutically acceptable salts.

[0092] Embodiment 5 provides a method for treating a disease or disorder in a patient, comprising administering to the patient a therapeutically effective amount of an agent to reduce the levels of IKZF1, Helios, Aeolus, and Eos proteins, wherein: (i) the level of the IKZF1 protein is reduced by at least 30%; (ii) the level of the Helios (IKZF2) protein is reduced by at least 60%; (iii) the level of the Aeolus (IKZF3) protein is reduced by at least 30%; and (iv) the level of the Eos (IKZF4) protein is reduced by at least 60%. This embodiment includes a method when the disease or disorder is cancer. This embodiment also includes a method when the disease or disorder is a viral infection. In addition, this embodiment includes a method when the agent is a compound of formula (I), its stereoisomers, tautomers, or pharmaceutically acceptable salts.

[0093] Embodiment 6 provides a method for treating a disease or disorder in a patient, comprising administering to the patient a therapeutically effective amount of an agent to reduce the levels of IKZF1, Helios, Aeolus, and Eos proteins, wherein: (i) the level of the IKZF1 protein is reduced by at least 40%; (ii) the level of the Helios (IKZF2) protein is reduced by at least 60%; (iii) the level of the Aeolus (IKZF3) protein is reduced by at least 40%; and (iv) the level of the Eos (IKZF4) protein is reduced by at least 60%. This embodiment includes a method when the disease or disorder is cancer. This embodiment also includes a method when the disease or disorder is a viral infection. In addition, this embodiment includes a method when the agent is a compound of formula (I), its stereoisomers, tautomers, or pharmaceutically acceptable salts.

[0094] Embodiment 7 provides a method for treating a disease or disorder in a patient, comprising administering to the patient a therapeutically effective amount of an agent to reduce the levels of IKZF1, Helios, Aeolus, and Eos proteins, wherein: (i) the level of the IKZF1 protein is reduced by at least 50%; (ii) the level of the Helios (IKZF2) protein is reduced by at least 60%; (iii) the level of the Aeolus (IKZF3) protein is reduced by at least 50%; and (iv) the level of the Eos (IKZF4) protein is reduced by at least 60%. This embodiment includes a method when the disease or disorder is cancer. This embodiment also includes a method when the disease or disorder is a viral infection. In addition, this embodiment includes a method when the agent is a compound of formula (I), its stereoisomers, tautomers, or pharmaceutically acceptable salts.

[0095] Embodiment 8 provides a method for treating a disease or disorder in a patient, comprising administering to the patient a therapeutically effective amount of an agent to reduce the levels of IKZF1, Helios, Aeolus, and Eos proteins, wherein: (i) the level of the IKZF1 protein is reduced by at least 60%; (ii) the level of the Helios (IKZF2) protein is reduced by at least 60%; (iii) the level of the Aeolus (IKZF3) protein is reduced by at least 60%; and (iv) the level of the Eos (IKZF4) protein is reduced by at least 60%. This embodiment includes a method when the disease or disorder is cancer. This embodiment also includes a method when the disease or disorder is a viral infection. In addition, this embodiment includes a method when the agent is a compound of formula (I), its stereoisomers, tautomers, or pharmaceutically acceptable salts.

[0096] Embodiment 9 provides a method for treating a disease or disorder in a patient, comprising administering to the patient a therapeutically effective amount of an agent to reduce the levels of IKZF1, Helios, Aeolus, and Eos proteins, wherein: (i) the level of the IKZF1 protein is reduced by at least 30%; (ii) the level of the Helios (IKZF2) protein is reduced by at least 70%; (iii) the level of the Aeolus (IKZF3) protein is reduced by at least 30%; and (iv) the level of the Eos (IKZF4) protein is reduced by at least 65%. This embodiment includes a method when the disease or disorder is cancer. This embodiment also includes a method when the disease or disorder is a viral infection. In addition, this embodiment includes a method when the agent is a compound of formula (I), its stereoisomers, tautomers, or pharmaceutically acceptable salts.

[0097] Embodiment 10 provides a method for treating a disease or disorder in a patient, comprising administering to the patient a therapeutically effective amount of an agent to reduce the levels of IKZF1, Helios, Aeolus, and Eos proteins, wherein: (i) the level of the IKZF1 protein is reduced by at least 40%; (ii) the level of the Helios (IKZF2) protein is reduced by at least 70%; (iii) the level of the Aeolus (IKZF3) protein is reduced by at least 40%; and (iv) the level of the Eos (IKZF4) protein is reduced by at least 65%. This embodiment includes a method when the disease or disorder is cancer. This embodiment also includes a method when the disease or disorder is a viral infection. In addition, this embodiment includes a method when the agent is a compound of formula (I), its stereoisomers, tautomers, or pharmaceutically acceptable salts.

[0098] Embodiment 11 provides a method for treating a disease or disorder in a patient, comprising administering to the patient a therapeutically effective amount of an agent to reduce the levels of IKZF1, Helios, Aeolus, and Eos proteins, wherein: (i) the level of the IKZF1 protein is reduced by at least 50%; (ii) the level of the Helios (IKZF2) protein is reduced by at least 70%; (iii) the level of the Aeolus (IKZF3) protein is reduced by at least 50%; and (iv) the level of the Eos (IKZF4) protein is reduced by at least 65%. This embodiment includes a method when the disease or disorder is cancer. This embodiment also includes a method when the disease or disorder is a viral infection. In addition, this embodiment includes a method when the agent is a compound of formula (I), its stereoisomers, tautomers, or pharmaceutically acceptable salts.

[0099] Embodiment 12 provides a method for treating a disease or disorder in a patient, comprising administering to the patient a therapeutically effective amount of an agent to reduce the levels of IKZF1, Helios, Aeolus, and Eos proteins, wherein: (i) the level of the IKZF1 protein is reduced by at least 60%; (ii) the level of the Helios (IKZF2) protein is reduced by at least 70%; (iii) the level of the Aeolus (IKZF3) protein is reduced by at least 60%; and (iv) the level of the Eos (IKZF4) protein is reduced by at least 65%. This embodiment includes a method when the disease or disorder is cancer. This embodiment also includes a method when the disease or disorder is a viral infection. In addition, this embodiment includes a method when the agent is a compound of formula (I), its stereoisomers, tautomers, or pharmaceutically acceptable salts.

[0100] Embodiment 13 provides a method for treating a disease or disorder in a patient, comprising administering to the patient a therapeutically effective amount of an agent to reduce the levels of IKZF1, Helios, Aeolus, and Eos proteins, wherein: (i) the level of the IKZF1 protein is reduced by at least 30%; (ii) the level of the Helios (IKZF2) protein is reduced by at least 80%; (iii) the level of the Aeolus (IKZF3) protein is reduced by at least 30%; and (iv) the level of the Eos (IKZF4) protein is reduced by at least 60%. This embodiment includes a method when the disease or disorder is cancer. This embodiment also includes a method when the disease or disorder is a viral infection. In addition, this embodiment includes a method when the agent is a compound of formula (I), its stereoisomers, tautomers, or pharmaceutically acceptable salts.

[0101] Embodiment 14 provides a method for treating a disease or disorder in a patient, comprising administering to the patient a therapeutically effective amount of an agent to reduce the levels of IKZF1, Helios, Aeolus, and Eos proteins, wherein: (i) the level of the IKZF1 protein is reduced by at least 40%; (ii) the level of the Helios (IKZF2) protein is reduced by at least 80%; (iii) the level of the Aeolus (IKZF3) protein is reduced by at least 40%; and (iv) the level of the Eos (IKZF4) protein is reduced by at least 60%. This embodiment includes a method when the disease or disorder is cancer. This embodiment also includes a method when the disease or disorder is a viral infection. In addition, this embodiment includes a method when the agent is a compound of formula (I), its stereoisomers, tautomers, or pharmaceutically acceptable salts.

[0102] Embodiment 15 provides a method for treating a disease or disorder in a patient, comprising administering to the patient a therapeutically effective amount of an agent to reduce the levels of IKZF1, Helios, Aeolus, and Eos proteins, wherein: (i) the level of the IKZF1 protein is reduced by at least 50%; (ii) the level of the Helios (IKZF2) protein is reduced by at least 80%; (iii) the level of the Aeolus (IKZF3) protein is reduced by at least 50%; and (iv) the level of the Eos (IKZF4) protein is reduced by at least 60%. This embodiment includes a method when the disease or disorder is cancer. This embodiment also includes a method when the disease or disorder is a viral infection. In addition, this embodiment includes a method when the agent is a compound of formula (I), its stereoisomers, tautomers, or pharmaceutically acceptable salts.

[0103] Embodiment 16 provides a method for treating a disease or disorder in a patient, comprising administering to the patient a therapeutically effective amount of an agent to reduce the levels of IKZF1, Helios, Aeolus, and Eos proteins, wherein: (i) the level of the IKZF1 protein is reduced by at least 60%; (ii) the level of the Helios (IKZF2) protein is reduced by at least 80%; (iii) the level of the Aeolus (IKZF3) protein is reduced by at least 60%; and (iv) the level of the Eos (IKZF4) protein is reduced by at least 60%. This embodiment includes a method when the disease or disorder is cancer. This embodiment also includes a method when the disease or disorder is a viral infection. In addition, this embodiment includes a method when the agent is a compound of formula (I), its stereoisomers, tautomers, or pharmaceutically acceptable salts.

[0104] Embodiment 17 provides a method for treating a disease or disorder in a patient, comprising administering to the patient a therapeutically effective amount of an agent to reduce the levels of IKZF1, Helios, Aeolus, and Eos proteins, wherein: (i) the level of the IKZF1 protein is reduced by at least 30%; (ii) the level of the Helios (IKZF2) protein is reduced by at least 85%; (iii) the level of the Aeolus (IKZF3) protein is reduced by at least 30%; and (iv) the level of the Eos (IKZF4) protein is reduced by at least 60%. This embodiment includes a method when the disease or disorder is cancer. This embodiment also includes a method when the disease or disorder is a viral infection. In addition, this embodiment includes a method when the agent is a compound of formula (I), its stereoisomers, tautomers, or pharmaceutically acceptable salts.

[0105] Embodiment 18 provides a method for treating a disease or disorder in a patient, comprising administering to the patient a therapeutically effective amount of an agent to reduce the levels of IKZF1, Helios, Aeolus, and Eos proteins, wherein: (i) the level of the IKZF1 protein is reduced by at least 40%; (ii) the level of the Helios (IKZF2) protein is reduced by at least 85%; (iii) the level of the Aeolus (IKZF3) protein is reduced by at least 40%; and (iv) the level of the Eos (IKZF4) protein is reduced by at least 60%. This embodiment includes a method when the disease or disorder is cancer. This embodiment also includes a method when the disease or disorder is a viral infection. In addition, this embodiment includes a method when the agent is a compound of formula (I), its stereoisomers, tautomers, or pharmaceutically acceptable salts.

[0106] Embodiment 19 provides a method for treating a disease or disorder in a patient, comprising administering to the patient a therapeutically effective amount of an agent to reduce the levels of IKZF1, Helios, Aeolus, and Eos proteins, wherein: (i) the level of the IKZF1 protein is reduced by at least 50%; (ii) the level of the Helios (IKZF2) protein is reduced by at least 85%; (iii) the level of the Aeolus (IKZF3) protein is reduced by at least 50%; and (iv) the level of the Eos (IKZF4) protein is reduced by at least 60%. This embodiment includes a method when the disease or disorder is cancer. This embodiment also includes a method when the disease or disorder is a viral infection. In addition, this embodiment includes a method when the agent is a compound of formula (I), its stereoisomers, tautomers, or pharmaceutically acceptable salts.

[0107] Embodiment 20 provides a method for treating a disease or disorder in a patient, comprising administering to the patient a therapeutically effective amount of an agent to reduce the levels of IKZF1, Helios, Aeolus, and Eos proteins, wherein: (i) the level of the IKZF1 protein is reduced by at least 60%; (ii) the level of the Helios (IKZF2) protein is reduced by at least 85%; (iii) the level of the Aeolus (IKZF3) protein is reduced by at least 60%; and (iv) the level of the Eos (IKZF4) protein is reduced by at least 60%. This embodiment includes a method when the disease or disorder is cancer. This embodiment also includes a method when the disease or disorder is a viral infection. In addition, this embodiment includes a method when the agent is a compound of formula (I), its stereoisomers, tautomers, or pharmaceutically acceptable salts.

[0108] Embodiment 21 provides a method for treating a disease or disorder in a patient, comprising administering to the patient a therapeutically effective amount of an agent to reduce the levels of IKZF1, Helios, Aeolus, and Eos proteins, wherein: (i) the level of the IKZF1 protein is reduced by at least 30%; (ii) the level of the Helios (IKZF2) protein is reduced by at least 90%; (iii) the level of the Aeolus (IKZF3) protein is reduced by at least 30%; and (iv) the level of the Eos (IKZF4) protein is reduced by at least 60%. This embodiment includes a method when the disease or disorder is cancer. This embodiment also includes a method when the disease or disorder is a viral infection. In addition, this embodiment includes a method when the agent is a compound of formula (I), its stereoisomers, tautomers, or pharmaceutically acceptable salts.

[0109] Embodiment 22 provides a method for treating a disease or disorder in a patient, comprising administering to the patient a therapeutically effective amount of an agent to reduce the levels of IKZF1, Helios, Aeolus, and Eos proteins, wherein: (i) the level of the IKZF1 protein is reduced by at least 40%; (ii) the level of the Helios (IKZF2) protein is reduced by at least 90%; (iii) the level of the Aeolus (IKZF3) protein is reduced by at least 40%; and (iv) the level of the Eos (IKZF4) protein is reduced by at least 60%. This embodiment includes a method when the disease or disorder is cancer. This embodiment also includes a method when the disease or disorder is a viral infection. In addition, this embodiment includes a method when the agent is a compound of formula (I), its stereoisomers, tautomers, or pharmaceutically acceptable salts.

[0110] Embodiment 23 provides a method for treating a disease or disorder in a patient, comprising administering to the patient a therapeutically effective amount of an agent to reduce the levels of IKZF1, Helios, Aeolus, and Eos proteins, wherein: (i) the level of the IKZF1 protein is reduced by at least 50%; (ii) the level of the Helios (IKZF2) protein is reduced by at least 90%; (iii) the level of the Aeolus (IKZF3) protein is reduced by at least 50%; and (iv) the level of the Eos (IKZF4) protein is reduced by at least 60%. This embodiment includes a method when the disease or disorder is cancer. This embodiment also includes a method when the disease or disorder is a viral infection. In addition, this embodiment includes a method when the agent is a compound of formula (I), its stereoisomers, tautomers, or pharmaceutically acceptable salts.

[0111] Embodiment 24 provides a method for treating a disease or disorder in a patient, comprising administering to the patient a therapeutically effective amount of an agent to reduce the levels of IKZF1, Helios, Aeolus, and Eos proteins, wherein: (i) the level of the IKZF1 protein is reduced by at least 60%; (ii) the level of the Helios (IKZF2) protein is reduced by at least 90%; (iii) the level of the Aeolus (IKZF3) protein is reduced by at least 60%; and (iv) the level of the Eos (IKZF4) protein is reduced by at least 60%. This embodiment includes a method when the disease or disorder is cancer. This embodiment also includes a method when the disease or disorder is a viral infection. In addition, this embodiment includes a method when the agent is a compound of formula (I), its stereoisomers, tautomers, or pharmaceutically acceptable salts.

[0112] Embodiment 25 provides a method for treating a disease or disorder in a patient, comprising administering to the patient a therapeutically effective amount of an agent to reduce the levels of IKZF1, Helios, Aeolus, and Eos proteins, wherein: (i) the level of the IKZF1 protein is reduced by at least 30%; (ii) the level of the Helios (IKZF2) protein is reduced by at least 90%; (iii) the level of the Aeolus (IKZF3) protein is reduced by at least 30%; and (iv) the level of the Eos (IKZF4) protein is reduced by at least 65%. This embodiment includes a method when the disease or disorder is cancer. This embodiment also includes a method when the disease or disorder is a viral infection. In addition, this embodiment includes a method when the agent is a compound of formula (I), its stereoisomers, tautomers, or pharmaceutically acceptable salts.

[0113] Embodiment 26 provides a method for treating a disease or disorder in a patient, comprising administering to the patient a therapeutically effective amount of an agent to reduce the levels of IKZF1, Helios, Aeolus, and Eos proteins, wherein: (i) the level of the IKZF1 protein is reduced by at least 40%; (ii) the level of the Helios (IKZF2) protein is reduced by at least 90%; (iii) the level of the Aeolus (IKZF3) protein is reduced by at least 40%; and (iv) the level of the Eos (IKZF4) protein is reduced by at least 65%. This embodiment includes a method when the disease or disorder is cancer. This embodiment also includes a method when the disease or disorder is a viral infection. In addition, this embodiment includes a method when the agent is a compound of formula (I), its stereoisomers, tautomers, or pharmaceutically acceptable salts.

[0114] Embodiment 27 provides a method for treating a disease or disorder in a patient, comprising administering to the patient a therapeutically effective amount of an agent to reduce the levels of IKZF1, Helios, Aeolus, and Eos proteins, wherein: (i) the level of the IKZF1 protein is reduced by at least 50%; (ii) the level of the Helios (IKZF2) protein is reduced by at least 90%; (iii) the level of the Aeolus (IKZF3) protein is reduced by at least 50%; and (iv) the level of the Eos (IKZF4) protein is reduced by at least 65%. This embodiment includes a method when the disease or disorder is cancer. This embodiment also includes a method when the disease or disorder is a viral infection. In addition, this embodiment includes a method when the agent is a compound of formula (I), its stereoisomers, tautomers, or pharmaceutically acceptable salts.

[0115] Embodiment 28 provides a method for treating a disease or disorder in a patient, comprising administering to the patient a therapeutically effective amount of an agent to reduce the levels of IKZF1, Helios, Aeolus, and Eos proteins, wherein: (i) the level of the IKZF1 protein is reduced by at least 60%; (ii) the level of the Helios (IKZF2) protein is reduced by at least 90%; (iii) the level of the Aeolus (IKZF3) protein is reduced by at least 60%; and (iv) the level of the Eos (IKZF4) protein is reduced by at least 65%. This embodiment includes a method when the disease or disorder is cancer. This embodiment also includes a method when the disease or disorder is a viral infection. In addition, this embodiment includes a method when the agent is a compound of formula (I), its stereoisomers, tautomers, or pharmaceutically acceptable salts.

[0116] Embodiment 29 provides a method for treating a disease or disorder in a patient, comprising administering to the patient a therapeutically effective amount of an agent to reduce the levels of IKZF1, Helios, Aeolus, and Eos proteins, wherein: (i) the level of the IKZF1 protein is reduced by 40–70%; (ii) the level of the Helios (IKZF2) protein is reduced by at least 50%; (iii) the level of the Aeolus (IKZF3) protein is reduced by 40–70%; and (iv) the level of the Eos (IKZF4) protein is reduced by at least 50%. This embodiment includes a method when the disease or disorder is cancer. This embodiment also includes a method when the disease or disorder is a viral infection. In addition, this embodiment includes a method when the agent is a compound of formula (I), its stereoisomers, tautomers, or pharmaceutically acceptable salts.

[0117] Embodiment 30 provides a method for treating a disease or disorder in a patient, comprising administering to the patient a therapeutically effective amount of an agent to reduce the levels of IKZF1, Helios, Aeolus, and Eos proteins, wherein: (i) the level of the IKZF1 protein is reduced by 40–70%; (ii) the level of the Helios (IKZF2) protein is reduced by at least 60%; (iii) the level of the Aeolus (IKZF3) protein is reduced by 40–70%; and (iv) the level of the Eos (IKZF4) protein is reduced by at least 60%. This embodiment includes a method when the disease or disorder is cancer. This embodiment also includes a method when the disease or disorder is a viral infection. In addition, this embodiment includes a method when the agent is a compound of formula (I), its stereoisomers, tautomers, or pharmaceutically acceptable salts.

[0118] Embodiment 31 provides a method for treating a disease or disorder in a patient, comprising administering to the patient a therapeutically effective amount of an agent to reduce the levels of IKZF1, Helios, Aeolus, and Eos proteins, wherein: (i) the level of the IKZF1 protein is reduced by 40–70%; (ii) the level of the Helios (IKZF2) protein is reduced by at least 70%; (iii) the level of the Aeolus (IKZF3) protein is reduced by 40–70%; and (iv) the level of the Eos (IKZF4) protein is reduced by at least 65%. This embodiment includes a method when the disease or disorder is cancer. This embodiment also includes a method when the disease or disorder is a viral infection. In addition, this embodiment includes a method when the agent is a compound of formula (I), its stereoisomers, tautomers, or pharmaceutically acceptable salts.

[0119] Embodiment 32 provides a method for treating a disease or disorder in a patient, comprising administering to the patient a therapeutically effective amount of an agent to reduce the levels of IKZF1, Helios, Aeolus, and Eos proteins, wherein: (i) the level of the IKZF1 protein is reduced by 40–70%; (ii) the level of the Helios (IKZF2) protein is reduced by at least 70%; (iii) the level of the Aeolus (IKZF3) protein is reduced by 40–70%; and (iv) the level of the Eos (IKZF4) protein is reduced by at least 70%. This embodiment includes a method when the disease or disorder is cancer. This embodiment also includes a method when the disease or disorder is a viral infection. In addition, this embodiment includes a method when the agent is a compound of formula (I), its stereoisomers, tautomers, or pharmaceutically acceptable salts.

[0120] Embodiment 33 provides a method for treating a disease or disorder in a patient, comprising administering to the patient a therapeutically effective amount of an agent to reduce the levels of IKZF1, Helios, Aeolus, and Eos proteins, wherein: (i) the level of the IKZF1 protein is reduced by 40–70%; (ii) the level of the Helios (IKZF2) protein is reduced by at least 80%; (iii) the level of the Aeolus (IKZF3) protein is reduced by 40–70%; and (iv) the level of the Eos (IKZF4) protein is reduced by at least 65%. This embodiment includes a method when the disease or disorder is cancer. This embodiment also includes a method when the disease or disorder is a viral infection. In addition, this embodiment includes a method when the agent is a compound of formula (I), its stereoisomers, tautomers, or pharmaceutically acceptable salts.

[0121] Embodiment 34 provides a method for treating a disease or disorder in a patient, comprising administering to the patient a therapeutically effective amount of an agent to reduce the levels of IKZF1, Helios, Aeolus, and Eos proteins, wherein: (i) the level of the IKZF1 protein is reduced by 40–70%; (ii) the level of the Helios (IKZF2) protein is reduced by at least 90%; (iii) the level of the Aeolus (IKZF3) protein is reduced by 40–70%; and (iv) the level of the Eos (IKZF4) protein is reduced by at least 65%. This embodiment includes a method when the disease or disorder is cancer. This embodiment also includes a method when the disease or disorder is a viral infection. In addition, this embodiment includes a method when the agent is a compound of formula (I), its stereoisomers, tautomers, or pharmaceutically acceptable salts.

[0122] Embodiment 35 provides a method for treating a disease or disorder in a patient, comprising administering to the patient a therapeutically effective amount of an agent to reduce the levels of IKZF1, Helios, Aeolus, and Eos proteins, wherein: (i) the level of the IKZF1 protein is reduced by 50–70%; (ii) the level of the Helios (IKZF2) protein is reduced by at least 50%; (iii) the level of the Aeolus (IKZF3) protein is reduced by 50–70%; and (iv) the level of the Eos (IKZF4) protein is reduced by at least 50%. This embodiment includes a method when the disease or disorder is cancer. This embodiment also includes a method when the disease or disorder is a viral infection. In addition, this embodiment includes a method when the agent is a compound of formula (I), its stereoisomers, tautomers, or pharmaceutically acceptable salts.

[0123] Embodiment 36 provides a method for treating a disease or disorder in a patient, comprising administering to the patient a therapeutically effective amount of an agent to reduce the levels of IKZF1, Helios, Aeolus, and Eos proteins, wherein: (i) the level of the IKZF1 protein is reduced by 50–70%; (ii) the level of the Helios (IKZF2) protein is reduced by at least 60%; (iii) the level of the Aeolus (IKZF3) protein is reduced by 50–70%; and (iv) the level of the Eos (IKZF4) protein is reduced by at least 60%. This embodiment includes a method when the disease or disorder is cancer. This embodiment also includes a method when the disease or disorder is a viral infection. In addition, this embodiment includes a method when the agent is a compound of formula (I), its stereoisomers, tautomers, or pharmaceutically acceptable salts.

[0124] Embodiment 37 provides a method for treating a disease or disorder in a patient, comprising administering to the patient a therapeutically effective amount of an agent to reduce the levels of IKZF1, Helios, Aeolus, and Eos proteins, wherein: (i) the level of the IKZF1 protein is reduced by 50–70%; (ii) the level of the Helios (IKZF2) protein is reduced by at least 70%; (iii) the level of the Aeolus (IKZF3) protein is reduced by 50–70%; and (iv) the level of the Eos (IKZF4) protein is reduced by at least 65%. This embodiment includes a method when the disease or disorder is cancer. This embodiment also includes a method when the disease or disorder is a viral infection. In addition, this embodiment includes a method when the agent is a compound of formula (I), its stereoisomers, tautomers, or pharmaceutically acceptable salts.

[0125] Embodiment 38 provides a method for treating a disease or disorder in a patient, comprising administering to the patient a therapeutically effective amount of an agent to reduce the levels of IKZF1, Helios, Aeolus, and Eos proteins, wherein: (i) the level of the IKZF1 protein is reduced by 50–70%; (ii) the level of the Helios (IKZF2) protein is reduced by at least 70%; (iii) the level of the Aeolus (IKZF3) protein is reduced by 50–70%; and (iv) the level of the Eos (IKZF4) protein is reduced by at least 70%. This embodiment includes a method when the disease or disorder is cancer. This embodiment also includes a method when the disease or disorder is a viral infection. In addition, this embodiment includes a method when the agent is a compound of formula (I), its stereoisomers, tautomers, or pharmaceutically acceptable salts.

[0126] Embodiment 39 provides a method for treating a disease or disorder in a patient, comprising administering to the patient a therapeutically effective amount of an agent to reduce the levels of IKZF1, Helios, Aeolus, and Eos proteins, wherein: (i) the level of the IKZF1 protein is reduced by 50–70%; (ii) the level of the Helios (IKZF2) protein is reduced by at least 80%; (iii) the level of the Aeolus (IKZF3) protein is reduced by 50–70%; and (iv) the level of the Eos (IKZF4) protein is reduced by at least 65%. This embodiment includes a method when the disease or disorder is cancer. This embodiment also includes a method when the disease or disorder is a viral infection. In addition, this embodiment includes a method when the agent is a compound of formula (I), its stereoisomers, tautomers, or pharmaceutically acceptable salts.

[0127] Embodiment 40 provides a method for treating a disease or disorder in a patient, comprising administering to the patient a therapeutically effective amount of an agent to reduce the levels of IKZF1, Helios, Aeolus, and Eos proteins, wherein: (i) the level of the IKZF1 protein is reduced by 50–70%; (ii) the level of the Helios (IKZF2) protein is reduced by at least 90%; (iii) the level of the Aeolus (IKZF3) protein is reduced by 50–70%; and (iv) the level of the Eos (IKZF4) protein is reduced by at least 90%. This embodiment includes a method when the disease or disorder is cancer. This embodiment also includes a method when the disease or disorder is a viral infection. In addition, this embodiment includes a method when the agent is a compound of formula (I), its stereoisomers, tautomers, or pharmaceutically acceptable salts.

[0128] In embodiments 1 to 40, the decrease in the protein levels of IKZF1-4 proteins can be measured using the following assays: (i) IKZF1: Human CD8+ T cell reprogramming assay; (ii) IKZF2: Jurkat cell degradation assay; (iii) IKZF3: Human CD8+ T cell reprogramming assay; and (iv) IKZF4: Human regulatory T cell reprogramming assay.

[0129] The types of cancer that can be treated with 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, hematological cancer, lung cancer, and bone cancer. Examples of such cancer types include neuroblastoma, rectal cancer, colon cancer and other intestinal cancers, anal cancer, familial adenomatous polypoid cancer and hereditary non-polypoid colorectal cancer, esophageal cancer, nasopharyngeal cancer, lip cancer, laryngeal cancer, hypopharyngeal cancer, tongue cancer, salivary gland cancer, thymic cancer, esophageal and gastric 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 such as glioblastoma, astrocytoma, meningioma, medulloblastoma, and peripheral ectodermal tumors. This includes Hodgkin lymphoma, non-Hodgkin lymphoma, Burkitt lymphoma, acute lymphoblastic 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 cancer, bronchial cancer, small cell lung cancer, non-small cell lung cancer, mesothelioma, multiple myeloma, basal cell tumor, teratoma, retinoblastoma, choroidal melanoma, seminomas, rhabdomyosarcoma, craniopharyngioma, osteosarcoma, chondrosarcoma, myosarcoma, liposarcoma, fibrosarcoma, Ewing's sarcoma, and plasmacytoma.

[0130] One embodiment provides a method for treating cancer in a patient, comprising administering to the patient a therapeutically effective amount of a compound relating to formula (I), or a stereoisomer, tautomer, or pharmaceutically acceptable salt thereof, wherein the cancer is melanoma.

[0131] One embodiment provides a method for treating cancer in a patient, comprising administering to the patient a therapeutically effective amount of a compound relating to formula (I), or a stereoisomer, tautomer, or pharmaceutically acceptable salt thereof, wherein the cancer is lung cancer, including small cell lung cancer (SCLC) and non-small cell lung cancer (NSCLC).

[0132] One embodiment provides a method for treating cancer in a patient, comprising administering to the patient a therapeutically effective amount of a compound relating to formula (I), or a stereoisomer, tautomer, or pharmaceutically acceptable salt thereof, wherein the cancer is mesothelioma.

[0133] One embodiment provides a method for treating cancer in a patient, comprising administering to the patient a therapeutically effective amount of a compound relating to formula (I), or a stereoisomer, tautomer, or pharmaceutically acceptable salt thereof, wherein the cancer is breast cancer, including ductal carcinoma, invasive ductal carcinoma, metastatic breast cancer, triple-negative breast cancer, human epidermal growth factor receptor 2 (HER2) positive breast cancer, estrogen receptor (ER) positive breast cancer, hormone receptor positive breast cancer, and hormone receptor negative breast cancer.

[0134] One embodiment provides a method for treating cancer in a patient, comprising administering to the patient a therapeutically effective amount of a compound relating to formula (I), or a stereoisomer, tautomer, or pharmaceutically acceptable salt thereof, wherein the cancer is prostate cancer, including prostate adenocarcinoma and castration-resistant prostate cancer.

[0135] One embodiment provides a method for treating cancer in a patient, comprising administering to the patient a therapeutically effective amount of a compound relating to formula (I), or a stereoisomer, tautomer, or pharmaceutically acceptable salt thereof, wherein the cancer is pancreatic cancer, including pancreatic adenocarcinoma, exocrine pancreatic cancer, and neuroendocrine pancreatic cancer.

[0136] One embodiment provides a method for treating cancer in a patient, comprising administering to the patient a therapeutically effective amount of a compound relating to formula (I), or a stereoisomer, tautomer, or pharmaceutically acceptable salt thereof, wherein the cancer is renal cancer, including renal cell carcinoma, clear cell renal cell carcinoma, and non-clear cell renal cell carcinoma, papillary renal cell carcinoma, Wilms tumor, and renal sarcoma.

[0137] One embodiment provides a method for treating cancer in a patient, comprising administering to the patient a therapeutically effective amount of a compound relating to formula (I), or a stereoisomer, tautomer, or pharmaceutically acceptable salt thereof, wherein the cancer is gastric cancer, including gastric carcinoma.

[0138] One embodiment provides a method for treating cancer in a patient, comprising administering to the patient a therapeutically effective amount of a compound relating to formula (I), or a stereoisomer, tautomer, or pharmaceutically acceptable salt thereof, wherein the cancer is renal cancer, including renal carcinoma and renal parenchymal carcinoma.

[0139] One embodiment provides a method for treating cancer in a patient, comprising administering to the patient a therapeutically effective amount of a compound relating to formula (I), or a stereoisomer, tautomer, or pharmaceutically acceptable salt thereof, wherein the cancer is liver cancer, including hepatocellular carcinoma.

[0140] One embodiment provides a method for treating cancer in a patient, comprising administering to the patient a therapeutically effective amount of a compound relating to formula (I), or a stereoisomer, tautomer, or pharmaceutically acceptable salt thereof, wherein the cancer is ovarian cancer, including ovarian carcinoma.

[0141] One embodiment provides a method for treating cancer in a patient, comprising administering to the patient a therapeutically effective amount of a compound relating to formula (I), or a stereoisomer, tautomer, or pharmaceutically acceptable salt thereof, wherein the cancer is a lymphoma, including Hodgkin lymphoma, non-Hodgkin lymphoma, Burkitt lymphoma, acute lymphoblastic leukemia (ALL), chronic lymphocytic leukemia (CLL), acute myeloid leukemia (AML), chronic myeloid leukemia (CML), adult T-cell leukemia, and diffuse large B-cell lymphoma (DLBCL).

[0142] One embodiment provides a method for treating cancer in a patient, comprising administering to the patient a therapeutically effective amount of a compound relating to formula (I), or a stereoisomer, tautomer, or pharmaceutically acceptable salt thereof, wherein the cancer is a lymphoma, including acute lymphoblastic leukemia (ALL), chronic lymphocytic leukemia (CLL), acute myeloid leukemia (AML), chronic myeloid leukemia (CML), adult T-cell leukemia, and diffuse large B-cell lymphoma (DLBCL).

[0143] One embodiment provides a method for treating cancer in a patient, comprising administering to the patient a therapeutically effective amount of a compound relating to formula (I), or a stereoisomer, tautomer, or pharmaceutically acceptable salt thereof, wherein the cancer is multiple myeloma.

[0144] Compounds of formula (I) and pharmaceutical compositions comprising compounds of formula (I) are useful for treating or preventing any disease or condition associated with the activity of the IKZF1-4 protein. These include viral and other infections (e.g., skin infections, GI infections, urinary tract infections, genitourinary tract infections, systemic infections), and proliferative disorders (e.g., cancer). The compound or pharmaceutical composition may be delivered to the patient by any method of administration. In certain embodiments, compounds of formula (I) or pharmaceutical compositions comprising compounds of formula (I) are administered orally. In other embodiments, compounds of formula (I) or pharmaceutical compositions comprising compounds of formula (I) are administered parenterally.

[0145] One embodiment provides a method for treating a viral infection in a patient, comprising administering to the patient a therapeutically effective amount of a compound relating to formula (I), or a stereoisomer, tautomer, or pharmaceutically acceptable salt thereof, wherein the viral infection is caused by exposure to HIV, hepatitis viruses (types A, B, or C), herpesviruses (e.g., VZV, HSV-1, HAV-6, HSV-II, and CMV, Epstein-Barr virus), adenovirus, influenza virus, flavivirus, echovirus, rhinovirus, coxsackivirus, coronavirus, respiratory syncytial virus, mumps virus, rotavirus, measles virus, rubella virus, parabovirus, vaccinia virus, HTLV virus, dengue virus, papillomavirus, molluscum contagiosum virus, poliovirus, rabies virus, JC virus, and arboviral encephalitis virus.

[0146] Compounds of formula (I) can selectively lower the protein levels of four IKZF1-4 proteins in cells, thereby regulating Treg differentiation and / or immunomodulatory status. For example, compounds of formula (I) can be selectively used by administering an effective amount of the compound of formula (I), or its stereoisomers, tautomers, or salts thereof, to lower the protein levels, decrease the activity levels, and / or inhibit the expression levels of each of the four IKZF1-4 proteins in cells or in individuals requiring a reduction in the protein levels, activity levels, and / or inhibition of the expression levels of each of the four IKZF1-4 proteins.

[0147] In one embodiment, the present invention provides a combination formulation of a compound of formula (I) and / or a pharmaceutically acceptable salt thereof with an additional therapeutic agent to be used simultaneously, separately, or sequentially in the treatment and / or prevention of a number of diseases or disorders associated with the activity of the IKZF1-4 protein. This combination formulation may be used to lower the protein levels of each of the four IKZF1-4 proteins, to reduce the level of their protein activity, and / or to inhibit their expression levels.

[0148] In one embodiment, the compound of formula (I) is administered sequentially before the administration of the immunotumor agent. In yet another embodiment, the compound of formula (I) is administered simultaneously with the immunotumor agent. In yet another embodiment, the compound of formula (I) is administered sequentially after the administration of the immunotumor agent. In another embodiment, the compound of formula (I) may be co-formulated with an immunotumor agent.

[0149] Immuno-oncological agents include, for example, small molecule drugs, antibodies, or other biological or small molecules. Examples of biological immuno-oncological 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 or human antibody.

[0150] In one embodiment, the immunotumor agent is either an (i) agonist of a stimulating (including costimulatory) receptor on a T cell, or an (ii) antagonist of an inhibitory (including coinhibitory) signal, both of which result in amplification of the antigen-specific T cell response (often also referred to as an immune checkpoint regulator).

[0151] Certain stimulant and inhibitory molecules are members of the immunoglobulin superfamily (IgSF). One important family of membrane-bound ligands that bind to co-stimulatory 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 co-stimulatory or co-inhibitory receptors is the TNF family of molecules that bind to the membranes of the congeneral 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, Examples include EDAR, EDA1, XEDAR, EDA2, TNFR1, lymphotoxin α / TNFβ, TNFR2, TNFα, LTβR, lymphotoxin α1β2, FAS, FASL, RELT, DR6, TROY, and NGFR.

[0152] In one embodiment, the T cell response may be stimulated by a combination of a compound of formula (I) and one or more (i) an antagonist of a protein that inhibits T cell activation (e.g., an immune checkpoint inhibitor), 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) an agonist of a protein that stimulates 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.

[0153] Other drugs that can be combined with the compound of formula (I) to treat cancer include inhibitory receptor antagonists or activating receptor agonists on NK cells. For example, the compound of formula (I) can be combined with KIR antagonists such as lirilumab. Other drugs in combination therapy include those that inhibit or deplete macrophages or monocytes, including, but not limited to, CSF-1R antagonists such as CSF-1R antagonist antibodies, including RG7155 (WO11 / 70024, WO11 / 107553, WO11 / 131407, WO13 / 87699, WO13 / 119716, WO13 / 132044) or FPA-008 (WO11 / 140249; WO13169264; WO14 / 036357).

[0154] In another embodiment, the compound of formula (I) may be used in conjunction with one or more agonists that link positive costimulatory receptors, blockers that attenuate signaling through inhibitory receptors, antagonists that systemically increase the frequency of antitumor T cells, and one or more agents that overcome different immunosuppressive pathways within the tumor microenvironment (e.g., agents that block the involvement of inhibitory receptors (e.g., PD-L1 / PD-1 interactions), agents that deplete or inhibit Tregs (e.g., using anti-CD25 monoclonal antibodies (e.g., daclizumab) or by ex vivo anti-CD25 bead depletion), agents that inhibit metabolic enzymes such as IDO, or agents that reverse / prevent T cell anergy or exhaustion), and agents that induce innate immune activation and / or inflammation at the tumor site.

[0155] In one embodiment, the immunotumor agent is a CTLA4 antagonist, such as an antagonistic CTLA-4 antibody. Suitable CTLA-4 antibodies include, for example, YERVOY (ipilimumab) or tremelimumab.

[0156] In another aspect, immunotumor agents are PD-1 antagonists, such as antagonistic PD-1 antibodies. Suitable PD-1 antibodies include, for example, OPDIVO (nivolumab), KEYTRUDA (pembrolizumab), MEDI-0680 (AMP-514; WO2012 / 145493), LIBTAYO (semiprimab), JEMPERLI (dostallimab), and ZYNYZ (retifanlimab). Immunotumor agents also include pizilizumab (CT-011), although its specificity for PD-1 binding is questionable. Another solution targeting the PD-1 receptor is a recombinant protein called AMP-224, which contains the extracellular domain of PD-L2 (B7-DC) fused with the Fc portion of IgG1.

[0157] In another aspect, the immunotumor agent is 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), MSB0010718C (WO2013 / 79174), TECENTRIQ (atezolizumab), and BAVENCIO (avelumab).

[0158] In another embodiment, the immunotumor agent is a LAG-3 antagonist, such as an antagonist 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).

[0159] In another aspect, the immunotumor agent is a CD137(4-1BB) agonist, such as an agonist CD137 antibody. Suitable CD137 antibodies include, for example, urelumab and PF-05082566(WO12 / 32433). In another embodiment, the immunotumor agent is a GITR agonist, such as an agonist-type GITR antibody. Suitable GITR antibodies include, for example, BMS-986153, BMS-986156, TRX-518 (WO06 / 105021, WO09 / 009116), and MK-4166 (WO11 / 028683).

[0160] In another aspect, immunotumor agents are IDO antagonists. 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). In another embodiment, the immunotumor agent is an OX40 agonist, such as an agonist OX40 antibody. Suitable OX40 antibodies include, for example, MEDI-6383 or MEDI-6469.

[0161] In another aspect, the immunotumor agent is an OX40L antagonist, such as an antagonist OX40 antibody. A suitable OX40L antagonist is, for example, RG-7888 (WO06 / 029879). In yet another embodiment, the immunotumor agent is a CD40 agonist, such as an agonist CD40 antibody. In yet another embodiment, the immunotumor agent is a CD40 antagonist, such as an antagonist CD40 antibody. Suitable CD40 antibodies include, for example, lucalumumab or dacetuzumab.

[0162] In another aspect, the immunotumor agent is a CD27 agonist, such as an agonist CD27 antibody. A suitable CD27 antibody is, for example, varylumab. In another aspect, the immunotumor agent is MGA271 (WO11 / 109400) (against B7H3).

[0163] In another aspect, immunotumor agents are anti-TIGIT agents. Suitable anti-TIGIT agents include BMS-986207, tilagolumab, or MK-7684. In another aspect, immunotumor agents are KRASG12C inhibitors. Suitable KRASG12C inhibitors include LUMAKRAS (sotrasib) or KRAZATI (adaglasib).

[0164] Combination therapy may involve administering these agents sequentially, i.e., by administering each therapeutic agent at different times, and by administering these therapeutic agents, or at least two therapeutic agents, substantially simultaneously. Substantially simultaneous administration may be achieved, for example, by administering a single dosage form containing each therapeutic agent in a fixed proportion, or by administering multiple single dosage forms of each therapeutic agent to a target. Sequential or substantially simultaneous administration of each therapeutic agent may be carried out by any appropriate route, including, but not limited to, oral, intravenous, intramuscular, and direct absorption via mucosal tissue. Therapeutics may be administered via the same route or different routes. For example, the first therapeutic agent of a selected combination may be administered by intravenous injection, while the other therapeutic agents of the combination may be administered orally. Alternatively, for example, all therapeutic agents may be administered orally, or all therapeutic agents may be administered by intravenous injection. Combination therapy may also involve administering the above-mentioned therapeutic agents in further combination with other bioactive components and non-pharmacological therapies (e.g., surgery or radiotherapy). If the combination therapy also includes non-pharmacological therapy, that non-pharmacological therapy can be administered at any appropriate time, as long as a beneficial effect is obtained from the combined action of the therapeutic agent and the non-pharmacological therapy. For example, in appropriate cases, a beneficial effect can still be obtained even if the non-pharmacological therapy is temporarily separated from the administration of the therapeutic agent, perhaps for several days or weeks.

[0165] For example, one or more additional therapeutic agents or methods, such as antiviral agents, chemotherapeutic agents or other anticancer agents, immunoenhancing agents, immunosuppressants, radiotherapy, antitumor and antiviral vaccines, cytokine therapies (e.g., IL-2 and GM-CSF), and / or tyrosine kinase inhibitors, may be used in combination with the compound of formula (I) to treat diseases, disorders, or symptoms associated with the IKZF1-4 protein, if desired. The agent may be combined with the compound of the present invention in a single dosage form, or the agents may be administered simultaneously or sequentially in separate dosage forms.

[0166] Appropriate chemotherapeutic agents or other anticancer agents include, for example, uracil mustard, chlormethine, cyclophosphamide (CYTOXAN®), ifosfamide, melphalan, chlorambucil, pipobromane, triethylene-melamine, triethylenethiophosphoramine, busulfan, carmustine, lomustine, streptozosin, dacarbazine, and alkylating agents (including, but not limited to, nitrogen mustard, ethyleneimine derivatives, alkyl sulfonates, nitrosourea, and triazenes) such as uracil mustard, chlormethine, cyclophosphamide (CYTOXAN®), ifosfamide, melphalan, chlorambucil, pipobromane, triethylene-melamine, triethylenethiophosphoramine, busulfan, carmustine, lomustine, streptozosin, dacarbazine, and temozolomide.

[0167] In the treatment of melanoma, suitable agents for use in combination with the compound of formula (I) include dacarbazine (DTIC) (which may be used with other chemotherapeutic agents such as carmustine (BCNU) and cisplatin); the “Dartmas regimen” consisting of DTIC, BCNU, cisplatin, and tamoxifen; and combinations of cissprine, vinblastine, and DTIC, temozolomide, or YERVOY®. The compound of formula (I) can also be used in combination with immunotherapeutic agents, including cytokines such as interferon alpha, interleukin-2, and tumor necrosis factor (TNF), in the treatment of melanoma.

[0168] The compound of formula (I) may also be used in combination with vaccine therapy in the treatment of melanoma. Antimelanoma vaccines are similar in some respects to antiviral vaccines used to prevent diseases caused by viruses such as polio, measles, and mumps. Attenuated melanoma cells or a portion of melanoma cells called antigens are injected into the patient to stimulate the body's immune system, which can recognize and destroy the melanoma cells.

[0169] Melanomas localized to the arm or leg may also be treated using a thermoisolated limb perfusion protocol with a combination of agents containing the compound of formula (I). This treatment protocol temporarily isolates the circulation of the affected leg from the rest of the body and injects a high dose of chemotherapeutic agents into the artery supplying the leg, thus delivering such doses to the tumor area without exposing the internal organs to these high doses, which could cause serious side effects. Typically, the fluid is subjected to heating to 38.9°C or 40°C. Melphalan is the most commonly used drug in this chemotherapy procedure. It may be administered together with another agent called tumor necrosis factor (TNF).

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

[0171] Appropriate chemotherapeutic agents or other anticancer agents further include certain natural products and their derivatives (e.g., vinca alkaloids, antitumor antibiotics, enzymes, lymphokines, and epipodophilotoxins), such as vinblastine, vincristine, vindesine, bleomycin, dactinomycin, daunorubicin, doxorubicin, epirubicin, idarubicin, ara-C, paclitaxel (Taxol), mithramycin, deoxyco-formycin, mitomycin-C, L-asparaginase, interferon (especially IFN-alpha), etoposide, and teniposide.

[0172] Other cytotoxic agents include navelben, CTP-11, anastrazole, letrazole, capecitabine, reloxafine, and doroxafine. Epidophyllotoxin; antitumor enzymes; topoisomerase inhibitors; procarbazine; mitoxantrone; platinum-coordinated complexes such as cisplatin and carboplatin; biological response modifiers; proliferation inhibitors; antihormone therapies; leucovorin; tegafur; and cytotoxic agents such as hematopoietic growth factors are also suitable.

[0173] Other anticancer agents include antibody therapies such as trastuzumab (HERCEPTIN®), antibodies against costimulatory molecules such as CTLA4, 4-1BB, and PD-1, or antibodies against cytokines (IL-10 or TGF-β). Other anticancer drugs also include those that block the migration of immune cells, such as antagonists to chemokine receptors, including CCR2 and CCR4.

[0174] Other anticancer drugs also include those that enhance the immune system, such as adjuvants or adoptive T-cell transfer. Anti-cancer vaccines include dendritic cell-based, synthetic peptide-based, DNA-based, and recombinant vaccines.

[0175] The pharmaceutical composition of the present invention may optionally contain at least one signaling inhibitor (STI). A "signaling inhibitor" is a drug that selectively inhibits one or more important steps in the signaling pathway in the normal function of cancer cells, thereby inducing apoptosis. Appropriate STIs include, but are not limited to, (i) bcr / abl kinase inhibitors such as STI571 (GLEEVEC®); (ii) epidermal growth factor (EGF) receptor inhibitors such as kinase inhibitors (IRESSA®, SSI-774) and antibodies (Imuclon: C225 [Goldstein et al., Clin. Cancer Res., 1:1311-1318 (1995)] and Abgenix: ABX-EGF); (iii) HER-2 / neu receptor inhibitors such as farnesyltransferase inhibitors such as L-744, 832 (Kohl et al., Nat. Med., 1(8):792-797 (1995)); and (iv) rapamycin (e.g., Sekulic et al., Cancer Res., Examples include (v) inhibitors of Akt family kinases or the Akt pathway, such as 60:3504-3513(200); (v) cell cycle kinase inhibitors, such as flavopyridol and UCN-01 (see, for example, Sausville, Curr. Med. Chem. Anti-Canc. Agents, 3:47-56(203)); and (vi) phosphatidylinositol kinase inhibitors, such as LY294002 (see, for example, Vlahos et al., J. Biol. Chem., 269:5241-5248(1994)). Alternatively, at least one STI and the compound of formula (I) may be incorporated into separate pharmaceutical compositions. In certain embodiments of the present invention, the compound of formula (I) and at least one STI may be administered to the patient simultaneously or sequentially. In other words, the compound of formula (I) may be administered first, or at least one STI may be administered first, or the compound of formula (I) and at least one STI may be administered simultaneously. In addition, when the compound of formula (I) and more than one STI are used, the compounds may be administered in any order.

[0176] The present invention further provides a pharmaceutical composition for treating chronic viral infections in patients, comprising a compound of formula (I) in a pharmaceutically acceptable carrier, and optionally comprising at least one chemotherapeutic agent and optionally at least one antiviral agent. Furthermore, a method for treating a patient's chronic viral infection is also provided, by administering the above-mentioned pharmaceutical composition in an effective amount.

[0177] In certain embodiments of the present invention, the compound of formula (I) and at least one chemotherapeutic agent are administered to the patient simultaneously or sequentially. In other words, the compound of formula (I) may be administered first, or at least one chemotherapeutic agent may be administered first, or the compound of formula (I) and at least one STI may be administered simultaneously. In addition, if multiple chemotherapeutic agents are used, the compound and the multiple chemotherapeutic agents may be administered in any order. Similarly, any antiviral agent or STI may be administered at any point in time compared to the compound of formula (I).

[0178] Chronic viral infections that can be treated with the combination therapy of this invention 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). Suitable antiviral agents that may be used in combination with the compound of formula (I) may include nucleoside and nucleotide reverse transcription inhibitors (NRTIs), non-nucleoside reverse transcription inhibitors (NNRTIs), protease inhibitors, and other antiviral agents.

[0179] Examples of appropriate non-reactive antibiotics (NRTIs) include zidovudine (AZT); didanosine (ddl); zalcitabine (ddC); stabudine (d4T); lamivudine (3TC); abacavir (1592U89); adefovir dipivoxil [bis(POM)-PMEA]; lobucavir (BMS-180194); BCH-I0652; emitricitabine [(-)-FTC]; beta-L-FD4 (also known as beta-L-D4C or beta-L-2',3'-dicreoxy-5-fluorocytidine); DAPD, ((-)-beta-D-2,6-diaminopurine dioxolane); and rhodenosine (FddA). Typical and appropriate NNRTIs include nevirapine (BI-RG-587); delavirazine (BHAP, U-90152); efavirenz (DMP-266); PNU-142721; AG-1549; MKC-442 (1-(ethoxymethyl)-5-(1-methylethyl)-6-(phenylmethyl)-(2,4(1H,3H)-pyrimidinedione); and (+)-calanolide A (NSC-675451) and B. Typical and appropriate protease inhibitors. The agents include saquinavir (Ro31-8959); ritonavir (ABT-538); indinavir (MK-639); nerufunavir (AG-1343); amprenavir (141W94); lasinavir (BMS-234475); DMP-450; BMS-2322623; ABT-378; and AG-1549. Other antiviral agents include hydroxyurea, ribavirin, IL-2, IL-12, pentafusid, and Yissum Project No. 11607.

[0180] Combination therapy encompasses administering these therapeutic agents in a sequential manner, i.e., administering each therapeutic agent at separate times, as well as administering these therapeutic agents, or at least two therapeutic agents, substantially simultaneously. Substantially simultaneous administration can be achieved, for example, by administering to the subject a single dosage form containing each therapeutic agent in a fixed proportion, or multiple single dosage forms for each therapeutic agent. Sequential or substantially simultaneous administration of each therapeutic agent can be carried out by any suitable route, including, but not limited to, oral, intravenous, intramuscular, and direct absorption through mucosal tissue. Therapeutics may be administered by the same or different routes. For example, the first therapeutic agent in a selected combination may be administered by intravenous injection, while the other therapeutic agents in the combination may be administered orally. Alternatively, for example, all therapeutic agents may be administered orally, or all therapeutic agents may be administered by intravenous injection. Combination therapy may also encompass administering the above therapeutic agents in further combination with other bioactive components and non-pharmacological therapies (e.g., surgery or radiation therapy). If the combination therapy further includes non-pharmacological treatments, the non-pharmacological treatments can be administered at any appropriate time, as long as beneficial effects derived from the synergistic effects of the combination of the therapeutic agent and the non-pharmacological treatment are obtained. For example, in appropriate cases, beneficial effects can still be obtained even if the non-pharmacological treatment is temporarily separated from the administration of the therapeutic agent, perhaps for several days or weeks.

[0181] Pharmaceutical composition The present invention also provides a pharmaceutical composition comprising a therapeutically effective amount of a compound of formula (I) formulated together with one or more pharmaceutically acceptable carriers (additives) and / or diluents, or optionally comprising one or more of the above-described therapeutic agents.

[0182] The compounds of formula (I) may be administered by any suitable route, preferably in the form of a pharmaceutical composition suitable for such route, in a dose effective for the intended treatment. For any of the uses described herein, compounds of formula (I) and compositions thereof may be administered by any suitable means, for example, orally, such as tablets, capsules (each including sustained-release or time-release formulations), pills, powders, granules, elixirs, tinctures, suspensions (including nanosuspensions, microsuspensions, spray-dried dispersions), syrups, and emulsions; sublingually; buccally; parenterally, such as by subcutaneous, intravenous, intramuscular, or intrasternal injection or infusion techniques (e.g., techniques such as sterile aqueous or non-aqueous solutions or suspensions for injection); transnasally, including administration to the nasal mucosa by inhalation spray; topically, such as in the form of a cream or ointment; or transrectally, such as in the form of a suppository. They may be administered alone, but generally will be administered based on the selected route of administration and standard pharmaceutical practice.

[0183] For oral administration, the pharmaceutical composition may be in the form of, for example, tablets, capsules, liquid capsules, suspensions, or liquids. The pharmaceutical composition is preferably manufactured in the form of dosage units containing a specific amount of the active ingredient. For example, the pharmaceutical composition may be provided as tablets or capsules containing a certain amount of the active ingredient in the range of about 0.1 to 1000 mg, preferably about 0.25 to 250 mg, and more preferably about 0.5 to 100 mg. An appropriate daily dose for humans or other mammals can be determined using conventional methods, but can vary considerably depending on the patient's condition and other factors.

[0184] Any pharmaceutical composition envisioned herein may be delivered orally, for example, via any acceptable and suitable oral formulation. Exemplary oral formulations include, but are not limited to, tablets, lozenges, aqueous and oily suspensions, dispersible powders or granules, emulsions, hard and soft capsules, liquid capsules, syrups, and elixirs. Pharmaceutical compositions intended for oral administration may be manufactured according to any method known in the art of manufacturing pharmaceutical compositions intended for oral administration. To provide a pharmaceutically palatable formulation, the pharmaceutical compositions according to the present invention may contain at least one agent selected from sweeteners, flavoring agents, colorants, lubricants, antioxidants, and preservatives.

[0185] Tablets may be manufactured, for example, by mixing a compound of formula (I) and / or at least one pharmaceutically acceptable salt thereof with at least one non-toxic, pharmaceutically acceptable excipient suitable for the manufacture of tablets. Exemplary excipients, for example, include, but are not limited to, inert diluents such as calcium carbonate, sodium carbonate, lactose, calcium phosphate, and sodium phosphate; granulating and disintegrating agents such as microcrystalline cellulose, croscarmellose sodium, corn starch, and alginic acid; binders such as starch, gelatin, polyvinylpyrrolidone, and acacia; and lubricants such as magnesium stearate, stearic acid, and talc. In addition, tablets may be uncoated or coated by known techniques, either to mask the unpleasant taste of an unpleasant-tasting drug or to slow the disintegration and absorption of the active ingredient in the gastrointestinal tract, thereby prolonging the effect of the active ingredient. Examples of water-soluble flavor-masking materials include, but are not limited to, hydroxypropyl-methylcellulose and hydroxypropyl-cellulose. Examples of time-delaying materials include, but are not limited to, ethylcellulose and cellulose acetate butyrate.

[0186] Hard gelatin capsules can be produced, for example, by mixing a compound of formula (I) and / or at least one salt thereof with at least one inert solid diluent, such as calcium carbonate; calcium phosphate; and kaolin. Soft gelatin capsules can be manufactured, for example, by mixing a compound of formula (I) and / or at least one pharmaceutically acceptable salt thereof with at least one water-soluble carrier, such as polyethylene glycol; and at least one oily medium, such as peanut oil, liquid paraffin, and olive oil.

[0187] An aqueous suspension may be prepared, for example, by mixing a compound of formula (I) and / or at least one pharmaceutically acceptable salt thereof with at least one excipient suitable for the preparation of an aqueous suspension. For example, exemplary excipients suitable for the preparation of aqueous suspensions include, but are not limited to, antiprecipitation agents such as sodium carboxymethylcellulose, methylcellulose, hydroxypropylmethylcellulose, sodium alginate, alginic acid, polyvinylpyrrolidone, tragacanth gum, and acacia gum; dispersants or wetting agents such as natural phosphatides, such as lecithin; condensation products of alkylene oxides such as polyoxyethylene stearate and fatty acids; condensation products of ethylene oxides such as heptadecaethylene-oxycetanol and long-chain aliphatic alcohols; condensation products of ethylene oxides such as polyoxyethylene sorbitol monooleate and partial esters derived from fatty acids and hexitol; and condensation products of ethylene oxides such as polyoxyethylene sorbitan monooleate and partial esters derived from fatty acids and anhydrous hexitol. The aqueous suspension may also contain, for example, at least one preservative such as ethyl p-hydroxybenzoate and n-propyl; at least one coloring agent; at least one flavoring agent; and / or at least one sweetener, but not limited to, sucrose, saccharin, and aspartame.

[0188] An oily suspension may be prepared, for example, by suspending a compound of formula (I) and / or at least one pharmaceutically acceptable salt thereof in a vegetable oil such as arachis oil; olive oil; sesame oil; and coconut oil; or in a mineral oil such as liquid paraffin. The oily suspension may also contain at least one thickener, such as beeswax; hard paraffin; and cetyl alcohol. To provide a palatable oily suspension, at least one sweetener and / or at least one flavoring, as already described above, may be added to the oily suspension. The oily suspension may further contain at least one preservative, including, but not limited to, butylated hydroxyanisole and antioxidants such as alpha-tocopherol.

[0189] Dispersible powders and granules may be prepared, for example, by mixing a compound of formula (I) and / or at least one pharmaceutically acceptable salt thereof with at least one dispersant and / or wetting agent; at least one anti-precipitation agent; and / or at least one preservative. Suitable dispersants, wetting agents, and anti-precipitation agents have already been described above. Exemplary preservatives include, but are not limited to, antioxidants such as ascorbic acid. In addition, dispersants and granules may also contain at least one excipient, but are not limited to, sweeteners; flavorings; and colorants.

[0190] Emulsions of compounds of formula (I) and / or pharmaceutically acceptable salts of at least one thereof may be prepared, for example, as oil-in-water emulsions. The oil phase of an emulsion containing a compound of formula (I) may consist of known components in known ways. The oil phase may be provided by, but is not limited to, vegetable oils such as olive oil and arachis oil; mineral oils such as liquid paraffin; and mixtures thereof. The phase may consist only of an emulsifier, or it may consist of at least one emulsifier and a fat or oil, or a mixture of both fat and oil. Suitable emulsifiers include, but are not limited to, natural phosphatides, such as soy lecithin; esters or partial esters derived from fatty acids such as polyoxyethylene sorbitan monooleate and anhydrous hexitol; and condensation products of partial esters such as polyoxyethylene sorbitan monooleate and ethylene oxide. Hydrophilic emulsifiers are preferably formulated together with lipophilic emulsifiers that act as stabilizers. Furthermore, it is preferable to include both oils and fats. The emulsifier, together with or without a stabilizer, constitutes a so-called emulsifying wax, which, together with the oils and fats, constitutes a so-called emulsifying ointment base, forming the oily dispersion phase of the cream formulation. The emulsion may also contain sweeteners, fragrances, preservatives, and / or antioxidants. Suitable emulsifiers and emulsifying stabilizers for use in the formulations of the present invention include Tween 60, Span 80, cetostearyl alcohol, myristyl alcohol, glyceryl monostearate, sodium lauryl sulfate, glyceryl distearate, either alone or together with wax, or other materials well known in the art.

[0191] Compounds of formula (I) and / or pharmaceutically acceptable salts of at least one thereof may also be delivered intravenously, subcutaneously, and / or intramuscularly, for example, via any pharmaceutically acceptable and suitable injection form. Exemplary injection forms, but not limited to, include sterile aqueous solutions; oil-in-water microemulsions; and aqueous or oily suspensions, each comprising an acceptable vehicle and solvent such as water, Ringer's solution, and isotonic sodium chloride solution.

[0192] Parenteral formulations 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 using one or more carriers or diluents mentioned for use in oral formulations, or by using other suitable dispersants or wetting agents and anti-precipitation 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, tragacanth gum, and / or various buffers. Other adjuvants and administration methods are well known in the pharmaceutical field. The active ingredient may also be administered by injection as a composition with a suitable carrier containing seline, dextrose, or water, or with a suitable carrier containing cyclodextrin (i.e., captisol), a co-solubilizer (i.e., polyethylene glycol), or a micelle solubilizer (i.e., Zween 80).

[0193] Sterile injectable formulations may also be sterile injectable solutions or suspensions in non-toxic, parenterally acceptable diluents or solvents, such as solutions in 1,3-butanediol. Among acceptable vehicles and solvents, those that can be used are water, Ringer's solution, and isotonic sodium chloride solution. In addition, sterile fixatives have conventionally been used as solvents or suspension media. Any bland fixative, including synthetic mono- or diglycerides, can be used for this purpose. Furthermore, fatty acids such as oleic acid find applications in the manufacture of injectable formulations.

[0194] Sterile oil-in-water microemulsions for injection can be produced, for example, by 1) dissolving the compound of formula (I) in an oil phase, such as a mixture of soybean oil and lecithin; 2) combining the oil phase containing the compound of formula (I) with a mixture of water and glycerol; and 3) processing the combination to form a microemulsion.

[0195] Sterile aqueous or oily suspensions can be prepared according to methods already known in the art. For example, a sterile aqueous solution or suspension can be prepared using a non-toxic, parenterally acceptable diluent or solvent, such as 1,3-butanediol; a sterile oily suspension can be prepared using, for example, a sterile fixative oil, a sterile, non-toxic, acceptable solvent or suspension medium, such as a synthetic mono or diglyceride, and a fatty acid, such as oleic acid.

[0196] Medicinally acceptable carriers are formulated according to a number of factors well within the knowledge of those skilled in the art. These factors include, but are not limited to, the type and nature of the activator being formulated; the target population to whom the composition containing the activator is intended to be administered; the intended route of administration of the composition; and the targeted therapeutic indications. Medicinally acceptable carriers include both aqueous and non-aqueous liquid media, as well as various solid and semi-solid dosage forms. Such carriers may contain many different components and additives in addition to the activator, and such additives are incorporated into the formulation for various reasons well known to those skilled in the art, such as activator stabilization or binding. A description of appropriate pharmaceutically acceptable carriers and the factors involved in their selection can be found in various readily available sources, such as Allen, LV Jr. et al., Remington: The Science and Practice of Pharmacy (2 Volumes), 22nd Edition (2012), Pharmaceutical Press.

[0197] pharmaceutically acceptable carriers, adjuvants, and vehicles that can be used in the pharmaceutical compositions of the present invention include, but are not limited to, ion exchangers, self-emulsifying drug delivery systems (SEDDS) such as alumina, aluminum stearate, lecithin, d-alpha-tocopherol polyethylene glycol 1000 succinate, surfactants used in pharmaceutical dosage forms such as Zween, polyethoxylated castor oil such as CREMOPHOR surfactant (BASF), or other similar polymer delivery matrices, serum proteins such as human serum albumin, buffers such as phosphates, glycine, sorbic acid, potassium sorbate, partial glyceride mixtures of saturated vegetable fatty acids, water, salts, or electrolytes such as protamine sulfate, disodium hydrogen phosphate, potassium hydrogen phosphate, hydrogen chloride, zinc salts, colloidal silica, magnesium trisilicate, polyvinylpyrrolidone, cellulosic substances, polyethylene glycol, sodium carboxymethylcellulose, polyacrylates, waxes, polyethylene-polyoxypropylene block polymers, polyethylene glycol, and lanolin. Chemically modified derivatives such as cyclodextrins including alpha-, beta-, and gamma-cyclodextrins, or hydroxyalkylcyclodextrins including 2- and 3-hydroxypropyl-cyclodextrins, or other solubilizing derivatives may also be advantageously used to enhance the delivery of the compounds represented by the formulas described herein.

[0198] The pharmaceutically active compounds of the present invention can be processed according to conventional pharmaceutical methods to produce drugs for administration to patients, including humans and other mammals. The pharmaceutical compositions may be subjected to conventional pharmaceutical operations such as sterilization and / or may contain conventional adjuvants such as preservatives, stabilizers, humectants, emulsifiers, buffers, etc. Tablets and pills may additionally be produced using enteric coatings. Such compositions may also contain adjuvants such as humectants, sweeteners, flavorings, and fragrances.

[0199] When intended for therapeutic purposes, the active compounds of the present invention are typically combined with one or more adjuvants suitable for the indicated route of administration. When administered orally, the compounds may be mixed with lactose, sucrose, starch powder, cellulose esters of alkanates, cellulose alkyl esters, talc, stearic acid, magnesium stearate, magnesium oxide, sodium and calcium salts of phosphoric acid and sulfate, gelatin, acacia gum, sodium alginate, polyvinylpyrrolidone, and / or polyvinyl alcohol, and then encapsulated or tableted for convenient administration. Such capsules or tablets may contain a controlled-release formulation, such as one in which the active compound is dispersed in hydroxypropyl methylcellulose.

[0200] The amount of compound administered and the dosage regimen for treating a condition with the compounds and / or compositions of the present invention depend on a variety of factors, including the subject's age, weight, sex, health status, disease type, disease severity, route and frequency of administration, and the specific compound used. Thus, the dosage regimen can vary widely but can be conventionally determined using standard methods. A daily dose of about 0.001 to 100 mg per kg of body weight, preferably about 0.0025 to 50 mg per kg of body weight, and most preferably about 0.005 to 10 mg per kg of body weight may be appropriate. The daily dose may be administered in 1 to 4 doses per day. Other dosage schedules include once-weekly administration and once-every-two-day cycles.

[0201] The pharmaceutical compositions of the present invention comprise a compound of formula (I) and / or a pharmaceutically acceptable salt thereof, and may optionally comprise an additive selected from any pharmaceutically acceptable carrier, adjuvant, and vehicle. Alternative compositions of the present invention comprise a compound of formula (I) as described herein, or its prodrug, and a pharmaceutically acceptable carrier, adjuvant, or vehicle.

[0202] The present invention also includes a pharmaceutical kit useful, for example, in the treatment or prevention of IKZF1-4 protein-related diseases or disorders and other diseases referred to herein, comprising one or more containers containing a pharmaceutical composition comprising a therapeutically effective amount of a compound of formula (I). Such a kit may further include one or more various conventional pharmaceutical kit components, such as further containers, for example, containers containing one or more pharmaceutically acceptable carriers, as would be readily understood by those skilled in the art. Instructions, either as an insert or as a label, indicating the amount of the component to be administered, guidelines for administration, and / or guidelines for mixing the components, may also be included in the kit.

[0203] The administration plan for the compounds of the present invention will, of course, vary depending on known factors such as the pharmacodynamic properties of the particular drug and its method and route of administration; the type, age, sex, health status, condition, and weight of the recipient; the nature and severity of symptoms; the type of combination therapy; the frequency of treatment; the route of administration; the patient's renal and hepatic function; and the desired effect.

[0204] According to general guidelines, the daily oral dose of each active ingredient, when used for a given effect, would be in the range of approximately 0.001 to 5000 mg per day, preferably approximately 0.01 to 1000 mg per day, and most preferably approximately 0.1 to 250 mg per day. For intravenous administration, the most preferred dose would be in the range of approximately 0.01 to 10 mg / kg (per minute of constant-rate infusion). The compound of formula (I) may be administered in a single dose per day, or the total daily dose may be divided into two, three, or four doses per day.

[0205] The compound is typically administered in mixture with a suitable pharmaceutical diluent, excipient, or carrier (collectively referred to herein as a pharmaceutical carrier), which is appropriately selected depending on the intended dosage form, e.g., oral tablets, capsules, elixirs, and syrups, and is not inconsistent with conventional pharmaceutical practices.

[0206] Suitable dosage forms (pharmaceutical compositions) may contain approximately 1 milligram to 200 milligrams of the active ingredient per dose. In these pharmaceutical compositions, the active ingredient will typically be present in an amount of approximately 0.1–95% by weight based on the total weight of the composition.

[0207] A typical capsule for oral administration contains the compound of formula (I) (250 mg), lactose (75 mg), and magnesium stearate (15 mg). The mixture is passed through a 60-mesh sieve and filled into No. 1 gelatin capsules. A typical injectable formulation is prepared by aseptically placing a compound of formula (I) (250 mg) into a vial, aseptically lyophilizing it, and sealing it.

[0208] The present invention encompasses, within its scope, pharmaceutical compositions comprising, as an active ingredient, a therapeutically effective amount of the compound of formula (I), either alone or in combination with a pharmaceutical carrier. Optionally, the compound of formula (I) may be used in combination with one or more other therapeutic agents, such as anticancer agents or other pharmaceutically active materials.

[0209] Regardless of the route of administration selected, the compounds of formula (I) and / or the pharmaceutical compositions of the present invention, which can be used in a properly hydrated form, are formulated into pharmaceutically acceptable dosage forms in a conventional manner known to those skilled in the art.

[0210] The actual dosage level of the active ingredient in the pharmaceutical composition of the present invention may be modified to obtain an amount of the active ingredient that is effective in achieving a therapeutic response without being toxic to the patient, for a particular patient, composition, and method of administration.

[0211] The dosage level selected will depend on a variety of factors including the activity of the compound of formula (I) or its ester, salt or amide utilized, the route of administration, the time of administration, the rate of excretion or metabolism of the particular compound utilized, the rate and extent of absorption, the duration of treatment, other drugs, compounds and / or materials used in combination with the particular compound utilized, the age, sex, weight, condition, health status, and prior history of the patient being treated, as well as similar factors well known in the medical arts.

[0212] 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 with a dosage of the compound of formula (I) utilized in the pharmaceutical composition at a level lower than that required to achieve a therapeutic effect and gradually increase the dosage until the therapeutic effect is achieved.

[0213] Generally, the appropriate daily dosage of the compound of formula (I) will be the amount of the compound that is the lowest dosage effective to obtain a therapeutic effect. Such effective dosage will generally depend on the factors described above. Generally, the oral, intravenous, intracerebroventricular and subcutaneous dosages of the compound of formula (I) for a patient will be in the range of about 0.01 to about 50 mg per kg of body weight per day.

[0214] If desired, the effective daily dosage of the active compound can be subdivided into dosages administered separately at appropriate intervals throughout the day, in dosages divided into 2, 3, 4, 5, 6 or more times, and if desired, administered in unit dosage forms. In certain embodiments of the invention, the administration is once a day. The compound of formula (I) can be administered alone, but the compound is preferably administered as a pharmaceutical formulation (composition).

[0215] When used in combination with the compound of formula (I), the other therapeutic agents described above may be used, for example, in the amounts indicated in the Physicians' Desk Reference (PDR), or as determined by those skilled in the art. In the method of the present invention, such other therapeutic agents may be administered before, simultaneously with, or after administering the compound of the present invention.

[0216] Manufacturing method The compounds of the present invention can be prepared by many methods well known to those skilled in the art of organic synthesis. The compounds of the present invention can be prepared by the following methods, in conjunction with or with modifications thereof as recognized by those skilled in the art. Preferred methods, but not limited to, include the following. All references cited herein are incorporated herein in their entirety with proper attribution.

[0217] The compounds of the present invention can 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 suitable for the deformation to occur. It should also be recognized that, in the description of the synthesis methods below, all proposed reaction conditions, including the choice of solvent, reaction environment, duration of the experiment, and workup procedures, are selected to be standard conditions for the reaction, and this should be readily apparent to those skilled in the art. Those skilled in the field of organic synthesis will understand that the functional groups present in various parts of the molecule must be compatible with the proposed reagents and reactions. Such restrictions on substituents compatible with the reaction conditions will be obvious to those skilled in the art, and in such cases, alternative methods must be used. This will sometimes require a decision to modify the order of the synthesis steps or to choose one particular process scheme over another in order to obtain the compounds of the present invention. It will also be understood that another important consideration in planning any synthesis route in this art is the careful selection of protecting groups used to protect the reactive functional groups present in the compounds described in the present invention. A highly authoritative work describing many alternatives for experienced professionals is Greene and Wuts (Protective Groups In Organic Synthesis, Fourth Edition, Wiley and Sons, 2007).

[0218] Examples The following examples illustrate specific embodiments of the present invention and do not limit the scope of the invention. Chemical and scientific abbreviations and symbols have their usual customary meanings unless otherwise noted. Further abbreviations used in the Examples section and elsewhere in this application are defined below. Compounds and intermediates in the Examples are identified by the Example and step in which they are produced (e.g., "1-A" indicates Example 1, step A), or by the Example alone if the compound is the nominated compound in the Example (e.g., "1" indicates the nominated compound in Example 1). In some cases, alternative preparations for intermediates or examples are described. A chemist skilled in the field of synthesis may frequently devise desirable alternative preparations based on one or more considerations such as reduced reaction time, lower starting material costs, ease of handling or isolation, improved yield, compliance with catalysts, avoidance of toxic reagents, accessibility to specialized equipment, and reduction in the number of linear steps. The intention in describing alternative preparations is to further facilitate the preparation of the Examples of the present invention. In some cases, some of the functional groups in the examples outlined and in the claims may be replaced by well-known biological equivalent substitutions known in the art, such as substituting a carboxylic acid group with a tetrazole or phosphate moiety.

[0219] Abbreviation ACN Acetonitrile BISPIN Bis (Pinacolato) Diboron CatacXium-PD-G3 Mesylate [(di(1-adamantyl)-n-butylphosphine)-2-(2'-amino-1,1'-biphenyl)]palladium(II) CDI (Carbonyldiimidazole) DBU 1,8-Diazabicyclo[5.4.0]Undeca-7-Ene DCM Dichloromethane DIPEA N,N-diisopropylethylamine DME (Dimethyl Ether) DMF Dimethylformamide DMSO (Dimethyl Sulfoxide) dppf bis(diphenylphosphino)ferrocene HCl ethyl acetate HPLC (High-Performance Liquid Chromatography) Me methyl MeOH methanol min mL (milliliter) NCS N-chlorosuccinimide NH4OAc (Ammonium acetate) NMP (N-methylpyrrolidinone) Pd(dppf)2Cl2 [1,1'-Bis(diphenylphosphino)ferrocene]dichloropalladium(II) SEM-Cl 2-Trimethylsilylethoxymethyl Chloride TEA (Triethylamine) TFA (Trifluoroacetic Acid) THF (Tetrahydrofuran)

[0220] LCMS analysis conditions Method A: ACQUITY ULC® BEH C18 (3.0 x 50 mm) 1.7 μm; Mobile phase A: 95:5 water:acetonitrile + 2.5 mM NH4OAc; Mobile phase B: 5:95 water:acetonitrile + 2.5 mM NH4OAc; Temperature: 40°C; Gradient: 20% B to 100% B over 2 minutes; Flow rate: 0.7 mL / min; Detection: MS and UV (220 nm) Method B: Column - Kinetic (Kinetex) XB-C18 (75x3mm-2.6μm); Mobile phase A: 10mM NH4COOH in water; Mobile phase B: Acetonitrile; Gradient: From 20%B to 100%B over 4.6 minutes; Flow rate: 1.0 mL / min

[0221] Intermediate A 3-(5-(2-fluoro-4-(4,4,5,5-tetramethyl-1,3,2-dioxaborolan-2-yl)phenyl)-2-oxoxazole-3(2H)-yl)-1-((2-(trimethylsilyl)ethoxy)methyl)piperidine-2,6-dione [Chemical formula] Production of Intermediate A1: 2-Bromo-1-(4-bromo-2-fluorophenyl)ethan-1-one [Chemical formula]

[0222] Copper(II) bromide (91 g, 405 mmol) was added to a stirred solution of 1-(4-bromo-2-fluorophenyl)ethan-1-one (44 g, 203 mmol) in EtOAc (1000 mL) at room temperature. The reaction mixture was heated at 60 °C for 16 h under argon, cooled to room temperature, and filtered through a pad of celite. The filtrate was concentrated under vacuum. The resulting crude product was purified by flash chromatography (SiO2, 330 g column, 0 - 10% EtOAc / petroleum ether) to give 2-bromo-1-(4-bromo-2-fluorophenyl)ethan-1-one (32.8 g, 51%). 1 1H-NMR (400 MHz, CDCl3): δ 4.50 (s, H), 7.40 - 7.48 (m, 2H), 7.83 - 7.87 (m, 1H)

[0223] Production of Intermediate A2: 3-((2-(4-bromo-2-fluorophenyl)-2-oxoethyl)amino)piperidine-2,6-dione [Chemical formula]

[0224] 3-aminopiperidine-2,6-dione·HCl (22.25 g, 135 mmol) was mixed in THF (200 mL) and K2CO3 (20 g, 145 mmol) was added. The reaction mixture was stirred under nitrogen for 15 minutes. 2-bromo-1-(4-bromo-2-fluorophenyl)ethane-1-one (20 g, 67.6 mmol) was added gradually, and the reaction mixture was heated at 70°C for 2 hours. The reaction mixture was cooled to room temperature, concentrated under vacuum, and diluted with water (200 mL). The solid was separated, filtered, and dried under vacuum to obtain 3-((2-(4-bromo-2-fluorophenyl)-2-oxoethyl)amino)piperidine-2,6-dione (19 g, 71%). LC-MS (Method A): Retention time 0.865 minutes, [M+H] + 344.0

[0225] Preparation of intermediate A3: 3-(5-(4-bromo-2-fluorophenyl)-2-oxoxazole-3(2H)-yl)piperidine 2,6-dione [ka]

[0226] To a stirred solution of 3-((2-(4-bromo-2-fluorophenyl)-2-oxoethyl)amino)piperidine-2,6-dione (30 g, 87 mmol) in THF (300 mL), K2CO3 (12.08 g, 87 mmol) and CDI (28.4 g, 175 mmol) were added at room temperature. The reaction mixture was stirred at the same temperature for 16 hours, concentrated under vacuum, and diluted with water (400 mL). The resulting solid precipitate was filtered through a Buchner funnel and dried under vacuum to obtain 3-(5-(4-bromo-2-fluorophenyl)-2-oxoxazole-3(2H)-yl)piperidine-2,6-dione (21.1 g, 50%). LC-MS (Method A): Retention time 1.79 minutes, [M+H] + 370.0

[0227] Preparation of intermediate A4: 3-(5-(4-bromo-2-fluorophenyl)-2-oxoxazole-3(2H)-yl)-1-((2-(trimethylsilyl)ethoxy)methyl)piperidine-2,6-dione [ka]

[0228] To a stirred solution of 3-(5-(4-bromo-2-fluorophenyl)-2-oxoxazole-3(2H)-yl)piperidine-2,6-dione (10 g, 27.1 mmol) in THF (100 mL), DBU (6.12 mL, 40.6 mmol) and SEM-Cl (5.77 mL, 32.5 mmol) were added at -50°C under nitrogen. The reaction was continued at the same temperature for 1 hour and stopped with water. The reaction mixture was extracted with ELISA (3x). The organic phases were combined, washed with brine, dried on anhydrous sodium sulfate, filtered, and concentrated under vacuum. The residue was purified by column chromatography (SiO2, 0-60% siRNA / petroleum ether) to obtain 3-(5-(4-bromo-2-fluorophenyl)-2-oxoxazole-3(2H)-yl)-1-((2-(trimethylsilyl)ethoxy)methyl)piperidine-2,6-dione (6.2 g, 45%). LC-MS (Method A): Retention time 1.68 minutes, [M+H] + 500.1

[0229] Manufacturing of intermediate A: To a stirred solution of 3-(5-(4-bromo-2-fluorophenyl)-2-oxoxazole-3(2H)-yl)-1-((2-(trimethylsilyl)ethoxy)methyl)piperidine-2,6-dione (7 g, 14.0 mmol) in anhydrous 1,4-dioxane (70 mL), BISPIN (5.34 g, 21.02 mmol) and potassium acetate (1.651 g, 16.82 mmol) were added at room temperature. The reaction mixture was purged with argon for 10 minutes, and Pd(dppf)Cl2·DCM complex (1.026 g, 1.40 mmol) was added under argon. The resulting mixture was heated at 80°C for 1 hour. The reaction mixture was cooled to room temperature, diluted with ELISA (70 mL), filtered through a Celite pad, and the filtrate was concentrated under reduced pressure. The residue was purified by flash chromatography (SiO2, 120 g column, 0-70% siRNA / petroleum ether). The product was isolated, stirred with diethyl ether for 1 hour, filtered, and dried under vacuum to obtain 3-(5-(2-fluoro-4-(4,4,5,5-tetramethyl-1,3,2-dioxaborolan-2-yl)phenyl)-2-oxoxazole-3(2H)-yl)-1-((2-(trimethylsilyl)ethoxy)methyl)piperidine-2,6-dione (4.85 g, 63%) as a white solid. LC-MS (Method A): Retention time 2.16 minutes, [MH] + 545.2; 1 H NMR (chloroform-d, 300MHz)δ 7.5-7.6(m,2H), 7.44(d,1H,J=11.7Hz), 6.90(d,1H,J=2.3Hz), 5.17(d,2H,J=12.5Hz), 4.82(dd,1H,J=5.9, 12.7Hz) , 3.56(t,2H, J=8.1Hz), 2.9-3.1(m,1H), 2.7-2.9(m,1H), 2.31(brs,2H), 1.28(s,12H), 0.8-0.9(m,2H), -0.07(s,9H)

[0230] Example 1 3-(5-(4-(6-amino-5-(difluoromethoxy)-4-methylpyridine-2-yl)-2-fluorophenyl)-2-oxoxazole-3(2H)-yl)piperidine-2,6-dione [Chemical]

[0231] Production of Intermediate 1A: 2,6-Dichloro-4-methylpyridin-3-ol [Chemical]

[0232] To a stirred solution of 4-methylpyridin-3-ol (12.5 g, 115 mmol) in acetonitrile (250 mL) was added NCS (33.6 g, 252 mmol) portionwise at room temperature. The reaction mixture was stirred at the same temperature for 16 h, filtered through celite, and concentrated under vacuum. The crude product was purified by flash chromatography (SiO2, 220 g column, 0-50% EtOAc / petroleum ether) to afford 2,6-dichloro-4-methylpyridin-3-ol (10 g, 49%) as an off-white solid. LCMS (Method A): retention time 1.07 min, [M+H] + 177.9; 1 H NMR (chloroform-d, 300 MHz) δ 7.08 (s, 1H), 5.54 (brs, 1H), 2.31 (s, 3H)

[0233] Production of Intermediate 1B: 2,6-Dichloro-3-(difluoromethoxy)-4-methylpyridine [Chemical]

[0234] To a stirred solution of 2,6-dichloro-4-methylpyridine-3-ol (7.0 g, 39.3 mmol) in DMF (70 mL), potassium carbonate (13.59 g, 98 mmol) was added, followed by sodium chlorodifluoroacetate (6.59 g, 43.3 mmol). The reaction mixture was stirred at 100 °C for 2 hours, cooled to room temperature, and partitioned between water and ethyl acetate. The layers were separated, and the aqueous layer was extracted with ethyl acetate (2x). The organic phases were combined, washed with brine, dried on anhydrous sodium sulfate, filtered, and concentrated under vacuum to obtain the crude material. This was purified by flash chromatography (SiO2, 220 g column, 0-20% ethyl acetate / petroleum ether) to obtain 2,6-dichloro-3-(difluoromethoxy)-4-methylpyridine (4.67 g, 52%) as a white amorphous solid. LCMS (Method A): Retention time 1.69 minutes, [M+H] + 227.9; 1 ¹H NMR (chloroform-d, 300MHz): δ 7.20 (s, 1H), 6.57 (brt, 1H, J=74.0Hz), 2.39 (s, 3H)

[0235] Preparation of intermediate 1C:6-chloro-3-(difluoromethoxy)-N-(2,4-dimethoxybenzyl)-4-methylpyridine-2-amine [ka]

[0236] To a stirred solution of 2,6-dichloro-3-(difluoromethoxy)-4-methylpyridine (2.2 g, 9.65 mmol) in NMP (8 mL), (2,4-dimethoxyphenyl)methanamine (1.936 g, 11.6 mmol) and DIPEA (3.37 mL, 19.3 mmol) were added at room temperature. The reaction mixture was heated in a microwave reactor at 170 °C for 4 hours. The reaction mixture was cooled to room temperature, poured into ice water, stirred for 5 minutes, and extracted with ELISA (30 mL x 3). The organic phases were combined, washed with 1N HCl solution and brine, dried over sodium sulfate, filtered, and concentrated under vacuum. The residue was first subjected to flash chromatography (SiO2, 40g column, 0-20% siRNA / petroleum ether), and then purified by reverse-phase flash chromatography (reverse-phase flash method: sample load: 6g; column size: C18-750g (Silicycle); buffer: neutral ammonium acetate; modifier: acetonitrile) to obtain 6-chloro-3-(difluoromethoxy)-N-(2,4-dimethoxybenzyl)-4-methylpyridine-2-amine (1.6g, 46%) as an off-white solid. LCMS (Method A): retention time 1.92 minutes, [M+H] + 359.0; 1 H NMR (400MHz, DMSO-d6) δ=7.05(d,1H,J=8Hz), 6.96(t,1H,J=76Hz), 6.75(brt,J=5.8Hz, 1H), 6.56(s,1H), 6.53(s,1H), 6.49-6.41(m,J=8.0Hz, 1H), 4.39(brd,J=5.5Hz, 2H), 3.74(s,3H), 3.32(s,3H), 2.17(s,3H)

[0237] Preparation of intermediate 1D: 3-(5-(4-(5-(difluoromethoxy)-6-((2,4-dimethoxybenzyl)amino)-4-methylpyridine-2-yl)-2-fluorophenyl)-2-oxoxazole-3(2H)-yl)-1-((2-(trimethylsilyl)ethoxy)methyl)piperidine-2,6-dione [ka]

[0238] To a stirred solution of 6-chloro-3-(difluoromethoxy)-N-(2,4-dimethoxybenzyl)-4-methylpyridine-2-amine (1.0 g, 2.79 mmol) in dioxane (14 mL) and water (0.1 mL), 3-(5-(2-fluoro-4-(4,4,5,5-tetramethyl-1,3,2-dioxaborolan-2-yl)phenyl)-2-oxoxazole-3(2H)-yl)-1-((2-(trimethylsilyl)ethoxy)methyl)piperidine-2,6-dione (1.385 g, 2.53 mmol), followed by cesium carbonate (1.24 g, 3.80 mmol), was added at room temperature. The reaction mixture was purged with argon for 10 minutes, CatacXium Pd G3 (92 mg, 0.131 mmol) was added under argon, and the reaction mixture was heated at 75°C for 2 hours. The reaction mixture was cooled to room temperature, diluted with ethyl acetate, dried on anhydrous Na2SO4, and filtered through a Celite pad. The filtrate was concentrated under reduced pressure, and the residue was purified by flash chromatography (SiO2, 80 g column, 0-3% MeOH / DCM) to obtain 3-(5-(4-(5-(difluoromethoxy)-6-((2,4-dimethoxybenzyl)amino)-4-methylpyridine-2-yl)-2-fluorophenyl)-2-oxoxazole-3(2H)-yl)-1-((2-(trimethylsilyl)ethoxy)methyl)piperidine-2,6-dione (1.1 g, 58%) as a pale amorphous solid. LCMS (Method A): Retention time 2.20 minutes, [M+H] + 743.3; 1H NMR (400MHz, DMSO-d6)δ 7.95-7.85(m,2H), 7.64-7.58(m,2H), 7.21-7.16(m,2H), 6.99(t,1H,J=76Hz), 6.64-6.5 5(m,2H), 6.43(dd,J=2.3, 8.3Hz, 1H), 5.27-5.15(m,1H), 5.12-5.04(m,2H), 4.56(d,J=6 .0Hz, 2H), 3.86(s,3H), 3.71(s,3H), 3.61-3.49(m,2H), 3.10-2.99(m,1H), 2.87-2.77(m ,1H), 2.66-2.53(m,1H), 2.25(s,3H), 2.22-2.14(m,1H), 0.92-0.82(m,2H), -0.01(s,9H)

[0239] Preparation of intermediate 1E: 3-(5-(4-(6-amino-5-(difluoromethoxy)-4-methylpyridine-2-yl)-2-fluorophenyl)-2-oxoxazole-3(2H)-yl)-1-(hydroxymethyl)piperidine-2,6-dione [ka]

[0240] To a stirred solution of 3-(5-(4-(5-(difluoromethoxy)-6-((2,4-dimethoxybenzyl)amino)-4-methylpyridine-2-yl)-2-fluorophenyl)-2-oxoxazole-3(2H)-yl)-1-((2-(trimethylsilyl)ethoxy)methyl)piperidine-2,6-dione (7.0 g, 9.42 mmol) in anhydrous DCM (60 ml), TFA (14.5 ml, 188 mmol) was added under nitrogen at 0°C. The reaction mixture was slowly heated to room temperature and stirred for 2 hours. The reaction mixture was concentrated under reduced pressure and co-evaporated with DME (3 times). The residue was stirred with diethyl ether at room temperature for 30 minutes, the solid was separated, filtered, washed with diethyl ether, and dried under vacuum to obtain 3-(5-(4-(6-amino-5-(difluoromethoxy)-4-methylpyridine-2-yl)-2-fluorophenyl)-2-oxoxazole-3(2H)-yl)-1-(hydroxymethyl)piperidine-2,6-dione, which was used directly in the next step. LCMS (Method A): Retention time 1.51 minutes, [M+H] + 493.1

[0241] Example 1: To a stirred solution of 3-(5-(4-(6-amino-5-(difluoromethoxy)-4-methylpyridine-2-yl)-2-fluorophenyl)-2-oxoxazole-3(2H)-yl)-1-(hydroxymethyl)piperidine-2,6-dione (492 mg, 1.0 mmol) in DME (5 mL), N,N'-dimethylethylenediamine (0.538 mL, 5.00 mmol) was added at 0°C. The reaction mixture was heated to room temperature and stirred for 1 hour. The reaction mixture was cooled to 0°C. Acetic acid (0.572 mL, 10 mmol) was added to stop the reaction. The reaction mixture was concentrated under reduced pressure. The residue was dissolved in DMSO and purified by reverse-phase preparative HPLC (column: SunFire C18 (150mm x 19mm ID, 5μm), mobile phase A = 10mM ammonium acetate in water, mobile phase B = acetonitrile, flow rate 20 mL / min, gradient -0 min: 40% B, 15 min: 50% B, 15.1 min: 100% B) to obtain 3-(5-(4-(6-amino-5-(difluoromethoxy)-4-methylpyridine-2-yl)-2-fluorophenyl)-2-oxoxazole-3(2H)-yl)piperidine-2,6-dione (110 mg, 23.8%) as a white solid. LC-MS (Method B): Retention time 1.29 minutes, [M+H] + 463.15; 1 H NMR (400MHz, DMSO-d6)δ 10.90(brs,1H), 7.96-7.93(m,1H), 7.92(dd,J=1.4, 7.1Hz, 1H), 7.67-7.61(m,2H), 7.20(s,1H), 7.13-6.73(m,1H), 6.0 9(s,2H), 5.07(dd,J=5.3, 13.0Hz, 1H), 2.94-2.82(m,1H), 2.66-2.54(m,2H), 2.25(s,3H), 2.13(td,J=5.0, 10.2Hz, 1H)

[0242] Comparative example compound A 3-(2-oxo-5-phenyloxazole-3(2H)-yl)piperidine-2,6-dione [ka] 3-(2-oxo-5-phenyloxazole-3(2H)-yl)piperidine-2,6-dione is disclosed as compound number I-33 in WO2019 / 060693A1.

[0243] Preparation of intermediate 3A: tert-butyl 5-amino-5-oxo-4-((2-oxo-2-phenylethyl)amino)pentanoate [ka]

[0244] 2-Bromo-1-phenylethane-1-one (200 mg, 1.0 mmol) and 4,5-diamino-5-oxopentanoate tert-butyl·HCl (360 mg, 1.5 mmol) were added to a stirred suspension in anhydrous acetonitrile (4.5 mL) with sodium iodide (181 mg, 1.21 mmol) under argon at 0°C. The reaction mixture was stirred at the same temperature for 5 minutes. DIPEA (351 μL, 2.01 mmol) was added dropwise to the reaction mixture. The reaction mixture was stirred at 0°C for 2 hours. The reaction mixture was heated to room temperature and stirred overnight. The reaction was stopped by adding a 10% sodium bisulfite solution. The mixture was extracted with DCM (3 x 10 mL). The organic phases were combined, washed with water and brine, dried over anhydrous Na2SO4, filtered, and concentrated under vacuum to obtain tert-butyl 5-amino-5-oxo-4-((2-oxo-2-phenylethyl)amino)pentanoate (322 mg, crude). LC-MS (Method A): Retention time 1.215 minutes, [M+H] + 321.1

[0245] Preparation of intermediate 3B: tert-butyl 5-amino-5-oxo-4-(2-oxo-5-phenyloxazole-3(2H)-yl)pentanoate [ka]

[0246] To a stirred solution of tert-butyl 5-amino-5-oxo-4-((2-oxo-2-phenylethyl)amino)pentanoate (322 mg, 1.0 mmol) in anhydrous DMF (7 mL), CDI (407 mg, 2.5 mmol) and triethylamine (420 μL, 3.0 mmol) were added under argon at 0°C. The reaction mixture was slowly heated to room temperature and stirred overnight. The reaction was stopped by adding ice water. The reaction mixture was extracted with ethyl acetate (3 x 15 mL). The organic phases were combined, washed with water and brine, dried over anhydrous Na₂SO₄, filtered, and concentrated under vacuum. The residue was purified by flash chromatography (SiO2, 24g column, 0-100% siRNA / petroleum ether) to obtain tert-butyl 5-amino-5-oxo-4-(2-oxo-5-phenyloxazole-3(2H)-yl)pentanoate (155 mg, 44%). LC-MS (Method A): Retention time 1.33 minutes, [M+Na] + 369.2; 1 H NMR (DMSO-d6, 300MHz)δ 7.75(s,2H), 7.5-7.6(m,2H), 7.42(t,2H, J=7.5Hz), 7.3-7.4(m,2H), 4. 50(dd,1H,J=4.3, 10.1Hz), 2.2-2.3(m,3H), 2.0-2.1(m,1H), 1.38(s,9H)

[0247] Comparative compound A: To a stirred solution of tert-butyl 5-amino-5-oxo-4-(2-oxo-5-phenyloxazole-3(2H)-yl)pentanoate (150 mg, 0.43 mmol) in acetonitrile (3.0 mL), methanesulfonic acid (42 μL, 0.65 mmol) was added at room temperature. The reaction mixture was heated at 90°C for 1 hour, cooled to room temperature, concentrated under vacuum, and the residue was purified by reverse-phase preparative HPLC (X-Bridge Phenyl C18 (250mm x 19mm) 5μm; mobile phase A: 10mM ammonium acetate in water, mobile phase B: ACN; flow rate: 20.0 mL / min; gradient time / %B: 0 / 30, 15 / 43, 15.1 / 100) to obtain 3-(2-oxo-5-phenyloxazole-3(2H)-yl)piperidine-2,6-dione (25 mg, 21%) as a white solid. LC-MS (Method A): Retention time 1.582 minutes, [M+H] + 273.20; 1 H NMR (400MHz, DMSO-d6) δ=11.02(brs,1H), 7.70(s,1H), 7.52-7.47(m,2H), 7.47-7.42(m,2H), 7.35-7.30(m,1 H), 5.03(dd,J=5.3, 13.3Hz, 1H), 2.95-2.84(m,1H), 2.68-2.60(m,1H), 2.48-2.35(m,1H), 2.21-2.11(m,1H)

[0248] Comparative example compound B 3-(5-(4-bromo-2-fluorophenyl)-2-oxoxazole-3(2H)-yl)piperidine-2,6-dione [ka] Preparation of intermediate 4A: 3-(5-(4-bromo-2-fluorophenyl)-2-oxoxazole-3(2H)-yl)-1-(hydroxymethyl)piperidine-2,6-dione [ka]

[0249] To a stirred solution of 3-(5-(4-bromo-2-fluorophenyl)-2-oxoxazole-3(2H)-yl)-1-((2-(trimethylsilyl)ethoxy)methyl)piperidine-2,6-dione (200 mg, 0.4 mmol) in anhydrous DCM (2.0 mL), TFA (0.154 mL, 2.0 mmol) was added at room temperature. The reaction mixture was stirred at room temperature for 2 hours, concentrated under vacuum, and the residue was subjected to co-evaporation with DME (4x) to obtain 3-(5-(4-bromo-2-fluorophenyl)-2-oxoxazole-3(2H)-yl)-1-(hydroxymethyl)piperidine-2,6-dione (150 mg, 94%). LCMS (Method A): Retention time 1.37 minutes, [M+Na] + 423.1

[0250] Comparative example compound B: To a stirred solution of 3-(5-(4-bromo-2-fluorophenyl)-2-oxoxazole-3(2H)-yl)-1-(hydroxymethyl)piperidine-2,6-dione (180 mg, 0.45 mmol) in anhydrous DME (2255 μL), N,N'-dimethylethane-1,2-diamine (199 mg, 2.26 mmol) was added under nitrogen at 0°C. The reaction mixture was heated to room temperature, stirred for 30 minutes, cooled to 0°C, acidified with acetic acid (258 μL, 4.5 mmol), and then concentrated under vacuum (bath temperature < 30°C). The residue was purified by reverse-phase preparative HPLC (Method: Column: X-bridge phenyl (19 mm x 250 mm x 5 μm); Mobile phase A: 10 mM ammonium acetate in water; Mobile phase B: ACN; Flow rate: 20 mL / min; Gradient conditions (time / %B): 0 / 30, 14 / 51, 14.1 / 100) to obtain 3-(5-(4-bromo-2-fluorophenyl)-2-oxoxazole-3(2H)-yl)piperidine-2,6-dione (67 mg, 40%). LC-MS (Method A): Retention time 2.155 minutes, [MH] + 366.8; 1H NMR (400MHz, DMSO-d6) δ=11.13(brs,1H), 7.75-7.70(m,1H), 7.66(d,J=3.0Hz, 1H), 7.53(s,1H), 7.52( d,J=5.1Hz, 1H), 5.06(dd,J=5.3, 13.3Hz, 1H), 2.94-2.80(m,1H), 2.70-2.54(m,2H), 2.16-2.08(m,1H)

[0251] LCMS analysis conditions Method A: Acquity UPLC® BEH C18 (3.0 x 50 mm) 1.7 μm; Mobile phase A: 95:5 water:acetonitrile + 2.5 mM NH4OAc; Mobile phase B: 5:95 water:acetonitrile + 2.5 mM NH4OAc; Temperature: 40°C; Gradient: 20% B to 100% B over 2 minutes; Flow rate: 0.7 mL / min; Detection: MS and UV (220 nm) Method B: Column-kinetic XB-C18 (75x3mm-2.6μm); Mobile phase A: 10mM NH4COOH in water; Mobile phase B: Acetonitrile; Gradient: From 20%B to 100%B over 4.6 minutes, flow rate: 1.0 mL / min

[0252] Biological assays The pharmacological properties of the compounds of the present invention can be confirmed by many biological assays. The following exemplary biological assays were performed using the compounds of the present invention.

[0253] JURKAT cytodegradation assay Jurkat cells were seeded at a rate of 80,000 cells / well in 40 μL RPMI + 10% FBS in a 384-well cell culture plate using an acoustic dispensing technique before adding the target compound. The cell culture was incubated at 37°C and 5% CO2 for 72 hours. To facilitate analysis, the cell culture was spun at 200 rpm for 5 minutes, and the supernatant was discarded. After shaking the plate to detach the cell pellet, the cells were resuspended in 50 μL of fixation buffer (eBioscience FoxP3 buffer set 00-5523-00) at room temperature for 60 minutes. After centrifugation and discarding the supernatant, the cells were permeabilized in 50 μL of permeabilization buffer (eBioscience FoxP3 buffer set 00-5523-00) at room temperature for 10 minutes. After permeabilization, the cells were spun, and the supernatant was replaced with 20 μL of fluorescently labeled antibodies against Helios, Icarus, and Aeolus or the corresponding isotype controls in 1x permeabilization buffer (Icarus-Alexa 488 [Biolegend, catalog no. 368408, 1:50], Helios-PE [CST, catalog no. 29360, 1:50], Aeolus-Alexa 647 [Biolegend, catalog no. 371106, 1:25]). The staining reaction was incubated at room temperature for 1 hour in the dark. Then, 30 μL of 1x permeabilization buffer was added, and the cells were centrifuged, discarding the supernatant. The stained cells were resuspended in 25 μL of flow cytometry staining buffer [PBS + 0.2% bovine serum albumin (BSA)] and analyzed using an Intellicyt Ique Plus flow cytometer.

[0254] Table A-1 Jurkat cytodegradation assay: Maximum degradation observed [Table 22]

[0255] Table A-1 lists the observed maximum degradation of IKZF1, IKZF2, and IKZF3 proteins as measured in the Jurkat cytodegradation assay. The results shown in Table A-1 are rounded to two digits. In the Jurkat cytodegradation assay, a value of 100% indicates that no detectable protein remains or that the protein is completely degraded; a value of 0% indicates that no protein degradation by the test compound was detected. In the tests reported in Table A-1, Example 1 was observed to degrade the IKZF2 (Helios) protein by more than 90%. In contrast, Comparative Compound A and Comparative Compound B were observed to degrade the IKZF2 protein by less than 30%.

[0256] Table A-2 Jurkat cytodegradation assay: DC 50 * [Table 23] * DC 50 This is defined as the concentration of a compound required to reduce the level of a given protein by 50% compared to treatment with DMSO alone.

[0257] Human regulatory T cell degradation assay Cryopreserved human regulatory T cells were thawed in RPMI + 10% FBS + 20 ng / mL IL-2. After spinning at 1200 rpm for 5 minutes, the cells were resuspended in RPMI + 10% FBS + 20 ng / mL IL-2 and allowed to stand at 37°C and 5% CO2 for 3 hours. Next, 40,000 cells / well were seeded in 40 μL RPMI + 10% FBS + 20 ng / mL IL-2 in a 384-well cell culture plate using an acoustic dispensing technique (ECHO555) before adding the target compound. The cell cultures were incubated at 37°C and 5% CO2 for 20 hours. To facilitate analysis, the cell cultures were spun at 1200 rpm for 5 minutes, and the supernatant was discarded using an EL406 plate washer. After washing the cell pellet three times with 70 μL of PBS, the cell pellet was resuspended in 50 μL of near-IR viability staining solution (Life Technologies, catalog no. L34975) and incubated on light-shielded ice for 30 minutes. The cells were washed three times with 70 μL of PBS + 0.5% BSA using an EL406 plate washer. After shaking the plate to detach the cell pellet, the cells were resuspended in 50 μL of fixation buffer (eBioScience FoxP3 buffer set 00-5523-00) at room temperature for 60 minutes. After centrifugation and discarding the supernatant, the cells were permeabilized in 50 μL of permeabilization buffer (eBioScience FoxP3 buffer set 00-5523-00) at room temperature for 10 minutes. After permeabilization, the cells were spun, and the supernatant was replaced with 30 μL of fluorescently labeled antibodies against the intracellular targets Helios (Helios-APC [BioLegend, catalog no. 137222, 1:50]), Aeolus, and Icarus 1x permeabilization buffer. The staining reaction was incubated at room temperature in the dark for 1 hour. Then, 30 μL of 1x permeabilization buffer was added, and the cells were centrifuged, discarding the supernatant. The stained cells were resuspended in 30 μL of flow cytometry staining buffer (PBS + 0.5% BSA) and analyzed using an Intellicyt Ique Plus flow cytometer.

[0258] Table B-1 Human regulatory T cell assay: Maximum degradation observed [Table 24]

[0259] Table B-1 lists the observed maximum degradation of IKZF1 and IKZF2 proteins as measured in human regulatory T cell degradation assays. The results shown in Table B-1 are rounded to two digits. In human regulatory T cell degradation assays, a value of 100% indicates that no detectable protein remains or that the protein is completely degraded; a value of 0% indicates that no protein degradation by the test compound is detected. In the tests reported in Table B-1, Example 1 was observed to degrade 46% of the IKZF1 (IKZF2) protein.

[0260] Table B-2 Human regulatory T cell degradation assay: DC50 * [Table 25] * DC 50 This is defined as the concentration of a compound required to reduce the level of a given protein by 50% compared to treatment with DMSO alone.

[0261] Human Regulatory T Cell Reprogramming Assay Human CD4 + T cells, RosetteSep HumanCD4 + Fresh healthy leukocyte packs (Stemcell Technologies) were isolated using a T-cell-rich cocktail (Stemcell Technologies) and Ficol density gradient centrifugation. The leukocyte packs were diluted with an equal volume of phosphate-buffered serine (PBS [Gibco]) supplemented with 2% fetal bovine serum (FBS, VWR Lifescience) and RosetteSep HumanCD4. +The cells were incubated with a T cell-rich cocktail for 20 minutes, then layered in Ficoll-Paque Plus solution (GE Health Care). The cell-rich interface layer was harvested and washed twice with PBS supplemented with 2% FBS. Regulatory T cells were then transferred to EasySep Human CD4. + CD127 low CD25 + Regulatory T cell isolation was performed manually using a Regulatory T cell isolation kit (Stemcell Technologies) according to the manufacturer's instructions. The cells were incubated overnight in Roswell Park Memorial Institute (RPMI) 1640 medium (Gibco) supplemented with 10% FBS, Pen / Strep (Gibco), MEM-NEAA (Gibco), and sodium pyruvate (Gibco) in a humidified incubator (37°C, 5% CO2). The cells were then stained with CD4 (clone: ​​RPA-T4, Biolegend), CD25 (clone: ​​2A3, BD Biosciences), and CD127 (clone: ​​hIL-7R-M21, BD Biosciences). + CD127 low CD25 + Cells were sorted to a purity of over 95% on a BD FACS Aria Fusion sorter. The sorted cells were either used immediately or cryopreserved for downstream assays.

[0262] Fresh or frozen, FACS-sorted CD4 + CD127 low CD25 +Treg cells were cultured at 25,000–50,000 cells / well in RPMI1640 medium (Gibco) supplemented with 10% FBS, Pen / Strep (Gibco), MEM-NEAA (Gibco), and sodium pyruvate (Gibco) in 96-well round-bottom plates. Cells were stimulated with Treg Xpander beads (Thermo Fisher) in the presence of 500 U / mL recombinant human IL-2 (Proleukin) at a cell:bead ratio of 1:4. Compounds were added at titration doses, and cells were incubated at 37°C and 5% CO2 for 12–13 days. Recombinant human IL-2 and compounds were replenished every 2–3 days throughout the culture period. On day 12 or 13, before proceeding to flow cytometry staining and analysis, cells were stimulated again with phorbol 12 myristate 13-acetate (PMA) and ionomycin in the presence of the protein transport inhibitors brefelzin A and monensin (eBioscience Cell Stimulation Cocktail + Protein Transport Inhibitor, 500x, catalog no. 00-4975-93).

[0263] For flow cytometry staining, cells were washed twice with flow cytometry staining buffer (Thermo Fisher), incubated with Human Tru-staining Fc blocker (Biolegend) for 10 minutes, and then treated with eFluor780 viability dye (Thermo Fisher) and a surface marker antibody cocktail at 4°C for 30 minutes. The cells were then fixed and permeabilized by incubation with FoxP3 transcription factor staining buffer (Thermo Fisher) at 4°C for 30 minutes, according to the kit manufacturer's instructions. The cells were washed twice with the kit's Perm / Wash buffer, according to the manufacturer's instructions, and incubated overnight at 4°C with an intracellular antibody cocktail (containing antibodies specific to the transcription factors shown in Table C). Before acquisition, the cells were washed twice with Perm / Wash buffer and resuspended in flow cytometry staining buffer (Thermo Fisher). Sample acquisition and analysis were performed using a BD LSRFortessa (BD Biosciences) flow cytometer. Single-stain controls for each fluorescent dye were prepared using UltraCompeBead Compensation Beads (Thermo Fisher). Data were analyzed using FlowJo version 10 and GraphPad Prism Software.

[0264] Table C Antibodies used for flow sorting and analysis [Table 26]

[0265] Table C-1 Human regulatory T cell reprogramming assay - Maximum degradation observed [Table 27]

[0266] Table C-1 lists the observed maximum degradation of IKZF2 and IKZF4 proteins as measured in the human regulatory T cell reprogramming assay. The results shown in Table C-1 are rounded to two digits. In the human regulatory T cell reprogramming assay, a value of 100% indicates that no detectable protein remains or that the protein is completely degraded; a value of 0% indicates that no protein degradation by the test compound was detected. In the tests reported in Table D-1, Example 1 was observed to degrade 86-92% of the IKZF2 (Helios) protein and 68-71% of the IKZF4 (Eos) protein. In contrast, Comparative Compound A and Comparative Compound B were observed to degrade less than 6% of the IKZF2 (Helios) protein and less than 11% of the IKZF4 (Eos) protein.

[0267] Table C-2 Human regulatory T cell reprogramming assay: DC 50 * [Table 28] * DC 50 This is defined as the concentration of a compound required to reduce the level of a given protein by 50% compared to treatment with DMSO alone.

[0268] Human CD8 + T-cell degradation assay Cryopreserved healthy donor human peripheral blood mononuclear cells (PBMCs) were thawed and seeded at 500,000 cells / well in 96-well round-bottom plates in RPMI1640 medium (Gibco) supplemented with 10% FBS, Pen / Strep (Gibco), MEM-NEAA (Gibco), and sodium pyruvate (Gibco). The cells were treated with titrated doses of compounds at 37°C and 5% CO2, and then subjected to flow cytometry analysis.

[0269] For flow cytometry staining, the cells were washed twice with flow cytometry staining buffer (Thermo Fisher), incubated with Human Tru-staining Fc blocker (Biolegend) for 10 minutes, and then added eFluor780 viability dye (Thermo Fisher) and a surface marker antibody cocktail (containing LD-eFluor780, CD3-BUV-395, CD4-BUV805, CD8-FITC, and CD25-BV605) at 4°C for 30 minutes. The cells were then fixed and subjected to permeabilization by incubation with permeabilization buffer (eBioscience FoxP3 buffer set 00-5523-00) at 4°C for 30 minutes, according to the kit manufacturer's instructions. Cells were washed twice with the Perm / Wash buffer provided in the kit, according to the manufacturer's instructions, and incubated overnight at 4°C with an intracellular antibody cocktail (containing antibodies specific to transcription factors, i.e., Foxp3-BV421, HELIOS-PE-Cy7, EOS-PE, IKAROS-PECF594, and AIOLOS-AF647). Before acquisition, cells were washed twice with Perm / Wash buffer and resuspended in flow cytometry staining buffer (Thermo Fisher). Sample acquisition and analysis were performed using a BD LSRFortessa (BD Biosciences) flow cytometer. Single-stain controls for each fluorescent dye were prepared using UltraCompeBead Compensation Beads (Thermo Fisher). Data were analyzed using FlowJo version 10 and GraphPad Prism Software.

[0270] Table D-1 Human CD8 + T cell reprogramming assay - maximal degradation observed [Table 29]

[0271] Table D-2 Human CD8+ T cell reprogramming assay: DC 50 * [Table 30] * DC 50 This is defined as the concentration of a compound required to reduce the level of a given protein by 50% compared to treatment with DMSO alone.

[0272] Table D-1 shows human CD8 + The observed maximum degradation of IKZF1 and IKZF3 proteins, as measured in T cell reprogramming assays, is listed below. The results shown in Table D-1 are rounded to two orders of magnitude. Human CD8 + In the T cell reprogramming assay, a value of 100% indicated that no detectable protein remained or that the protein was completely degraded; a value of 0% indicated that no protein degradation by the test compound was detected. Example 1 was observed to degrade 68-69% of the IKZF1 (Ikaros) protein and 69-74% of the IKZF3 protein.

[0273] Table E Maximum decomposition observed for IKZF1-4: Comparison of compounds A and B in Example 1 and the comparative example. [Table 31] * IKZF1 and IKZF3: Human CD8 + T-cell reprogramming assay (D-1) ** IKZF2 and IKZF4: Human Regulatory T Cell Reprogramming Assay (C-1)

[0274] Example 1 was found to have particular advantages compared to Comparative Compound A and Comparative Compound B disclosed in WO2019 / 060693A1. Example 1 has the unexpected advantage of reducing the levels of the four IKZF1-4 proteins: Ikaros, Helios, Aeolus, and Eos. As shown in Tables C-1 and D-1 in the reported tests, (i) Example 1 reduced the level of IKZF1 (Ikaros) by 69% (Table D-1); (ii) Example 1 reduced the level of the IKZF2 (Helios) protein by 86-92% (Table C-1); (iii) Example 1 reduced the level of IKZF3 (Aeolus) by 69-74% (Table D-1); and (iv) Example 1 reduced the level of IKZF4 (Eos) by 68-71% (Table C-1). In contrast, in similar tests, comparative compound A and comparative compound B reduced the level of IKZF2 (Helios) protein by less than 6% (Table C-1) and the level of IKZF4 (Eos) by less than 11% (Table C-1).

[0275] The present invention satisfies the above requirements by providing compounds that are useful in reducing the levels of Ikaros, Helios, Aeolus, and Eos of four types of IKZF1-4 proteins.

Claims

1. Equation (I): 【Chemistry 1】 A compound, or its stereoisomer, tautomer, or salt, as indicated by [the symbol].

2. The compound described in claim 1, or its stereoisomer or tautomer.

3. A salt of the compound described in claim 1, or a stereoisomer or tautomer thereof.

4. A pharmaceutical salt of the compound described in claim 1, or a stereoisomer or tautomer thereof.

5. Structural formula: 【Chemistry 2】 The compound according to claim 1, or its stereoisomer, tautomer, or salt.

6. A pharmaceutical composition comprising a compound according to any one of claims 1 to 5, or a stereoisomer, tautomer, or pharmaceutically acceptable salt thereof; and a pharmaceutically acceptable carrier.

7. A pharmaceutical composition comprising the compound described in claim 2, or a stereoisomer or tautomer; and a pharmaceutically acceptable carrier.

8. A pharmaceutical composition comprising a pharmaceutically acceptable salt of the compound described in claim 4, or a stereoisomer or tautomer thereof; and a pharmaceutically acceptable carrier.

9. Use of a compound according to any one of claims 1 to 5, or a stereoisomer, tautomer, or pharmaceutically acceptable salt thereof, for the treatment of cancer.

10. The use according to claim 9, wherein the cancer is selected from colon or stomach cancer, pancreatic cancer, breast cancer, prostate cancer, lung cancer, ovarian cancer, cervical cancer, kidney cancer, head and neck cancer, lymphoma, leukemia, and melanoma.

11. A method for treating cancer in a patient, comprising administering to the patient a therapeutically effective amount of a compound according to any one of claims 1 to 5, or a stereoisomer, tautomer, or pharmaceutically acceptable salt thereof.

12. The method according to claim 11, wherein the cancer is selected from colon or stomach cancer, pancreatic cancer, breast cancer, prostate cancer, lung cancer, ovarian cancer, cervical cancer, kidney cancer, head and neck cancer, lymphoma, leukemia, and melanoma.

13. The method according to claim 11, wherein the cancer is selected from lymphoma, leukemia, and multiple myeloma.

14. The method according to claim 11, further comprising administering to a patient a therapeutically effective amount of a second agent before, simultaneously with, or after administering the compound, wherein the second agent is selected from a PD1 / PD-L1 antagonist, a CTLA4 antagonist, a chemotherapeutic agent, radiation, or an antitumor vaccine.

15. A method for treating cancer in a patient, comprising administering to the patient a therapeutically effective amount of an agent to reduce the levels of Icarus, Helios, Aeolus, and Eos proteins, wherein the agent is a compound according to any one of claims 1 to 5, or a stereoisomer, tautomer, or pharmaceutically acceptable salt thereof.

16. a) The Icarus protein is an amino acid sequence encoded by SEQ ID NO: 1, 2, 3, 4, 5, or 6; b) The helios protein is an amino acid sequence encoded by SEQ ID NO: 7, 8, 9, 10, or 11; c) The Aeolus protein is an amino acid sequence encoded by SEQ ID NOs: 12, 13, 14, 15, 16, 17, 18 or 19; and d) The Eos protein is the amino acid sequence encoded by SEQ ID NO: 20 or 21. The method according to claim 15.

17. a) Icarus protein levels decreased by at least 30%; b) Helios protein levels decreased by at least 50%; c) Aeolus protein levels decreased by at least 30%; and d) Eos protein levels decrease by at least 50% The method according to claim 15.

18. The method according to claim 15, further comprising administering to a patient a therapeutically effective amount of a second agent before, simultaneously with, or after administering the compound, wherein the second agent is selected from a PD1 / PD-L1 antagonist, a CTLA4 antagonist, a chemotherapeutic agent, radiation, or an antitumor vaccine.

19. A method for reducing the levels of Icarus, Helios, Aeolus, and Eos proteins in cells, comprising contacting the cells with a compound according to any one of claims 1 to 5, or a stereoisomer, tautomer, or pharmaceutically acceptable salt thereof.

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