RU2844734C1 - Preferential therapies for ikaros or aiolos mediated disorders - Google Patents

Abstract

FIELD: medicine.

SUBSTANCE: disclosed is use of a compound in a dose of not more than about 500 micrograms (mcg) once a day (QD) or twice a day (BID), wherein the compound is selected from the group of compounds that bind with high affinity to cereblon E3 ligase for the treatment of Ikaros and/or Aiolos mediated cancer. Also disclosed is the use of said compounds or a pharmaceutically acceptable salt thereof in a combination with a BTK inhibitor or with a proteasome inhibitor, and where the patient's blood or tissue sample has a statistically lower or higher concentration of one or more biomarkers than in a healthy individual. Methods of treating cancer are disclosed. Invention provides treatment of cancer mediated by Ikaros and/or Aiolos, using said compounds.

EFFECT: high binding affinity of said compounds for cereblon provides fast, deep and prolonged destruction of IKZF1/3 which leads to intense activity in cancer cells.

78 cl, 64 dwg, 23 tbl, 46 ex

Description

Link to related applications

This application claims priority from U.S. Provisional Patent Application No. 63/063,011 filed August 7, 2020, U.S. Provisional Patent Application No. 63/173,160 filed April 9, 2021, and U.S. Provisional Patent Application No. 63/212,463 filed June 18, 2021. The contents of these applications are incorporated herein by reference for all purposes.

Field of the invention

The present invention relates to therapeutic compositions, combinations and their use for the treatment of disorders mediated by the transcription factors Ikaros (IKZF1) and/or Aiolos (IKZF3) proteins by degrading these proteins via the ubiquitin-proteasome pathway.

Prior art of the present invention

The Ikaros family is a series of zinc finger protein transcription factors that are important for certain physiological processes, especially hematopoietic cell and lymphocyte development (see Fan, Y. and Lu, D. "The Ikaros family of zinc-finger proteins" Acta Pharmaceutica Sinica B, 2016, 6:513-521). Ikaros (IKZF1) was first discovered in 1992 (see Georgopoulos, K. et al. "Ikaros, an early lymphoid-specific transcription factor and a putative mediator for T cell commitment" Science, 1992, 258:802-812), and four more homologues were identified over the next two decades: Helios (IKZF2), Aiolos (IKZF3), Eos (IKZF4), and Pegasus (IKZF5) (see John, L.B., and Ward, A.C. "The Ikaros gene family: transcriptional regulators of hematopoiesis and immunity" Mol Immunol, 2011, 48:1272-1278). The distribution of different members of the Ikaros protein family in the body varies considerably.

Ikaros, Helios and Aiolos are found primarily in lymphoid cells and their corresponding precursors, with Ikaros additionally found in the brain and Ikaros and Helios found in erythroid cells. Eos and Pegasus are more widely distributed and are found in skeletal muscle, liver, brain, and heart (see Perdomo, J. et al. "Eos and Pegasus, two members of the Ikaros family of proteins with distinct DNA binding activities: J Biol Chem, 2000, 275:38347-38354, Schmitt, C. et al. "Aiolos and Ikaros: regulators of lymphocyte development, homeostasis and lymphoproliferation" Apoptosis, 2002, 7:277-284, Yoshida, T. and Georgopoulos, K. "Ikaros fingers on lymphocyte differentiation" Int J Hematol, 2014, 100:220-229).

Ikaros is essential for proper lymphocyte development. Deletion of the exons encoding the first three N-terminal zinc fingers results in mice lacking T cells, B cells, NK cells and their precursors. Genetic alterations in Ikaros correlate with poor treatment outcome in acute lymphoblastic leukemia (ALL). Ikaros and Aiolos are involved in the proliferation of multiple myeloma and lymphoma cells.

Multiple myeloma (MM) is a malignancy of plasma cells typically characterized by abnormal monoclonal immunoglobulin production, bone marrow involvement, renal dysfunction, immune dysfunction, and skeletal damage. In the United States, MM accounts for nearly 1.8% of all new cancer cases. Although outcomes for patients with MM have improved significantly over the past several decades, the disease remains incurable, with a currently projected five-year relative survival rate of 53.9%. Because available treatments are not curative, nearly all patients eventually progress.

Treatment of MM has historically involved chemotherapy, including alkylating agents together with corticosteroids. Treatment was further advanced in the 1980s with autologous stem cell transplantation. In the 1990s, the discovery of the efficacy of thalidomide (a first-in-class immunomodulatory imide drug, IMiD®) in myeloma led to significant changes in treatment regimens and improved patient outcomes. Subsequently approved IMiD®s, lenalidomide and pomalidomide, are now widely used to treat MM, as is the first-in-class drug thalidomide. Agents in this class bind to the E3 ligase substrate recognition adaptor protein cereblon (CRBN) and promote the degradation of Ikaros (IKZF1) and Aiolos (IKZF3), resulting in antitumor effects, tumor microenvironment targeting, and immunomodulation leading to T-cell priming and antitumor activity. Many patients with MM are treated with multiple regimens containing one of these IMiDs. These agents are currently considered the standard of care for MM in multiple lines of therapy in combination with agents including dexamethasone, anti-cluster of differentiation 38 (CD38) antibodies, and proteasome inhibitors such as bortezomib. Although these agents have been successful in prolonging progression-free survival in patients with MM, patients typically relapse with shorter progression-free survival periods following each relapse. Recent studies with new agents, belantamab and selinexor, have shown improved outcomes in multigrade refractory myeloma, leading to accelerated FDA approval, however, response rates were low (26-31%) and progression-free survival intervals were short (3.7-4.9 months), highlighting the remaining unmet medical need for these patients.

Non-Hodgkin lymphoma (NHL) is a heterogeneous group of lymphoid malignancies originating from T, B, or NK cells. It includes diffuse large B-cell lymphoma, anaplastic large cell lymphoma, Burkitt lymphoma, lymphoblastic lymphoma, mantle cell lymphoma, nodular T-cell lymphoma, follicular lymphoma, cutaneous T-cell lymphoma, lymphoplasmacytic lymphoma, marginal zone B-cell lymphoma, MALT lymphoma, and small lymphocytic lymphoma.

B-cell NHL predominates, while T-cell lymphomas are less common. Among newly diagnosed patients with aggressive NHL, chemotherapeutic regimens including cyclophosphamide, vincristine, prednisone, and daunorubicin (known as CHOP) remain the mainstay of therapy. In B-cell aggressive lymphomas, rituximab in combination with CHOP is the mainstay of therapy given to newly diagnosed patients. In some patients, particularly with T-cell NHL, initial chemotherapy is followed by autologous stem cell therapy. For relapsed and refractory patients, various targeted agents have been developed, improving treatment options for several NHL subtypes; however, these treatment options are generally not curative. Furthermore, NHL subtypes are biologically heterogeneous, limiting the development of therapeutic agents for a wide range of indications.

In newly diagnosed patients with aggressive lymphoma, initial treatment is often intensive and curative. Other than the addition of rituximab to CHOP in B-cell NHL and brentuximab to CHOP in anaplastic large cell lymphoma (ALCL), no other new targeted agent has demonstrated a survival benefit, and therefore no other drugs have been approved for treatment of previously untreated patients. Recent therapeutic advances in relapsed NHL include Bruton tyrosine kinase (BTK) inhibitors, particularly for mantle cell lymphoma (MCL) and more advanced forms of NHL, chimeric antigen receptor T-cell (CAR-T) therapies, which have been approved for diffuse large B-cell lymphoma (DBLCL) and MCL, and novel antibody-drug conjugates such as polatuzumab, belantamab, or tafasitamab, which have been approved for the treatment of DLBCL. Drugs recently approved for the treatment of T-cell NHL include romidepsin, belinostat, and brentuximab. Regarding the NHL population studied in this protocol, lenalidomide has demonstrated clinical activity in both B-cell and T-cell NHL, including MCL, DLBCL, and nodular T-cell lymphoma (PTCL). Lenalidomide has been studied in patients with relapsed/refractory (r/r) MCL and was approved by the Food and Drug Administration (FDA) in June 2013 based on results from the phase II EMERGE trial, which investigated the efficacy and safety of lenalidomide in y/y subjects with MCL following bortezomib (overall response rate [ORR] 28%, median duration of response [DOR] 16.6 months). Lenalidomide is also active in DLBCL and PTCL. While lenalidomide and other newer targeted therapies have moderate to good response rates, response duration is generally short in most NHL subtypes. Once patients relapse after 1-2 regimens, median duration of response tends to be low and relies on patients having sufficient performance status and organ function to tolerate these treatments. Thus, there remains an unmet medical need for patients with r/r NHL.

Protein degradation is a tightly regulated and essential process that maintains cellular homeostasis. Selective identification and removal of damaged, misfolded, or redundant proteins is achieved via the ubiquitin-proteasome pathway (UPP). The UPP plays a central role in the regulation of nearly all cellular processes, including antigen processing, apoptosis, organelle biogenesis, cell cycle, transcription and DNA repair, differentiation and development, immune response and inflammation, neural and muscular degeneration, morphogenesis of neural networks, modulation of cell surface receptors, ion channels, and the secretory pathway, response to stress and extracellular modulators, ribosome biogenesis, and viral infection. The covalent attachment of multiple ubiquitin molecules by E3 ubiquitin ligase to the terminal lysine residue marks the protein for proteasomal degradation, where the protein is broken down into small peptides and ultimately into its constituent amino acids, which serve as building blocks for new proteins. Defective proteasomal degradation has been linked to a variety of clinical disorders, including Alzheimer's disease, Parkinson's disease, Huntington's disease, muscular dystrophies, cardiovascular disease, and cancer, among others.

Patent applications describing certain protein disrupting agents include WO 2020/210630, WO 2020/006262, WO 2020/010227, and WO 2020/010177.

Patent applications filed by C4 Therapeutics, Inc. that describe compounds capable of binding to an E3 ubiquitin ligase and a target protein for degradation include: WO/2021/127561 entitled "Isoindolinone And Indazole Compounds For The Degradation Of EGFR", WO/2021/086785 entitled "Bifunctional Compounds", WO/2021/083949 entitled "Bifunctional Compounds for the Treatment of Cancer", WO/2020/210630 entitled "Tricyclic Degraders of Ikaros and Aiolos", WO/2020/181232 entitled "Heterocyclic Compounds for Medical Treatment", WO/2020/132561 entitled "Targeted Protein Degradation", WO/2019/236483 with the title "Spirocyclic Compounds", WO2020/051235 with the title "Compounds for the degradation of BRD9 or MTH1", WO/2019/191112 with the title "Cereblon binders for the Degradation of Ikaros", WO/2019/204354 with the title "Spirocyclic Compounds", WO/2019/099868 with the title "Degraders and Degrons for Targeted Protein Degradation", WO/2018/237026 with the title "N/O-Linked Degrons and Degronimers for Protein Degradation", WO 2017/197051 with the title "Amine-Linked C3-Glutarimide Degronimers for Target Protein" Degradation", WO 2017/197055 with the title "Heterocyclic Degronimers for Target Protein Degradation", WO 2017/197036 entitled "Spirocyclic Degronimers for Target Protein Degradation", WO 2017/197046 entitled "C3-Carbon Linked Glutarimide Degronimers for Target Protein Degradation" and WO 2017/197056 entitled "Bromodomain Targeting Degronimers for Target Protein Degradation."

The object of the present invention is to provide new compositions of matter, combinations, formulations and their uses, as well as methods for preparing compounds for the treatment of diseases mediated by Ikaros or Aiolos.

Brief summary of the present invention

Compound 1 is a small molecule anticancer agent that binds with high affinity to the cereblon E3 ligase, thereby creating a novel surface on cereblon that interacts with IKZF1 and IKZF3 (see WO 2020/210630). As a result, IKZF1 and IKZF3 are efficiently ubiquitinated by the cereblon E3 ligase and degraded by the proteasome. The high binding affinity of Compound 1 for cereblon enables rapid, deep, and sustained degradation of IKZF1/3, resulting in potent activity in cancer cells, including, for example, but not limited to, hematopoietic cancers such as multiple myeloma and multiple types of non-Hodgkin's lymphoma.

According to the present invention, it has been found that Compound 1 and other compounds described herein can be administered for the treatment of Ikaros (IKZF1) and/or Aiolos (IKZF3) mediated disorders in a low dose once or twice daily regimen, optionally with drug holidays, according to highly effective treatment regimens. For example, it has been found that anti-cancer treatment can be effective in a patient using one of the compounds described herein at a dose of no more than about 500, 450, 400, 350, 325, 300, 275, 250, 225, 200, 175, 150, 125, or even 100, 75, 50, 45, 40, 35, 30, 25, 20, 15, 10, 5, or 1 microgram (mcg) once per day (QD) or twice per day (BID). In certain embodiments, the patient is an adult (typically a human weighing at least 100 pounds or more, and sometimes even 125 or 150 pounds, such as 70 kg or more, and at least 18 years of age or more). In an alternative embodiment, the patient is a pediatric patient (and may weigh less than 100, 125, or 150 pounds and is typically less than 18 years of age). It has been found that Compound 1 is not only effective as a low dose therapy, but can also be delivered with a drug holiday, which is preferred by the patient. For example, Compound 1 can be delivered once or twice daily for 21 days, followed by a drug holiday of 7 days. Alternative dosing regimens are also useful, including, for example, but not limited to, those that provide for increasing or decreasing drug holidays of 1, 2, 3, 4, 5, 6, or 7 days.

In certain embodiments, Compound 1 or another compound described herein modulates the immune activity of a patient. For example, Compound 1 has been found to activate the proliferation of cytotoxic T cells, which may be critical for cancer therapy.

Compound 1 or another compound described herein, in a non-limiting embodiment, can be administered daily or intermittently with dexamethesone. In certain embodiments, dexamethesone or another corticosteroid or other immunosuppressant or anti-inflammatory agent is administered every day without a drug holiday for a 28-day cycle. In other embodiments, dexamethesone or another corticosteroid or other immunosuppressant or anti-inflammatory agent is administered with a drug holiday that can be the same as or different from the drug holiday for Compound 1 or another compound described herein.

According to one aspect of the present invention, the Ikaros (IKZF1) and/or Aiolos (IKZF3) degrading compound at a low dose is Compound 1 or a pharmaceutically acceptable salt thereof.

Compound 1 has high binding affinity for cereblon (dissociation constant K _d = 0.9 nM). Compound 1 promotes degradation of >75% of IKZF1 at steady state in multiple myeloma cells within 1.5 hours at 0.3 nM. The high binding affinity and ability to catalyze degradation of Compound 1 provide potent growth inhibition in both naive NCIH929 multiple myeloma cell lines (maximum growth inhibition 96%, mean IC ₅₀ 0.071 nM) and in NCIH929 cells resistant to both lenalidomide and pomalidomide (maximum growth inhibition 70%, mean IC ₅₀ 2.3 nM).

The compounds described herein can be used to treat cancer cells that are resistant, refractory, or unresponsive to standard therapy for IKZF1 or IKZF3 mediated cancer, including IMiDs, including pomalidomide, or any of those described in the background or detailed disclosure section.

As shown in Example 12, Compound 1 demonstrated potent anticancer activity in a panel of multiple myeloma cell lines (eight out of twelve showed a response, average IC _{50 of} 0.3 nM among responding lines). The anticancer activity of Compound 1 was also demonstrated in several cell line models of different subtypes of non-Hodgkin's lymphoma, including mantle cell lymphoma (MCL) (four of six lines tested responded, mean IC50 _of 13 nM among responding lines), diffuse large B-cell lymphoma (DLBCL) (six of eleven germinal center B-cell lymphoma DLBCL lines and three of six activated B-cell DLBCL lines responded, with mean IC50 _of 12 nM and 1.6 nM, respectively, among responding lines), anaplastic large cell lymphoma (ALCL) (four of six cell lines responded, mean _{IC50 of} 1.7 nM among responding lines), and cutaneous T-cell lymphoma (CTCL) (three of four cell lines tested responded, mean _{IC50 of} 30 nM among responding lines). In mouse tumor xenograft models, Compound 1 demonstrated dose-dependent efficacy at doses ranging from 3 mcg/kg/day to 100 mcg/kg/day (see Example 13). For several tumor xenografts tested, daily administration of Compound 1 at doses ranging from 30 mcg/kg/day to 100 mcg/kg/day resulted in robust tumor regression. As shown in Fig. 45A, Fig. 45B, Fig. 45C, Fig. 52, and others, Compound 1 is more than 100-fold more potent than pomalidomide in various cancer studies.

In mouse xenograft models, Compound 1 has measurable plasma and tumor concentrations for a longer time than CC-92480 (currently in human clinical trials by Celgene, a subsidiary of Bristol Myers Squibb), despite being administered at a 10-fold lower dose. It also takes significantly longer for IKZF3 levels to recover after treatment with Compound 1 than with pomalidomide or CC-92480. For example, in the NCI-H929 tumor model, it takes more than 48 hours for IKZF3 levels to reach 50% of their pre-treatment levels following administration of Compound 1, whereas both CC-92480 (10-fold higher dose) and pomalidomide (30-fold higher dose) result in pre-treatment IKZF3 levels being achieved within 48 hours post-treatment.

As a result of the marked efficacy of Compound 1 and other compounds described herein, novel preferred treatments for Ikaros (IKZF1) and/or Aiolos (IKZF3) mediated disorders have been identified. In non-limiting embodiments of the present invention, the Ikaros (IKZF1) and/or Aiolos (IKZF3) depleting compounds described herein can be used, according to the following non-limiting examples:

1. Treatment of a disorder mediated by Ikaros (IKZF1) and/or Aiolos (IKZF3), wherein the compound is administered at a low dose. For example, the dose is no more than about 500, 450, 400, 350, 325, 300, 275, 250, 225, 200, 175, 150, 125, 100, 95, 90, 85, 80, 75, 70, 65, 60, 55, 50, 45, 40, 35, 30, 25, 20, 15, 10, 5, or 1 microgram (mcg) once daily (QD) or twice daily (BID), optionally with drug holidays.

2. Treatment of a disorder mediated by Ikaros (IKZF1) and/or Aiolos (IKZF3), wherein a compound described herein is administered in an effective amount according to a dosing regimen that includes drug holidays, such as holidays of 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 11, 12, 13 or 14 days over a 28 day treatment cycle.

3. Treatment of an Ikaros (IKZF1) and/or Aiolos (IKZF3)-mediated disorder, wherein a blood or tissue sample is first collected from the patient and the concentration of one or more biomarkers is determined, such as a marker of anti-tumor immunity (e.g., a tumor infiltrating lymphocyte cytokine, T cell activation and/or proliferation, or a B cell marker such as BCMA or M protein, or combinations thereof), an apoptosis marker (e.g., total and/or cleavage caspase-1, caspase-3, caspase-7, PARP, BIM, or survivin, or combinations thereof), or a zinc finger protein (e.g., IKZF1, IKZF3, ZFP91, WIZ, or SALL4, or combinations thereof), wherein if the patient exhibits a statistically different concentration of the biomarker, including but not limited to about 5, 10, 15, or 20% different than the concentration of a healthy individual, then Compound 1 or a pharmaceutically acceptable salt thereof or another compound described herein is administered to the patient.

4. Treatment of a disorder mediated by Ikaros (IKZF1) and/or Aiolos (IKZF3), wherein a blood or tissue sample is first taken from a patient and the concentration of one or more biomarkers, such as IRF-1, caspase-3, IL-2 and/or IFN-γ, is determined, and wherein if the patient has a statistically lower concentration of the biomarker than a healthy individual, including without limitation up to about 5, 10, 15 or 20% lower, then Compound 1 or a pharmaceutically acceptable salt thereof or another compound described herein is administered to the patient.

5. Treatment of a disorder mediated by Ikaros (IKZF1) and/or Aiolos (IKZF3), wherein a blood or tissue sample is first taken from a patient and the concentration of one or more biomarkers, such as cyclin D1, E2F1, ZFP91, SALL4, IRF-4, BLIMP1 and/or MYC, is determined, and wherein if the patient has a statistically higher concentration of the biomarker, including without limitation up to about 5, 10, 15 or 20% higher than a healthy individual, then Compound 1 or a pharmaceutically acceptable salt thereof or another compound described herein is administered to the patient.

6. Treatment of a disorder mediated by Ikaros (IKZF1) and/or Aiolos (IKZF3), wherein Compound 1 or a pharmaceutically acceptable salt thereof or another compound described herein is administered to a patient, and then a blood or tissue sample is taken from the patient and the concentration of one or more biomarkers, such as IRF-1, caspase-3, IL-2 and/or IFN-γ, is determined, wherein if the concentration of the biomarker is slightly increased, such as by at least about 1.25, 1.5, 1.75 or 2-fold, then the dose of the compound is increased. Treatment of a disorder mediated by Ikaros (IKZF1) and/or Aiolos (IKZF3), wherein Compound 1 or a pharmaceutically acceptable salt thereof or another compound described herein is administered to a patient, and then a blood or tissue sample is taken from the patient and the concentration of one or more biomarkers, such as cyclin D1, E2F1, ZFP91, SALL4, IRF-4, BLIMP 1 and/or MYC, is determined, wherein if the concentration of the biomarker is slightly decreased, such as by about 1.25, 1.5, 1.75 or 2-fold, then the dose of the compound is increased.

7. Treatment of a disorder mediated by Ikaros (IKZF1) and/or Aiolos (IKZF3), wherein Compound 1 or a pharmaceutically acceptable salt thereof or another compound described herein is administered to a patient, and then a blood or tissue sample is collected from the patient and the concentration of one or more biomarkers, such as a marker of anti-tumor immunity (e.g., a cytokine, tumor infiltrating lymphocyte, T cell activation and/or proliferation, or a B cell marker such as BCMA or M protein, or combinations thereof), an apoptosis marker (e.g., total and/or cleavage caspase-1, caspase-3, caspase-7, PARP, BIM, or survivin, or combinations thereof), or a zinc finger protein (e.g., IKZF1, IKZF3, ZFP91, WIZ, or SALL4, or combinations thereof), is determined, wherein if the concentration of the biomarker is not significantly altered, such as by at least about 1.25, 1.5, 1.75 or 2 times, then the dose of the compound is increased.

8. Treatment of activated diffuse large B-cell lymphoma, diffuse large germinal center B-cell lymphoma, extranodal natural killer (NK) T-cell lymphoma, or extranodal T-cell lymphoma with an effective amount of a compound described herein or a pharmaceutically acceptable salt thereof.

9. Treatment of a disorder mediated by Ikaros (IKZF1) and/or Aiolos (IKZF3), wherein the compound is administered at a low dose. For example, the dose is no more than about 500, 450, 400, 350, 325, 300, 275, 250, 225, 200, 175, 150, 125 or even 100, 75, 50 or 25 micrograms (mcg) once daily (QD) or twice daily (BID), optionally with drug holidays.

10. Treatment of a disorder mediated by Ikaros (IKZF1) and/or Aiolos (IKZF3), wherein a compound as described herein is used in combination with a BTK inhibitor, such as a BTK inhibitor selected from acalabrutinib, spebrutinib, zanubrutinib, LOXO-305, evobrutinib, TG-1701, tolebrutinib, BIIB091, DZD-9008, HZ-A-018, orelabrutinib, AC0058TA, SN1011, rilzabrutinib, ARQ 531, DTRMWXHS-12, JNJ-64264681, branebrutinib, ibrutinib and fenebrutinib.

11. Treatment of a disorder mediated by Ikaros (IKZF1) and/or Aiolos (IKZF3), wherein the compound described herein is used in combination with an anti-CD38 antibody, such as an anti-CD38 antibody selected from felzartamab, daratumumab, GBR 1342, TAK-573, CID-103, OKT10, STI-6129, SGX301, TAK-079 and mesagitamab.

12. Treatment of a disorder mediated by Ikaros (IKZF1) and/or Aiolos (IKZF3), wherein a compound as described herein is used in combination with a proteasome inhibitor, such as a proteasome inhibitor selected from bortezomib, carfilzomib, ixazomib citrate, oprozomib, delanozomib, lactacystin, epoxomicin, MG132, MG-262, CEP-18770, NEOSH101, TQB3602, and KZR-616.

13. Treatment of a disorder mediated by Ikaros (IKZF1) and/or Aiolos (IKZF3), wherein the compound described herein is used in combination with an IMiD, for example, an IMiD selected from thalidomide, pomalidomide, lenalidomide, ibertomide, CC-92480, CC-90009 and CC-99282.

14. Treatment of a disorder mediated by Ikaros (IKZF1) and/or Aiolos (IKZF3), wherein a compound as described herein is used in combination with an HDAC inhibitor, e.g., an HDAC inhibitor selected from trapoxin B, sodium phenylbutyrate, tacedinalin, mocetinostat, BRD73954, BG45, domatinostat, cay10603, HPOV, TMP269, nexturastat A, Santacruzamate A, splitomycin, LMK-235, sodium butyrate, pivaloyloxymethylbutyrate, pyroxamide, abexinostat, resminostat, givinostat, quisinostat, Psammapline A, KD5170, 1-Alanine chlamidocin, depudecin, panobinostat, ricolinostat, vorinostat and CUDC-101.

15. Treatment of a disorder mediated by Ikaros (IKZF1) and/or Aiolos (IKZF3), wherein a compound as described herein is used in combination with another compound, such as a compound selected from selinexor, oxafenamide, belantamab mafodotin, denosumab, zoledronic acid, plerixafor, eltrombopag, ipilimumab, palbociclib, ricolinostat, afuresertib, dinaciclib, filanesib, indatuximab ravtansine, masitinib, sonidegib, sotatercept, ulocuplumab and urelumab.

16. Treatment of Ikaros (IKZF1) and/or Aiolos (IKZF3) mediated cancer comprising administering an effective amount of Compound 1 to activate T cell proliferation.

These new treatments offer advantages over currently approved cancer treatments. For example, a compound described herein can be administered at a lower dose than first-generation IMiDs, can penetrate the blood-brain barrier to treat cancers involving the central nervous system (CNS), such as lymphoma involving the CNS, can treat cancers that have relapsed or are refractory to standard treatment regimens, including first-generation IMiDs, and/or can provide longer progression-free survival than first-generation IMiDs, such as thalidomide, pomalidomide, and lenalidomide. In certain embodiments, a compound described herein is about 30, 40, 50, 60, 70, 80, 90, 100, 500, or even 1000 times more potent in vivo than thalidomide, pomalidomide, lenalidomide, or CC-92480. In certain embodiments, a compound described herein causes sustained degradation of IKZF1 and/or IKZF3 (e.g., IKZF1 and/or IKZF3 levels require 12 hours, 18 hours, 24 hours, 36 hours, 48 hours, or more to return to pre-treatment levels). This sustained degradation is a result of high in vivo potency, metabolic stability, and/or selectivity.

According to one aspect, the compound used for the treatments described herein is selected from:

or a pharmaceutically acceptable salt thereof.

The compound described herein modulates the activity of the immune system, for example by activating IFN-alpha, IFN-beta or IFN-gamma. By activating the immune system, the compound may lead to more effective cancer treatment. This immunomodulatory activity increases the effectiveness of the compound described herein with another anticancer agent, such as daratumumab.

In certain embodiments, the destruction of IKZF1 and IKZF3 from malignant B or T cells results in tumor cell death, and their removal from the tumor microenvironment results in T cell activation.

The compound may be, for example, intended for oral, parenteral or local delivery. According to certain embodiments, the compound is presented as a solid, gel or liquid dosage form for oral administration or can be administered intravenously. According to some embodiments, the selected compound is presented as a soft-shell capsule or a solid dosage form of a tablet for oral administration. According to certain embodiments, the dose of the pharmaceutical composition is about 1 mcg, 5 mcg, 10 mcg, 15 mcg, 20 mcg, 25 mcg, 50 mcg, 75 mcg, 100 mcg, 125 mcg, 150 mcg, 175 mcg, 200 mcg, 225 mcg, 250 mcg, 275 mcg, 300 mcg, 325 mcg, 350 mcg, 375 mcg, 400 mcg, 425 mcg, 450 mcg, 475 mcg, 500 mcg, 525 mcg, 550 mcg, 575 mcg, 600 mcg, 625 mcg, 650 mcg, 675 mcg, 700 mcg, 725 mcg, 750 mcg, 775 mcg, or 800 mcg, which, according to one non-limiting aspect, can be administered once daily (QD) or twice daily (BID) on days 1-21 of a 28-day cycle.

The compound may be, for example, intended for oral or parenteral delivery. In certain embodiments, the compound is provided as a solid, gel or liquid dosage form for oral administration or may be administered intravenously. In some embodiments, the selected compound is provided as a soft-shell capsule or a solid tablet dosage form for oral administration. In certain embodiments, the dose of the solid or gel dosage form is 25 mcg, 50 mcg, 100 mcg, 200 mcg, 300 mcg or 400 mcg, which in one non-limiting aspect may be administered once daily (QD) on days 1-21 of a 28-day cycle.

According to a general aspect, the compound used for the treatments described herein is Compound 1 or a pharmaceutically acceptable salt thereof.

In certain embodiments, any of the compounds described herein has at least one desired isotopic substitution of an atom in an amount close to the natural abundance of the isotope, i.e., enriched. In certain embodiments, the compound includes a deuterium atom or multiple deuterium atoms.

Other features and advantages of the present invention will be apparent from the following detailed description and claims.

Brief description of the drawings

Fig. 1A is a dose response curve describing the effect of Compound 1 on H929 cell viability compared to pomalidomide as described in Example 9. The x-axis represents the concentration of Compound 1 or pomalidomide in nM and the y-axis represents the viability of H929 cells at 96 hours.

Fig. 1B shows a dose response curve describing the effect of Compound 1 on IKZF1 degradation compared to pomalidomide as described in Example 9. The x-axis represents the concentration of Compound 1 or pomalidomide in nM and the y-axis represents the % of IKZF1 remaining after 1.5 hours.

Figure 2A shows a scatter plot of tumor lysates analyzed by multiplex quantitative proteomics from mice treated with Compound 1 for 4 hours. Tumor lysates were analyzed by multiplex quantitative proteomics, and fold changes in relative abundance comparing Compound 1 treatment to DMSO control treatment are shown in the scatter plot. Log2 fold changes are shown on the x-axis, and negative Log10 adjusted P values are shown on the y-axis. The horizontal dotted line indicates statistical significance (adjusted P ≤ 0.01), and the vertical line indicates fold change ≥ 2. IKZF1 and IKZF3 were the only proteins significantly down-regulated, with fold changes of 5.5 and 4.1, respectively. The data shown are from biological replicates measured in a single 10-plex TMT experiment, in which a total of 7903 proteins were quantified. P values were derived from moderated t-statistics and adjusted for multiple hypothesis testing using the Benjamini-Hochberg approach. The experimental procedure is presented in Example 10.

Figure 2B shows a scatter plot of tumor lysates analyzed by multiplex quantitative proteomics from mice treated with Compound 1 for 24 hours. Tumor lysates were analyzed by multiplex quantitative proteomics, and fold changes in relative abundance comparing Compound 1 treatment to DMSO control treatment are shown in the scatter plot. Log2 fold changes are shown on the x-axis, and negative Log10 corrected P values are shown on the y-axis. The horizontal dotted line indicates statistical significance (corrected P ≤ 0.01), and the vertical line indicates fold change ≥ 2. The experimental procedure is presented in Example 10.

Figure 3 shows a dose response curve describing the effect of Compound 1, with or without bortezomib (a proteasome inhibitor) or MLN-4924 (a neddylation inhibitor), on the destruction of Ikaros. As described in Example 11, Ki-JK cells were exposed to Compound 1 with or without bortezomib or MLN-4924, and the concentration of IKZF1 was determined by flow cytometry at 1.5, 3, and 6 hours for Compound 1 and at 6 hours for the combination of Compound 1 and bortezomib or MLN-4924. The x-axis shows the concentration of Compound 1 alone or in combination with bortezomib or MLN-4924. The y-axis shows the remaining concentration of IKZF1-488 measured as a percentage relative to DMSO.

Fig. 4 shows a graph of the effect of Compound 1 in mice bearing NCI-H929 multiple myeloma cells as described in Example 13. Compound 1 was administered orally (PO) every day (QD) at four different concentrations (3 μg/kg, 10 μg/kg, 30 μg/kg, and 100 μg/kg) and compared to pomalidomide. The x-axis represents time measured in days and the y-axis represents NCI-H929 tumor volume measured in mm ³ .

Fig. 5 shows a graph of the effect of Compound 1 in mice bearing RPMI-8226 multiple myeloma cells as described in Example 13. Compound 1 was administered orally (PO) every day (QD) at four different concentrations (3 μg/kg, 10 μg/kg, 30 μg/kg, and 100 μg/kg) and compared to pomalidomide. The x-axis shows time measured in days and the y-axis shows RPMI-8226 tumor volume measured in mm ³ .

Figure 6 shows images comparing mice treated with Compound 1 and vehicle. Mice were imaged using IVIS Lumina II as described in Example 14.

Fig. 7 shows a graph of the bioluminescence signal of mice administered Compound 1 orally (PO) every day (QD) or vehicle and visualized with IVIS Lumina II as described in Example 14. The x-axis shows time measured in days and the y-axis shows the systemic BLI of MM1S-Luc (total bioluminescence signal) measured in photons/10 ⁶ .

Fig. 8 is a graph of tumor volume in mice injected with pomalidomide-resistant NCI-H929 cells and treated with Compound 1 (100 μg/kg) or pomalidomide (3000 μg/kg) orally (PO) every day (QD) as described in Example 15. The x-axis represents time measured in days and the y-axis represents tumor volume measured in mm ³ .

Fig. 9 is a graph of tumor volume in mice injected with the refractory multiple myeloma cell line RPMI-8226. As described in Example 15, mice were injected with RPMI-8226 tumors and the tumors were allowed to grow to a volume of 109-158 mm ³ (28 days after implantation). Vehicle-treated animals continued treatment until tumors reached an MTV (mean tumor volume) of 2211 mm ³ . Pomalidomide-treated animals were switched to Compound 1 (100 μg/kg/day) on day 17 for 21 additional days. The x-axis represents time in days and the y-axis represents tumor volume in mm ³ .

Fig. 10 is a graph of tumor volume in mice injected with REC1 mantle cell lymphoma cells and treated with Compound 1 or pomalidomide as described in Example 16. Compound 1 was administered at four different concentrations (3 μg/kg, 10 μg/kg, 30 μg/kg, or 100 μg/kg) and pomalidomide was administered at a dose of 3000 μg/kg. Both agents were administered orally (PO) every day (QD). The x-axis represents time measured in days and the y-axis represents tumor volume measured in mm ³ .

Fig. 11 is a graph of tumor volume in mice inoculated with DLBCL TMD8 lymphoma and administered Compound 1 (100 μg/kg) or pomalidomide (3000 μg/kg) orally (PO) every day (QD) as described in Example 16. The x-axis represents time measured in days and the y-axis represents tumor volume measured in mm ³ .

Fig. 12 is a graph of tumor volume in mice bearing ALCL KI-JK tumors and treated with Compound 1 at either 30 μg/kg or 100 μg/kg PO QD, pomalidomide (3000 μg/kg, QD PO), or CC-92480 (1000 μg/kg, QD PO), as described in Example 17. Mice were dosed for 21 days. The x-axis is time measured in days, and the y-axis is tumor volume measured in mm ³ .

Fig. 13A is a bar graph of IKZF1 protein expression in mice inoculated with ALCL KI-JK tumors and treated with Compound 1 or pomalidomide as described in Example 17. Mice were sacrificed and tumors were collected at 6 and 24 hours after a single dose. The x-axis represents the time after dosing when the animal was sacrificed, and the y-axis represents the percentage of IKZF1 protein compared to vehicle control, normalized to GAPDH.

Figure 13B is a bar graph of IRF4 protein expression in mice bearing ALCL KI-JK tumors and treated with Compound 1 or pomalidomide as described in Example 17. Mice were sacrificed and tumors were collected at 6 and 24 hours after a single dose. The x-axis represents the time after dosing when the animal was sacrificed, and the y-axis represents the percentage of IRF4 protein compared to vehicle control, normalized to GAPDH.

Fig. 14 is a graph of tumor volume in mice inoculated with ALCL DL-40 tumors and administered Compound 1, pomalidomide, or CC-92480 as described in Example 18. Compound 1 was administered at three different concentrations (10 μg/kg, 30 μg/kg, or 100 μg/kg) orally (PO) every day (QD). Pomalidomide was administered at 3000 μg/kg orally every day. CC-92480 was administered at 300 μg/kg orally every day. The x-axis represents time measured in days and the y-axis represents tumor volume measured in mm ³ .

Figure 15A is a graph of IKZF1 protein expression in mice inoculated with ALCL DL-40 tumors and treated with Compound 1 or pomalidomide as described in Example 18. Mice were sacrificed and tumors were collected at 1, 4, and 24 hours after a single dose. The x-axis represents the time after dosing that the animal was sacrificed, and the y-axis represents the percentage of IKZF1 protein compared to vehicle control, normalized to GAPDH.

Fig. 15B shows a Western blot measuring the concentration of IKZF1 and IKZF3 in mice inoculated with ALCL DL-40 tumors and administered Compound 1 or pomalidomide as described in Example 18. The concentration was determined when mice were sacrificed at 1, 4, and 24 hours after a single dose.

Fig. 16 is a graph of tumor volume in mice inoculated with ALCL DL-40 tumors and administered Compound 1 or pomalidomide as described in Example 19. Compound 1 was administered at five different concentrations (3 μg/kg, 10 μg/kg, 30 μg/kg, 100 μg/kg, or 300 μg/kg) orally (PO) every day (QD). Pomalidomide was administered at a dose of 3000 μg/kg orally every day. The x-axis represents time measured in days and the y-axis represents tumor volume measured in mm ³ .

Fig. 17 is a graph of tumor volume in mice inoculated with ALCL DL-40 tumors and treated with Compound 1 as described in Example 19. Compound 1 was administered at three different concentrations (30 μg/kg, 100 μg/kg, or 300 μg/kg) orally (PO) every day (QD). The x-axis represents time in days and the y-axis represents change in body weight in percent.

Fig. 18 is a graph of tumor volume in mice inoculated with ALCL KI-JK tumors and treated with Compound 1 (100 μg/kg orally (PO) every day (QD)) as described in Example 20. The x-axis represents time measured in days and the y-axis represents tumor volume measured in mm ³ .

Fig. 19 is a graph of the protein expression of IKZF1, IKZF3, IRF-4, caspase-3, and IRF-1 in mice bearing ALCL KI-JK tumors and treated with Compound 1 as described in Example 20. The x-axis represents the time after dosing when the animal was sacrificed, and the y-axis represents the percentage of protein compared to vehicle control, normalized to GAPDH.

Fig. 20 is a graph comparing the effect of Compound 1, ibrutinib, and the combination of Compound 1 and ibrutinib on TMD8 tumor volume in mice as described in Example 13. Compound 1 was administered at a dose of 50 μg/kg orally every day and ibrutinib was administered at a dose of 12.5 mg/kg orally every day. The combination was dosed at the appropriate dose of each agent. The x-axis represents time measured in days and the y-axis represents tumor volume measured in mm ³ .

Fig. 21 is a graph of the percent survival of DLBCL TMD8 mice administered Compound 1, ibrutinib, and the combination of Compound 1 and ibrutinib as described in Example 21. Compound 1 was administered at a dose of 50 μg/kg orally every day and ibrutinib was administered at a dose of 12.5 mg/kg orally every day. The combination was dosed at the appropriate dose of each agent. The x-axis represents days after dosing and the y-axis represents percent survival.

Fig. 22 is a graph comparing the effect of Compound 1, dexamethasone, and a combination of Compound 1 and dexamethasone on RPMI-8226 multiple myeloma tumor volume in mice as described in Example 22. Dexamethasone was administered at a dose of 5 mg/kg intravenously (IV) every week (QW) and Compound 1 was administered at a dose of 10 μg/kg orally every day. Compound 1 was dosed in combination with dexamethasone at the appropriate dose levels and schedules for each agent. The x-axis represents time measured in days and the y-axis represents tumor volume measured in mm ³ .

Fig. 23 is a graph of the percent survival of mice bearing RPMI-8226 multiple myeloma tumors that were administered Compound 1, dexamethesone, and a combination of Compound 1 and dexamethesone as described in Example 22. Dexamethesone was administered at a dose of 5 mg/kg intravenously (IV) every week (QW) and Compound 1 was administered at a dose of 10 μg/kg orally every day. Compound 1 was dosed in combination with dexamethesone at the appropriate dose levels and schedules for each agent. The x-axis represents days after dosing and the y-axis represents percent survival.

Fig. 24 is a graph of the measurement of IKZF1 and IKZF3 concentrations in monkeys administered Compound 1. As described in Example 23, one monkey was administered Compound 1 and blood was collected at 0, 4, and 24 hours post-dose to determine the concentration of IKZF1 and IKZF3. The x-axis represents the time post-dose measured in hours, and the y-axis represents the average fluorescence intensity of IKZF1 and IKZF3.

Fig. 25A is a graph of the plasma concentration of Compound 1 or Compound 14 following administration of Compound 1 (60 μg/kg or 100 μg/kg) PO to monkeys as described in Example 24. The x-axis represents time measured in hours and the y-axis represents plasma concentration measured in ng/mL.

Fig. 25B is a graph of the plasma concentration of Compound 1 or Compound 14 following administration of Compound 1 (30 mg/kg) PO to rats as described in Example 24. The x-axis represents time measured in hours and the y-axis represents plasma concentration measured in ng/mL.

Fig. 26A is a graph of the effect of Compound 1 (100 μg/kg, QD PO) versus vehicle on tumor volume in mice inoculated with H929 tumors as described in Example 25. The x-axis represents the duration of treatment in days and the y-axis represents the H929 tumor volume measured in mm ³ .

Fig. 26B is a graph measuring the effect of Compound 2 (100 μg/kg, QD PO) versus vehicle on tumor volume in mice inoculated with H929 tumors as described in Example 25. The x-axis represents the duration of treatment in days and the y-axis represents the H929 tumor volume measured in mm ³ .

Fig. 26C is a graph measuring the effect of Compound 3 (100 μg/kg, QD PO) versus vehicle on tumor volume in mice inoculated with H929 tumors as described in Example 25. The x-axis represents the duration of treatment in days and the y-axis represents the H929 tumor volume measured in mm ³ .

Fig. 26D is a graph of the effect of Compound 4 (100 μg/kg, QD PO) versus vehicle on tumor volume in mice inoculated with H929 tumors as described in Example 25. The x-axis represents the duration of treatment in days and the y-axis represents the H929 tumor volume measured in mm ³ .

Fig. 26E is a graph measuring the effect of Compound 5 (100 μg/kg, QD PO) versus vehicle on tumor volume in mice inoculated with H929 tumors as described in Example 25. The x-axis represents the duration of treatment in days and the y-axis represents the H929 tumor volume measured in mm ³ .

Fig. 26F is a graph of the effect of Compound 6 (100 μg/kg, QD PO) versus vehicle on tumor volume in mice inoculated with H929 tumors as described in Example 25. The x-axis represents the duration of treatment in days and the y-axis represents the H929 tumor volume measured in mm ³ .

Fig. 26G is a graph of the effect of Compound 7 (100 μg/kg, QD PO) versus vehicle on tumor volume in mice inoculated with H929 tumors as described in Example 25. The x-axis represents the duration of treatment in days and the y-axis represents the H929 tumor volume measured in mm ³ .

Fig. 26H is a graph measuring the effect of Compound 8 (100 μg/kg, QD PO) versus vehicle on tumor volume in mice inoculated with H929 tumors as described in Example 25. The x-axis represents the duration of treatment in days and the y-axis represents the H929 tumor volume measured in mm ³ .

Fig. 26I is a graph of the effect of Compound 9 (100 μg/kg, QD PO) versus vehicle on tumor volume in mice inoculated with H929 tumors as described in Example 25. The x-axis represents the duration of treatment in days and the y-axis represents the H929 tumor volume measured in mm ³ .

Fig. 26J is a graph measuring the effect of Compound 10 (100 μg/kg, QD PO) versus vehicle on tumor volume in mice inoculated with H929 tumors as described in Example 25. The x-axis represents the duration of treatment in days and the y-axis represents the H929 tumor volume measured in mm ³ .

Fig. 26K is a graph measuring the effect of Compound 11 (100 μg/kg, QD PO) versus vehicle on tumor volume in mice inoculated with H929 tumors as described in Example 25. The x-axis represents the duration of treatment in days and the y-axis represents the H929 tumor volume measured in mm ³ .

Fig. 26L is a graph of the effect of Compound 12 (100 μg/kg, QD PO) versus vehicle on tumor volume in mice inoculated with DL-40 ALCL tumors as described in Example 25. The x-axis represents the duration of treatment in days and the y-axis represents the H929 tumor volume measured in mm ³ .

Fig. 26M is a graph measuring the effect of Compound 13 (100 μg/kg, QD PO) versus vehicle on tumor volume in mice inoculated with H929 tumors as described in Example 25. The x-axis represents the duration of treatment in days and the y-axis represents the H929 tumor volume measured in mm ³ .

Figure 27 is a graph showing the in vitro binding of Compound 1 to purified cereblon-DDB1 measured by a fluorescence polarization experiment. Compound 1 (circles) or the reference compound (pomalidomide) competes with the Alexa-647-based fluorescent probe for binding to cereblon-DDB1. Probe bias results in a decrease in the level of fluorescence polarization, which is used to calculate the fraction of probe bound using signals from positive and negative controls. Error bars represent standard deviation (SD). The measured Kd values are as follows: Compound 1: _Kd = 0.9 ± 0.5 nM, Pomalidomide _Kd = 712 ± 140 nM. The fluorescence polarization experiment is described in Example 26.

Figure 28 shows a dose response curve describing the displacement of a cereblon-binding tracer molecule by Compound 1 or pomalidomide in 293T cells expressing the cereblon-NanoLuc fusion as described in Example 27. The x-axis shows the concentration of Compound 1 or pomalidomide in nM and the y-axis shows the % NanoBRET signal relative to cells treated with the cereblon-binding tracer alone (100%) or without tracer (0%). 50% of the tracer was displaced by Compound 1 with an IC ₅₀ of 0.4 nM and by pomalidomide with an IC ₅₀ of 644 nM.

Fig. 29 shows a dose response curve describing the effect of Compound 1 on the destruction of IKZF1 compared to pomalidomide, as described in Example 9. The x-axis shows the concentration of Compound 1 or pomalidomide in nM and the y-axis shows the % of IKZF1 remaining after treatment with Compound 1 or pomalidomide for 1 or 2 hours.

Fig. 30 is a Western blot showing the levels of Aiolos and Ikaros remaining in H929 cells after 4 hours of treatment with Compound 1 or pomalidomide in response to the dose. The method for this experiment is described in Example 28.

Fig. 31 shows a dose response curve describing the effect of Compound 1 on caspase 3/7 activity in the multiple myeloma cell line NCIH929 compared to pomalidomide after 72 hours of treatment as described in Example 29. The x-axis shows the concentration of Compound 1 or pomalidomide in nM and the y-axis shows the luminescence signal (RLU) measured after addition of caspase 3/7 substrate reagents. The method for this experiment is described in Example 29.

Fig. 32 is a graph showing the effect of Compound 1 on the growth of eight multiple myeloma cell lines compared to pomalidomide as described in Example 30. The x-axis is the concentration of Compound 1 or pomalidomide in nM that inhibited cell growth at 96 hours by 50% (IC ₅₀ ), and the y-axis is the cell line tested.

Fig. 33 is a graph showing the in vivo efficacy of Compound 1 and pomalidomide in treating female NOD SCID mice bearing xenografted NCI-H929 multiple myeloma tumors. Mice were treated with vehicle control, Compound 1 in a dose-response relationship (3, 10, 30, and 100 μg/kg/day), or 3000 μg/kg/day pomalidomide for 21 days. All compounds were administered orally (PO) daily (QD). After 21 days of dosing, tumors were monitored for regrowth. The arrow indicates re-administration of 30 μg/kg/day Compound 1 beginning on day 40. The x-axis represents time measured in days, and the y-axis represents NCI-H929 tumor volume measured in mm ³ . This assay is described in Example 31.

Fig. 34 shows a graph of the bioluminescence signal of mice administered Compound 1 orally (PO) every day (QD) or vehicle, as imaged using IVIS Lumina II as described in Example 14. The x-axis shows time measured in days, and the y-axis shows the MMIS-Luc systemic BL (total bioluminescence signal) measured in photons/106.

Figure 35 shows the in vivo efficacy of Compound 1 and pomalidomide in female CB17 SCID mice bearing RPM-8226 xenografted multiple myeloma tumors. Mice were treated with vehicle control, dose (3, 10, 30, and 100 μg/kg/day) of Compound 1, or 3000 μg/kg/day of pomalidomide for 21 days. All compounds were administered orally (PO) daily (QD). Tumors were plotted individually and are presented as percentages of their original volume as shown on the y-axis.

Fig. 36 shows a graph of IKZF3 protein expression in mice inoculated with RPMI-8226 tumors and treated with Compound 1 in a dose-response relationship as described in Example 32. The x-axis shows the time after dosing when the animal was sacrificed, and the y-axis shows the percentage of protein compared to vehicle control, normalized to GAPDH.

Fig. 37 is a graph of tumor protein expression of IKZF1, IKZF3, and IRF-4 in mice bearing RPMI-8226 MM tumors that were administered Compound 1 as described in Example 32. The x-axis represents the time after dosing when the animal was sacrificed, and the y-axis represents the percentage of protein compared to vehicle control, normalized to GAPDH.

Figure 38 shows a Western blot assessing the levels of cereblon, IKZF1, and IKZF3 in untreated (parental) H929 cells treated with DMSO (LTC, long-term culture), lenalidomide, or pomalidomide for four months. Vinculin was used as a loading control. Western blotting was performed using the method described in Example 33.

Fig. 39 depicts the effect of Compound 1 on the growth of IMiD-resistant NCIH929 multiple myeloma cells compared to pomalidomide, as described in Example 34. The x-axis shows the concentration of Compound 1 or pomalidomide in nM, and the y-axis shows the % viability of IMiD-resistant NCIH929 cells at 96 hours relative to untreated cells.

Fig. 40 is a graph of tumor volume in mice injected with the refractory multiple myeloma cell line RPMI-8226. Mice were orally administered vehicle control, pomalidomide (3000 μg/kg/day), or Compound 1 (100 μg/kg/day) daily for 35 days, except for the pomalidomide-treated group. For this group, pomalidomide dosing was discontinued on day 17 and replaced with Compound 1 dosing (100 μg/kg/day) for the remainder of the study. This experiment is described in more detail in Example 15.

Fig. 41 is a graph comparing the effect of Compound 1, dexamethesone, and a combination of Compound 1 and dexamethesone on RPMI-8226 multiple myeloma tumor volume in mice as described in Example 22. Dexamethesone was administered at a dose of 5 mg/kg intravenously (IV) every week (QW) and Compound 1 was administered at a dose of 10 μg/kg or 100 μg/kg orally every day. Compound 1 at 10 μg/kg was dosed in combination with dexamethesone at the appropriate dose levels and schedules for each agent. The x-axis represents time measured in days and the y-axis represents tumor volume measured in mm ³ .

Fig. 42 is a graph comparing the effect of Compound 1, dexamethesone, and a combination of Compound 1 and dexamethesone on RPMI-8226 multiple myeloma tumor volume in mice as described in Example 22. Dexamethesone was administered at a dose of 5 mg/kg intravenously (IV) every week (QW) and Compound 1 was administered at a dose of 10 μg/kg orally every day. Compound 1 was dosed in combination with dexamethesone at the appropriate dose levels and schedules for each agent. When a tumor volume of 1000 mm ³ was reached, the animal was withdrawn from the study and death was recorded. The y-axis represents the probability of an animal surviving a given treatment, and the x-axis represents the number of days the animal survived.

Fig. 43 shows a graph of tumor protein expression of IKZF1, IKZF3, and IRF-4 in mice bearing REC1 MCL tumors that were administered Compound 1 as described in Example 35. The x-axis shows the time after dosing when the animal was sacrificed, and the y-axis shows the percentage of protein compared to vehicle control, normalized to GAPDH.

Fig. 44 shows a graph of E2F1 and Cyclin D1 protein expression in mice inoculated with REC1 MCL tumors and treated with Compound 1 as described in Example 35. The x-axis shows the time after dosing when the animal was sacrificed, and the y-axis shows the percentage of protein compared to vehicle control, normalized to GAPDH.

Fig. 45A, Fig. 45B, and Fig. 45C are dot plots depicting the effect of Compound 1 on the growth of NHL cell lines compared to pomalidomide as described in Example 37. The x-axis represents the concentration of Compound 1 or pomalidomide in nM that inhibits cell growth by 50% at 96 hours (IC ₅₀ ), and the y-axis represents the cell lines tested. Open symbols indicate that growth was not inhibited by more than 50% at the highest concentration tested (100 nM or 10 μM for Compound 1, 10 μM for pomalidomide, and 10 μM for CC-92480) and therefore the IC _{50 's} were not determined.

Figure 46 shows a Western blot of mice bearing established KI-JK xenografts that were treated with a single dose of Compound 1 (100 μg/kg) or daily for 5 days. Tumors were harvested at 4 and 24 hours after the single dose and at 24 hours after 5 daily doses. Tumors were analyzed by Western blot for IKZF1 and IRF-4 levels. The experimental procedure is described in Example 38.

Fig. 47 is a line graph showing the change in tumor volume in mice bearing Mino xenograft tumors treated with Compound 1 (100 μg/kg) once daily p.o., rituximab once weekly IV (10 mg/kg), or a combination of the two at their respective doses. Data are presented as mean tumor volume ± SEM. The experimental procedure is described in Example 39.

Fig. 48 is a line graph showing the concentration over time after treatment of mice bearing established NCI-H929 xenograft tumors treated with a single dose of Compound 1 (100 μg/kg) or CC-92480 (1000 μg/kg). Plasma and tumor samples were collected at 1, 4, 24, and 48 hours after a single dose and analyzed by LC-MS/MS. Data are expressed as total compound concentration values ± SEM. The experimental procedure is described in Example 40.

Figure 49 is a line graph showing the change in concentration over time after treatment of mice bearing established NCI-H929 xenograft tumors treated with a single dose of Compound 1 (100 μg/kg), CC-92480 (1000 μg/kg), or pomalidomide (3000 μg/kg). Tumor samples were collected at 1, 4, 24, and 48 hours after a single dose and analyzed by Western blotting for IKZF3 levels. Data are expressed as the percentage of IKZF3 remaining compared to vehicle control and normalized to GAPDH ± SEM values. The experimental procedure is described in Example 40.

Fig. 50 is a line graph showing the change in tumor volume over time after treatment of mice bearing established NCI-H929 xenograft tumors treated with Compound 1 (100 μg/kg), CC-92480 (1000 μg/kg), or pomalidomide (3000 μg/kg) orally daily for 18 days. Tumor volume and body weight were measured twice weekly. Data are presented as mean tumor volume ± SEM. The experimental procedure is described in Example 40.

Fig. 51 shows a dose response curve describing the effect of Compound 1 on caspase 3/7 activity in the TMD8 cell line compared to pomalidomide after 48 hours of treatment as described in Example 41. The x-axis shows the concentration of Compound 1 or pomalidomide in nM and the y-axis shows the luminescence signal (RLU) relative to DMSO-treated controls measured after addition of caspase 3/7 substrate reagents.

Fig. 52 shows a dose response curve describing the effect of Compound 1 on TMD8 cell viability compared to pomalidomide after 96 hours of treatment as described in Example 42. The x-axis represents the concentration of Compound 1 or pomalidomide in nM and the y-axis represents the % cell viability relative to DMSO-treated controls measured after addition of CellTiter Glo reagents.

Fig. 53 is a line graph showing the effect of Compound 1, ibrutinib, and the combination of Compound 1 and ibrutinib on Mino mantle cell lymphoma xenograft tumors. Mice were administered Compound 1 at 30 μg/kg/day, ibrutinib at 25 mg/kg, the combination of ibrutinib and Compound 1 at their respective dose levels, or vehicle control orally daily for 34 days. The x-axis represents time measured in days, and the y-axis represents tumor volume measured in mm ³ . Statistics were performed using the paired t-test in GraphPad Prism software. The experimental procedure is described in Example 44.

Fig. 54 is a line graph showing the effect of Compound 1, CC-92480, pomalidomide, or vehicle control on RPMI-8226 multiple myeloma xenograft tumors. Mice were administered Compound 1 at 100 μg/kg/day, CC-92480 at 1000 μg/kg/day, or pomalidomide at 3000 μg/kg/day orally daily for 19 days. The x-axis represents time in days and the y-axis represents tumor volume in mm ³ . Statistics were performed using paired t-test in GraphPad Prism software. The experimental procedure is described in Example 45.

Figure 55 is a line graph showing the effect of Compound 1, CC-92480, pomalidomide, or vehicle control on IKZF3 levels in RPMI-8226 multiple myeloma xenograft tumors. Mice were administered Compound 1 at 100 μg/kg/day, CC-92480 at 1000 μg/kg/day, or pomalidomide at 3000 μg/kg/day orally daily for 7 days. Tumor samples were collected at 4 and 24 hours after a single dose and at 24 hours after 3, 5, and 7 daily doses. Data are presented as percent target present in the vehicle control and are normalized for total protein. Error bars represent values ± SEM. The experimental procedure is described in Example 45.

Figure 5B is a line graph showing the effect of Compound 1, bortezomib, and the combination of Compound 1 and bortezomib on NCI-H929 xenografts. Mice bearing established NCI-H929 xenografts were dosed with Compound 1 (10 μg/kg), bortezomib (0.5 mg/kg), the combination of Compound 1 and bortezomib, or vehicle control for 14 days. Compound 1 was administered orally daily, while bortezomib was administered intravenously every two weeks. Statistical analysis was performed on day 14, the last day all animals were in the study, using a two-way ANOVA using GraphPad Prism software. Data are expressed as mean tumor volumes ± SEM.

Figure 57 is a line graph showing the effect of Compound 1, bortezomib, and the combination of Compound 1 and bortezomib on NCI-H929 xenografts. Mice bearing established NCI-H929 xenografts were dosed with Compound 1 (10 μg/kg), bortezomib (0.25 mg/kg), the combination of Compound 1 and bortezomib, or vehicle control for 14 days. Compound 1 was administered orally daily, while bortezomib was administered intravenously twice weekly. Statistical analysis was performed on day 14, the last day all animals were in the study, using a two-way ANOVA using GraphPad Prism software. Data are expressed as mean tumor volumes ± SEM.

Fig. 58 is a line graph showing the effect of Compound 1, Compound 2, Compound 15, CC-92480, or Pomalidomide on the concentration of IL2 secreted by anti-CD3-stimulated T cells after a six-day incubation. The x-axis represents the concentration of Compound in nM and the y-axis represents the fold change relative to DMSO-treated control wells. The experimental procedure is described in Example 46.

Fig. 59 is a line graph showing the effect of Compound 1, Compound 2, Compound 15, CC-92480, or pomalidomide on the concentration of IFNg secreted by anti-CD3-stimulated T cells after a six-day incubation. The x-axis represents the concentration of Compound in nM and the y-axis represents the fold change relative to DMSO-treated control wells. The experimental procedure is described in Example 46.

Fig. 60 is a line graph showing the effect of Compound 1, Compound 2, Compound 15, CC-92480, or pomalidomide on the concentration of IL21 secreted by anti-CD3-stimulated T cells after a six-day incubation. The x-axis represents the concentration of Compound in nM and the y-axis represents the fold change relative to DMSO-treated control wells. The experimental procedure is described in Example 46.

Fig. 61 is a line graph showing the effect of Compound 1, Compound 2, Compound 15, CC-92480, or pomalidomide on the concentration of IL9 secreted by anti-CD3-stimulated T cells after a six-day incubation. The x-axis represents the concentration of Compound in nM and the y-axis represents the fold change relative to DMSO-treated control wells. The experimental procedure is described in Example 46.

Fig. 62 is a line graph showing the effect of Compound 1, Compound 2, Compound 15, CC-92480, or pomalidomide on the concentration of IL4 secreted by anti-CD3-stimulated T cells after a six-day incubation. The x-axis represents the concentration of Compound in nM and the y-axis represents the fold change relative to DMSO-treated control wells. The experimental procedure is described in Example 46.

Fig. 63 is a line graph showing the effect of Compound 1, Compound 2, Compound 15, CC-92480, or pomalidomide on the concentration of TNF-alpha secreted by anti-CD3-stimulated T cells after a six-day incubation. The x-axis represents the concentration of Compound in nM and the y-axis represents the fold change relative to DMSO-treated control wells. The experimental procedure is described in Example 46.

Fig. 64 is a line graph showing the effect of Compound 1, Compound 2, Compound 15, CC-92480, or pomalidomide on the concentration of IL17A secreted by anti-CD3-stimulated T cells after a six-day incubation. The x-axis represents the concentration of Compound in nM and the y-axis represents the fold change relative to DMSO-treated control wells. The experimental procedure is described in Example 46.

Detailed disclosure of the present invention

Novel treatments, such as low-dose regimens, are based on the discovery that the compounds described are unusually potent Ikaros and Aiolos disruptors. Compound 1 is believed to be the most potent IKZF1/3 disruptor publicly disclosed to date. The data described herein and by other means demonstrate a significantly improved antitumor effect of the combination of Compound 1 and dexamethasone. The compounds described herein, and especially Compound 1, are significantly more potent than pomalidomide in NHL models, including PTCL and MCL, both in vitro and in vivo. Thus, these compounds may be used instead of thalidomide, lenalidomide and pomalidomide in NHL subtypes for which classical IMiDs have demonstrated clinical activity but have not been widely adopted as standard of care (MCL, DLBCL, PTCL, etc.).

It has been found that the compounds described herein can be administered for the treatment of Ikaros (IKZF1) and/or Aiolos (IKZF3) mediated disorders in a low dose regimen once or twice daily, optionally with drug holidays, according to highly effective treatment regimens. For example, it has been found that anti-cancer treatment can be effective in a patient using one of the compounds described herein at a dose of no more than about 500, 450, 400, 350, 325, 300, 275, 250, 225, 200, 175, 150, 125, or even 100, 75, 50, 45, 40, 35, 30, 25, 20, 15, 10, 5, or 1 microgram (mcg) once per day (QD) or twice per day (BID). In certain embodiments, the patient is an adult (typically a human weighing at least 100 pounds or more, and sometimes even 125 or 150 pounds, such as 70 kg or more, and at least 18 years of age or more). In an alternative embodiment, the patient is pediatric (and may weigh less than 100, 125, or 150 pounds and is typically less than 18 years of age).

According to certain embodiments, the dosing includes a drug holiday. A drug holiday is a period of time during which the patient is not administered the active compound. Treatment cycles for the purposes of the present disclosure are typically based on a 28-day cycle. For example, the patient may be administered the active compound or a pharmaceutically acceptable salt thereof for 21 consecutive days and not administered the chemotherapeutic agent for 7 days during a 28-day cycle, and then optionally the regimen is repeated one or more times. According to certain examples, one of the compounds described herein may be administered once or twice daily for at least 13, 14, 15, 16, 17, 18, 19, 20, 21, 22, 23, 24, 25, 26, or 27 consecutive days, followed by a holiday until the next 28-day cycle. According to some embodiments, the compound is administered once or twice daily for at least 20, 21, 22, 23 or 24 consecutive days, followed by a drug holiday until the end of a 28-day cycle. According to another embodiment, the drug is administered every day without a holiday, achieving continuous dosing during the period of the dosing regimen, which can be 1, 2, 3 or 4 weeks, or even 1, 2, 3, 4, 5 or 6 or more continuous or intermittent months. According to another embodiment, the use of a compound described herein eliminates the need for an off-cycle period, a drug holiday or a reduction in the concentration of the co-administered antitumor compound during treatment. According to another embodiment, the cycle period is more than 28 days, such as more than 30 or 35 days, and the appropriate on-cycle and off-cycle regimen is determined by the patient's treating physician.

In certain embodiments, the Ikaros (IKZF1) and/or Aiolos (IKZF3) disruptor is administered in combination with one or more additional therapeutic agents. Non-limiting examples of therapeutic agents that may be used in combination with the disruptor described herein include:

1. A BTK inhibitor selected from acalabrutinib, spebrutinib, zanubrutinib, LOXO-305, evobrutinib, TG-1701, tolebrutinib, BIIB091, DZD-9008, HZ-A-018, orelabrutinib, AC0058TA, SN1011, rilzabrutinib, ARQ 531, DTRMWXHS-12, JNJ-64264681, branebrutinib, ibrutinib, and fenebrutinib.

2. An anti-CD38 antibody selected from felzartamab, daratumumab, GBR 1342, TAK-573, CID-103, OKT10, STI-6129, SGX301, TAK-079, and mesagitamab.

3. A proteasome inhibitor selected from ixazomib citrate, oprozomib, delanozomib, lactacystin, bortezomib, carfilzomib, VLX1570, epoxomicin, MG132, MG-262, CEP-18770, NEOSH101, TQB3602, and KZR-616.

4. An IMiD selected from pomalidomide, lenalidomide, thalidomide, iberdomide, CC-92480, CC-90009, and CC-99282.

5. An HDAC inhibitor selected from trapoxin B, sodium phenylbutyrate, tacedinalin, mocetinostat, BRD73954, BG45, domatinostat, sau10603, HPOV, TMP269, nexturastat A, santacruzamate A, splitomycin, LMK-235, sodium butyrate, pivaloyloxymethylbutyrate, pyroxamide, abexinostat, resminostat, givinostat, quisinostat, psammapline A, KD5170, 1-Alanine chlamidocin, depudecin, and CUDC-101.

6. A compound selected from selinexor, oxafenamide, belantamab mafodotin, denosumab, zoledronic acid, plerixafor, eltrombopag, ipilimumab, palbociclib, ricolinostat, afuresertib, dinaciclib, filanesib, indatuximab ravtansine, masitinib, sonidegib, sotatercept, ulocuplumab, and urelumab.

I. Definitions

Unless defined otherwise, all technical and scientific terms used herein have the same meaning as commonly understood by one of ordinary skill in the art to which this invention belongs. In the description, the singular forms "a" and "an" also include the plural unless the context clearly dictates otherwise. Although methods and materials similar or equivalent to those described herein can be used in the practice and testing of the present invention, suitable methods and materials are described below. All publications, patent applications, patents, and other references mentioned herein are incorporated by reference. References cited herein are not considered prior art to the present invention. In case of conflict, the present description, including definitions, will control. Furthermore, the materials, methods, and examples are illustrative only and are not intended to be limiting.

The compounds are described using standard nomenclature. Unless otherwise defined, all technical and scientific terms used herein have the same meaning as commonly understood by one of ordinary skill in the art to which this invention belongs.

According to certain embodiments of each compound described herein, the compound can be in the form of a racemate, an enantiomer, a mixture of enantiomers, a diastereomer, a mixture of diastereomers, a tautomer, an N-oxide, or an isomer such as a rotamer, as if each were specifically described, unless specifically excluded by context.

The singular form does not imply a limitation of quantity, but rather denotes the presence of at least one of the recited elements. The term "or" means "and/or." Recitation of ranges of values is intended only to serve as a shorthand method of referring to each individual value falling within the range, unless otherwise indicated herein, and each individual value is included in the description as if it were individually recited herein. The endpoints of all ranges are included within the range and may be combined independently of one another. All methods and treatments described herein may be performed in a suitable order, unless otherwise indicated herein or otherwise clearly contradicted by context. The use of examples or illustrative language (e.g., "such as") is intended only to better illustrate the present invention and does not impose a limitation on the scope of the present invention unless otherwise claimed.

The present invention includes compounds described herein with at least one desired isotopic substitution of an atom in an amount greater than the natural abundance of the isotope, i.e. enriched. Isotopes are atoms with the same atomic number but different mass numbers, i.e. with the same number of protons but different numbers of neutrons. If isotopic substitutions are used, a common substitution is the replacement of hydrogen with at least one deuterium.

More generally, examples of isotopes that can be included in the compounds of the present invention include isotopes of hydrogen, carbon, nitrogen, oxygen, and fluorine, such as ^2H , ^3H , ^11C , ^13C , ^14C , 15N, ^17O , ^18O , and ^18F , respectively. According to one non ^- limiting embodiment, isotopically labeled compounds can be used in metabolic studies (e.g., with ^14C ), reaction kinetics studies (e.g., with ^2H or ^3H ), in detection or imaging methods such as positron emission tomography (PET) or single photon emission computed tomography (SPECT), including drug or substrate tissue distribution analysis, or in radioactive treatment of patients. Additionally, any hydrogen atom present in a compound of the present invention can be replaced with an ^18F atom, a substitution that may be particularly desirable for PET or SPECT studies. The isotopically labeled compounds of the present invention and prodrugs thereof can generally be prepared by carrying out the procedures described in the Schemes or in the Examples and Preparations described below, by replacing the non-isotopically labeled reagent with a readily available isotopically labeled reagent.

By way of general example and without limitation, hydrogen isotopes such as deuterium ( ^2H ) and tritium ( ^3H ) may be used anywhere in the described structures to achieve the desired result. Carbon isotopes such as ^13C and ^14C may be used alternatively or in addition.

Isotopic substitutions, such as deuterium substitutions, may be partial or complete. Partial deuterium substitution means that at least one hydrogen is replaced by deuterium. In certain embodiments, the isotope is enriched by 90, 95, or 99% or more in the isotope at any location of interest. In one non-limiting embodiment, deuterium is enriched by 90, 95, or 99% at the desired location.

In one non-limiting embodiment, the substitution of a hydrogen atom with a deuterium atom can be in any compound described herein. For example, when any of the groups is methyl or ethyl, the alkyl residue can be deuterated ₍ in non _- limiting embodiments _{CDH2 , CD2H} , CD3, _CH2CD3 , _CD2CD3 , CHDCH2D, _{CH2CD3 ,} CHDCHD2 _, etc.) _.

The compounds of the present invention may form solvates with a solvent (including water). Thus, in one non-limiting embodiment, the present invention includes a solvated form of the compounds described herein. The term "solvate" refers to a molecular complex of a compound of the present invention (including a salt thereof) with one or more solvent molecules. Non-limiting examples of solvents include water, ethanol, isopropanol, dimethyl sulfoxide, acetone and other common organic solvents. The term "hydrate" refers to a molecular complex comprising a compound of the present invention and water. Pharmaceutically acceptable solvates according to the present invention include solvates in which the solvent may be isotopically substituted, for example, D ₂ O, d ₆ -acetone, d ₆ -DMSO. The solvate may be in liquid or solid form.

"Dosage form" means a unit of administration of an active agent. Examples of dosage forms include tablets, capsules, injections, suspensions, liquids, emulsions, implants, particles, pellets, creams, ointments, suppositories, inhalants, transdermals, buccal, sublingual, topical, gel, mucosal, etc. "Dosage form" may also include an implant, such as an optical implant.

In the context of the present invention, the term "endogenous" refers to any substance from or produced within an organism, cell, tissue or system.

In the context of the present invention, the term "exogenous" refers to any substance introduced from within or obtained from outside an organism, cell, tissue or system.

By the term "modulating" in the context of the present invention is meant mediating a detectable increase or decrease in the level of response in a patient compared to the level of response in a patient in the absence of treatment or compound, and/or compared to the level of response in an identical but untreated patient. The term includes disruption and/or influence on a native signal or response, thereby mediating a beneficial therapeutic response in a patient, preferably a human.

"Parenteral" administration of a compound includes, for example, subcutaneous (s.c.), intravenous (i.v.), intramuscular (i.m.), or intrasternal injection or infusion.

In the context of the present invention, the term "pharmaceutical compositions" is a composition comprising at least one active agent, such as a selected active compound described herein, and at least one other substance, such as a carrier. "Pharmaceutical combinations" are combinations of at least two active agents that may be combined into a single dosage form or supplied together in separate dosage forms with instructions that the active agents are to be used together to treat any disorder described herein.

In the context of the present invention, the term "pharmaceutically acceptable salt" is a derivative of the disclosed compound in which the parent compound is modified by preparing its addition salts with inorganic and organic acids or bases with a biologically acceptable lack of toxicity. Salts of the compounds of the present invention can be synthesized from the parent compound, which contains a basic or acidic moiety, using conventional chemical methods. Typically, such salts can be prepared by reacting the free acid forms of these compounds with a stoichiometric amount of an appropriate base (e.g., Na, Ca, Mg or K hydroxide, carbonate, bicarbonate, etc.) or by reacting the free base forms of these compounds with a stoichiometric amount of an appropriate acid. Such reactions are typically carried out in water or in an organic solvent or a mixture of both. Typically, where possible, non-aqueous media such as ether, ethyl acetate, ethanol, isopropanol or acetonitrile are typical. Salts of the compounds of the present invention further include solvates of the compounds and salts of the compounds.

Examples of pharmaceutically acceptable salts include, but are not limited to, mineral or organic acid salts of basic residues such as amines, alkali or organic salts of acidic residues such as carboxylic acids, and the like. Pharmaceutically acceptable salts include conventional non-toxic salts and quaternary ammonium salts of the parent compound formed, for example, from non-toxic inorganic or organic acids. For example, conventional salts of non-toxic acids include salts derived from inorganic acids such as hydrochloric, hydrobromic, sulfuric, sulfamic, phosphoric, nitric acid and the like, and salts derived from organic acids such as acetic, propionic, succinic, glycolic, stearic, lactic, malic, tartaric, citric, ascorbic, pamoic, maleic, hydroxymaleic, phenylacetic, glutamic, benzoic, salicylic, mesyl, esyl, besyl, sulfanilic, 2-acetoxybenzoic, fumaric, toluenesulfonic, methanesulfonic, ethanedisulfonic, oxalic, isethionic acid, HOOC-( _CH2 ) _n- COOH, where n is 0-4, and the like, or using another acid that forms the same counterion. Lists of additional suitable salts can be found, for example, in Remington's Pharmaceutical Sciences, ^17th ed, Mack Publishing Company, Easton, Pa., p. 1418 (1985).

The term "carrier" means a diluent, excipient, or vehicle in which the active agent is employed or delivered.

"Pharmaceutically acceptable excipient" means an excipient that can be used in the preparation of a pharmaceutical composition/combination that is generally safe and is neither biologically nor otherwise unsuitable for administration to a host, typically a human. In certain embodiments, an excipient that is suitable for veterinary use is used.

"Patient" or "host" is a human or non-human animal in need of treatment for any of the disorders specifically described herein. Typically, the host is a human. Alternatively, "host" may refer to, for example, a mammal, a primate (e.g., a human), a cow, a sheep, a goat, a horse, a dog, a cat, a rabbit, a rat, a mouse, a fish, a bird, etc.

A "therapeutically effective amount" of a pharmaceutical composition/combination according to the present invention means an amount effective when administered to a host to provide a therapeutic effect, such as alleviation of symptoms or reduction or amelioration of the disease itself.

II. Compounds according to the present invention

According to certain embodiments, a compound used in accordance with the present invention is

or a pharmaceutically acceptable salt thereof.

According to certain embodiments, a compound used in accordance with the present invention is

or a pharmaceutically acceptable salt thereof.

According to certain embodiments, a compound used in accordance with the present invention is

or a pharmaceutically acceptable salt thereof.

According to certain embodiments, a compound used in accordance with the present invention is

or a pharmaceutically acceptable salt thereof.

According to certain embodiments, a compound used in accordance with the present invention is

or a pharmaceutically acceptable salt thereof.

According to certain embodiments, a compound used in accordance with the present invention is

or a pharmaceutically acceptable salt thereof.

According to certain embodiments, a compound used in accordance with the present invention is

or a pharmaceutically acceptable salt thereof.

According to certain embodiments, a compound used in accordance with the present invention is

or a pharmaceutically acceptable salt thereof.

According to certain embodiments, a compound used in accordance with the present invention is

or a pharmaceutically acceptable salt thereof.

According to certain embodiments, a compound used in accordance with the present invention is

or a pharmaceutically acceptable salt thereof.

According to certain embodiments, a compound used in accordance with the present invention is

or a pharmaceutically acceptable salt thereof.

According to certain embodiments, a compound used in accordance with the present invention is

or a pharmaceutically acceptable salt thereof.

According to certain embodiments, a compound used in accordance with the present invention is

or a pharmaceutically acceptable salt thereof.

III. Embodiments of the present invention.

1. According to certain embodiments, the present invention relates to the treatment of a disorder mediated by Ikaros or Aiolos comprising administering to a host in need thereof a low dose according to a treatment regimen comprising administering a dose of no more than about 500, 450, 400, 350, 300, 250, 200, 150, or even 100 micrograms (mcg) once daily (QD) or twice daily (BID) of a compound selected from Compound 1, Compound 2, Compound 3, Compound 4, Compound 5, Compound 6, Compound 7, Compound 8, Compound 9, Compound 10, Compound 11, Compound 12, or Compound 13, or a pharmaceutically acceptable salt thereof.

2. Treatment according to embodiment 1, wherein the treatment regimen includes drug holidays.

3. Treatment according to embodiment 2, wherein the drug holidays are carried out according to the treatment regimen once or twice a day for at least 13, 14 15, 16, 17, 18, 19, 20, 21, 22, 23, 24, 25, 26 or 27 consecutive days, followed by holidays until the next cycle of 28 days.

4. Treatment according to embodiment 3, wherein the therapy is administered once or twice daily for 21 days followed by a 7-day rest period.

5. Treatment according to any one of embodiments 1-4, wherein the dose is less than about 400 mcg.

6. Treatment according to any one of embodiments 1-4, wherein the dose is less than about 300 mcg.

7. Treatment according to any one of embodiments 1-4, wherein the dose is less than about 200 mcg.

8. Treatment according to any one of embodiments 1-4, wherein the dose is less than about 100 mcg.

9. Treatment according to any one of embodiments 1-4, wherein the dose is less than about 50 or 25 mcg.

10. Treatment according to any one of embodiments 1-9, wherein the disorder mediated by Ikaros or Aiolos is diffuse large B-cell lymphoma.

11. The treatment of embodiment 10, wherein the diffuse large B-cell lymphoma is activated B-cell lymphoma.

12. The treatment of embodiment 10, wherein the diffuse large B-cell lymphoma is germinal center B-cell lymphoma.

13. Treatment according to embodiment 1-9, wherein the disorder mediated by Ikaros or Aiolos is anaplastic large cell lymphoma.

14. The treatment of any one of embodiments 1-9, wherein the disorder mediated by Ikaros or Aiolos is cutaneous T-cell lymphoma.

15. The treatment of any one of embodiments 1-9, wherein the disorder mediated by Ikaros or Aiolos is mantle cell lymphoma.

16. The treatment of any one of embodiments 1-9, wherein the disorder mediated by Ikaros or Aiolos is multiple myeloma.

17. The treatment according to any one of embodiments 1-16, wherein the disorder is resistant to treatment with first generation IMiD drugs.

18. The treatment of embodiment 17, wherein the disorder is resistant to treatment with thalidomide.

19. Treatment according to embodiment 17, wherein the disorder is resistant to treatment with pomalidomide.

20. Treatment according to embodiment 17, wherein the disorder is resistant to treatment with lenalidomide.

21. Treatment according to embodiment 17, wherein the disorder is resistant to treatment with iberdomide.

22. Treatment of a disorder mediated by Ikaros (IKZF1) and/or Aiolos (IKZF3), wherein a blood or tissue sample is taken from a patient and the concentration of one or more biomarkers, such as IRF-1, caspase-3, IL-2 and/or IFN-γ, is determined, wherein if the patient has a statistically lower concentration, including without limitation up to about 5, 10, 15 or 20% lower, of the biomarker than a healthy individual, then Compound 1 or a pharmaceutically acceptable salt thereof or another compound described herein is administered to the patient.

23. Treatment of a disorder mediated by Ikaros (IKZF1) and/or Aiolos (IKZF3), wherein a blood or tissue sample is taken from a patient and the concentration of one or more biomarkers, such as cyclin D1, E2F1, ZFP91, SALL4, IRF-4, BLIMP 1 and/or MYC, is determined, wherein if the patient has a statistically higher concentration, including without limitation up to about 5, 10, 15 or 20% higher, of the biomarker than a healthy individual, then Compound 1 or a pharmaceutically acceptable salt thereof or another compound described herein is administered to the patient.

24. Treatment of a disorder mediated by Ikaros (IKZF1) and/or Aiolos (IKZF3), wherein Compound 1 or a pharmaceutically acceptable salt thereof or another compound described herein is administered to a patient, and then a blood or tissue sample is taken from the patient and the concentration of one or more biomarkers, such as IRF-1, caspase-3, IL-2 and/or IFN-γ, is determined, wherein if the concentration of the biomarker is slightly increased, such as by at least about 1.25, 1.5, 1.75 or 2-fold, then the dose of the compound is increased.

25. Treatment of a disorder mediated by Ikaros (IKZF1) and/or Aiolos (IKZF3), wherein Compound 1 or a pharmaceutically acceptable salt thereof or another compound described herein is administered to a patient, and then a blood or tissue sample is taken from the patient and the concentration of one or more biomarkers, e.g., cyclin D1, E2F1, ZFP91, SALL4, IRF-4, BLIMP 1 and/or MYC, is determined, wherein if the concentration of the biomarker is slightly decreased, e.g., by at least about 1.25-fold, 1.5-fold, 1.75-fold, or 2-fold, then the dose of the compound is increased.

26. Treatment according to any one of embodiments 22-25, wherein the dose is increased if the concentration is less than 150%.

27. Treatment according to any one of embodiments 22-25, wherein the dose is increased if the concentration is less than 160%.

28. Treatment according to any one of embodiments 22-25, wherein the dose is increased if the concentration is less than 170%.

29. Treatment according to any one of embodiments 22-25, wherein the dose is increased if the concentration is less than 180%.

30. Treatment of a disorder mediated by Ikaros or Aiolos, comprising administering an effective amount of a compound selected from Compound 1, Compound 2, Compound 3, Compound 4, Compound 5, Compound 6, Compound 7, Compound 8, Compound 9, Compound 10, Compound 11, Compound 12 and Compound 13, or a pharmaceutically acceptable salt thereof, to a patient in need thereof, wherein the disorder mediated by Ikaros or Aiolos is activated diffuse large B-cell lymphoma or germinal center large B-cell lymphoma.

31. Treatment of a disorder mediated by Ikaros (IKZF1) and/or Aiolos (IKZF3), wherein a compound selected from Compound 1, Compound 2, Compound 3, Compound 4, Compound 5, Compound 6, Compound 7, Compound 8, Compound 9, Compound 10, Compound 11, Compound 12, and Compound 13, or a pharmaceutically acceptable salt thereof, is used in combination with a BTK inhibitor selected from acalabrutinib, spebrutinib, zanubrutinib, LOXO-305, evobrutinib, TG-1701, tolebrutinib, BIIB091, DZD-9008, HZ-A-018, orelabrutinib, AC0058TA, SN1011, rilzabrutinib, ARQ 531, DTRMWXHS-12, JNJ-64264681, branebrutinib and fenebrutinib.

32. Treatment of a disorder mediated by Ikaros (IKZF1) and/or Aiolos (IKZF3), wherein a compound selected from Compound 1, Compound 2, Compound 3, Compound 4, Compound 5, Compound 6, Compound 7, Compound 8, Compound 9, Compound 10, Compound 11, Compound 12 and Compound 13 or a pharmaceutically acceptable salt thereof, is used in combination with an anti-CD38 antibody selected from felzartamab, GBR 1342, TAK-573, CID-103, OKT10, STI-6129, SGX301, TAK-079 and mesagitamab.

33. Treatment of a disorder mediated by Ikaros (IKZF1) and/or Aiolos (IKZF3), wherein a compound selected from Compound 1, Compound 2, Compound 3, Compound 4, Compound 5, Compound 6, Compound 7, Compound 8, Compound 9, Compound 10, Compound 11, Compound 12 and Compound 13, or a pharmaceutically acceptable salt thereof, is used in combination with a proteasome inhibitor selected from ixazomib citrate, oprosomib, delanozomib, lactacystin, epoxomicin, MG132, MG-262, CEP-18770, NEOSH101, TQB3602 and KZR-616.

34. Treatment of a disorder mediated by Ikaros (IKZF1) and/or Aiolos (IKZF3), wherein a compound selected from Compound 1, Compound 2, Compound 3, Compound 4, Compound 5, Compound 6, Compound 7, Compound 8, Compound 9, Compound 10, Compound 11, Compound 12 and Compound 13, or a pharmaceutically acceptable salt thereof, is used in combination with an IMiD selected from CC-92480, CC-90009 and CC-99282.

35. Treatment of a disorder mediated by Ikaros (IKZF1) and/or Aiolos (IKZF3), wherein a compound selected from Compound 1, Compound 2, Compound 3, Compound 4, Compound 5, Compound 6, Compound 7, Compound 8, Compound 9, Compound 10, Compound 11, Compound 12, and Compound 13, or a pharmaceutically acceptable salt thereof, is used in combination with an HDAC inhibitor selected from trapoxin B, sodium phenylbutyrate, tacedinalin, mocetinostat, BRD73954, BG45, domatinostat, cay10603, HPOV, TMP269, nexturastat A, Santacruzamate A, splitomycin, LMK-235, sodium butyrate, pivaloyloxymethylbutyrate, pyroxamide, abexinostat, resminostat, givinostat, quisinostat, Psammapline A, KD5170, 1-Alanine chlamydocin, depudecin and CUDC-101.

36. Treatment of a disorder mediated by Ikaros (IKZF1) and/or Aiolos (IKZF3), wherein a compound selected from Compound 1, Compound 2, Compound 3, Compound 4, Compound 5, Compound 6, Compound 7, Compound 8, Compound 9, Compound 10, Compound 11, Compound 12 and Compound 13, or a pharmaceutically acceptable salt thereof, is administered in combination with a compound selected from selinexor, oxafenamide, belantamab mafodotin, denosumab, zoledronic acid, plerixafor, eltrombopag, ipilimumab, palbociclib, ricolinostat, afuresertib, dinaciclib, filanesib, indatuximab ravtansine, masitinib, sonidegib, sotatercept, ulocuplumab and urelumab.

37. The treatment according to any one of embodiments 1-36, wherein the compound is

or a pharmaceutically acceptable salt thereof.

38. The treatment according to any one of embodiments 1-36, wherein the compound is

or a pharmaceutically acceptable salt thereof.

39. The treatment according to any one of embodiments 1-36, wherein the compound is

or a pharmaceutically acceptable salt thereof.

40. The treatment according to any one of embodiments 1-36, wherein the compound is

or a pharmaceutically acceptable salt thereof.

41. The treatment according to any one of embodiments 1-36, wherein the compound is

or a pharmaceutically acceptable salt thereof.

42. The treatment according to any one of embodiments 1-36, wherein the compound is

or a pharmaceutically acceptable salt thereof.

43. The treatment according to any one of embodiments 1-36, wherein the compound is

or a pharmaceutically acceptable salt thereof.

44. The treatment according to any one of embodiments 1-36, wherein the compound is

or a pharmaceutically acceptable salt thereof.

45. The treatment according to any one of embodiments 1-36, wherein the compound is

or a pharmaceutically acceptable salt thereof.

46. The treatment according to any one of embodiments 1-36, wherein the compound is

or a pharmaceutically acceptable salt thereof.

47. The treatment according to any one of embodiments 1-36, wherein the compound is

or a pharmaceutically acceptable salt thereof.

48. The treatment according to any one of embodiments 1-36, wherein the compound is

or a pharmaceutically acceptable salt thereof.

49. The treatment according to any one of embodiments 1-36, wherein the compound is

or a pharmaceutically acceptable salt thereof.

50. Treatment according to any one of embodiments 1-49, wherein the compound is administered in combination with a Bruton tyrosine kinase inhibitor.

51. Treatment according to embodiment 50, wherein the Bruton tyrosine kinase inhibitor is ibrutinib.

52. Treatment according to any one of embodiments 1-49, wherein the compound is administered in combination with a corticosteroid.

53. Treatment according to embodiment 52, wherein the corticosteroid is dexamethesone.

54. The treatment according to any one of embodiments 1-49, wherein the compound is administered in combination with CAR T cell therapy.

55. The treatment according to any one of embodiments 1-49, wherein the compound is administered in combination with an antibody-drug conjugate.

56. The treatment according to any one of embodiments 1-49, wherein the compound is administered in combination with BiTE therapy.

57. The treatment according to any one of embodiments 1-49, wherein the compound is administered in combination with a bispecific antibody.

58. The treatment according to any one of embodiments 1-49, wherein the compound is administered in combination with a monoclonal antibody.

59. The treatment of any one of embodiments 1-49, wherein the compound is administered in combination with a BTK inhibitor selected from acalabrutinib, spebrutinib, zanubrutinib, LOXO-305, evobrutinib, TG-1701, tolebrutinib, BIIB091, DZD-9008, HZ-A-018, orelabrutinib, AC0058TA, SN1011, rilzabrutinib, ARQ 531, DTRMWXHS-12, JNJ-64264681, branebrutinib, ibrutinib, and fenebrutinib.

60. Treatment according to any one of embodiments 1-49, wherein the compound is administered in combination with an anti-CD38 antibody selected from felzartamab, daratumumab, GBR 1342, TAK-573, CID-103, OKT10, STI-6129, SGX301, TAK-079, and mesagitamab.

61. The treatment according to any one of embodiments 1-49, wherein the compound is administered in combination with a proteasome inhibitor selected from ixazomib citrate, oprozomib, delanozomib, lactacystin, bortezomib, carfilzomib, VLX1570, epoxomicin, MG132, MG-262, CEP-18770, NEOSH101, TQB3602, and KZR-616.

62. The treatment according to any one of embodiments 1-49, wherein the compound is administered in combination with an IMiD selected from pomalidomide, lenalidomide, thalidomide, iberdomide, CC-92480, CC-90009, and CC-99282.

63. The treatment of any one of embodiments 1-49, wherein the compound is administered in combination with an HDAC inhibitor selected from trapoxin B, sodium phenylbutyrate, tacedinalin, mocetinostat, BRD73954, BG45, domatinostat, cay10603, HPOV, TMP269, nexturastat A, Santacruzamate A, splitomycin, LMK-235, sodium butyrate, pivaloyloxymethylbutyrate, pyroxamide, abexinostat, resminostat, givinostat, quisinostat, Psammapline A, KD5170, 1-Alanine chlamidocin, depudecin, and CUDC-101.

64. The treatment according to any one of embodiments 1-49, wherein the compound is administered in combination with a compound selected from selinexor, oxafenamide, belantamab mafodotin, denosumab, zoledronic acid, plerixafor, eltrombopag, ipilimumab, palbociclib, ricolinostat, afuresertib, dinaciclib, filanesib, indatuximab ravtansine, masitinib, sonidegib, sotatercept, ulocuplumab, and urelumab.

According to certain embodiments, the present invention provides a treatment as described above, wherein the dose of Compound 1 is less than or equal to about 5 mcg.

According to certain embodiments, the present invention provides a treatment as described above, wherein the dose of Compound 1 is less than or equal to about 10 mcg.

Additional embodiments

1. According to certain embodiments, the present invention relates to a method of treating a disorder mediated by Ikaros and/or Aiolos, comprising administering a dose of no more than about 500 micrograms (mcg) once daily (QD) or twice daily (BID) of a compound selected from

or a pharmaceutically acceptable salt thereof.

2. The method according to embodiment 1, wherein the compound is administered at a dose of from about 500 micrograms to 1 microgram.

3. The method according to embodiment 1 or according to embodiment 2, wherein the compound is administered over several days with drug holidays between subsequent treatment cycles.

4. The method according to embodiment 3, wherein the compound is administered once or twice daily for at least 13, 14, 15, 16, 17, 18, 19, 20, 21, 22, 23, 24, 25, 26 or 27 consecutive days, followed by a drug holiday until the next cycle of 28 days.

5. The method according to embodiment 3, wherein the compound is administered once or twice daily for 21 days followed by a 7-day rest period.

6. The method according to any one of embodiments 1-5, wherein the dose is less than or equal to about 400 mcg.

7. The method according to any one of embodiments 1-5, wherein the dose is less than or equal to about 300 mcg.

8. The method according to any one of embodiments 1-5, wherein the dose is less than or equal to about 200 mcg.

9. The method according to any one of embodiments 1-5, wherein the dose is less than or equal to about 100 mcg.

10. The method according to any one of embodiments 1-5, wherein the dose is less than or equal to about 50 mcg.

11. The method according to any one of embodiments 1-5, wherein the dose is less than or equal to about 25 mcg.

12. The method according to any one of embodiments 1-5, wherein the dose is less than or equal to about 10 mcg.

13. The method according to any one of embodiments 1-5, wherein the dose is less than or equal to about 5 mcg.

14. The method according to any one of embodiments 1-5, wherein the dose is about 50 mcg.

15. The method according to any one of embodiments 1-5, wherein the dose is about 25 mcg.

16. The method according to any one of embodiments 1-5, wherein the dose is about 10 mcg.

17. The method according to any one of embodiments 1-5, wherein the dose is about 5 mcg.

18. The method according to any one of embodiments 1-17, wherein the disorder is diffuse large B-cell lymphoma.

19. The method of embodiment 18, wherein the diffuse large B-cell lymphoma is activated B-cell lymphoma.

20. The method of embodiment 18, wherein the diffuse large B-cell lymphoma is germinal center B-cell lymphoma.

21. The method according to any one of embodiments 1-17, wherein the disorder is anaplastic large cell lymphoma.

22. The method according to any one of embodiments 1-17, wherein the disorder is cutaneous T-cell lymphoma.

23. The method according to any one of embodiments 1-17, wherein the disorder is mantle cell lymphoma.

24. The method according to any one of embodiments 1-17, wherein the disorder is multiple myeloma.

25. The method according to any one of embodiments 1-24, wherein the disorder is resistant to treatment with first generation IMiD drugs.

26. The method of embodiment 25, wherein the disorder is resistant to treatment with thalidomide.

27. The method of embodiment 25, wherein the disorder is resistant to treatment with pomalidomide.

28. The method of embodiment 25, wherein the disorder is resistant to treatment with lenalidomide.

29. The method of embodiment 25, wherein the disorder is resistant to treatment with iberdomide.

30. According to certain embodiments, the present invention relates to a method of treating a disorder mediated by Ikaros (IKZF1) and/or Aiolos (IKZF3), wherein a blood or tissue sample is taken from a patient and the concentration of one or more biomarkers selected from IRF-1, caspase-3, IL-2 and IFN-γ is determined, wherein if the patient has a statistically lower concentration of the biomarker(s) than a healthy individual, then a compound selected from Compound 1, Compound 2, Compound 3, Compound 4, Compound 5, Compound 6, Compound 7, Compound 8, Compound 9, Compound 10, Compound 11, Compound 12 and Compound 13, or a pharmaceutically acceptable salt thereof, is administered to the patient.

31. The method of embodiment 30, wherein the statistically lower concentration of the biomarker(s) is 5% lower than the average for a healthy individual.

32. The method of embodiment 30, wherein the statistically lower concentration of the biomarker(s) is 20% lower than the average for a healthy individual.

33. A method for treating a disorder mediated by Ikaros (IKZF1) and/or Aiolos (IKZF3), wherein a blood or tissue sample is taken from a patient and the concentration of one or more biomarkers selected from cyclin D1, E2F1, ZFP91, SALL4, IRF-4, BLIMP 1 and MYC is determined, wherein if the patient has a statistically higher concentration of the biomarker(s) than a healthy individual, then a compound selected from Compound 1, Compound 2, Compound 3, Compound 4, Compound 5, Compound 6, Compound 7, Compound 8, Compound 9, Compound 10, Compound 11, Compound 12 and Compound 13 or a pharmaceutically acceptable salt thereof is administered to the patient.

34. The method of embodiment 33, wherein the statistically higher concentration of the biomarker(s) is 5% higher than the average for a healthy individual.

35. The method of embodiment 33, wherein the statistically higher concentration of the biomarker(s) is 20% higher than the average for a healthy individual.

36. According to certain embodiments, the present invention relates to a method of treating a disorder mediated by Ikaros (IKZF1) and/or Aiolos (IKZF3), wherein a patient is administered a compound selected from Compound 1, Compound 2, Compound 3, Compound 4, Compound 5, Compound 6, Compound 7, Compound 8, Compound 9, Compound 10, Compound 11, Compound 12, and Compound 13, or a pharmaceutically acceptable salt thereof, and then a blood or tissue sample is taken from the patient and the concentration of one or more biomarkers selected from IRF-1, caspase-3, IL-2, and IFN-γ is determined, wherein if the concentration of the biomarker(s) is(are) slightly increased, then the dose of the compound is increased.

37. The method of embodiment 36, wherein the dose of the compound is increased if the concentration of the biomarker(s) is not increased by at least about 25%.

38. The method of embodiment 36, wherein the dose of the compound is increased if the concentration of the biomarker(s) is not increased by at least about 100%.

39. The method of embodiment 36, wherein the dose of the compound is increased if the concentration of the biomarker(s) is not increased by at least about 200%.

40. According to certain embodiments, the present invention relates to a method of treating a disorder mediated by Ikaros (IKZF1) and/or Aiolos (IKZF3), wherein a patient is administered a compound selected from Compound 1, Compound 2, Compound 3, Compound 4, Compound 5, Compound 6, Compound 7, Compound 8, Compound 9, Compound 10, Compound 11, Compound 12, and Compound 13, or a pharmaceutically acceptable salt thereof, and then a blood or tissue sample is collected from the patient and the concentration of one or more biomarkers selected from cyclin D1, E2F1, ZFP91, SALL4, IRF-4, BLIMP1, and MYC is determined, wherein if the concentration of the biomarker(s) is(are) not significantly decreased, then the dose of the compound is increased.

41. The method of embodiment 40, wherein the dose of the compound is increased if the concentration of the biomarker(s) is not reduced by at least about 10%.

42. The method of embodiment 40, wherein the dose of the compound is increased if the concentration of the biomarker(s) is not reduced by at least about 25%.

43. The method of embodiment 40, wherein the dose of the compound is increased if the concentration of the biomarker(s) is not reduced by at least about 50%.

44. According to certain embodiments, the present invention relates to a method of treating a disorder mediated by Ikaros and/or Aiolos, comprising administering an effective amount of a compound selected from Compound 1, Compound 2, Compound 3, Compound 4, Compound 5, Compound 6, Compound 7, Compound 8, Compound 9, Compound 10, Compound 11, Compound 12, and Compound 13, or a pharmaceutically acceptable salt thereof, to a patient in need thereof, wherein the disorder mediated by Ikaros or Aiolos is activated diffuse large B-cell lymphoma or germinal center large B-cell lymphoma.

45. According to certain embodiments, the present invention provides a method of treating a disorder mediated by Ikaros and/or Aiolos, comprising administering an effective amount of a compound selected from Compound 1, Compound 2, Compound 3, Compound 4, Compound 5, Compound 6, Compound 7, Compound 8, Compound 9, Compound 10, Compound 11, Compound 12, and Compound 13, or a pharmaceutically acceptable salt thereof, wherein the patient further receives a BTK inhibitor selected from acalabrutinib, spebrutinib, zanubrutinib, LOXO-305, evobrutinib, TG-1701, tolebrutinib, BIIB091, DZD-9008, HZ-A-018, orelabrutinib, AC0058TA, SN1011, rilzabrutinib, ARQ 531, DTRMWXHS-12, JNJ-64264681, branebrutinib and fenebrutinib.

46. According to certain embodiments, the present invention relates to a method of treating a disorder mediated by Ikaros and/or Aiolos, comprising administering an effective amount of a compound selected from Compound 1, Compound 2, Compound 3, Compound 4, Compound 5, Compound 6, Compound 7, Compound 8, Compound 9, Compound 10, Compound 11, Compound 12, and Compound 13, or a pharmaceutically acceptable salt thereof, wherein the patient also receives an anti-CD38 antibody selected from felzartamab, GBR 1342, TAK-573, CID-103, OKT10, STI-6129, SGX301, TAK-079, and mesagitamab.

47. According to certain embodiments, the present invention relates to a method of treating a disorder mediated by Ikaros and/or Aiolos, comprising administering an effective amount of a compound selected from Compound 1, Compound 2, Compound 3, Compound 4, Compound 5, Compound 6, Compound 7, Compound 8, Compound 9, Compound 10, Compound 11, Compound 12, and Compound 13, or a pharmaceutically acceptable salt thereof, wherein the patient also receives a proteasome inhibitor selected from ixazomib citrate, oprozomib, delanozomib, lactacystin, epoxomicin, MG132, MG-262, CEP-18770, NEOSH101, TQB3602HKZR-616.

48. According to certain embodiments, the present invention relates to a method of treating a disorder mediated by Ikaros and/or Aiolos, comprising administering an effective amount of a compound selected from Compound 1, Compound 2, Compound 3, Compound 4, Compound 5, Compound 6, Compound 7, Compound 8, Compound 9, Compound 10, Compound 11, Compound 12, and Compound 13, or a pharmaceutically acceptable salt thereof, to a patient in need thereof, wherein the patient also receives an IMiD selected from CC-92480, CC-90009, and CC-99282.

49. According to certain embodiments, the present invention provides a method of treating a disorder mediated by Ikaros and/or Aiolos comprising administering an effective amount of a compound selected from Compound 1, Compound 2, Compound 3, Compound 4, Compound 5, Compound 6, Compound 7, Compound 8, Compound 9, Compound 10, Compound 11, Compound 12, and Compound 13, or a pharmaceutically acceptable salt thereof, to a patient in need thereof, wherein the patient also receives an HDAC inhibitor selected from trapoxin B, sodium phenylbutyrate, tacedinalin, mocetinostat, BRD73954, BG45, domatinostat, cay10603, HPOV, TMP269, nexturastat A, Santacruzamate A, splitomycin, LMK-235, sodium butyrate, pivaloyloxymethylbutyrate, pyroxamide, abexinostat, resminostat, givinostat, quisinostat, psammapline A, KD5170, 1-Alanine chlamidocin, depudecin hCUDC-101.

50. According to certain embodiments, the present invention provides a method of treating a disorder mediated by Ikaros and/or Aiolos, comprising administering an effective amount of a compound selected from Compound 1, Compound 2, Compound 3, Compound 4, Compound 5, Compound 6, Compound 7, Compound 8, Compound 9, Compound 10, Compound 11, Compound 12, and Compound 13, or a pharmaceutically acceptable salt thereof, to a patient in need thereof, wherein the patient further receives a compound selected from selinexor, oxafenamide, belantamab mafodotin, denosumab, zoledronic acid, plerixafor, eltrombopag, ipilimumab, palbociclib, ricolinostat, afuresertib, dinaciclib, filanesib, indatuximab ravtansine, masitinib, sonidegib, sotatercept, ulocuplumab and urelumab.

51. The method according to any one of embodiments 1-50, wherein the compound is

or a pharmaceutically acceptable salt thereof.

52. The method according to any one of embodiments 1-50, wherein the compound is

or a pharmaceutically acceptable salt thereof.

53. The method according to any one of embodiments 1-50, wherein the compound is

or a pharmaceutically acceptable salt thereof.

54. The method according to any one of embodiments 1-50, wherein the compound is

or a pharmaceutically acceptable salt thereof.

55. The method according to any one of embodiments 1-50, wherein the compound is

or a pharmaceutically acceptable salt thereof.

56. The method according to any one of embodiments 1-50, wherein the compound is

or a pharmaceutically acceptable salt thereof.

57. The method according to any one of embodiments 1-50, wherein the compound is

or a pharmaceutically acceptable salt thereof.

58. The method according to any one of embodiments 1-50, wherein the compound is

or a pharmaceutically acceptable salt thereof.

59. The method according to any one of embodiments 1-50, wherein the compound is

or a pharmaceutically acceptable salt thereof.

60. The method according to any one of embodiments 1-50, wherein the compound is

or a pharmaceutically acceptable salt thereof.

61. The method according to any one of embodiments 1-50, wherein the compound is

or a pharmaceutically acceptable salt thereof.

62. The method according to any one of embodiments 1-50, wherein the compound is

or a pharmaceutically acceptable salt thereof.

63. The method according to any one of embodiments 1-50, wherein the compound is

or a pharmaceutically acceptable salt thereof.

64. The method according to any one of embodiments 1-63, wherein the Bruton tyrosine kinase inhibitor is also administered to the patient.

65. The method of embodiment 64, wherein the Bruton tyrosine kinase inhibitor is ibrutinib.

66. The method according to any one of embodiments 1-65, wherein the corticosteroid is also administered to the patient.

67. The method of embodiment 66, wherein the corticosteroid is dexamethesone.

68. The method according to any one of embodiments 1-67, wherein the CAR T cell therapy is also administered to the patient.

69. The method according to any one of embodiments 1-67, wherein the antibody-drug conjugate is also administered to the patient.

70. The method according to any one of embodiments 1-67, wherein the BiTE therapy is also administered to the patient.

71. The method according to any one of embodiments 1-67, wherein the bispecific antibody is also administered to the patient.

72. The method according to any one of embodiments 1-67, wherein the monoclonal antibody is also administered to the patient.

73. The method according to any one of embodiments 1-67, wherein a BTK inhibitor selected from acalabrutinib, spebrutinib, zanubrutinib, LOXO-305, evobrutinib, TG-1701, tolebrutinib, BIIB091, DZD-9008, HZ-A-018, orelabrutinib, AC0058TA, SN1011, rilzabrutinib, ARQ 531, DTRMWXHS-12, JNJ-64264681, branebrutinib, ibrutinib, and fenebrutinib is also administered to the patient.

74. The method according to any one of embodiments 1-67, wherein an anti-CD38 antibody selected from felzartamab, daratumumab, GBR 1342, TAK-573, CID-103, OKT10, STI-6129, SGX301, TAK-079, and mesagitamab is also administered to the patient.

75. The method according to any one of embodiments 1-67, wherein a proteasome inhibitor selected from ixazomib citrate, oprozomib, delanozomib, lactacystin, bortezomib, carfilzomib, VLX1570, epoxomicin, MG132, MG-262, CEP-18770, NEOSH101, TQB3602, and KZR-616 is also administered to the patient.

76. The method according to any one of embodiments 1-67, wherein an IMiD selected from pomalidomide, lenalidomide, thalidomide, iberdomide, CC-92480, CC-90009, and CC-99282 is also administered to the patient.

77. The method according to any one of embodiments 1-67, wherein an HDAC inhibitor selected from trapoxin B, sodium phenylbutyrate, tacedinalin, mocetinostat, BRD73954, BG45, domatinostat, cay10603, HPOV, TMP269, nexturastat A, Santacruzamate A, splitomycin, LMK-235, sodium butyrate, pivaloyloxymethylbutyrate, pyroxamide, abexinostat, resminostat, givinostat, quisinostat, Psammapline A, KD5170, 1-Alanine chlamidocin, depudecin, and CUDC-101 is also administered to the patient.

78. The method according to any one of embodiments 1-67, wherein a compound selected from selinexor, oxafenamide, belantamab mafodotin, denosumab, zoledronic acid, plerixafor, eltrombopag, ipilimumab, palbociclib, ricolinostat, afuresertib, dinaciclib, filanesib, indatuximab ravtansine, masitinib, sonidegib, sotatercept, ulocuplumab, and urelumab is also administered to the patient.

79. The method according to any one of embodiments 1-78, wherein the compound selected from Compound 1, Compound 2, Compound 3, Compound 4, Compound 5, Compound 6, Compound 7, Compound 8, Compound 9, Compound 10, Compound 11, Compound 12, and Compound 13, or a pharmaceutically acceptable salt thereof, is administered once daily.

80. The method according to any one of embodiments 1-78, wherein the compound selected from Compound 1, Compound 2, Compound 3, Compound 4, Compound 5, Compound 6, Compound 7, Compound 8, Compound 9, Compound 10, Compound 11, Compound 12, and Compound 13, or a pharmaceutically acceptable salt thereof, is administered twice daily.

81. The method according to any one of embodiments 1-80, wherein the disorder is non-Hodgkin lymphoma.

82. The method according to any one of embodiments 1-80, wherein the disorder is multiple myeloma.

83. The method according to any one of embodiments 1-82, wherein the disorder is recurrent.

84. The method according to any one of embodiments 1-82, wherein the violation is non-responsive.

85. The method according to any one of embodiments 1-82, wherein the disorder is recurrent and refractory.

IV. Treatment of disorders mediated by Ikaros and/or Aiolos

The present invention relates to advantageous treatments for disorders mediated by Ikaros and/or Aiolos. In certain embodiments, the treatment comprises administering a low dose dosage form once or twice daily, optionally with a drug holiday, which is a highly effective method of treatment. For example, it has been found that treatment can be effective for a patient using one of the compounds described herein at a dose of no more than about 500, 450, 400, 350, 325, 300, 275, 250, 225, 200, 175, 150, 125 or even 100, 75, 50 or 25 micrograms (mcg) once daily (QD) or twice daily (BID), optionally with a treatment holiday. In some embodiments, the patient is an adult (typically a person weighing at least 100 pounds or more and typically 18 years of age or older). In an alternative embodiment, the patient is a pediatric patient (and may weigh less than 100 pounds and typically under 18 years of age).

The selected compound, for example Compound 1, can be administered as a monotherapy or it can be combined with a standard therapy for the target tumor or cancer, including but not limited to any of those described in the "prior art of the present invention" section, or such as a proteasome inhibitor and/or an anti-CD38 monoclonal antibody (mAb). In MCL, the selected compound can be used, for example, in combination with a Bruton tyrosine kinase (BTK) inhibitor or anra-CD20 monoclonal antibody. In PTCL, in particular ALCL, the selected compound can be administered, for example, in combination with an anti-CD30 or anti-CD38 monoclonal antibody.

In certain embodiments, Compound 1 is used to treat a disorder mediated by Ikaros or Aiolos according to a treatment regimen described herein.

In certain embodiments, Compound 2 is used to treat a disorder mediated by Ikaros or Aiolos according to a treatment regimen described herein.

In certain embodiments, Compound 3 is used to treat a disorder mediated by Ikaros or Aiolos according to a treatment regimen described herein.

In certain embodiments, Compound 4 is used to treat a disorder mediated by Ikaros or Aiolos according to the treatment regimen described herein.

In certain embodiments, Compound 5 is used to treat a disorder mediated by Ikaros or Aiolos according to the treatment regimen described herein.

In certain embodiments, Compound 6 is used to treat a disorder mediated by Ikaros or Aiolos according to the treatment regimen described herein.

In certain embodiments, Compound 7 is used to treat a disorder mediated by Ikaros or Aiolos according to the treatment regimen described herein.

In certain embodiments, Compound 8 is used to treat a disorder mediated by Ikaros or Aiolos according to the treatment regimen described herein.

In certain embodiments, Compound 9 is used to treat a disorder mediated by Ikaros or Aiolos according to the treatment regimen described herein.

In certain embodiments, Compound 10 is used to treat a disorder mediated by Ikaros or Aiolos according to the treatment regimen described herein.

In certain embodiments, Compound 11 is used to treat a disorder mediated by Ikaros or Aiolos according to a treatment regimen described herein.

In certain embodiments, Compound 12 is used to treat a disorder mediated by Ikaros or Aiolos according to the treatment regimen described herein.

In certain embodiments, Compound 13 is used to treat a disorder mediated by Ikaros or Aiolos according to the treatment regimen described herein.

In certain embodiments, a compound described herein, such as Compound 1, is used to treat cancer that has metastasized. In certain embodiments, a compound described herein, such as Compound 1, is used to treat cancer that has metastasized to the brain.

Treatment cycles

In certain embodiments, dosing includes a drug holiday. A drug holiday is a period of time during which the patient is not administered the active compound. For example, the patient may be administered the active compound or a pharmaceutically acceptable salt thereof for 21 consecutive days and not administered the chemotherapeutic agent for 7 days during a 28-day treatment cycle, and then optionally the regimen is repeated one, several, or more times. In certain examples, one of the compounds described herein may be administered once or twice daily for at least 18, 19, 20, 21, 22, 23, 24, 25, 26, or 27 consecutive days, followed by a drug holiday until the next 28-day cycle. In some embodiments, the compound is administered once or twice daily for at least 20, 21, 22, 23, or 24 consecutive days, followed by a drug holiday until the end of the 28-day cycle. According to another embodiment, the drug is administered every day without a holiday, achieving continuous dosing during the period of the dosing regimen, which can be 2, 3 or 4 weeks, or even 1, 2, 3, 4, 5 or 6 or more consecutive months. According to another embodiment, the use of a compound described herein eliminates the need for an off-cycle period, drug holidays or a decrease in the concentration of the co-administered antitumor compound during treatment. According to another embodiment, the cycle period is more than 28 days, such as more than 30 or 35 days.

In certain embodiments, a compound of the present invention is administered for 10, 11, 12, 13, 14, 15, 16, 17, 18, 19, 20, 21, 22, 23, 24, 25, 26, 27, or 28 days of a 28-day treatment cycle. In certain embodiments, a compound of the present invention is administered for 14 consecutive days followed by a dosing break of 14 days. In certain embodiments, a compound of the present invention is administered for 21 consecutive days followed by a dosing break of 7 days.

In certain embodiments, a compound of the present invention is administered once daily for 10, 11, 12, 13, 14, 15, 16, 17, 18, 19, 20, 21, 22, 23, 24, 25, 26, 27, or 28 days of a 28-day treatment cycle. In certain embodiments, a compound of the present invention is administered daily for 14 consecutive days, followed by a dosing break of 14 days. In certain embodiments, a compound of the present invention is administered daily for 21 consecutive days, followed by a dosing break of 7 days.

In certain embodiments, a compound of the present invention is administered twice daily for 10, 11, 12, 13, 14, 15, 16, 17, 18, 19, 20, 21, 22, 23, 24, 25, 26, 27, or 28 days of a 28-day treatment cycle. In certain embodiments, a compound of the present invention is administered twice daily for 14 consecutive days, followed by a dosing break of 14 days. In certain embodiments, a compound of the present invention is administered twice daily for 21 consecutive days, followed by a dosing break of 7 days.

In certain embodiments, Compound 1 is administered orally daily for 21 days followed by a 7 day rest period during each 28 day treatment cycle.

Administration of doses

In certain alternative embodiments, a compound of the present invention is administered at a dose of about 800 μg. In certain embodiments, a compound of the present invention is administered at a dose of less than about 600 μg. In certain embodiments, a compound of the present invention is administered at a dose of less than about 400 μg. In certain embodiments, a compound of the present invention is administered at a dose of less than about 300 μg. In certain embodiments, a compound of the present invention is administered at a dose of less than about 200 μg. In certain embodiments, a compound of the present invention is administered at a dose of less than about 100 μg.

In certain embodiments, a compound of the present invention is administered at a dose of less than about 50 mcg. In certain embodiments, a compound of the present invention is administered at a dose of less than about 25 mcg.

In certain embodiments, a compound of the present invention is administered at a dose of about 50 mcg.

In certain embodiments, a compound of the present invention is administered at a dose of about 45 mcg.

In certain embodiments, a compound of the present invention is administered at a dose of about 40 mcg.

In certain embodiments, a compound of the present invention is administered at a dose of about 35 mcg.

In certain embodiments, a compound of the present invention is administered at a dose of about 30 mcg.

In certain embodiments, a compound of the present invention is administered at a dose of about 25 mcg.

In certain embodiments, a compound of the present invention is administered at a dose of about 20 mcg.

In certain embodiments, a compound of the present invention is administered at a dose of about 15 mcg.

In certain embodiments, a compound of the present invention is administered at a dose of about 10 mcg.

In certain embodiments, a compound of the present invention is administered at a dose of about 5 mcg.

In certain embodiments, a compound of the present invention is administered at a dose of about 1 mcg.

In certain embodiments, Compound 1 is administered at a dose of less than about 800 mcg. In certain embodiments, Compound 1 is administered at a dose of less than about 600 mcg. In certain embodiments, Compound 1 is administered at a dose of less than about 400 mcg. In certain embodiments, Compound 1 is administered at a dose of less than about 300 mcg. In certain embodiments, Compound 1 is administered at a dose of less than about 200 mcg. In certain embodiments, Compound 1 is administered at a dose of less than about 100 mcg.

In certain embodiments, Compound 1 is administered at a dose of about 800 μg. In certain embodiments, Compound 1 is administered at a dose of about 600 μg. In certain embodiments, Compound 1 is administered at a dose of about 400 μg. In certain embodiments, Compound 1 is administered at a dose of about 300 μg. In certain embodiments, Compound 1 is administered at a dose of about 200 μg. In certain embodiments, Compound 1 is administered at a dose of about 100 μg.

According to certain embodiments, Compound 1 is administered at a dose of at least about or between 25, 50, 75, 100, 125, 150, 175, 200, 225, 250, 275, 300, 325, 350, 375, 400, 425, 450, 475, 500, 525, 550, 575, 600, 625, 650, 675, 700, 725, 750, 775, 800, 825, 850, 875, 900, 925, 950, 975, or 1000 mcg.

In certain embodiments, Compound 1 is administered at a dose of about 25 μg. In certain embodiments, Compound 1 is administered at a dose of about 50 or 75 μg. In certain embodiments, Compound 1 is administered at a dose of about 100 or 150 μg. In certain embodiments, Compound 1 is administered at a dose of about 175 or 200 μg. In certain embodiments, Compound 1 is administered at a dose of about 225 or 250 μg. In certain embodiments, Compound 1 is administered at a dose of about 275, 300, 325, or 350 μg. In certain embodiments, Compound 1 is administered at a dose of about 400 or 450 μg. In certain embodiments, Compound 1 is administered at a dose of about 550 μg. In certain embodiments, Compound 1 is administered at a dose of about 650 mcg. In certain embodiments, Compound 1 is administered at a dose of about 725 mcg. In certain embodiments, Compound 1 is administered at a dose of about 800 mcg.

The present invention includes at least the following low dose features:

(a) a low dose treatment regimen for a disorder mediated by Ikaros or Aiolos in a host comprising administering a dose of no more than 500, 450, 400, 350, 300, 250, 200, 150 or even 100 micrograms (mcg) once daily (QD) or twice daily (BID) to a patient in need thereof of a compound selected from Compound 1, Compound 2, Compound 3, Compound 4, Compound 5, Compound 6, Compound 7, Compound 8, Compound 9, Compound 10, Compound 11, Compound 12 and Compound 13,

(b) treatment according to (a), where the treatment regimen includes a drug holiday,

(c) treatment according to (b), where the drug holiday is carried out according to the treatment regimen once or twice daily for at least 13, 14, 15, 16, 17, 18, 19, 20, 21, 22, 23, 24, 25, 26 or 27 consecutive days, followed by a holiday until the next cycle of 28 days,

(d) treatment according to (c), where the therapy is administered once or twice daily for 21 days followed by a rest period of 7 days,

(e) treatment according to (a) to (d), where the single dose is not more than 400 micrograms,

(f) treatment according to (a) to (d), where the single dose is not more than 300, 200 or 100 micrograms,

(g) treatment according to (a) to (d), where the single dose is not more than 25, 50 or 75 micrograms,

(h) treatment under (b), which may be for 2, 3, 4, 5 or 6 weeks or even 1, 2, 3, 4, 5 or 6 or more months with staggered holiday periods,

(i) treatment under (a) which does not include the holiday period,

(j) treatment under (a) where the cycle period is more than 28 days, for example more than 30 or 35 days, and includes drug holidays,

(k) the treatment of (a)-(j), wherein the disorder is selected from diffuse large B-cell lymphoma, anaplastic large cell lymphoma, cutaneous T-cell lymphoma, mantle cell lymphoma and multiple myeloma, comprising administering an effective amount of Compound 1, Compound 2, Compound 3, Compound 4, Compound 5, Compound 6, Compound 7, Compound 8, Compound 9, Compound 10, Compound 11, Compound 12 or Compound 13 to a patient in need thereof,

(1) the treatment of (a)-(j), where the disorder is refractory to treatment with other cereblon ligands, comprising administering an effective amount of Compound 1, Compound 2, Compound 3, Compound 4, Compound 5, Compound 6, Compound 7, Compound 8, Compound 9, Compound 10, Compound 11, Compound 12 or Compound 13 to a patient in need thereof,

(m) treatment according to (a)-(j), where the patient suffers from a disorder mediated by cereblon, comprising monitoring the concentration of one or more biomarkers selected from IRF-1 and caspase-3,

(n) the treatment of (a)-(j), wherein the combination treatment of a patient suffering from a cereblon-mediated disorder comprises administering an effective amount of Compound 1, Compound 2, Compound 3, Compound 4, Compound 5, Compound 6, Compound 7, Compound 8, Compound 9, Compound 10, Compound 11, Compound 12 or Compound 13 to a patient in need thereof, in combination or in alternation with a Bruton's tyrosine kinase inhibitor, a corticosteroid, CAR T-cell therapy, an antibody-drug conjugate, BiTE therapy, a bispecific antibody or a monoclonal antibody,

(o) a low dose pharmaceutical composition of a compound described herein or a pharmaceutically acceptable salt, isotopic derivative (including a deuterated derivative) or prodrug thereof in a pharmaceutically acceptable carrier,

(p) treatment according to (a)-(j) of any disorder described herein, comprising administering an effective amount of Compound 1, Compound 2, Compound 3, Compound 4, Compound 5, Compound 6, Compound 7, Compound 8, Compound 9, Compound 10, Compound 11, Compound 12 or Compound 13 to a patient in need thereof,

(q) a compound as described herein, or a pharmaceutically acceptable salt, isotopic derivative or prodrug thereof, for the treatment of (a)-(j) of a disorder mediated by Ikaros or Aiolos,

(r) the use of a compound described herein, or a pharmaceutically acceptable salt, isotopic derivative or prodrug thereof, in an effective amount for the treatment according to (a)-(j) of a patient, typically a human, suffering from any of the disorders described herein, including those mediated by Ikaros or Aiolos,

(s) a method for producing a low dose medicament for treating a disorder described herein in a host, characterized in that the compound described herein is used to produce the low dose amounts specified herein,

(t) the use of a compound described herein, or a pharmaceutically acceptable salt, isotopic derivative or prodrug thereof, for the preparation of a medicament for the treatment of (a)-(j) of cancer, including any of the cancers described herein,

(u) a method for producing a medicament for the treatment of (a)-(j) of cancer in a host, including any of the cancers described herein, characterized in that the compound described herein is used in the production,

(v) a compound described herein, or a pharmaceutically acceptable salt, isotopic derivative or prodrug thereof, for the treatment of (a)-(j) a tumor in a host, including any of the tumors described herein,

(w) the use of a compound described herein, or a pharmaceutically acceptable salt, isotopic derivative or prodrug thereof, for the preparation of a medicament for the treatment according to (a)-(j) of a tumor, including any of the tumors described herein,

(x) a method for producing a medicament for the treatment according to (a)-(j) of a tumor in a host, including any of the tumors described herein, characterized in that the compound described herein is used in the production,

(y) a compound as described herein, or a pharmaceutically acceptable salt, isotopic derivative or prodrug thereof, for the treatment according to (a)-(j) of an immune, autoimmune or inflammatory disorder in a host,

(z) the use of a compound described herein, or a pharmaceutically acceptable salt, isotopic derivative or prodrug thereof, for the preparation of a medicament for the treatment according to (a)-(j) of an immune, autoimmune or inflammatory disorder,

(aa) a method for producing a medicament for the treatment according to (a)-(j) of an immune, autoimmune or inflammatory disease in a host, characterized in that the compound described herein is used in the production,

(bb) a compound as described herein, or a pharmaceutically acceptable salt, isotopic derivative or prodrug thereof, for the treatment of (a)-(j) a hematological malignancy such as multiple myeloma, leukemia, lymphoblastic leukemia, chronic lymphocytic leukemia, Hodgkin's lymphoma or non-Hodgkin's lymphoma,

(cc) the use of a compound as described herein, or a pharmaceutically acceptable salt, isotopic derivative or prodrug thereof, for the preparation of a medicament for the treatment of (a) to (j) a hematological malignancy such as multiple myeloma, leukemia, lymphoblastic leukemia, chronic lymphocytic leukemia, Hodgkin's lymphoma or non-Hodgkin's lymphoma,

(dd) a method for producing a medicament for the treatment of (a) to (j) of a hematological malignancy such as multiple myeloma, leukemia, lymphoblastic leukemia, chronic lymphocytic leukemia, Hodgkin's lymphoma or non-Hodgkin's lymphoma, characterized in that the compound described herein is used in the production,

(ee) a pharmaceutical composition comprising an effective amount for treating a host according to (a)-(j) of a compound described herein, or a pharmaceutically acceptable salt, isotopic derivative or prodrug thereof, with a pharmaceutically acceptable carrier or diluent,

(ff) treatment according to (a)-(j), wherein the compound described herein is a mixture of enantiomers or diastereomers (respectively), including a racemate,

(gg) the treatment according to (a)-(j), wherein the compound described herein is in an enantiomerically or diastereomerically (as appropriate) enriched form, including an isolated enantiomer or diastereomer (i.e., with a purity of greater than 85, 90, 95, 97, or 99%), and

(hh) a method for producing therapeutic products that contain an effective amount of a low dose of the compound described herein.

In certain alternative embodiments, a compound of the present invention is administered at a dose of less than about 800 mg. In certain embodiments, a compound of the present invention is administered at a dose of less than about 600 mg. In certain embodiments, a compound of the present invention is administered at a dose of less than about 400 mg. In certain embodiments, a compound of the present invention is administered at a dose of less than about 300 mg. In certain embodiments, a compound of the present invention is administered at a dose of less than about 200 mg. In certain embodiments, a compound of the present invention is administered at a dose of less than about 100 mg.

According to certain embodiments, a compound of the present invention is administered at a dose of about 1000, 950, 900, 850, 800, 750, 700, 650, 600, 550, 500, 475, 450, 425, 400, 375, 350, 325, 300, 275, 250, 225, 200, 175, 150, 125, 100, 90, 80, 75, 70, 65, 60, 55, 50, 45, 40, 35, 30, 25, 20, 15, 10, 5, or 1 mg.

According to certain embodiments, a compound of the present invention is administered at a dose of about 1000, 950, 900, 850, 800, 750, 700, 650, 600, 550, 500, 475, 450, 425, 400, 375, 350, 325, 300, 275, 250, 225, 200, 175, 150, 125, 100, 90, 80, 75, 70, 65, 60, 55, 50, 45, 40, 35, 30, 25, 20, 15, 10, 5, or 1 mcg.

According to certain embodiments, a compound of the present invention is administered at a dose of at least about 1000, 950, 900, 850, 800, 750, 700, 650, 600, 550, 500, 475, 450, 425, 400, 375, 350, 325, 300, 275, 250, 225, 200, 175, 150, 125, 100, 90, 80, 75, 70, 65, 60, 55, 50, 45, 40, 35, 30, 25, 20, 15, 10, 5, or 1 mcg.

According to certain embodiments, a compound of the present invention is administered at a dose of less than at least about 1000, 950, 900, 850, 800, 750, 700, 650, 600, 550, 500, 475, 450, 425, 400, 375, 350, 325, 300, 275, 250, 225, 200, 175, 150, 125, 100, 90, 80, 75, 70, 65, 60, 55, 50, 45, 40, 35, 30, 25, 20, 15, 10, 5, or 1 mcg.

Disorders mediated by Ikaros and/or Aiolos

According to one aspect of the present invention, a disorder mediated by Ikaros or Aiolos is treated with Compound 1 or a pharmaceutically acceptable salt thereof or another compound described herein. In certain embodiments, the cancer is a hematopoietic cancer. In some embodiments, the cancer is a lymphoma, leukemia, or myeloma. In certain aspects, the cancer is a non-Hodgkin's lymphoma or Hodgkin's lymphoma.

According to certain embodiments, a compound of the present invention is administered to a patient in need thereof in an amount effective to treat diffuse large B-cell lymphoma. In certain embodiments, the diffuse large B-cell lymphoma is an activated B-cell lymphoma or a germinal center B-cell lymphoma. In certain embodiments, the diffuse large B-cell lymphoma is a cancer that carries a BCL2/6 translocation. In certain embodiments, the diffuse large B-cell lymphoma is a cancer that carries a double hit. In certain embodiments, the diffuse large B-cell lymphoma is a wild-type BCL2/6 MUS cancer.

In certain embodiments, a compound of the present invention increases the concentration of caspase-3 and/or caspase-7. In certain embodiments, the increased concentration of caspase-3 and/or caspase-7 causes cancer cell death, such as by mediating the treatment of diffuse large B-cell lymphoma.

According to certain embodiments, a compound of the present invention is administered to a patient in need thereof in an amount effective for the treatment of anaplastic large cell lymphoma.

According to certain embodiments, a compound of the present invention is administered to a patient in need thereof in an amount effective for the treatment of cutaneous T-cell lymphoma.

According to certain embodiments, a compound of the present invention is administered to a patient in need thereof in an amount effective to treat mantle cell lymphoma.

According to certain embodiments, a compound of the present invention is administered to a patient in need thereof in an amount effective for the treatment of multiple myeloma.

In certain embodiments, a compound of the present invention is administered to a patient in need thereof in an amount effective to treat a disorder refractory to treatment with other cereblon ligands. In certain embodiments, a compound of the present invention is used to treat an IMiD-refractory disorder.

In certain embodiments, Compound 1 is administered to a patient in need thereof in an amount effective to treat diffuse large B-cell lymphoma. In certain embodiments, the diffuse large B-cell lymphoma is an activated B-cell lymphoma or a germinal center B-cell lymphoma. In certain embodiments, the diffuse large B-cell lymphoma is a cancer that harbors a BCL2/6 translocation. In certain embodiments, the diffuse large B-cell lymphoma is a cancer that harbors a double hit. In certain embodiments, the diffuse large B-cell lymphoma is a BCL2/6 MYC wild-type cancer.

According to certain embodiments, Compound 1 is administered to a patient in need thereof in an amount effective to treat anaplastic large cell lymphoma.

According to certain embodiments, Compound 1 is administered to a patient in need thereof in an amount effective to treat cutaneous T-cell lymphoma.

According to certain embodiments, Compound 1 is administered to a patient in need thereof in an amount effective to treat mantle cell lymphoma.

According to certain embodiments, Compound 1 is administered to a patient in need thereof in an amount effective to treat multiple myeloma.

In certain embodiments, Compound 1 is administered to a patient in need thereof in an amount effective to treat a disorder refractory to treatment with other cereblon ligands. In certain embodiments, Compound 1 is administered to a patient in need thereof in an amount effective to treat a disorder refractory to IMiD.

In one aspect, an effective amount of a compound described herein, or a pharmaceutically acceptable salt thereof, is administered to a patient for the treatment of a cancer involving the CNS, such as a lymphoma involving the CNS. In certain embodiments, the patient suffering from a cancer involving the CNS, such as a lymphoma involving the CNS, is also administered one or more additional therapeutic agents, such as ibrutinib or rituximab. In certain embodiments, a compound described herein is used to treat lymphoma of the peripheral and central nervous system.

In certain embodiments, Compound I is administered to treat PTCL-NOS (i.e., PTCL not elsewhere specified). In other embodiments, Compound 1 is administered to treat PTCL of a specified subtype, such as anaplastic large cell lymphoma (ALCL), angioimmunoblastic T-cell lymphoma (AITL), enteropathic type T-cell lymphoma, extranodal natural killer (NK) T-cell lymphoma, or extranodal T-cell lymphoma.

In certain embodiments, the disorder to be treated by a compound of the present invention is a disorder associated with an immunomodulatory effect. In certain embodiments, the disorder to be treated by a compound of the present invention is mediated by angiogenesis. In certain embodiments, the disorder to be treated by a compound of the present invention is associated with the lymphatic system.

According to certain embodiments, a compound of the present invention or a pharmaceutically acceptable salt thereof, optionally in a pharmaceutical composition as described herein, is administered to a patient in need thereof in an amount effective to destroy Ikaros or Aiolos, which is a mediator of a disorder affecting the patient, such as a human. The control of protein levels provided by any of the compounds of the present invention provides for treatment of a disease state or condition that is modulated by Ikaros or Aiolos, by reducing the level of the protein in a cell, such as a patient's cell, or by reducing the level of downstream proteins in the cell. According to certain embodiments, the treatment comprises administering an effective amount of a compound as described herein, optionally including a pharmaceutically acceptable excipient, carrier, adjuvant (i.e., a pharmaceutically acceptable composition), optionally in combination or alternating with another biologically active agent or combination of agents.

In certain embodiments, a compound of the present invention is administered to a patient in need thereof in an amount effective to treat a disorder, including, but not limited to, a benign tumor, neoplasm, tumor, cancer, abnormal cell proliferation, immune system disorder, inflammatory disorder, graft-versus-host disease, viral infection, bacterial infection, amyloid proteinopathy, proteinopathy, or fibrotic disorder.

The term "disease state" or "condition" when used in connection with any of the compounds means any disease state or condition mediated by Ikaros or Aiolos, such as cell proliferation, or proteins downstream of Ikaros or Aiolos, and wherein the destruction of such protein in a patient can provide a beneficial treatment or symptomatic relief to a patient in need thereof. In certain instances, the disease state or condition can be cured.

According to certain embodiments, a compound or its corresponding pharmaceutically acceptable salt, isotopic derivative or prodrug as described herein can be administered in an effective amount to treat a host, such as a human, suffering from lymphoma or a disorder or abnormality of lymphocyte or myelocyte proliferation. For example, a compound as described herein can be administered to a host suffering from Hodgkin's lymphoma or non-Hodgkin's lymphoma. For example, the host may have non-Hodgkin's lymphoma such as, but not limited to, AIDS-related lymphoma, anaplastic large cell lymphoma, angioimmunoblastic lymphoma, blastic NK cell lymphoma, Burkitt's lymphoma, Burkitt-like lymphoma (diffuse small cleaved cell), diffuse small cleaved cell lymphoma (DSCCL), chronic lymphocytic leukemia, small lymphocytic lymphoma, non-Hodgkin's lymphoma NOS, cutaneous T-cell lymphoma, diffuse large B-cell lymphoma, enteropathic type T-cell lymphoma, follicular lymphoma, hepatosplenic gamma-delta T-cell lymphoma, lymphoblastic lymphoma, mantle cell lymphoma, marginal zone cell lymphoma, nasal T-cell lymphoma, pediatric lymphoma, peripheral lymphoma, nodular T-cell lymphoma, primary central nervous system lymphoma, T-cell leukemia, transformed lymphomas, treatment-related T-cell lymphomas, Langerhans cell histiocytosis, or Waldenstrom's macroglobulinemia.

According to another embodiment, a compound or a corresponding pharmaceutically acceptable salt, isotopic derivative, or prodrug thereof, as described herein, can be administered in an effective amount to treat a host, e.g., a human, suffering from Hodgkin's lymphoma, such as, but not limited to, nodular sclerosis, classical Hodgkin's lymphoma (CHL), mixed cell-rich CHL, lymphocyte-depleted CHL, lymphocyte-rich CHL, lymphocyte-predominant Hodgkin's lymphoma, or nodular lymphocyte-predominant Hodgkin's lymphoma.

According to another embodiment, a compound or its corresponding pharmaceutically acceptable salt, isotopic derivative or prodrug as described herein can be administered in an effective amount to treat a host, such as a human, suffering from a condition associated with an immunomodulatory effect. Non-limiting examples of a condition associated with an immunomodulatory effect include arthritis, lupus, celiac disease, Sjogren's syndrome, polymyalgia rheumatica, multiple sclerosis, ankylosing spondylitis, type 1 diabetes, focal alopecia, vasculitis and temporal arteritis.

In certain embodiments, the condition to be treated with a compound of the present invention is a disorder associated with abnormal cell proliferation. Abnormal cell proliferation, namely hyperproliferation, can result from a wide range of factors, including genetic mutation, infection, exposure to toxins, immune system disorders, and the induction of a benign or malignant tumor.

Abnormal proliferation of B cells, T cells and/or NK cells can lead to a wide range of diseases, such as cancer, proliferative disorders and inflammatory/immune diseases. A host, such as a human, suffering from any of these disorders can be treated with an effective amount of a compound as described herein to achieve a reduction in symptoms (palliative) or a reduction in the underlying disease (disease-modifying agent).

In certain embodiments, a compound or a corresponding pharmaceutically acceptable salt, isotopic derivative, or prodrug thereof as described herein can be administered in an effective amount to treat a host, e.g., a human, suffering from a specific B-cell lymphoma or proliferative disorder, such as, but not limited to, multiple myeloma, diffuse large B-cell lymphoma, follicular lymphoma, mucosa-associated lymphoid tissue (MALT) lymphoma, small lymphocytic lymphoma, diffuse low-grade lymphocytic lymphoma, mediastinal large B-cell lymphoma, nodular marginal zone B-cell lymphoma (NMZL), splenic marginal zone lymphoma (SMZL), intravascular large B-cell lymphoma, primary effusion lymphoma or lymphogranulomatosis, B-cell prolymphocytic leukemia, hairy cell leukemia, splenic lymphoma/leukemia, unclassifiable, diffuse splenic small B-cell lymphoma with red pulp, hairy cell leukemia variant, lymphoplasmacytic lymphoma, heavy chain diseases such as alpha heavy chain disease, gamma heavy chain disease, mu heavy chain disease, plasma cell myeloma, solitary plasmacytoma of bone, extraosseous plasmacytoma, primary cutaneous follicular center cell lymphoma, T-cell/histiocyte-rich large B-cell lymphoma, DLBCL associated with chronic inflammation, Epstein-Barr virus (EBV)+ DLBCL of the elderly, primary mediastinal (thymic) large B-cell lymphoma, primary cutaneous DLBCL, leg type, ALK+ large B-cell lymphoma, plasmablastic lymphoma, large B-cell lymphoma arising in HHV8-associated multicentric neoplasm, Castleman disease, unclassifiable B-cell lymphoma with features intermediate to diffuse large B-cell lymphoma, or unclassifiable B-cell lymphoma with features intermediate to diffuse large B-cell lymphoma and classical Hodgkin lymphoma.

According to certain embodiments, a compound or a corresponding pharmaceutically acceptable salt, isotopic derivative or prodrug thereof as described herein can be administered in an effective amount to treat a host, e.g., a human, suffering from a T-cell or NK cell lymphoma, such as, but not limited to: anaplastic lymphoma kinase (ALK) positive, ALK negative anaplastic large cell lymphoma or primary cutaneous anaplastic large cell lymphoma, angioimmunoblastic lymphoma, cutaneous T-cell lymphoma, e.g., mycosis fungoides, Sezary syndrome, primary cutaneous anaplastic large cell lymphoma, primary cutaneous CD30+ T-cell lymphoproliferative disorder, primary cutaneous aggressive epidermotropic CD8+ cytotoxic T-cell lymphoma, primary cutaneous gamma-delta T-cell lymphoma, primary cutaneous small/intermediate CD4+ T-cell lymphoma and lymphomatoid papulosis, adult T-cell leukemia/lymphoma (ATLL), blastic NK-cell lymphoma, enteropathic type T-cell lymphoma, hepatosplenic gamma-delta T-cell lymphoma, lymphoblastic lymphoma, nasal NK/T-cell lymphomas, treatment-related T-cell lymphomas such as those that occur after solid organ or bone marrow transplantation, T-cell prolymphocytic leukemia, large granular lymphocyte T-cell leukemia, NK cell chronic lymphoproliferative disorder, aggressive NK-cell leukemia, systemic EBV+ T-cell lymphoproliferative disorder of childhood (associated with chronic acute EBV infection), light pox-like lymphoma, T-cell leukemia/lymphoma adults, enteropathy-associated T-cell lymphoma, hepatosplenic T-cell lymphoma, or cutaneous cellulitis-like T-cell lymphoma.

In certain embodiments, a compound or a corresponding pharmaceutically acceptable salt, isotopic derivative, or prodrug thereof as described herein can be administered to treat a host, such as a human, suffering from leukemia. For example, the host can suffer from an acute or chronic leukemia of lymphocytic or myelogenous origin, such as, but not limited to, acute lymphoblastic leukemia (ALL), acute myelocytic leukemia (AML), chronic lymphocytic leukemia (CLL), chronic myelogenous leukemia (CML), juvenile myelomonocytic leukemia (JMML), hairy cell leukemia (HCL), acute promyelocytic leukemia (a subtype of AML), large granular lymphocyte leukemia, or adult T-cell chronic leukemia. In certain embodiments, the patient has an acute myelogenous leukemia, such as undifferentiated AML (M0), myeloblastic leukemia (M1, with/without minimal cell maturation), myeloblastic leukemia (M2, with cell maturation), promyelocytic leukemia (M3 variant or M3 [M3V]), myelomonocytic leukemia (M4 variant or M4 with eosinophilia [M4E]), monocytic leukemia (M5), erythroleukemia (M6), or megakaryoblastic leukemia (M7).

There are a number of skin diseases associated with cellular hyperproliferation. Psoriasis, for example, is a benign skin disease of humans, typically characterized by plaques covered with thickened scales. The disease is caused by increased proliferation of epidermal cells of unknown cause. Chronic eczema is also associated with significant hyperproliferation of the epidermis. Other diseases caused by hyperproliferation of skin cells include atopic dermatitis, lichen planus, warts, pemphigus vulgaris, actinic keratosis, basal cell carcinoma, and squamous cell carcinoma.

Other cell hyperproliferation disorders include angiogenesis disorders, fibrotic disorders, autoimmune disorders, graft-versus-host disease, tumors, and cancer.

Proliferative disorders of blood vessels include angiogenic and vasculogenic disorders. Proliferation of smooth muscle cells during plaque development in vascular tissue causes, for example, restenosis, retinopathy, and atherosclerosis. Both cell migration and cell proliferation play a role in the formation of atherosclerotic lesions.

Fibrotic disorders are often associated with abnormal formation of the extracellular matrix. Examples of fibrotic disorders include liver cirrhosis and mesangial proliferative cell disorders. Liver cirrhosis is characterized by an increase in extracellular matrix components, leading to the formation of a liver scar. Liver cirrhosis can cause diseases such as liver cirrhosis. Increased extracellular matrix leading to liver scarring can also be caused by viral infections such as hepatitis. Adipocytes have been shown to play an important role in the development of liver cirrhosis.

Mesangial disorders are caused by abnormal proliferation of mesangial cells. Mesangial hyperproliferative cell disorders include various human kidney diseases such as glomerulonephritis, diabetic nephropathy, malignant nephrosclerosis, thrombotic microangiopathy syndromes, transplant rejection, and glomerulopathies.

Another disease with a proliferative component is rheumatoid arthritis. Rheumatoid arthritis is generally considered an autoimmune disease, which is thought to be associated with the activity of autoreactive T cells and is caused by autoantibodies produced against collagen and IgE.

Other disorders that may include an abnormal cellular proliferative component include Behcet's disease, acute respiratory distress syndrome (ARDS), coronary artery disease, postdialysis syndrome, leukemia, acquired immunodeficiency syndrome, vasculitis, lipid histiocytosis, septic shock, and inflammation in general.

A compound or a pharmaceutically acceptable salt, isotopic analog or prodrug thereof as described herein can be administered in an effective amount to treat a host, e.g., a human, suffering from a proliferative condition such as myeloproliferative disorder (MPD), polycythemia vera (PV), essential thrombocythemia (ET), myeloid metaplasia with myelofibrosis (MMF), chronic myelomonocytic leukemia (CMML), hypereosinophilic syndrome (HES), systemic mastocytosis (SMCD), and the like. According to another embodiment, a compound of the present invention is useful for the treatment of primary myelofibrosis, myelofibrosis due to polycythemia vera, myelofibrosis due to essential thrombocythemia and secondary acute myelocytic leukemia.

In certain embodiments, a compound or a pharmaceutically acceptable salt, isotopic analog, or prodrug thereof as described herein can be administered in an effective amount to treat a host, e.g., a human, suffering from a myelodysplastic syndrome (MDS), such as, but not limited to, refractory cytopenia with unilineage dysplasia, refractory anemia with ringed sideroblasts (RARS), refractory anemia with ringed sideroblast thrombocytosis (RARS-t), refractory cytopenia with multilineage dysplasia (RCMD), including RCMD with multilineage dysplasia and ringed sideroblasts (RCMD-RS), refractory anemia with excess blasts I (RAEB-I) and II (RAEB-II), 5q- syndrome, refractory cytopenia of childhood, and the like.

In certain embodiments, a compound of the present invention may provide a therapeutic effect through direct disruption of Ikaros or Aiolos, which may alter the transcriptional regulation of a downstream protein relative to Ikaros or Aiolos.

The term "neoplasia" or "cancer" is used to denote a pathological process that results in the formation and growth of a cancerous or malignant neoplasm, i.e., abnormal tissue that grows by cellular proliferation, often faster than normal, and continues to grow after the stimuli that caused the new growth have ceased. Malignant neoplasms exhibit a partial or complete lack of structural organization and functional coordination with normal tissue and in most cases invade surrounding tissues, metastasize to multiple sites, and are likely to recur after attempted removal and cause death of the patient if adequate treatment is not performed. In the context of the present invention, the term "neoplasia" is used to describe all cancerous disease states and encompasses or includes the pathological process associated with malignant hematogenous, ascitic and solid tumors. Typical cancers that can be treated with the compounds of the present invention either alone or in combination with at least one additional anticancer agent include squamous cell carcinoma, basal cell carcinoma, adenocarcinoma, hepatocellular carcinoma and renal cell carcinoma, cancer of the bladder, bowel, breast, cervix, colon, esophagus, head, kidney, liver, lung, neck, ovary, pancreas, prostate and stomach, leukemias, benign and malignant lymphomas, especially Burkitt's lymphoma and non-Hodgkin's lymphoma, benign and malignant melanomas, myeloproliferative diseases, sarcomas including Ewing's sarcoma, hemangiosarcoma, Kaposi's sarcoma, liposarcoma, myosarcoma, peripheral neuroepithelioma, synovial sarcoma, gliomas, astrocytomas, oligodendrogliomas, ependymomas, glioblastomas, neuroblastomas, ganglioneuromas, gangliogliomas, medulloblastomas, pineal gland tumors, neurofibromas, meningeofibromas and schwannomas, colorectal cancer, breast cancer, prostate cancer, cervical cancer, uterine cancer, lung cancer, ovarian cancer, testicular cancer, thyroid cancer, astrocytoma, esophageal cancer, pancreatic cancer, gastric cancer, liver cancer, colon cancer, melanoma, carcinosarcoma, Hodgkin's disease, Wilms' tumor and teratocarcinomas. Additional cancers that can be treated using the compounds of the present invention include, for example, T-lineage acute lymphoblastic leukemia (T-ALL), T-lineage lymphoblastic lymphoma (T-LL), nodular T-cell lymphoma, adult T-cell leukemia, Pre-B ALL, Pre-B lymphomas, large B-cell lymphoma, Burkitt's lymphoma, B-cell ALL, Philadelphia chromosome positive ALL, and Philadelphia chromosome positive CML.

Additional cancers that can be treated using the disclosed compounds of the present invention include, for example, acute granulocytic leukemia, acute lymphocytic leukemia (ALL), acute myelogenous leukemia (AML), adenocarcinoma, adenosarcoma, adrenal cancer, adrenocortical carcinoma, anal cancer, anaplastic astrocytoma, angiosarcoma, appendiceal cancer, astrocytoma, basal cell carcinoma, B-cell lymphoma, bile duct cancer, bladder cancer, bone cancer, bone marrow cancer, colorectal cancer, brain cancer, brain stem glioma, breast cancer, triple (estrogen, progesterone, and HER-2) negative breast cancer, double negative breast cancer (negative for two of estrogen, progesterone, and HER-2), single negative (negative for one of estrogen, progesterone, and HER-2), estrogen receptor positive, HER2-negative breast cancer, estrogen receptor-negative breast cancer, estrogen receptor-positive breast cancer, metastatic breast cancer, luminal A breast cancer, luminal B breast cancer, HER2-negative breast cancer, HER2-positive or negative breast cancer, progesterone receptor-negative breast cancer, progesterone receptor-positive breast cancer, recurrent breast cancer, carcinoid tumors, cervical cancer, cholangiocarcinoma, chondrosarcoma, chronic lymphocytic leukemia (CLL), chronic myelogenous leukemia (CML), colon cancer, colorectal cancer, craniopharyngioma, cutaneous lymphoma, cutaneous melanoma, diffuse astrocytoma, ductal carcinoma in situ (DCIS), endometrial cancer, ependymoma, epithelioid sarcoma, esophageal cancer, Ewing's sarcoma, extrahepatic bile duct cancer, eye cancer, fallopian tube cancer, fibrosarcoma, gallbladder cancer, stomach cancer, gastrointestinal cancer, gastrointestinal carcinoid cancer, gastrointestinal stromal tumors (GIST), glioblastoma germ cell multiforme (GBM), glioma, hairy cell leukemia, head and neck cancer, hemangioendothelioma, Hodgkin's lymphoma, hypopharyngeal cancer, infiltrating ductal carcinoma (IDC), infiltrating lobular carcinoma (ILC), inflammatory breast cancer (IBC), colorectal cancer, intrahepatic bile duct cancer, invasive/infiltrating breast cancer, islet cell cancer, jaw cancer, Kaposi's sarcoma, kidney cancer, laryngeal cancer, leiomyosarcoma, leptomeningeal metastasis, leukemia, lip cancer, liposarcoma, liver cancer, lobular carcinoma in situ, low-grade astrocytoma, lung cancer, lymph node cancer, lymphoma, male breast cancer, medullary carcinoma, medulloblastoma, melanoma, meningioma, Merkel cell carcinoma, mesenchymal chondrosarcoma, mesenchymal mesothelioma metastatic breast cancer, metastatic melanoma, metastatic squamous cell carcinoma of the neck, mixed gliomas, monodermal teratoma, oral cancer, mucinous carcinoma, mucosal melanoma, multiple myeloma, mycosis fungoides, myelodysplastic syndrome, nasal cavity cancer, nasopharyngeal cancer, neck cancer, neuroblastoma, neuroendocrine tumors (NETs), non-Hodgkin lymphoma, non-small cell lung cancer (NSCLC), oat cell carcinoma, eye cancer, ocular melanoma, oligodendroglioma, oral cancer, oral cavity cancer, oropharyngeal cancer, osteosarcoma, osteosarcoma, ovarian cancer, epithelial ovarian cancer, ovarian embryonal cell tumor, primary ovarian peritoneal carcinoma, ovarian sex cord stromal tumor, Paget's disease, pancreatic cancer, papillary carcinoma, paranasal sinus cancer, parathyroid cancer, pelvic cancer, penile cancer, peripheral nerve cancer, peritoneal cancer, pharyngeal cancer, pheochromocytoma, pilocytic astrocytoma, pineal tumor, pineoblastoma, pituitary cancer, primary central nervous system (CNS) lymphoma, prostate cancer, rectal cancer, renal cell carcinoma, cancer renal pelvis, rhabdomyosarcoma, salivary gland cancer, soft tissue sarcoma, bone sarcoma, sarcoma, paranasal sinus cancer, skin cancer, small cell lung cancer (SCLC), small bowel cancer, spine cancer, spinal column cancer, spinal cord cancer, squamous cell carcinoma, gastric cancer, synovial sarcoma, T-cell lymphoma, testicular cancer, throat cancer, thymoma/thymus cancer, thyroid cancer, tongue cancer, tonsil cancer, transitional cell carcinoma, tubal cancer, tubular cancer, undiagnosed cancer, ureteral cancer, urethral cancer, uterine adenocarcinoma, uterine cancer, uterine sarcoma, vaginal cancer, vulvar cancer, T-lineage acute lymphoblastic leukemia (T-ALL), T-lineage lymphoblastic lymphoma (T-LL), nodular T-cell lymphoma, Adult T-cell leukemia, Pre-B ALL, Pre-B lymphomas, Large B-cell lymphoma, Burkitt lymphoma, B-cell ALL, Philadelphia chromosome-positive ALL and Philadelphia chromosome-positive CML, Juvenile myelomonocytic leukemia (JMML), Acute promyelocytic leukemia (AML subtype), Large granular lymphocyte leukemia, Adult chronic T-cell leukemia, Diffuse large B-cell lymphoma, Follicular lymphoma, Mucosa-associated lymphoid tissue (MALT) lymphoma, Small lymphocytic lymphoma, Mediastinal large B-cell lymphoma, Nodal marginal zone B-cell lymphoma (NMZL), Splenic marginal zone lymphoma (SMZL), Intravascular large B-cell lymphoma, Primary effusion lymphoma or lymphogranulomatosis, B-cell prolymphocytic leukemia, splenic lymphoma/leukemia unclassifiable, diffuse small B-cell lymphoma of the spleen with red pulp, lymphoplasmacytic lymphoma, heavy chain diseases such as alpha heavy chain disease, gamma heavy chain disease, mu heavy chain disease, plasma cell myeloma, solitary plasmacytoma of bone, extraosseous plasmacytoma, primary cutaneous follicular center cell lymphoma, T-cell/histiocyte-rich large B-cell lymphoma, DLBCL associated with chronic inflammation, Epstein-Barr virus (EBV)+ DLBCL of the elderly, primary mediastinal (thymic) large B-cell lymphoma, primary cutaneous DLBCL, leg-type, ALK+ large B-cell lymphoma, plasmablastic lymphoma, large B-cell lymphoma arising in HHV8-associated multicentric tumor, Castleman disease, B-cell lymphoma, unclassifiable with features intermediate between diffuse large B-cell lymphoma, or B-cell lymphoma, unclassifiable with features intermediate between diffuse large B-cell lymphoma and classical Hodgkin lymphoma. In certain embodiments, the disorder is adenoid cystic carcinoma. In certain embodiments, the disorder is midline carcinoma having a NUT rearrangement.

According to another embodiment, a compound or a pharmaceutically acceptable salt, isotopic derivative or prodrug thereof, as described herein, can be used in an effective amount to treat a host, such as a human, suffering from an autoimmune disorder. Examples include, but are not limited to, acute disseminated encephalomyelitis (ADEM), Addison's disease, agammaglobulinemia, alopecia areata, amyotrophic lateral sclerosis (also Lou Gehrig's disease, motor neuron disease), Strumpell-Marie ankylosing spondylitis, antiphospholipid syndrome, antisynthetase syndrome, atopic allergy, atopic dermatitis, autoimmune aplastic anemia, autoimmune arthritis, autoimmune cardiomyopathy, autoimmune enteropathy, autoimmune granulocytopenia, autoimmune hemolytic anemia, autoimmune hepatitis, autoimmune hypoparathyroidism, autoimmune inner ear disease, autoimmune lymphoproliferative syndrome, autoimmune myocarditis, autoimmune pancreatitis, autoimmune peripheral neuropathy, autoimmune ovarian insufficiency, autoimmune polyendocrine syndrome, autoimmune progesterone dermatitis, autoimmune thrombocytopenic purpura, autoimmune thyroid disease, autoimmune urticaria, autoimmune uveitis, autoimmune vasculitis, Balo disease/Balo concentric sclerosis, Behcet's disease, Berger's disease, Bickerstaff encephalitis, Blau syndrome, pemphigus, cancer, Castleman disease, celiac disease, Chagas disease, chronic inflammatory demyelinating polyneuropathy, chronic inflammatory demyelinating polyneuropathy, chronic obstructive pulmonary disease, chronic relapsing multiple osteomyelitis, Churg-Strauss syndrome, cicatricial pemphigoid, Cogan syndrome, Cold agglutinin disease, complement component 2 deficiency, contact dermatitis, temporal arteritis, CREST syndrome, Crohn's disease, Cushing's syndrome, cutaneous leukocytoclastic vasculitis, Dego's disease, morbid obesity, dermatitis herpetiformis, dermatomyositis, diabetes mellitus type 1, diffuse cutaneous systemic sclerosis, discoid lupus erythematosus, Dressler's syndrome, drug-induced lupus syndrome, atopic dermatitis, endometriosis, enthesitis-associated arthritis, atonic fasciitis, eosinophilic gastroenteritis, eosinophilic pneumonia, acquired epidermolysis bullosa, erythema nodosum, erythroblastosis fetalis, primary cryoglobulinemia of the mixed type, Evan's syndrome, external and internal airway irritation (asthma), fibrodysplasia ossificans progressiva, fibrosing alveolitis (or Hamman-Rich syndrome), gastritis, gastrointestinal pemphigoid, glomerulonephritis, Goodpasture syndrome, Graves' disease, acute idiopathic polyneuritis (GBS), Hashimoto's encephalopathy, Hashimoto's thyroiditis, hemolytic anemia, Henoch-Schonlein disease, gestational herpes (gestational pemphigoid), hidradenitis suppurativa, Hughes-Stovin syndrome, agammaglobulinemia, idiopathic inflammatory demyelinating diseases, Hamman-Rich syndrome, idiopathic thrombocytopenic purpura, IgA nephropathy, immune glomerulonephritis, immune nephritis, immune pneumonitis, inclusion body myositis, inflammatory bowel disease, interstitial cystitis, juvenile idiopathic arthritis, also called Still's disease, Kawasaki syndrome, Lambert-Eaton syndrome, necrotizing vasculitis, lichen planus, lichen sclerosus, IgA-dependent linear dermatosis (LAD), lupus hepatitis, also called aggressive hepatitis, erythematosus, Majid syndrome, microscopic polyangiitis, Miller-Fisher syndrome, mixed connective tissue disorder, localized scleroderma, Mucha-Habermann disease, also called lichenoid acute parapsoriasis, multiple sclerosis, myasthenia gravis, myositis, Meniere's disease, narcolepsy, neuromyelitis optica (also Devic's disease), neuromyotonia, ocular cicatricial pemphigoid, opsome-myoclonal syndrome, atrophic form of autoimmune thyroiditis, migratory arthritis, PANDAS (pediatric autoimmune neuropsychiatric disorders associated with streptococcus), paraneoplastic degeneration cerebellum, paroxysmal nocturnal hemoglobinuria (PNH), Parry-Romberg syndrome, pars planitis, neuralgic amyotrophy, vulgar pemphigus, paravenous encephalomyelitis, pernicious anemia, POEMS syndrome, polyarteritis nodosa, polymyalgia rheumatica, polymyositis, primary biliary cirrhosis of the liver, primary sclerosing cholangitis, progressive inflammatory neuropathy, psoriasis, psoriatic arthritis, true red cell aplasia, toxic-allergic pustular ulcerative dermatitis, Rasmussen's encephalitis, Raynaud's disease, Reiter's syndrome, Meyenburg-Altherr-Uhlinger syndrome, restless legs syndrome, retroperitoneal fibrosis, rheumatic polyarthritis, rheumatoid arthritis, sarcoidosis, schizophrenia, Schmidt syndrome, Schnitzler syndrome, scleritis, scleroderma, sclerosing cholangitis, serum sickness, Sjogren's syndrome, spondyloarthropathy, rigid person syndrome, Still's disease, subacute septic endocarditis (SBE), Susac syndrome, Sweet syndrome, Sydenham disease, metastatic ophthalmia, systemic lupus erythematosus, Takayasu syndrome, temporal arteritis (also known as "giant cell arteritis"), thrombocytopenia, Tolosa-Hunt syndrome, transverse myelitis, ulcerative colitis, undifferentiated connective tissue disease, undifferentiated spondyloarthropathy, urticarial vasculitis, vasculitis, vitiligo, viral diseases such as Epstein-Barr virus (EBV), hepatitis B, hepatitis C, HIV, HTLV 1, varicella-zoster virus (VZV) and human papillomavirus (HPV) or Wegener's syndrome. According to some embodiments, the autoimmune disease is an allergic condition, including those caused by asthma, food allergies, atopic dermatitis, chronic pain, and rhinitis.

Cutaneous contact hypersensitivity and asthma are just two examples of immune reactions that can be associated with significant morbidity. Others include atopic dermatitis, eczema, Sjogren's syndrome including keratoconjunctivitis sicca secondary to Sjogren's syndrome, alopecia areata, allergic reactions to arthropod bites, Crohn's disease, aphthous ulcers, iritis, conjunctivitis, keratoconjunctivitis, ulcerative colitis, cutaneous lupus erythematosus, scleroderma, vaginitis, proctitis, and drug rash. These conditions may result in one or more of the following symptoms or signs: itching, swelling, redness, blisters, crusting, ulceration, pain, scaling, cracking, hair loss, scarring, or fluid discharge from the skin, eyes, or mucous membranes.

In atopic dermatitis and eczema in general, immunologically mediated leukocyte infiltration (especially infiltration of mononuclear cells, lymphocytes, neutrophils, and eosinophils) into the skin plays an important role in the pathogenesis of these diseases. Chronic eczema is also associated with significant hyperproliferation of the epidermis. Immunologically mediated leukocyte infiltration also occurs in sites other than the skin, such as the airways in asthma and the lacrimal gland of the eye in keratoconjunctivitis sicca.

A compound or a pharmaceutically acceptable salt, isotopic variant or prodrug thereof as described herein can be administered in an effective amount to treat a host, such as a human, suffering from a skin disorder such as psoriasis (e.g., psoriasis vulgaris), atopic dermatitis, skin rash, skin irritation, skin sensitization (e.g., contact dermatitis or allergic contact dermatitis). For example, some substances, including some pharmaceuticals, can cause skin sensitization when applied topically. In some embodiments, a skin disorder is treated by topical administration of compounds known in the art in combination with the compounds described herein. According to one non-limiting embodiment, the compounds of the present invention are used as topical agents for the treatment of contact dermatitis, atopic dermatitis, eczematous dermatitis, psoriasis, Sjogren's syndrome, including keratoconjunctivitis sicca secondary to Sjogren's syndrome, focal alopecia, allergic reactions caused by reactions to arthropod bites, Crohn's disease, aphthous ulcer, iritis, conjunctivitis, keratoconjunctivitis, ulcerative colitis, asthma, allergic asthma, cutaneous lupus erythematosus, scleroderma, vaginitis, proctitis, and drug rash.

Disease conditions that can be treated using the compounds of the present invention include, for example, asthma, autoimmune diseases such as multiple sclerosis, various types of cancer, ciliopathies, cleft palate, diabetes, heart disease, hypertension, inflammatory bowel disease, mental retardation, mood disorder, obesity, refractive error, infertility, Angelman syndrome, Canavan disease, celiac disease, Charcot-Marie-Tooth disease, cystic fibrosis, Duchenne muscular dystrophy, hemochromatosis, hemophilia, Klinefelter syndrome, neurofibromatosis, Felling's disease, polycystic kidney disease 1 (PKD1) or 2 (PKD2), Prader-Willi syndrome, sickle cell anemia, Tay-Sachs disease, sexogenic dwarfism.

Other disease states or conditions that can be treated with the compounds of the present invention include Alzheimer's disease, amyotrophic lateral sclerosis (Lou Gehrig's disease), anorexia nervosa, anxiety disorder, atherosclerosis, attention deficit hyperactivity disorder, autism, bipolar disorder, chronic fatigue syndrome, chronic obstructive pulmonary disease, Crohn's disease, ischemic heart disease, dementia, depression, type 1 diabetes mellitus, type 2 diabetes mellitus, epilepsy, acute idiopathic polyneuritis, irritable bowel syndrome, lupus, metabolic syndrome, multiple sclerosis, myocardial infarction, obesity, obsessive-compulsive disorder, panic disorder, Parkinson's disease, psoriasis, rheumatoid arthritis, sarcoidosis, schizophrenia, stroke, thromboangiitis obliterans, Tourette's, vasculitis.

Other disease states or conditions that can be treated with the compounds of the present invention include aceruloplasminemia, achondrogenesis type II, achondroplasia, acrocephaly, Gaucher disease type 2, acute intermittent porphyria, Canavan disease, adenomatous polyposis coli, ALA dehydratase deficiency, adenylosuccinate lyase deficiency, congenital adrenal hyperplasia, adrenoleukodystrophy, ALA-D porphyria, ALA dehydratase deficiency, alkaptonuria, Alexander disease, alkaptonuric ochronosis, alpha 1-antitrypsin deficiency, alpha-1 proteinase inhibitor, emphysema, amyotrophic lateral sclerosis, Alstrom syndrome, Alexander disease, amelogenesis imperfecta, ALA dehydratase deficiency, Anderson-Fabry disease, androgen insensitivity syndrome, diffuse angiokeratoma of the body, angiomatosis retinalis (Von Hippel-Lindau disease), Apert syndrome, arachnodactyly (Marfan syndrome), Stickler syndrome, congenital multiple arthrochalasis (Ehlers-Danlos syndrome, arthrochalasia type), Louis-Bar syndrome, Rett syndrome, primary pulmonary hypertension, Sandhoff disease, neurofibromatosis type II, cutis gyrus Weare Stevenson syndrome, Mediterranean fever, hereditary, Benjamin syndrome, beta-thalassemia, bilateral acoustic neurofibromatosis (neurofibromatosis type II), factor V Leiden thrombophilia, Bloch-Sulzberger syndrome (incontinentia pigmenti syndrome), Bloom syndrome, X-linked sideroblastic anemia, Bonnevie-Ulrich syndrome (sexogenic dwarfism), Bourneville disease (tuberous sclerosis), prion disease, Birt-Hogg-Dube syndrome, osteogenesis imperfecta (osteogenesis imperfecta), broad thumb syndrome (Rubinstein-Taybi syndrome), hemochromatosis/bronze cirrhosis (hemochromatosis), bulbospinal muscular atrophy (spinobulbar muscular atrophy), Buerger-Grutz disease (lipoprotein lipase deficiency), CGD chronic granulomatous disease, campomelic syndrome, biotinidase deficiency, cardiomyopathy (Noonan syndrome), cri du chat syndrome, CAVD (congenital absence of vas deferens), velo-cardiofacial syndrome (CBAVD), CEP (congenital porphyria), cystic fibrosis, congenital hypothyroidism, chondroplasia, otospondylomegaepiphyseal dysplasia, Lesch-Nyhan syndrome, galactosemia, Ehlers-Danlos syndrome, thanatophoric dysplasia, Coffin-Lowry syndrome, Cockayne syndrome (familial adenomatous polyposis), erythropoietic uroporphyria, congenital heart disease, methemoglobinemia/congenital methemoglobinemia, achondroplasia, X-linked sideroblastic anemia, collagenosis, cardiofacial syndrome, Cooley's anemia (beta-thalassemia), copper storage syndrome (Wilson's disease), copper transport disorder (Menkes syndrome), hereditary coproporphyria, Cowden syndrome, craniofacial dysarthrosis (Crouzon disease), Creutzfeldt-Jakob disease (prion disease), Cockayne syndrome, Cowden syndrome, Curschmann-Batten-Steinert disease (myotonic dystrophy), Beare-Stevenson syndrome of folded skin, primary hyperoxaluria, spondyloepimetaphyseal dysplasia (Strudwick type), muscular dystrophy, Duchenne and Becker types (DBMD), Ucher syndrome, degenerative nerve diseases including de Grouchy syndrome and Dejerine-Sottas syndrome, developmental disability, distal spinal muscular atrophy type V, androgen insensitivity syndrome, diffuse globoid sclerosis (Krabbe disease), DiGeorge syndrome, dihydrotestosterone receptor deficiency, androgen insensitivity syndrome, Down syndrome, dwarfism, erythropoietic protoporphyria, erythroid 5-aminolevulinate synthase deficiency, erythropoietic porphyria, erythropoietic protoporphyria, erythropoietic uroporphyria, Friedreich's ataxia - familial relapsing polyserositis, hemochromatosis, familial pressure-sensitive neuropathy, primary pulmonary hypertension (PPH), cystic fibrosis of the pancreas, fragile X syndrome, galactosemia, genetic disorders of the brain, congenital giant cell hepatitis (neonatal hemochromatosis), Grenblad-Strandberg syndrome (pseudoxanthoma elasticum), Gunther's disease (erythropoietic uroporphyria), hemochromatosis, Usher syndrome, sickle cell anemia, hemophilia, hepatoerythropoietic porphyria (HEP), multiple angioreticulomatosis (von Hippel-Lindau disease), Huntington's disease, Hutchinson-Gilford progeria syndrome (progeria), hyperandrogenism, hypochondroplasia, hypochromic anemia, immune system disorders including X-linked severe combined immunodeficiency, Insley-Astley syndrome, Jackson-Weiss syndrome, Joubert syndrome, Lesch-Nyhan syndrome, Jackson-Weiss syndrome, kidney disease including hyperoxaluria, Klinefelter syndrome, Kniest dysplasia, lacunar dementia, Langer-Saldino achondrogenesis, ataxia-telangiectasia syndrome, Lynch syndrome, lysyl hydroxylase deficiency, Machado-Joseph disease, metabolic diseases including Kniest dysplasia, Marfan syndrome, movement disorders, Mowat-Wilson syndrome, cystic fibrosis, Muenke syndrome, multiple neurofibromatosis, Nancy-Insley syndrome, Nancy-Sweeney chondrodysplasia, lipoid histiocytosis, Noack syndrome (Pfeiffer syndrome), Rendu-Weber-Osler disease, Peutz-Jeghers syndrome, polycystic kidney disease, disseminated fibrous osteitis (pseudohypoparathyroid syndrome), Peutz-Jeghers syndrome, Prader-Labhart-Willi syndrome, hemochromatosis, primary hyperuricemia syndrome (Lesch-Nyhan syndrome), primary pulmonary hypertension, primary senile degenerative dementia, prion disease, progeria (Hutchinson-Gilford progeria syndrome), developmental chorea, chronic hereditary (Huntington) (Huntington's disease), progressive muscular dystrophy, spinal muscular atrophy, propionic acidemia, protoporphyria, proximal myotonic dystrophy, pulmonary arterial hypertension, ECE (elastic pseudoxanthoma), Rb (retinoblastoma), Recklinghausen's disease (neurofibromatosis type I), recurrent polyserositis, retinal disorders, retinoblastoma, Rett syndrome, RFALS type 3, Rieger syndrome, Riley-Day syndrome, Roussy-Levy disease, severe achondroplasia with developmental delay and acanthosis nigricans (SADDAN), Li-Fraumeni syndrome, sarcoma of the breast, leukemia, and adrenal gland (SBLA) syndrome, tuberous sclerosis (tuberous sclerosis), SDAT, SED congenita (spondyloepiphyseal dysplasia congenita), SED Strudwick's (spondyloepimetaphyseal dysplasia, Strudwick type), SEDc (spondyloepiphyseal dysplasia congenita) SEMD, Strudwick's type (spondyloepimetaphyseal dysplasia, type Strudwick syndrome), cardiofacial syndrome, skin pigmentation disorders, Smith-Lemli-Opitz syndrome, South African genetic porphyria (variegate porphyria of the South African type), early-onset ascending hereditary spastic paralysis, speech and communication disorders, sphingolipidosis, Tay-Sachs disease, spinocerebellar ataxia, Stickler syndrome, stroke, androgen insensitivity syndrome, tetrahydrobiopterin deficiency, beta-thalassemia, thyroid disease, tomacular neuropathy (hereditary neuropathy with liability to pressure palsies), Treacher Collins syndrome, Triplo X syndrome (trisomy X), trisomy 21 (Down syndrome), trisomy X, VHL syndrome (von Hippel-Lindau disease), visual impairment and blindness (Alstrom syndrome), Vrolik disease, Waardenburg, Warburg-Sjo Fedelius syndrome, Wolff-Hirschhorn syndrome, Wolff syndrome in children, Weissenbacher-Zweimüller syndrome and xeroderma pigmentosum, among others.

According to certain embodiments, the present invention relates to a treatment for the treatment of multiple myeloma, comprising administering to a patient an effective amount of a compound described herein, or a pharmaceutically acceptable salt, isotopic analog, or prodrug thereof, optionally in a pharmaceutically acceptable carrier to form a composition. According to another embodiment, a compound described herein, or a pharmaceutically acceptable salt, isotopic analog, or prodrug thereof, optionally in a pharmaceutically acceptable carrier to form a composition, is for use in the treatment of multiple myeloma, wherein the treatment comprises administering the compound to a patient.

According to certain embodiments, the present invention relates to a treatment for controlling the progression of multiple myeloma, comprising administering to a patient an effective amount of a compound described herein, or a pharmaceutically acceptable salt, isotopic analog, or prodrug thereof, optionally in a pharmaceutically acceptable carrier to form a composition. According to another embodiment, a compound described herein, or a pharmaceutically acceptable salt, isotopic analog, or prodrug thereof, optionally in a pharmaceutically acceptable carrier to form a composition, is for use in a treatment for controlling the progression of multiple myeloma, wherein the treatment comprises administering the compound to a patient.

Treatments are also intended for patients who have previously received treatment for multiple myeloma but do not respond to standard therapy, in addition to those who have not previously received treatment. Additional treatments are intended for patients who have undergone surgery in an attempt to treat multiple myeloma, in addition to those who have not undergone surgery. Treatments are also intended for patients who have previously undergone transplant therapy, in addition to those who have not.

In certain embodiments, the disorder to be treated according to the present invention is a wild-type cancer, where the term "wild-type" refers to a cancer that has not developed resistance to previous effective treatment (i.e., a recurrent cancer) and does not have any mutations that confer resistance (i.e., a refractory cancer). In certain embodiments, the disorder to be treated according to the present invention is a recurrent cancer. In certain embodiments, the disorder to be treated according to the present invention is a refractory cancer. In certain embodiments, the disorder to be treated according to the present invention is a recurrent and refractory cancer.

A compound described herein, such as Compound 1 or a pharmaceutically acceptable salt thereof, can be administered to treat or control multiple myeloma or non-Hodgkin's lymphoma that is relapsed, refractory or resistant. In some embodiments, the disorder is a primary, secondary, tertiary, fourth or fifth relapse. In certain embodiments, the compounds described herein can be used to reduce, maintain or eliminate minimal residual disease (MRD).

Types of multiple myeloma that can be treated with the compounds described herein include, but are not limited to, monoclonal gammopathy of undetermined significance (MGUS), low-risk, intermediate-risk, or high-risk multiple myeloma, newly diagnosed multiple myeloma, including newly diagnosed low-risk, intermediate-risk, or high-risk multiple myeloma, transplant-eligible and transplant-ineligible multiple myeloma, indolent (slowly progressing) multiple myeloma (including low-risk, intermediate-risk, or high-risk indolent multiple myeloma), active multiple myeloma, solitary plasmacytoma, plasma cell leukemia, multiple myeloma of the central nervous system, light chain myeloma, non-secreting myeloma, immunoglobulin D myeloma, and immunoglobulin E myeloma.

According to certain embodiments, the present invention relates to a treatment for the control of multiple myeloma comprising administering to a patient an effective amount of a compound described herein, or a pharmaceutically acceptable salt, isotopic analog, or prodrug thereof, optionally in a pharmaceutically acceptable carrier to form a composition, as an induction therapy.

According to certain embodiments, the present invention relates to a treatment for the treatment or management of multiple myeloma comprising administering to a patient an effective amount of a compound described herein, or a pharmaceutically acceptable salt, isotopic analog or prodrug thereof, optionally in a pharmaceutically acceptable carrier to form a composition, as a consolidation therapy.

According to certain embodiments, the present invention relates to a method of treating or managing multiple myeloma comprising administering to a patient an effective amount of a compound described herein, or a pharmaceutically acceptable salt, isotopic analog, or prodrug thereof, optionally in a pharmaceutically acceptable carrier to form a composition, as maintenance therapy.

In certain embodiments, multiple myeloma is plasma cell leukemia.

In certain embodiments, the multiple myeloma is high-risk multiple myeloma. In certain embodiments, the high-risk multiple myeloma is relapsed or refractory. In certain embodiments, the high-risk multiple myeloma has relapsed within 12 months of the first treatment. In another embodiment, the high-risk multiple myeloma is characterized by genetic abnormalities, such as one or more of del(17/17p) and t(14,16)(q32,q32). In some embodiments, the high-risk multiple myeloma is relapsed or refractory to one, two, or three prior treatments.

In some embodiments, the multiple myeloma is newly diagnosed multiple myeloma that is transplantable. In other embodiments, the multiple myeloma is newly diagnosed multiple myeloma that is not transplantable.

In some embodiments, the multiple myeloma shows early progression (e.g., less than 12 months) after initial treatment. In other embodiments, the multiple myeloma shows early progression (e.g., less than 12 months) after autologous stem cell transplantation. In another embodiment, the multiple myeloma is refractory to lenalidomide. In another embodiment, the multiple myeloma is refractory to pomalidomide. In some such embodiments, the multiple myeloma is predicted to be refractory to pomalidomide (e.g., by molecular characterization). According to another embodiment, the multiple myeloma is relapsed or refractory to 3 or more treatments and has been treated with a proteasome inhibitor (e.g., bortezomib, carfilzomib, ixazomib, oprozomib, or marizomib) and an immunomodulatory compound (e.g., thalidomide, lenalidomide, pomalidomide, iberdomide, or avadomide), or is doubly refractory to a proteasome inhibitor and an immunomodulatory compound. In other embodiments, the multiple myeloma is relapsed or refractory to 3 or more prior therapies, including, for example, an anti-CD38 monoclonal antibody (CD38 mAb, for example, daratumumab or isatuximab), a proteasome inhibitor (for example, bortezomib, carfilzomib, ixazomib, or marizomib), and an immunomodulatory compound (for example, thalidomide, lenalidomide, pomalidomide, iberdomide, or avadomide), or is doubly refractory to a proteasome inhibitor or immunomodulatory compound and a CD38 mAb. In other embodiments, the multiple myeloma is triply refractory, such as the multiple myeloma is refractory to a proteasome inhibitor (e.g., bortezomib, carfilzomib, ixazomib, oprozomib, or marizomib), an immunomodulatory compound (e.g., thalidomide, lenalidomide, pomalidomide, iberdomide, or avadomide), and one other active agent as described herein.

According to certain embodiments, the present invention relates to a treatment for the control of relapsed or refractory multiple myeloma in patients with renal impairment or symptom thereof, comprising administering to the patient an effective amount of a compound described herein, or a pharmaceutically acceptable salt, isotopic analog or prodrug thereof, optionally in a pharmaceutically acceptable carrier to form a composition.

According to another embodiment, the present invention relates to a treatment for the control of relapsed or refractory multiple myeloma in frail patients, comprising administering to the patient an effective amount of a compound described herein, or a pharmaceutically acceptable salt, isotopic analogue or prodrug thereof, optionally in a pharmaceutically acceptable carrier to form a composition, wherein the frail patient is ineligible for induction therapy or intolerant of dexamethasone treatment. According to other embodiments, the frail patient is elderly, such as over 65 years of age.

According to another embodiment, the present invention relates to a treatment for the control of multiple myeloma, relapsed or refractory to fourth-line treatment, comprising administering to a patient an effective amount of a compound described herein, or a pharmaceutically acceptable salt, isotopic analog or prodrug thereof, optionally in a pharmaceutically acceptable carrier to form a composition.

According to another embodiment, the present invention relates to a treatment for the control of newly diagnosed multiple myeloma that does not allow transplantation, comprising administering to a patient an effective amount of a compound described herein, or a pharmaceutically acceptable salt, isotopic analog or prodrug thereof, optionally in a pharmaceutically acceptable carrier to form a composition.

According to another embodiment, the present invention relates to a treatment for the control of newly diagnosed multiple myeloma that does not allow transplantation, comprising administering to a patient an effective amount of a compound described herein, or a pharmaceutically acceptable salt, isotopic analog or prodrug thereof, optionally in a pharmaceutically acceptable carrier to form a composition, as maintenance therapy following other therapy or transplantation.

According to another embodiment, the present invention relates to a treatment for the control of high-risk multiple myeloma that is relapsed or refractory to one, two or three prior treatments, comprising administering to a patient an effective amount of a compound described herein, or a pharmaceutically acceptable salt, isotopic analog or prodrug thereof, optionally in a pharmaceutically acceptable carrier to form a composition.

In certain embodiments, the disorder to be treated according to the present invention is relapsed and/or refractory non-Hodgkin lymphoma. In certain embodiments, the disorder to be treated according to the present invention is relapsed non-Hodgkin lymphoma. In certain embodiments, the disorder to be treated according to the present invention is refractory non-Hodgkin lymphoma. In certain embodiments, the disorder to be treated according to the present invention is relapsed and refractory non-Hodgkin lymphoma.

In certain embodiments, the disorder to be treated according to the present invention is relapsed and/or refractory multiple myeloma. In certain embodiments, the disorder to be treated according to the present invention is relapsed multiple myeloma. In certain embodiments, the disorder to be treated according to the present invention is refractory multiple myeloma. In certain embodiments, the disorder to be treated according to the present invention is relapsed and refractory multiple myeloma.

In certain embodiments, the disorder treated according to the present invention is relapsed and/or refractory nodal T-cell lymphoma. In certain embodiments, the disorder treated according to the present invention is relapsed nodal T-cell lymphoma. In certain embodiments, the disorder treated according to the present invention is refractory nodal T-cell lymphoma. In certain embodiments, the disorder treated according to the present invention is relapsed and refractory nodal T-cell lymphoma.

In certain embodiments, Compound 1 is administered to treat PTCL-NOS (i.e., PTCL not elsewhere specified). In other embodiments, Compound 1 is administered to treat PTCL of a specified subtype, such as anaplastic large cell lymphoma (ALCL), angioimmunoblastic T-cell lymphoma (AITL), enteropathic type T-cell lymphoma, or extranodal natural killer (NK)/T-cell lymphoma.

In certain embodiments, the disorder treated according to the present invention is relapsed and/or refractory systemic anaplastic large cell lymphoma (ALK ⁺ ). In certain embodiments, the disorder treated according to the present invention is relapsed systemic anaplastic large cell lymphoma (ALK ⁺ ). In certain embodiments, the disorder treated according to the present invention is refractory systemic anaplastic large cell lymphoma (ALK ⁺ ). In certain embodiments, the disorder treated according to the present invention is relapsed and refractory systemic anaplastic large cell lymphoma (ALK ⁺ ).

In certain embodiments, the disorder treated according to the present invention is relapsed and/or refractory systemic anaplastic large cell lymphoma ( ^ALK- ). In certain embodiments, the disorder treated according to the present invention is relapsed systemic anaplastic large cell lymphoma ( ^ALK- ). In certain embodiments, the disorder treated according to the present invention is refractory systemic anaplastic large cell lymphoma ( ^ALK- ). In certain embodiments, the disorder treated according to the present invention is relapsed and refractory systemic anaplastic large cell lymphoma ( ^ALK- ).

In certain embodiments, the disorder to be treated according to the present invention is relapsed and/or refractory angioimmunoblastic T-cell lymphoma. In certain embodiments, the disorder to be treated according to the present invention is relapsed angioimmunoblastic T-cell lymphoma.

In certain embodiments, the disorder to be treated according to the present invention is refractory angioimmunoblastic T-cell lymphoma. In certain embodiments, the disorder to be treated according to the present invention is relapsed and refractory angioimmunoblastic T-cell lymphoma.

In certain embodiments, the disorder treated according to the present invention is relapsed and/or refractory anaplastic large cell lymphoma. In certain embodiments, the disorder treated according to the present invention is relapsed anaplastic large cell lymphoma. In certain embodiments, the disorder treated according to the present invention is refractory anaplastic large cell lymphoma. In certain embodiments, the disorder treated according to the present invention is relapsed and refractory anaplastic large cell lymphoma.

In certain embodiments, the disorder treated according to the present invention is relapsed and/or refractory mantle cell lymphoma. In certain embodiments, the disorder treated according to the present invention is relapsed mantle cell lymphoma. In certain embodiments, the disorder treated according to the present invention is refractory mantle cell lymphoma. In certain embodiments, the disorder treated according to the present invention is relapsed and refractory mantle cell lymphoma.

In certain embodiments, the disorder to be treated according to the present invention is relapsed and/or refractory follicular lymphoma. In certain embodiments, the disorder to be treated according to the present invention is relapsed follicular lymphoma. In certain embodiments, the disorder to be treated according to the present invention is refractory follicular lymphoma. In certain embodiments, the disorder to be treated according to the present invention is relapsed and refractory follicular lymphoma.

In certain embodiments, the disorder treated according to the present invention is relapsed and/or refractory follicular T-cell lymphoma. In certain embodiments, the disorder treated according to the present invention is relapsed follicular T-cell lymphoma. In certain embodiments, the disorder treated according to the present invention is refractory follicular T-cell lymphoma. In certain embodiments, the disorder treated according to the present invention is relapsed and refractory follicular T-cell lymphoma.

In certain embodiments, the disorder treated according to the present invention is relapsed and/or refractory nodal T-cell lymphoma. In certain embodiments, the disorder treated according to the present invention is relapsed nodal T-cell lymphoma. In certain embodiments, the disorder treated according to the present invention is refractory nodal T-cell lymphoma. In certain embodiments, the disorder treated according to the present invention is relapsed and refractory nodal T-cell lymphoma.

In certain embodiments, the disorder treated according to the present invention is relapsed and/or refractory diffuse large B-cell lymphoma. In certain embodiments, the disorder treated according to the present invention is relapsed diffuse large B-cell lymphoma. In certain embodiments, the disorder treated according to the present invention is refractory diffuse large B-cell lymphoma. In certain embodiments, the disorder treated according to the present invention is relapsed and refractory diffuse large B-cell lymphoma.

In certain embodiments, Compound 1 is administered to a patient in need thereof in an effective amount to treat follicular T-cell lymphoma. In certain embodiments, Compound 1 is administered to a patient in need thereof in an effective amount to treat angioimmunoblastic T-cell lymphoma. In certain embodiments, Compound 1 is administered to a patient in need thereof in an effective amount to treat systemic anaplastic large cell lymphoma ( ^ALK- ). In certain embodiments, Compound 1 is administered to a patient in need thereof in an effective amount to treat systemic anaplastic large cell lymphoma (ALK ⁺ ).

Additional examples of non-Hodgkin lymphomas that can be treated with the compound described herein include double-hit lymphoma, triple-hit lymphoma, extranodal marginal zone B-cell lymphoma of MALT cell type, extranodal NK/T-cell lymphoma, and lymphoma of myeloid origin.

Pharmacodynamic dose modification and biomarkers

According to certain embodiments, a compound of the present invention is administered in an effective amount to treat a patient suffering from a cereblon-mediated disorder, comprising administering to the patient an effective amount of the compound and monitoring the concentration of a biomarker selected from IRF-1, caspase-1, caspase-3, caspase-7, cyclin D1, E2F1, ZFP91, SALL4, IRF-4, BLIMP1, MYC, IL-2, T-cell activation and/or proliferation, BCMA, M-protein, PARP, BIM, survivin, IKZF1, IKZF3, ZFP91, WIZ and/or IFN-γ, or a combination thereof.

In certain embodiments, the concentration of IRF-1 and/or caspase 3 increases when a patient is treated with a compound described herein. The amount of increase can be used to determine whether or not the dose of the compound described herein should be increased, decreased, or maintained the same. For example, if the concentration of IRF-1 and/or caspase 3 increases by less than 1.25-fold, 1.5-fold, 1.75-fold, or 2-fold, then the physician may increase the dose of Compound 1 administered to the patient being treated for lymphoma.

In certain embodiments, the concentration of cyclin D and/or E2F1 decreases when a patient is treated with a compound described herein. The amount of decrease can be used to determine whether or not the dose of the compound described herein should be increased, decreased, or maintained the same. For example, if the concentration of cyclin D and E2F1 decreases by less than 1.25, 1.5, 1.75, or 2-fold, the physician may increase the dose of Compound 1 administered to the patient being treated for lymphoma.

In certain embodiments, the concentration of cyclin D, E2F1, ZFP91, SALL4, IRF-4, BLIMP 1 and/or MYC is reduced when a patient is treated with a compound described herein. In certain embodiments, the patient has lymphoma. The amount of reduction can be used to determine whether or not the dose of the compound described herein should be increased, decreased, or maintained at the same level.

In certain embodiments, the concentration of IL-2 and/or IFN-γ increases when a patient is treated with a compound described herein. In certain embodiments, the patient has myeloma. The magnitude of the increase can be used to determine whether or not the dose of the compound described herein should be increased, decreased, or maintained at the same level.

In certain embodiments, the concentration of the biomarker is increased by about 3, 4, 5, 6, 7, or 8-fold upon delivery of an effective dose of a compound described herein, such as Compound 1. For example, as shown in Fig. 51, the level of caspase-3 and caspase-7 activity is increased by greater than 800% upon treatment with Compound 1. In certain embodiments, the level of caspase 3/7 activity is assessed, and if it is increased by less than about 2, 3, 4, 5, 6, 7, or 8-fold, the dose of Compound 1 is increased.

In certain embodiments, the biomarker is STAT3.

In certain embodiments, the biomarker is Ki67.

In certain embodiments, the concentration of the biomarker is reduced by about 3, 4, 5, 6, 7, or 8 times upon delivery of an effective dose of a compound described herein, such as Compound 1.

According to certain embodiments, a patient to be treated with a compound selected from Compound 1, Compound 2, Compound 3, Compound 4, Compound 5, Compound 6, Compound 7, Compound 8, Compound 9, Compound 10, Compound 11, Compound 12, and Compound 13 is selected based on the concentration of a biomarker. For example, a patient to be treated with Compound 1 can be selected based on the concentration of a biomarker.

According to certain embodiments, the biomarker is selected from IKZF1, IKZF3, MYC, il2, INFy, TNFa, sFLC, and sBCMA.

In certain embodiments, the biomarker is selected from IRF-1, caspase-3, cyclin D1, E2F1, ZFP91, SALL4, IRF-4, BLIMP1, MYC, IL-2 and/or IFN-γ.

In certain embodiments, the biomarker is a marker of anti-tumor immunity (e.g., a cytokine, tumor infiltrating lymphocyte, T cell activation and/or proliferation, and B cell markers such as BCMA or M protein, or a combination thereof).

In certain embodiments, the biomarker is an apoptosis marker (e.g., total and/or cleavage caspase-1, caspase-3, caspase-7, PARP, BIM, or survivin, or a combination thereof).

In certain embodiments, the biomarker is a zinc finger protein (e.g., IKZF1, IKZF3, ZFP91, WIZ, or SALL4, or a combination thereof).

Benefits of treatment

According to one aspect of the present invention, the treatment described herein has one or more advantages over currently approved cancer treatments, such as multiple myeloma or non-Hodgkin's lymphoma. For example, Compound 1 or a pharmaceutically acceptable salt thereof, when administered using the treatment methods described herein, has a better outcome in one or more of the indicators described below than currently approved treatments, such as thalidomide, pomalidomide or lenalidomide (see Examples 9, 12, 13, 15-19, 26-31 and 35, demonstrating the superior efficacy of Compound 1 in multiple myeloma and non-Hodgkin's lymphoma models).

In certain embodiments, an advantage provided by Compound 1 over currently known treatments is a reduced tendency to develop resistance. For example, mice that had developed resistance to pomalidomide treatment still responded rapidly to treatment with Compound 1 (see Fig. 40). Furthermore, when the mice were off treatment for a long enough period of time for their tumors to regrow, the tumors still shrank rapidly when re-treated with Compound 1 (see Fig. 33).

In other embodiments, an advantage over currently approved therapies is the ability to treat refractory tumors. For example, in mice, doses as low as 30 μg/kg of Compound 1 were effective in treating NCI-H929 tumors that were unresponsive to 3000 μg/kg (see Fig. 33). Additionally, in a panel of cell lines, Compound 1 was consistently 2-3 magnitudes more potent than pomalidomide and even demonstrated efficacy against cells that were refractory to pomalidomide (see Fig. 32). This effect was also demonstrated by monitoring the concentration of biomarkers such as caspase 3 (see Fig. 31).

In other embodiments, the advantage over currently approved therapies is a more rapid degradation of IKZF1 and IKZF3 and thus a more rapid treatment of cancer. For example, Compound 1 degrades more IKZF1 in one hour than pomalidomide degrades in two hours when dosed at the same concentration (see Fig. 29).

According to certain embodiments, the present invention relates to a treatment for inducing a therapeutic response as assessed by the International Uniform Response Criteria (IURC) for multiple myeloma (described in Durie B. G. M, et al. "International uniform response criteria for multiple myeloma. Leukemia 2006, 10(10): 1-7) in a patient suffering from multiple myeloma, comprising administering to the patient an effective amount of a compound described herein, or a pharmaceutically acceptable salt, isotopic analog, or prodrug thereof, optionally in a pharmaceutically acceptable carrier to form a composition.

According to another embodiment, the present invention relates to a treatment to achieve a strong complete response, a complete response or a very good partial response as assessed by IURC for multiple myeloma in a patient suffering from multiple myeloma, comprising administering to the patient an effective amount of a compound described herein or a pharmaceutically acceptable salt, isotopic analog or prodrug thereof, optionally in a pharmaceutically acceptable carrier to form a composition.

According to another embodiment, the present invention relates to a treatment to achieve an increase in overall survival, progression-free survival, event-free survival, time to process, or disease-free survival in a patient suffering from multiple myeloma, comprising administering to the patient an effective amount of a compound described herein, or a pharmaceutically acceptable salt, isotopic analog, or prodrug thereof, optionally in a pharmaceutically acceptable carrier to form a composition (see Examples 21 and 22, which demonstrate a dose-dependent increase in overall survival in animal models).

The benefits of Compound 1 can be further enhanced by the administration of an additional biologically active agent in combination therapy. For example, in mouse studies, when Compound 1 was administered according to a treatment regimen that included weekly dexamethasone, it was significantly more effective than the same dose of Compound 1 or dexamethasone alone (see Fig. 41).

Treatment of IKZF1/IKZF3-mediated cancers with high-grade mutations

In certain embodiments, a compound described herein is administered in an effective amount to treat a cancer mediated by a protein with one or more mutations, such as multiple myeloma mediated by a protein with one or more mutations.

In some embodiments, the compounds described herein can be administered in an effective amount to treat or manage multiple myeloma characterized by a genetic abnormality, such as, but not limited to, cyclin D translocations (e.g., t(11,14)(q13,q32), t(6,14)(p21,32), t(12,14)(p13,q32), or t(6,20),), MMSET translocations (e.g., t(4,14)(p16,q32), MAF translocations (e.g., t(14,16)(q32,a32), t(20,22), t(16,22)(q11,q13), or t(14,20)(q32,q11), or another chromosomal factor (e.g., deletion of 17p13 or chromosome 13, del(17/17p), nonhyperdiploidy and addition (1q)).

In certain embodiments, the multiple myeloma has a p53 mutation. In certain embodiments, the p53 mutation is the Q331 mutation. In certain embodiments, the p53 mutation is the R273H mutation. In certain embodiments, the p53 mutation is the K132 mutation. In certain embodiments, the p53 mutation is the K132N mutation. In certain embodiments, the p53 mutation is the R337 mutation. In certain embodiments, the p53 mutation is the R337L mutation. In certain embodiments, the p53 mutation is the W146 mutation. In certain embodiments, the p53 mutation is the S261 mutation. In certain embodiments, the p53 mutation is the S261T mutation. In certain embodiments, the p53 mutation is the E286 mutation. In certain embodiments, the p53 mutation is the E286K mutation. In certain embodiments, the p53 mutation is the R175 mutation. In certain embodiments, the p53 mutation is the R175H mutation. In certain embodiments, the p53 mutation is the E258 mutation. In certain embodiments, the p53 mutation is the E258K mutation. In certain embodiments, the p53 mutation is the A161 mutation. In certain embodiments, the p53 mutation is the A161T mutation.

In certain embodiments, the multiple myeloma has a homozygous deletion of p53. In certain embodiments, the multiple myeloma has a homozygous deletion of wild-type p53. In certain embodiments, the multiple myeloma has wild-type p53.

In certain embodiments, multiple myeloma exhibits activation of one or more oncogenic factors. In certain embodiments, the one or more oncogenic factors are selected from the group consisting of C-MAF, MAFB, FGFR3, MMset, cyclin D1, and cyclin D. In certain embodiments, multiple myeloma exhibits activation of C-MAF. In certain embodiments, multiple myeloma exhibits activation of MAFB. In certain embodiments, multiple myeloma exhibits activation of FGFR3 and MMset. In certain embodiments, multiple myeloma exhibits activation of C-MAF, FGFR3, and MMset. In certain embodiments, multiple myeloma exhibits activation of Cyclin D1. In certain embodiments, multiple myeloma exhibits activation of MAFB and Cyclin D1. In certain embodiments, multiple myeloma exhibits activation of Cyclin D.

In certain embodiments, the multiple myeloma has one or more chromosomal translocations. In certain embodiments, the chromosomal translocation is t(14,16). In certain embodiments, the chromosomal translocation is t(14,20). In certain embodiments, the chromosomal translocation is t(4,14). In certain embodiments, the chromosomal translocations are t(4,14) and t(14,16). In certain embodiments, the chromosomal translocation is t(11,14). In certain embodiments, the chromosomal translocation is t(6,20). In certain embodiments, the chromosomal translocation is t(20,22). In certain embodiments, the chromosomal translocations are t(6,20) and t(20,22). In certain embodiments, the chromosomal translocation is t(16,22). In certain embodiments, the chromosomal translocations are t(14,16) and t(16,22). In certain embodiments, the chromosomal translocations are t(14,20) and t(11,14).

In certain embodiments, the multiple myeloma has a p53 Q331 mutation, C-MAF activation, and a chromosomal translocation at t(14, 16). In certain embodiments, the multiple myeloma has a homozygous p53 deletion, C-MAF activation, and a chromosomal translocation at t(14, 16). In certain embodiments, the multiple myeloma has a p53 K132N mutation, MAFB activation, and a chromosomal translocation at t(14,20). In certain embodiments, the multiple myeloma has wild-type p53, FGFR3 and MMset activation, and a chromosomal translocation at t(4, 14). In certain embodiments, the multiple myeloma has wild-type p53, C-MAF activation, and a chromosomal translocation at t(14, 16). In certain embodiments, the multiple myeloma has a homozygous deletion of p53, activation of FGFR3, MMset, and C-MAF, and chromosomal translocations at t(4,14) and t(14,16). In certain embodiments, the multiple myeloma has a homozygous deletion of p53, activation of Cyclin D1, and a chromosomal translocation at t(11,14). In certain embodiments, the multiple myeloma has a p53 R337L mutation, activation of Cyclin D1, and a chromosomal translocation at t(11,14). In certain embodiments, the multiple myeloma has a p53 W146 mutation, activation of FGFR3 and MMset, and a chromosomal translocation at t(4,14). In certain embodiments, the multiple myeloma has a p53 S261T mutation, MAFB activation, and chromosomal translocations at t(6,20) and t(20,22). In certain embodiments, the multiple myeloma has a p53 E286K mutation, FGFR3 and MMset activation, and a chromosomal translocation at t(4,14). In certain embodiments, the multiple myeloma has a p53 R175H mutation, FGFR3 and MMset activation, and a chromosomal translocation at t(4,14). In certain embodiments, the multiple myeloma has a p53 E258K mutation, C-MAF activation, and chromosomal translocations at t(14,16) and t(16,22). In certain embodiments, the multiple myeloma has wild-type p53, MAFB and Cyclin D1 activation, and chromosomal translocations at t(14,20) and t(11,14). In certain embodiments, the multiple myeloma has a p53 A161T mutation, Cyclin D activation, and a chromosomal translocation at t(11,14).

Patient selection

In certain embodiments, a patient treated with a compound described herein has previously received treatment with an IMiD, such as thalidomide, lenalidomide, or pomalidomide.

In certain embodiments, a patient treated with a compound described herein has previously been treated with an anti-CD20 antibody, such as rituximab, ocrelizumab, obinutuzumab, ofatumumab, ibritumomab, tositumomab, or ublituximab.

In certain embodiments, a patient receiving treatment with a compound described herein has previously received treatment with an anti-CD38 antibody, such as daratumumab or isatuximab.

In certain embodiments, a patient receiving treatment with a compound described herein has previously received treatment with an anti-CD30 antibody, such as brentuximab.

In certain embodiments, a patient treated with a compound described herein has previously been treated with a BTK inhibitor, such as ibrutinib, acalabrutinib, or zanubrutinib.

In certain embodiments, the patient being treated with a compound described herein has previously been treated with an alkylating agent, such as cyclophosphamide, melfan, melphalan flufenamide, or bendamustine.

In certain embodiments, a patient receiving treatment with a compound described herein has previously received treatment with a proteasome inhibitor, such as bortezomib, carfilzomib, or ixazomib.

In certain embodiments, a patient receiving treatment with a compound described herein has previously received treatment with a glucocorticoid, such as dexamethasone, predinisone, or methylprednisolone.

In certain embodiments, a patient treated with a compound described herein has previously received treatment with an osteomodifying agent, such as denosumab, zoledronic acid, or pamidronate.

In certain embodiments, a patient treated with a compound described herein has previously received treatment with an HDAC inhibitor, such as panobinostat.

In certain embodiments, a patient receiving treatment with a compound described herein has previously received treatment with a nuclear export inhibitor, such as selinexor.

In certain embodiments, a patient receiving treatment for multiple myeloma has received at least 1, 2, 3, or 4 prior myeloma or lymphoma treatment regimens, such as lenalidomide, pomalidomide, a proteasome inhibitor, a glucocorticoid, or an anti-CD38 antibody. For example, a patient who has previously received at least 3 myeloma treatment regimens, including at least two consecutive cycles of lenalidomide, pomalidomide, a proteasome inhibitor, a glucocorticoid, or an anti-CD38 antibody.

In certain embodiments, a patient receiving treatment for multiple myeloma has an M-protein level of ≥ about 5 g/dL as determined by serum protein electrophoresis (sPEP), ≥ 200 mg/24-hour urine collection as determined by urine protein electrophoresis (uPEP), serum free light chain (FLC) levels > 100 mg/L, including the light chain, and an abnormal kappa/lambda (κ/λ) ratio in subjects without measurable serum or urine M-protein levels and/or with a serum IgA level of ≥ 0.50 g/dL.

In certain embodiments, the patient receiving treatment has nodular T-cell lymphoma and has received at least one prior alkylating agent-based chemotherapy treatment.

In certain embodiments, the patient receiving treatment has anaplastic large cell lymphoma (ALCL) and has received at least one prior alkylating agent-based chemotherapy and has also received anti-CD30 antibody therapy.

In certain embodiments, the patient receiving treatment has mantle cell lymphoma and has received at least two prior lines of therapy, including an anti-CD20 antibody and an alkylating chemotherapeutic agent, as well as a Bruton tyrosine kinase inhibitor.

In certain embodiments, the patient receiving treatment has follicular lymphoma and has received at least two prior lines of therapy, including a line with an anti-CD20 antibody and an alkylating chemotherapeutic agent.

In certain embodiments, the patient receiving treatment has diffuse large B-cell lymphoma and has received at least two prior lines of therapy, including anti-CD20 antibody therapy, and has previously received an autologous bone marrow transplant (or is not suitable for a bone marrow transplant).

In certain embodiments, a patient receiving treatment for non-Hodgkin lymphoma has a lesion that is measurable in at least two dimensions using PET-CT, such as a lesion with a minimum dimension of at least about 15 mm in greatest diameter.

In some embodiments, the patient to be treated with one of the compounds described herein has not been treated with a therapy for multiple myeloma prior to administration. In some embodiments, the patient to be treated with one of the compounds described herein has been treated with a therapy for multiple myeloma prior to administration. In some embodiments, the patient to be treated with one of the compounds described herein has acquired resistance to a drug for the therapy for multiple myeloma. In some embodiments, a patient to be treated with one of the compounds described herein has acquired resistance to one, two, or three therapies for multiple myeloma, wherein the therapies are selected from an anti-CD38 antibody (CD38 mAB, e.g., daratumumab or isatuximab), a proteasome inhibitor (e.g., bortezomib, carfilzomib, ixazomib, or marizomib), and an immunomodulatory compound (e.g., thalidomide, lenalidomide, pomalidomide, iberdomide, or avadomide).

The compounds described herein can be administered in an effective amount to treat a patient, regardless of the age of the patient. In some embodiments, the patient is 18 years of age or older. In other embodiments, the patient is 18, 25, 35, 40, 45, 50, 55, 60, 65, or 70 years of age. In other embodiments, the patient is under 65 years of age. In other embodiments, the patient is over 65 years of age. In certain embodiments, the patient is an elderly patient with multiple myeloma, such as a patient over 65 years of age. In certain embodiments, the patient is an elderly patient suffering from multiple myeloma, such as a patient over 75 years of age.

V. Combination therapy

Any of the compounds described herein can be administered in an effective amount alone or in combination to treat a host, such as a human, suffering from a disorder described herein. In certain embodiments, a compound described herein is administered with an additional biologically active agent.

The term "biologically active agent" is used to describe an agent other than a compound of the present invention that can be administered in combination or in alternation with a compound of the present invention to achieve a desired therapeutic outcome. In certain embodiments, a compound of the present invention and a biologically active agent are administered such that they are active in vivo for overlapping periods of time, such as having overlapping periods of Cmax, Tmax, AUC, or other pharmacokinetic parameter. In another embodiment, a compound of the present invention and a biologically active agent are administered to a host in need thereof, which do not have overlapping pharmacokinetic parameters, but one has a therapeutic effect on the therapeutic efficacy of the other.

In the context of the present invention, when a compound is administered in combination with another, the combination may be in the form of a single dosage form or several dosage forms for administration at the same time or at different times. In addition, the compounds administered in combination may be administered according to different dosage regimens. For example, a combination of Compound 1 with dexamethasone includes a treatment regimen in which Compound 1 is administered once a day for 21 consecutive days in a 28-day treatment cycle, and dexamethasone is administered once a week during the treatment cycle.

According to certain embodiments, Compound 1 is administered to a patient in need thereof in an effective amount in combination with one or more additional therapeutic agents described herein.

According to certain embodiments, Compound 2 is administered to a patient in need thereof in an effective amount in combination with one or more additional therapeutic agents described herein.

According to certain embodiments, Compound 3 is administered to a patient in need thereof in an effective amount in combination with one or more additional therapeutic agents described herein.

According to certain embodiments, Compound 4 is administered to a patient in need thereof in an effective amount in combination with one or more additional therapeutic agents described herein.

According to certain embodiments, Compound 5 is administered to a patient in need thereof in an effective amount in combination with one or more additional therapeutic agents described herein.

According to certain embodiments, Compound 6 is administered to a patient in need thereof in an effective amount in combination with one or more additional therapeutic agents described herein.

According to certain embodiments, Compound 7 is administered to a patient in need thereof in an effective amount in combination with one or more additional therapeutic agents described herein.

According to certain embodiments, Compound 8 is administered to a patient in need thereof in an effective amount in combination with one or more additional therapeutic agents described herein.

According to certain embodiments, Compound 9 is administered to a patient in need thereof in an effective amount in combination with one or more additional therapeutic agents described herein.

According to certain embodiments, Compound 10 is administered to a patient in need thereof in an effective amount in combination with one or more additional therapeutic agents described herein.

According to certain embodiments, Compound 11 is administered to a patient in need thereof in an effective amount in combination with one or more additional therapeutic agents described herein.

According to certain embodiments, Compound 12 is administered to a patient in need thereof in an effective amount in combination with one or more additional therapeutic agents described herein.

According to certain embodiments, Compound 13 is administered to a patient in need thereof in an effective amount in combination with one or more additional therapeutic agents described herein.

Corticosteroids

In certain embodiments, a compound of the present invention is administered in combination with a corticosteroid. In certain embodiments, the corticosteroid is dexamethasone. In certain embodiments, Compound 1 is administered with a corticosteroid. In certain embodiments, Compound 1 is administered with dexamethasone, such as according to the dosing regimen shown below:

In certain aspects, Compound 1 is administered QD according to a 21/7 dosing schedule for each 28-day cycle in combination with another biologically active agent, such as dexamethasone once weekly. In certain embodiments, the dose of dexamethasone for an adult ≤ 75 years old is 40 mg QW on days 1, 8, 15, and 22 of a 28-day cycle. In certain embodiments, the dose of dexamethasone for an adult > 75 years old is 20 mg QW on days 1, 8, 15, and 22 of a 28-day cycle.

In certain embodiments, a compound described herein is administered in combination with a corticosteroid. Non-limiting examples of corticosteroids include dexamethasone, prednisone, fludrocortisone, hydrocortisone, cortisone, betamethasone, methylprednisolone.

In certain embodiments, Compound 1 is administered in combination with a corticosteroid. In certain embodiments, Compound 2 is administered in combination with a corticosteroid. In certain embodiments, Compound 3 is administered in combination with a corticosteroid. In certain embodiments, Compound 4 is administered in combination with a corticosteroid. In certain embodiments, Compound 5 is administered in combination with a corticosteroid. In certain embodiments, Compound 6 is administered in combination with a corticosteroid. In certain embodiments, Compound 7 is administered in combination with a corticosteroid. In certain embodiments, Compound 8 is administered in combination with a corticosteroid. In certain embodiments, Compound 9 is administered in combination with a corticosteroid. In certain embodiments, Compound 10 is administered in combination with a corticosteroid. In certain embodiments, Compound 11 is administered in combination with a corticosteroid. In certain embodiments, Compound 12 is administered in combination with a corticosteroid. In certain embodiments, Compound 13 is administered in combination with a corticosteroid. In certain embodiments, the corticosteroid is dexamethesone.

Additional non-limiting examples of corticosteroids include corticosterone, aldosterone, prednisolone, triamcinolone, budesonide, deflazacort, flugestone, fluorometholone, medrysone, prebedilone acetate, chlorprednisone, cloprednol, difluprednate, fluocinolone, fluperolone, fluperolone acetate, fluprednisolone, loteprednol, prednicarbate, tixocortol, alclomethasone, alclomethasone dipropionate, beclomethasone, clobetasol, clobetasone, clocortolone, desoximetasone, diflorasone, diflorasone diacetate, difluocortolone, difluocortolone valerate, fluprednide, fluprednide acetate, fluticasone, fluticasone furoate, halomethasone, meprednisone, mometasone, mometasone furoate, paramethasone, prednylidene, rimexolone, ulobetazole, amcinonide, ciclesonide, desonide, formocortal, fluclorolone, fluclorolone acetonide, fludroxycortide, flunisolide, fluocinolone, fluocinolone acetonide, fluocinonide, halcinonide, triamcinolone, triamcinolone acetonide, cortivazole, RU-28362, dexamethasone acefurate, dexamethasone acetate, dexamethasone cypionate, dexamethasone diethylamino acetate, dexamethasone dipropionate, dexamethasone isonicotinate, dexamethasone linoleate, dexamethasone metasulfobenzoate, dexamethasone palmitate, dexamethasone phosphate, dexamethasone pivalate, dexamethasone succinate, dexamethasone sulfate, dexamethasone tebutate, dexamethasone troxundate and dexamethasone valerate.

Kinase inhibitors

In certain embodiments, the biologically active agent is a kinase inhibitor, such as a Bruton tyrosine kinase (BTK) inhibitor. In certain embodiments, the kinase inhibitor is selected from a phosphoinositide 3-kinase (PI3K) inhibitor, a Bruton tyrosine kinase (BTK) inhibitor, or a spleen tyrosine kinase (Syk) inhibitor, or a combination thereof.

In certain embodiments, a compound of the present invention is administered in combination with a BTK inhibitor. In certain embodiments, the BTK inhibitor is ibrutinib. In certain embodiments, the BTK inhibitor is acalabrutinib. In certain embodiments, Compound 1 is administered in combination with a BTK inhibitor. In certain embodiments, Compound 1 is administered in combination with the BTK inhibitor zanubrutinib. In certain embodiments, with ibrutinib. In certain embodiments, Compound 1 is administered in combination with zanubrutinib. In certain embodiments, Compound 1 is administered in combination with acalabrutinib.

In certain embodiments, Compound 2 is administered in combination with a BTK inhibitor. In certain embodiments, Compound 3 is administered in combination with a BTK inhibitor. In certain embodiments, Compound 4 is administered in combination with a BTK inhibitor. In certain embodiments, Compound 5 is administered in combination with a BTK inhibitor. In certain embodiments, Compound 6 is administered in combination with a BTK inhibitor. In certain embodiments, Compound 7 is administered in combination with a BTK inhibitor. In certain embodiments, Compound 8 is administered in combination with a BTK inhibitor. In certain embodiments, Compound 9 is administered in combination with a BTK inhibitor. In certain embodiments, Compound 10 is administered in combination with a BTK inhibitor. In certain embodiments, Compound 11 is administered in combination with a BTK inhibitor. In certain embodiments, Compound 12 is administered in combination with a BTK inhibitor. In certain embodiments, Compound 13 is administered in combination with a BTK inhibitor. In certain embodiments, the BTK inhibitor is selected from Ibrutinib, Zanubrutinib, and Acalabrutinib.

Examples of BTK inhibitors include ibrutinib (also known as PCI-32765) (Imbruvica™) (1-[(3R)-3-[4-amino-3-(4-phenoxy-phenyl)pyrazolo[3,4-d]pyrimidin-1-yl]piperidin-1-yl]prop-2-en-1-one), dianilinopyrimidine-based inhibitors such as AVL-101 and AVL-291/292 (N-(3-((5-fluoro-2-((4-(2-methoxyethoxy)phenyl)amino)pyrimidin-4-yl)amino)phenyl)acrylamide) (Avila Therapeutics) (see U.S. Patent Publication No. 2011/0117073, incorporated herein in its entirety), dasatinib ([N-(2-chloro-6-methylphenyl)-2-(6-(4-(2-hydroxyethyl)piperazin-1-yl)-2-methylpyrimidin-4-ylamino)thiazole-5-carboxamide], LFM-A13 (alpha-cyano-beta-hydroxy-beta-methyl-N-(2,5-dibromophenyl)propenamide), GDC-0834 ([R-N-(3-(6-(4-(1,4-dimethyl-3-oxopiperazin-2-yl)phenylamino)-4-methyl-5-oxo-4,5-dihydropyrazin-2-yl)-2-methylphenyl)-4,5,6,7-tetrahydrobenzo[b]thiophene-2-carboxamide], CGI-560 4-(tert-butyl)-N-(3-(8-(phenylamino)imidazo[1,2-a]pyrazin-6-yl)phenyl)benzamide, CGI-1746 (4-(tert-butyl)-N-(2-methyl-3-(4-methyl-6-((4-(morpholine-4-carbonyl)phenyl)amino)-5-oxo-4,5-dihydropyrazin-2-yl)phenyl)benzamide), CNX-774 (4-(4-((4-((3-acrylamidophenyl)amino)-5-fluoropyrimidin-2-yl)amino)phenoxy)-N-methylpicolinamide), STA056 (7-benzyl-1-(3-(piperidin-1-yl)propyl)-2-(4-(pyridin-4-yl)phenyl)-1H-imidazo[4,5-g]quinoxalin-6(5H)-one), GDC-0834 ((R)-N-(3-(6-((4-(1,4-dimethyl-3-oxopiperazin-2-yl)phenyl)amino)-4-methyl-5-oxo-4,5-dihydropyrazin-2-yl)-2-methylphenyl)-4,5,6,7-tetrahydrobenzo[b]thiophene-2-carboxamide), GDC-0837 ((R)-N-(3-(6-((4-(1,4-dimethyl-3-oxopiperazin-2-yl)phenyl)amino)-4-methyl-5-oxo-4,5-dihydropyrazin-2-yl)-2-methylphenyl)-4,5,6,7-tetrahydrobenzo[b]thiophene-2-carboxamide), NM-71224, ACP-196, ONO-4059 (Ono Pharmaceuticals), PRT062607 (4-((3-(2H-1,2,3-triazol-2-yl)phenyl)amino)-2-(((1R,2S)-2-aminocyclohexyl)amino)pyrimidine-5-carboxamide hydrochloride), QL-47 (1-(1-acryloylindolin-6-yl)-9-(1-methyl-1H-pyrazol-4-yl)benzo[h][1,6]naphthyridin-2(1H)-one) and RN486 (6-cyclopropyl-8-fluoro-2-(2-hydroxymethyl-3-{1-methyl-5-[5-(4-methyl-piperazin-1-yl)-pyridin-2-ylamino]-6-oxo-1,6-dihydro-pyridin-3-yl}-phenyl)-2H-isoquinolin-1-one), and other molecules capable of inhibiting BTK activity, such as those BTK inhibitors disclosed in Akinleye et al., Journal of Hematology & Oncology, 2013, 6:59, the contents of which are incorporated herein by reference.

According to certain embodiments, the BTK inhibitor is selected from acalabrutinib, spebrutinib, zanubrutinib, LOXO-305, evobrutinib, TG-1701, tolebrutinib, BIIB091, DZD-9008, HZ-A-018, orelabrutinib, AC0058TA, SN1011, rilzabrutinib, ARQ 531, DTRMWXHS-12, JNJ-64264681, branebrutinib, and fenebrutinib. According to certain embodiments, Compound 1 is administered in combination with a BTK inhibitor selected from acalabrutinib, spebrutinib, zanubrutinib, LOXO-305, evobrutinib, TG-1701, tolebrutinib, BIIB091, DZD-9008, HZ-A-018, orelabrutinib, AC0058TA, SN1011, rilzabrutinib, ARQ 531, DTRMWXHS-12, JNJ-64264681, branebrutinib, and fenebrutinib.

Examples of PI3 kinase inhibitors include, but are not limited to, wortmannin, demethoxyviridine, perifosine, idelalisib, pictilisib, palomid 529, ZSTK474, PWT33597, CUDC-907 and AEZS-136, duvelisib, GS-9820, BKM120, GDC-0032 (taselisib) (2-[4-[2-(2-isopropyl-5-methyl-1,2,4-triazol-3-yl)-5,6-dihydroimidazo[1,2-d][1,4]benzoxazepin-9-yl]pyrazol-1-yl]-2-methylpropanamide), MLN-1117 ((2R)-1-phenoxy-2-butanylhydro(S)-methylphosphonate or methyl(oxo){[(2R)-1-phenoxy-2-butanyl]oxy}phosphonium)), BYL-719 ((2S)-N1-[4-methyl-5-[2-(2,2,2-trifluoro-1,1-dimethylethyl)-4-pyridinyl]-2-thiazolyl]-1,2-pyrrolidinedicarboxamide), GSK2126458 (2,4-difluoro-N-{2-(methyloxy)-5-[4-(4-pyridazinyl)-6-quinolinyl]-3-pyridinyl}benzenesulfonamide)(omipalisib), TGX-221 ((±)-7-methyl-2-(morpholin-4-yl)-9-(1-phenylaminoethyl)-pyrido[1,2-a]-pyrimidin-4-one), GSK2636771 (2-methyl-1-(2-methyl-3-(trifluoromethyl)benzyl)-6-morpholino-1H-benzo[d]imidazole-4-carboxylic acid dihydrochloride), KIN-193 ((R)-2-((1-(7-methyl-2-morpholino-4-oxo-4H-pyrido[1,2-a]pyrimidin-9-yl)ethyl)amino)benzoic acid), TGR-1202/RP5264, GS-9820 ((S)-1-(4-((2-(2-aminopyrimidin-5-yl)-7-methyl-4-mohydroxypropan-1-one), GS-1101 (5-fluoro-3-phenyl-2-([S)]-1-[9H-purin-6-ylamino]propyl)-3H-quinazolin-4-one), AMG-319, GSK-2269557, SAR245409 (N-(4-(N-(3-((3,5-dimethoxyphenyl)amino)quinoxalin-2-yl)sulfamoyl)phenyl)-3-methoxy-4 methylbenzamide), BAY80-6946 (2-amino-N-(7-methoxy-8-(3-morpholinopropoxy)-2,3-dihydroimidazo[1,2-c]quinase), AS 252424 (5-[1-[5-(4-fluoro-2-hydroxy-phenyl)-furan-2-yl]-meth-(Z)-ylidene]-thiazolidine-2,4-dione), CZ 24832 (5-(2-amino-8-fluoro-[1,2,4]triazolo[1,5-a]pyridin-6-yl)-N-tert-butylpyridine-3-sulfonamide), buparlisib (5-[2,6-di(4-morpholinyl)-4-pyrimidinyl]-4-(trifluoromethyl)-2-pyridinamine), GDC-0941 (2-(1H-indazol-4-yl)-6-[[4-(methylsulfonyl)-1-piperazinyl]methyl]-4-(4-morpholinyl)thieno[3,2-d]pyrimidine), GDC-0980 ((S)-1-(4-((2-(2-aminopyrimidin-5-yl)-7-methyl-4-morpholinothieno[3,2-d]pyrimidin-6-yl)methyl)piperazin-1-yl)-2-hydroxypropan-1-one (also known as RG7422)), SF1126 ((8S,14S,17S)-14-(carboxymethyl)-8-(3-guanidinopropyl)-17-(hydroxymethyl)-3,6,9,12,15-pentaoxo-1-(4-(4-oxo-8-phenyl-4H-chromen-2-yl)morpholin-4-ium)-2-oxa-7,10,13,16-tetraazaoctadecane-18-oate), PF-05212384 (N-[4-[[4-(dimethylamino)-1-piperidinyl]carbonyl]phenyl]-N'-[4-(4,6-di-4-morpholinyl-1,3,5-triazin-2-yl)phenyl]urea) (gedatolisib), LY3023414, BEZ235 (2-methyl-2-{4-[3-methyl-2-oxo-8-(quinolin-3-yl)-2,3-dihydro-1H-imidazo[4,5-c]quinolin-1-yl]phenyl}propanenitrile) (dactolisib), XL-765 (N-(3-(N-(3-(3,5-dimethoxyphenylamino)quinoxalin-2-yl)sulfamoyl)phenyl)-3-methoxy-4-methylbenzamide), and GSK1059615 (5-[[4-(4-pyridinyl)-6-quinolinyl]methylene]-2,4-thiazolidenedione), PX886 ([(3aR,6E,9S,9aR,10R,11aS)-6-[[bis(prop-2-enyl)amino]methylidene]-5-hydroxy-9-(methoxymethyl)-9a,11a-dimethyl-1,4,7-trioxo-2,3,3a,9,10,11-hexahydroindeno[4,5h]isochromen-10-yl] acetate (also known as sonolisib)), LY294002, AZD8186, PF-4989216, pilaralisib, GNE-317, PI-3065, PI-103, NU7441 (KU-57788), HS 173, VS-5584 (SB2343), CZC24832, TG100-115, A66, YM201636, CAY10505, PIK-75, PIK-93, AS-605240, BGT226 (NVP-BGT226), AZD6482, voxtalisib, alpelisib, IC-87114, TGI100713, CH5132799, PKI-402, copanlisib (BAY 80-6946), XL 147, PIK-90, PIK-293, PIK-294, 3-MA (3-methyladenine), AS-252424, AS-604850, apitolisib (GDC-0980, RG7422) and the structure described in WO2014/071109.

Syk inhibitors include, for example, cerdulatinib (4-(cyclopropylamino)-2-((4-(4-(ethylsulfonyl)piperazin-1-yl)phenyl)amino)pyrimidine-5-carboxamide), entospletinib (6-(1H-indazol-6-yl)-N-(4-morpholinophenyl)imidazo[1,2-a]pyrazin-8-amine), fostamatinib ([6-({5-fluoro-2-[(3,4,5-trimethoxyphenyl)amino]-4-pyrimidinyl}amino)-2,2-dimethyl-3-oxo-2,3-dihydro-4H-pyrido[3,2-b][1,4]oxazin-4-yl]methyl dihydrogen phosphate), fostamatinib disodium salt (sodium (6-((5-fluoro-2-((3,4,5-trimethoxyphenyl)amino)pyrimidin-4-yl)amino)-2,2-dimethyl-3-oxo-2H-pyrido[3,2-b][1,4]oxazin-4(3H)-yl)methyl phosphate), BAY 61-3606 (2-(7-(3,4-dimethoxyphenyl)imidazo[1,2-c]pyrimidin-5-ylamino)-nicotinamide HCl), RO9021 (6-[(1R,2S)-2-amino-cyclohexylamino]-4-(5,6-dimethyl-pyridin-2-ylamino)-pyridazine-3-carboxylic acid amide), imatinib (Gleevac, 4-[(4-methylpiperazin-1-yl)methyl]-N-(4-methyl-3-{[4-(pyridin-3-yl)pyrimidin-2-yl]amino}phenyl)benzamide), staurosporine, GSK143 (2-(((3R,4R)-3-aminotetrahydro-2H-pyran-4-yl)amino)-4-(p-tolylamino)pyrimidine-5-carboxamide), PP2 (1-(tert-butyl)-3-(4-chlorophenyl)-1H-pyrazolo[3,4-d]pyrimidin-4-amine), PRT-060318(2-(((1H,2S)-2-aminocyclohexyl)amino)-4-(m-tolylamino)pyrimidine-5-carboxamide), PRT-062607 (4-((3-(2H-1,2,3-triazol-2-yl)phenyl)amino)-2-(((1R,2S)-2-aminocyclohexyl)amino)pyrimidine-5-carboxamide hydrochloride), R112 (3,3'-((5-fluoropyrimidine-2,4-diyl)bis(azanediyl))diphenol), R348 (3-ethyl-4-methylpyridine), R406 (6-((5-fluoro-2-((3,4,5-trimethoxyphenyl)amino)pyrimidin-4-yl)amino)-2,2-dimethyl-2H-pyrido[3,2-b][1,4]oxazin-3(4H)-one), piceatannol (3-hydroxyresveratrol), YM193306 (see Singh et al. Discovery and Development of Spleen Tyrosine Kinase (SYK) Inhibitors, J. Med. Chem. 2012, 55, 3614-3643), 7-azaindole, piceatannol, ER-27319 (see Singh et al. Discovery and Development of Spleen Tyrosine Kinase (SYK) Inhibitors, J. Med. Chem. 2012, 55, 3614-3643, incorporated herein in its entirety), Compound D (see Singh et al. Discovery and Development of Spleen Tyrosine Kinase (SYK) Inhibitors, J. Med. Chem. 2012, 55, 3614-3643, incorporated herein in its entirety), PRT060318 (see Singh et al. Discovery and Development of Spleen Tyrosine Kinase (SYK) Inhibitors, J. Med. Chem. 2012, 55, 3614-3643, incorporated herein in its entirety), luteolin (see Singh et al. Discovery and Development of Spleen Tyrosine Kinase (SYK) Inhibitors, J. Med. Chem. 2012, 55, 3614-3643, incorporated herein in its entirety), apigenin (see Singh et al. Discovery and Development of Spleen Tyrosine Kinase (SYK) Inhibitors, J. Med. Chem. 2012, 55, 3614-3643, incorporated herein in its entirety), quercetin (see Singh et al. Discovery and Development of Spleen Tyrosine Kinase (SYK) Inhibitors, J. Med. Chem. 2012, 55, 3614-3643, incorporated herein in its entirety in its entirety), fisetin (see Singh et al. Discovery and Development of Spleen Tyrosine Kinase (SYK) Inhibitors, J. Med. Chem. 2012, 55, 3614-3643, incorporated herein in its entirety), myricetin (see Singh et al. Discovery and Development of Spleen Tyrosine Kinase (SYK) Inhibitors, J. Med. Chem. 2012, 55, 3614-3643, incorporated herein in its entirety), morin (see Singh et al. Discovery and Development of Spleen Tyrosine Kinase (SYK) Inhibitors, J. Med. Chem. 2012, 55, 3614-3643, incorporated herein in its entirety).

Proteasome inhibitors

In certain embodiments, a compound of the present invention is administered in combination with a proteasome inhibitor. In certain embodiments, the proteasome inhibitor is Bortezomib. In certain embodiments, the proteasome inhibitor is Ixazomib. In certain embodiments, the proteasome inhibitor is Carfilzomib. In certain embodiments, Compound 1 is administered in combination with a proteasome inhibitor. In certain embodiments, Compound 1 is administered in combination with Bortezomib. In certain embodiments, Compound 1 is administered in combination with Ixazomib. In certain embodiments, Compound 1 is administered in combination with Carfilzomib. In certain embodiments, Compound 1 is administered in combination with Carfilzomib. In certain embodiments, Compound 1 is administered in combination with carfilzomib and daratumumab.

In certain embodiments, Compound 2 is administered in combination with a proteasome inhibitor. In certain embodiments, Compound 3 is administered in combination with a proteasome inhibitor. In certain embodiments, Compound 4 is administered in combination with a proteasome inhibitor. In certain embodiments, Compound 5 is administered in combination with a proteasome inhibitor. In certain embodiments, Compound 6 is administered in combination with a proteasome inhibitor. In certain embodiments, Compound 7 is administered in combination with a proteasome inhibitor. In certain embodiments, Compound 8 is administered in combination with a proteasome inhibitor. In certain embodiments, Compound 9 is administered in combination with a proteasome inhibitor. In certain embodiments, Compound 10 is administered in combination with a proteasome inhibitor. In certain embodiments, Compound 11 is administered in combination with a proteasome inhibitor. In certain embodiments, Compound 12 is administered in combination with a proteasome inhibitor. In certain embodiments, Compound 13 is administered in combination with a proteasome inhibitor. In certain embodiments, the proteasome inhibitor is selected from Bortezomib, Ixazomib, VLX1570, and Carfilzomib.

Additional examples of proteasome inhibitors include ixazomib citrate, oprozomib, delanzomib, lactacystin, epoxomicin, MG132, MG-262, CEP-18770, NEOSH101, TQB3602, and KZR-616. In certain embodiments, Compound 1 is administered in combination with a proteasome inhibitor selected from ixazomib citrate, oprozomib, delanzomib, lactacystin, epoxomicin, MG132, MG-262, CEP-18770, NEOSH101, TQB3602, VLX1570, and KZR-616.

HDAC inhibitors

In certain embodiments, a compound of the present invention is administered in combination with an HDAC inhibitor. In certain embodiments, the HDAC inhibitor is Vorinostat. In certain embodiments, the HDAC inhibitor is Romidepsin. In certain embodiments, the HDAC inhibitor is Panobinostat. In certain embodiments, the HDAC inhibitor is Belinostat. In certain embodiments, Compound 1 is administered in combination with an HDAC inhibitor. In certain embodiments, Compound 1 is administered in combination with Vorinostat. In certain embodiments, Compound 1 is administered in combination with Romidepsin. In certain embodiments, Compound 1 is administered in combination with Panobinostat. In certain embodiments, Compound 1 is administered in combination with Belinostat.

In certain embodiments, Compound 2 is administered in combination with an HDAC inhibitor. In certain embodiments, Compound 3 is administered in combination with an HDAC inhibitor. In certain embodiments, Compound 4 is administered in combination with an HDAC inhibitor. In certain embodiments, Compound 5 is administered in combination with an HDAC inhibitor. In certain embodiments, Compound 6 is administered in combination with an HDAC inhibitor. In certain embodiments, Compound 7 is administered in combination with an HDAC inhibitor. In certain embodiments, Compound 8 is administered in combination with an HDAC inhibitor. In certain embodiments, Compound 9 is administered in combination with an HDAC inhibitor. In certain embodiments, Compound 10 is administered in combination with an HDAC inhibitor. In certain embodiments, Compound 11 is administered in combination with an HDAC inhibitor. In certain embodiments, Compound 12 is administered in combination with an HDAC inhibitor. In certain embodiments, Compound 13 is administered in combination with an HDAC inhibitor. In certain embodiments, the HDAC inhibitor is selected from Vorinostat, Romidepsin, Panobinostat, and Belinostat.

According to certain embodiments, the HDAC inhibitor is selected from trapoxin B, sodium phenylbutyrate, tacedinalin, mocetinostat, BRD73954, BG45, domatinostat, cay10603, HPOV, TMP269, nexturastat A, Santacruzamate A, splitomycin, LMK-235, sodium butyrate, pivaloyloxymethylbutyrate, pyroxamide, abexinostat, resminostat, givinostat, quisinostat, Psammapline A, KD5170, 1-Alanine chlamidocin, depudecin, and CUDC-101. According to certain embodiments, Compound 1 is administered in combination with an HDAC inhibitor selected from trapoxin B, sodium phenylbutyrate, tacedinalin, mocetinostat, BRD73954, BG45, domatinostat, cautinoside, HPOV, TMP269, nexturastat A, Santacruzamate A, splitomycin, LMK-235, sodium butyrate, pivaloyloxymethylbutyrate, pyroxamide, abexinostat, resminostat, givinostat, quisinostat, Psammapline A, KD5170, 1-Alanine chlamidocin, depudecin, and CUDC-101.

IMiD

In certain embodiments, a compound of the present invention is administered in combination with an IMiD. In certain embodiments, the IMiD is thalidomide. In certain embodiments, the IMiD is lenalidomide. In certain embodiments, the IMiD is pomalidomide. In certain embodiments, Compound 1 is administered in combination with thalidomide. In certain embodiments, Compound 1 is administered in combination with lenalidomide. In certain embodiments, Compound 1 is administered in combination with pomalidomide.

In certain embodiments, Compound 2 is administered in combination with an IMiD. In certain embodiments, Compound 3 is administered in combination with an IMiD. In certain embodiments, Compound 4 is administered in combination with an IMiD. In certain embodiments, Compound 5 is administered in combination with an IMiD. In certain embodiments, Compound 6 is administered in combination with an IMiD. In certain embodiments, Compound 7 is administered in combination with an IMiD. In certain embodiments, Compound 8 is administered in combination with an IMiD. In certain embodiments, Compound 9 is administered in combination with an IMiD. In certain embodiments, Compound 10 is administered in combination with an IMiD. In certain embodiments, Compound 11 is administered in combination with an IMiD. In certain embodiments, Compound 12 is administered in combination with an IMiD. In certain embodiments, Compound 13 is administered in combination with an IMiD. In certain embodiments, the IMiD is selected from pomalidomide, thalidomide, and lenalidomide.

In certain embodiments, the IMiD is CC-90009. In certain embodiments, the IMiD is CC-99282. In certain embodiments, the IMiD is CC-92480. In certain embodiments, Compound 1 is administered in combination with CC-90009. In certain embodiments, Compound 1 is administered in combination with CC-99282. In certain embodiments, Compound 1 is administered in combination with CC-92480.

In certain embodiments, Compound 2, Compound 3, Compound 4, Compound 5, Compound 6, Compound 7, Compound 8, Compound 9, Compound 10, Compound 11, Compound 12, or Compound 13 is administered in combination with an IMiD. In certain embodiments, the IMiD is selected from CC-90009, CC-99282, and CC-92480.

Antibodies

In certain embodiments, a compound of the present invention is administered in combination with an antibody that targets CD20, CD30, or CD38. In certain embodiments, the targeting antibody is Rituximab. In certain embodiments, the targeting antibody is Daratumumab. In certain embodiments, the targeting antibody is Elotuzumab. In certain embodiments, the targeting antibody is Isatuximab. In certain embodiments, Compound 1 is administered in combination with an antibody that targets CD20, CD30, or CD38. In certain embodiments, Compound 1 is administered in combination with Rituximab. In certain embodiments, Compound 1 is administered in combination with Daratumumab. In certain embodiments, Compound 1 is administered in combination with Elotuzumab. In certain embodiments, Compound 1 is administered in combination with Isatuximab.

In certain embodiments, Compound 2 is administered in combination with an antibody targeting CD20, CD30, or CD38. In certain embodiments, Compound 3 is administered in combination with an antibody targeting CD20, CD30, or CD38. In certain embodiments, Compound 4 is administered in combination with an antibody targeting CD20, CD30, or CD38. In certain embodiments, Compound 5 is administered in combination with an antibody targeting CD20, CD30, or CD38. In certain embodiments, Compound 6 is administered in combination with an antibody targeting CD20, CD30, or CD38. In certain embodiments, Compound 7 is administered in combination with an antibody targeting CD20, CD30, or CD38. In certain embodiments, Compound 8 is administered in combination with an antibody targeting CD20, CD30, or CD38. In certain embodiments, Compound 9 is administered in combination with an antibody targeting CD20, CD30, or CD38. In certain embodiments, Compound 10 is administered in combination with an antibody targeting CD20, CD30, or CD38. In certain embodiments, Compound 11 is administered in combination with an antibody targeting CD20, CD30, or CD38. In certain embodiments, Compound 12 is administered in combination with an antibody targeting CD20, CD30, or CD38. In certain embodiments, Compound 13 is administered in combination with an antibody targeting CD20, CD30, or CD38. In certain embodiments, the targeted antibody is selected from Rituximab, Daratumumab, Elotuzumab, and Isatuximab.

In certain embodiments, a compound of the present invention is administered in combination with an antibody drug conjugate. In certain embodiments, the antibody drug conjugate is Brentuximab vedotin. In certain embodiments, the antibody drug conjugate is Ibritutomab tiuxetan. In certain embodiments, the antibody drug conjugate is Mogamulizumab. In certain embodiments, the antibody drug conjugate is Obinutuzumab. In certain embodiments, the antibody drug conjugate is Polatuzumab vedotin. In certain embodiments, the antibody drug conjugate is Belantumab mafodotin (GSK2857916). In certain embodiments, the antibody drug conjugate is MEDI2228. In certain embodiments, the antibody drug conjugate is CC-99712. In certain embodiments, Compound 1 is administered in combination with the antibody drug conjugate. In certain embodiments, Compound 1 is administered in combination with Brentuximab vedotin. In certain embodiments, Compound 1 is administered in combination with Ibritutomab tiuxetan. In certain embodiments, Compound 1 is administered in combination with Mogamulizumab. In certain embodiments, Compound 1 is administered in combination with Obinutuzumab. In certain embodiments, Compound 1 is administered in combination with Polatuzumab vedotin. In certain embodiments, Compound 1 is administered in combination with Belantumab mafodotin (GSK2857916). In certain embodiments, Compound 1 is administered in combination with MEDI2228. In certain embodiments, Compound 1 is administered in combination with CC-99712. In certain embodiments, Compound 1 is administered in combination with tafasitamab.

In certain embodiments, Compound 2 is administered in combination with an antibody drug conjugate. In certain embodiments, Compound 3 is administered in combination with an antibody drug conjugate. In certain embodiments, Compound 4 is administered in combination with an antibody drug conjugate. In certain embodiments, Compound 5 is administered in combination with an antibody drug conjugate. In certain embodiments, Compound 6 is administered in combination with an antibody drug conjugate. In certain embodiments, Compound 7 is administered in combination with an antibody drug conjugate. In certain embodiments, Compound 8 is administered in combination with an antibody drug conjugate. In certain embodiments, Compound 9 is administered in combination with an antibody drug conjugate. In certain embodiments, Compound 10 is administered in combination with an antibody drug conjugate. In certain embodiments, Compound 11 is administered in combination with an antibody drug conjugate. In certain embodiments, Compound 12 is administered in combination with an antibody drug conjugate. In certain embodiments, Compound 13 is administered in combination with an antibody drug conjugate. In certain embodiments, the antibody drug conjugate is selected from Brentuximab vedotin, Ibritutomab tiuxetan, Mogamulizumab, Obinutuzumab, Polatuzumab vedotin, Belantumab mafodotin (GSK2857916), MEDI2228, and CC-99712.

In certain embodiments, a compound of the present invention is administered in combination with a bispecific antibody. In certain embodiments, the bispecific antibody is PF-06863135. In certain embodiments, the bispecific antibody is TNB-383B. In certain embodiments, the bispecific antibody is REGN5458. In certain embodiments, the bispecific antibody is JNJ-64007957. In certain embodiments, Compound 1 is administered in combination with a bispecific antibody. In certain embodiments, Compound 1 is administered in combination with PF-06863135. In certain embodiments, Compound 1 is administered in combination with TNB-383B. In certain embodiments, Compound 1 is administered in combination with REGN5458. In certain embodiments, Compound 1 is administered in combination with JNJ-64007957.

In certain embodiments, Compound 2 is administered in combination with a bispecific antibody. In certain embodiments, Compound 3 is administered in combination with a bispecific antibody. In certain embodiments, Compound 4 is administered in combination with a bispecific antibody. In certain embodiments, Compound 5 is administered in combination with a bispecific antibody. In certain embodiments, Compound 6 is administered in combination with a bispecific antibody. In certain embodiments, Compound 7 is administered in combination with a bispecific antibody. In certain embodiments, Compound 8 is administered in combination with a bispecific antibody. In certain embodiments, Compound 9 is administered in combination with a bispecific antibody. In certain embodiments, Compound 10 is administered in combination with a bispecific antibody. In certain embodiments, Compound 11 is administered in combination with a bispecific antibody. In certain embodiments, Compound 12 is administered in combination with a bispecific antibody. In certain embodiments, Compound 13 is administered in combination with a bispecific antibody. In certain embodiments, the bispecific antibody is selected from PF-06863135, TNB-383B, REGN5458, and JNJ-64007957.

In certain embodiments, a compound of the present invention is administered in combination with a naked monoclonal antibody (mAb). In certain embodiments, the naked mAb is SEA-BCMA. In certain embodiments, Compound 1 is administered in combination with a naked mAb. In certain embodiments, Compound 1 is administered in combination with SEA-BCMA.

In certain embodiments, Compound 2 is administered in combination with a naked mAb. In certain embodiments, Compound 3 is administered in combination with a naked mAb. In certain embodiments, Compound 4 is administered in combination with a naked mAb. In certain embodiments, Compound 5 is administered in combination with a naked mAb. In certain embodiments, Compound 6 is administered in combination with a naked mAb. In certain embodiments, Compound 7 is administered in combination with a naked mAb. In certain embodiments, Compound 8 is administered in combination with a naked mAb. In certain embodiments, Compound 9 is administered in combination with a naked mAb. In certain embodiments, Compound 10 is administered in combination with a naked mAb. In certain embodiments, Compound 11 is administered in combination with a naked mAb. In certain embodiments, Compound 12 is administered in combination with a naked mAb. In certain embodiments, Compound 13 is administered in combination with a naked mAb. In certain embodiments, the naked mAb is SEA-BCMA.

Additional non-limiting examples of anti-CD38 antibodies include felzartamab, GBR 1342, TAK-573, CID-103, OKT10, STI-6129, SGX301, and mezagitamab. In certain embodiments, Compound 1 is administered in combination with an anti-CD38 antibody selected from felzartamab, GBR 1342, TAK-573, CID-103, OKT10, STI-6129, SGX301, TAK-079, and mezagitamab.

CAR T-cell therapy

In certain embodiments, a compound of the present invention is administered in combination with a CAR T cell therapy. In certain embodiments, the CAR T cell therapy is Axicabtagene ciloleucel. In certain embodiments, the CAR T cell therapy is Tisagenleucel. In certain embodiments, the CAR T cell therapy is Idecabtagene vicleucel (ide-cel, bb2121). In certain embodiments, the CAR T cell therapy is LCAR-B38M (JNJ-4528, JNJ-68284528). In certain embodiments, the CAR T cell therapy is P-BCMA-101. In certain embodiments, the CAR T cell therapy is PBCAR269A. In certain embodiments, the CAR T cell therapy is bb21217. In certain embodiments, the CAR T cell therapy is JCARK125 (orva-cel, orvacabtagene autoleucel). In certain embodiments, the CAR T cell therapy is ALLO-715. In certain embodiments, the CAR T cell therapy is Descartes-08. In certain embodiments, the CAR T cell therapy is FCARH143. In certain embodiments, the CAR T cell therapy is CT053.

In certain embodiments, Compound 1 is administered in combination with a CAR T cell therapy. In certain embodiments, Compound 1 is administered in combination with Axicabtagene ciloleucel. In certain embodiments, Compound 1 is administered in combination with Tisagenleucel. In certain embodiments, Compound 1 is administered in combination with Idecabtagene vicoleucel (ide-cel, bb2121). In certain embodiments, Compound 1 is administered in combination with LCAR-B38M (JNJ-4528, JNJ-68284528). In certain embodiments, Compound 1 is administered in combination with P-BCMA-101. In certain embodiments, Compound 1 is administered in combination with PBCAR269A. In certain embodiments, Compound 1 is administered in combination with bb21217. In certain embodiments, Compound 1 is administered in combination with JCARK125 (orva-cel, orvacabtagene autoleucel). In certain embodiments, Compound 1 is administered in combination with ALLO-715. In certain embodiments, Compound 1 is administered in combination with Descartes-08. In certain embodiments, Compound 1 is administered in combination with FCARH143. In certain embodiments, Compound 1 is administered in combination with CT053.

In certain embodiments, Compound 2 is administered in combination with a CAR T cell therapy. In certain embodiments, Compound 3 is administered in combination with a CAR T cell therapy. In certain embodiments, Compound 4 is administered in combination with a CAR T cell therapy. In certain embodiments, Compound 5 is administered in combination with a CAR T cell therapy. In certain embodiments, Compound 6 is administered in combination with a CAR T cell therapy. In certain embodiments, Compound 7 is administered in combination with a CAR T cell therapy. In certain embodiments, Compound 8 is administered in combination with a CAR T cell therapy. In certain embodiments, Compound 9 is administered in combination with a CAR T cell therapy. In certain embodiments, Compound 10 is administered in combination with a CAR T cell therapy. In certain embodiments, Compound 11 is administered in combination with a CAR T cell therapy. In certain embodiments, Compound 12 is administered in combination with a CAR T cell therapy. In certain embodiments, Compound 13 is administered in combination with a CAR T cell therapy. In certain embodiments, the CAR T cell therapy is selected from Axicabtagene ciloleucel, Tisagenleucel, Idecabtagene vicleucel (ide-cel, bb2121), LCAR-B38M (JNJ-4528, JNJ-68284528), and P-BCMA-101. In certain embodiments, the CAR T cell therapy is selected from PBCAR269A, bb21217, JCARK125 (oral-cel, orvacabtagene autoleucel), ALLO-715, Descartes-08, FCARH143, and CT053.

In certain embodiments, the CAR T cell therapy is selected from ALLO-715, bb21217, BCMA CAR-T, CD138 CAR-T, CD19 CAR-T, ciltacabtagene autoleucel, CS1 (SLAMF7) CAR-T, CT053, Descartes-11, idecabtagene viclasel, NKG2D CAR-T, orvacabtagene autoleucel, P-BCMA-101, and UCARTCS1.

Cell therapy

In certain embodiments, a compound of the present invention is administered in combination with a cell therapy. In certain embodiments, Compound 1 is used in combination with a cell therapy.

Non-limiting examples of cell therapies include allo-HSCT, allo-NKT, auto-HSCT, and auto-NKT.

T cell recruiters

In certain embodiments, a compound of the present invention is administered in combination with bispecific T cell recruiters (BiTEs). In certain embodiments, the BiTE is Blinatumomab. In certain embodiments, the BiTE is CC-93268. In certain embodiments, the BiTE is AMG 420. In certain embodiments, the BiTE is AMG 701. In certain embodiments, Compound 1 is administered in combination with bispecific T cell recruiters (BiTEs). In certain embodiments, Compound 1 is administered in combination with Blinatumomab. In certain embodiments, Compound 1 is administered in combination with AMG 420. In certain embodiments, Compound 1 is administered in combination with CC-93269. In certain embodiments, Compound 1 is administered in combination with AMG 701.

In certain embodiments, Compound 2 is administered in combination with BiTE. In certain embodiments, Compound 3 is administered in combination with BiTE. In certain embodiments, Compound 4 is administered in combination with BiTE. In certain embodiments, Compound 5 is administered in combination with BiTE. In certain embodiments, Compound 6 is administered in combination with BiTE. In certain embodiments, Compound 7 is administered in combination with BiTE. In certain embodiments, Compound 8 is administered in combination with BiTE. In certain embodiments, Compound 9 is administered in combination with BiTE. In certain embodiments, Compound 10 is administered in combination with BiTE. In certain embodiments, Compound 11 is administered in combination with BiTE. In certain embodiments, Compound 12 is administered in combination with BiTE. In certain embodiments, Compound 13 is administered in combination with BiTE. In certain embodiments, BiTE is selected from Blinatumomab, AMG 420, CC-93269, and AMG 4701.

According to certain embodiments, Compound 1 is used in combination with a bispecific antibody selected from AMG 420, AMG 701, BFCR4350A, blinatumomab, CC-93269, elranatamab, EM801, REGN5458, talquetamab, teclistamab, and TNB-383B.

Immunomodulators immune checkpoint inhibitors

In certain embodiments, a compound of the present invention is administered in combination with an immune checkpoint inhibitor. In certain embodiments, a compound of the present invention is administered in combination with a PD-1 immune checkpoint inhibitor. In certain embodiments, a compound of the present invention is administered in combination with a PD-L1 immune checkpoint inhibitor. In certain embodiments, a compound of the present invention is administered in combination with an IFNAR immune checkpoint inhibitor. In certain embodiments, the immune checkpoint inhibitor is Nivolumab. In certain embodiments, the immune checkpoint inhibitor is Pembrolizumab. In certain embodiments, the immune checkpoint inhibitor is Interferon alfa-2b. In certain embodiments, Compound 1 is administered in combination with an immune checkpoint inhibitor. In certain embodiments, Compound 1 is administered in combination with a PD-1 immune checkpoint inhibitor. In certain embodiments, Compound 1 is administered in combination with a PD-L1 immune checkpoint inhibitor. In certain embodiments, Compound 1 is administered in combination with an IFNAR immune checkpoint inhibitor. In certain embodiments, Compound 1 is administered in combination with Nivolumab. In certain embodiments, Compound 1 is administered in combination with Pembrolizumab. In certain embodiments, Compound 1 is administered in combination with Interferon alpha-2b.

In certain embodiments, Compound 2 is administered in combination with an immune checkpoint inhibitor. In certain embodiments, Compound 3 is administered in combination with an immune checkpoint inhibitor. In certain embodiments, Compound 4 is administered in combination with an immune checkpoint inhibitor. In certain embodiments, Compound 5 is administered in combination with an immune checkpoint inhibitor. In certain embodiments, Compound 6 is administered in combination with an immune checkpoint inhibitor. In certain embodiments, Compound 7 is administered in combination with an immune checkpoint inhibitor. In certain embodiments, Compound 8 is administered in combination with an immune checkpoint inhibitor. In certain embodiments, Compound 9 is administered in combination with an immune checkpoint inhibitor. In certain embodiments, Compound 10 is administered in combination with an immune checkpoint inhibitor. In certain embodiments, Compound 11 is administered in combination with an immune checkpoint inhibitor. In certain embodiments, Compound 12 is administered in combination with an immune checkpoint inhibitor. In certain embodiments, Compound 13 is administered in combination with an immune checkpoint inhibitor. In certain embodiments, the immune checkpoint inhibitor is a PD-1 immune checkpoint inhibitor. In certain embodiments, the immune checkpoint inhibitor is a PD-L1 immune checkpoint inhibitor. In certain embodiments, the immune checkpoint inhibitor is an IFNAR immune checkpoint inhibitor. In certain embodiments, the immune checkpoint inhibitor is selected from Nivolumab, Pembrolizumab, and Interferon alfa-2b.

PD-1 inhibitors that block the interaction of PD-1 and PD-L1 by binding to the PD-1 receptor and, in turn, inhibit immunosuppression include, for example, nivolumab (Opdivo), pembrolizumab (Keytruda), pidilizumab, AMP-224 (AstraZeneca and MedImmune), PF-06801591 (Pfizer), MEDI0680 (AstraZeneca), PDR001 (Novartis), REGN2810 (Regeneron), SHR-12-1 (Jiangsu Hengrui Medicine Company and Incyte Corporation), TSR-042 (Tesaro), and the PD-L1/VISTA inhibitor CA-170 (Curis Inc.). PD-L1 inhibitors, which block the interaction of PD-1 and PD-L1 by binding to the PD-L1 receptor and in turn inhibit immunosuppression, include, for example, atezolizumab (Tecentriq), durvalumab (AstraZeneca and MedImmune), KN035 (Alphamab), and BMS-936559 (Bristol-Myers Squibb). CTLA-4 immune checkpoint inhibitors, which bind to CTLA-4 and inhibit immunosuppression, include, but are not limited to, ipilimumab, tremelimumab (AstraZeneca and MedImmune), AGEN1884, and AGEN2041 (Agenus). LAG-3 immune checkpoint inhibitors include, but are not limited to, BMS-986016 (Bristol-Myers Squibb), GSK2831781 (GlaxoSmithKline), IMP321 (Prima BioMed), LAG525 (Novartis), and the dual PD-1 and LAG-3 inhibitor MGD013 (MacroGenics). An example of a TIM-3 inhibitor is TSR-022 (Tesaro).

In certain embodiments, the immune checkpoint inhibitor is selected from nivolumab/OPDIVO®, pembrolizumab/KEYTRUDA® and pidilizumab/CT-011, MPDL3280A/RG7446, MEDI4736, MSB0010718C, BMS 936559, a PDL2/Ig fusion protein such as AMP 224, or an inhibitor of the B7-H3 family of ligands (e.g., MGA271), B7-H4, BTLA, HVEM, TIM3, GAL9, LAG 3, VISTA, KIR, 2B4, CD160, CGEN-15049, CHK1, CHK2, A2aR, B-7, or a combination thereof.

In certain embodiments, the PD-1 inhibitor is BGB-A317. In certain embodiments, the PD-L1 inhibitor is MED14736. In certain embodiments, the PD-L2 inhibitor is rHIgM12B7A.

In certain embodiments, the immune checkpoint inhibitor is a B7 inhibitor, such as a B7-H3 inhibitor or a B7-H4 inhibitor. In certain embodiments, the B7-H3 inhibitor is MGA271.

In certain embodiments, the immune checkpoint inhibitor is an OX40 agonist. In certain embodiments, the immune checkpoint inhibitor is an anti-OX40 antibody, such as anti-OX-40 or MEDI6469.

In certain embodiments, the immune checkpoint inhibitor is a GITR agonist. In certain embodiments, the GITR agonist is an anti-GITR antibody, such as TRX518.

In certain embodiments, the immune checkpoint inhibitor is a CD137 agonist. In certain embodiments, the CD137 agonist is an anti-CD137 antibody, such as PF-05082566.

In certain embodiments, the immune checkpoint inhibitor is a CD40 agonist. In certain embodiments, the CD40 agonist is an anti-CD40 antibody, such as CF-870,893.

In certain embodiments, the immune checkpoint inhibitor is an IDO inhibitor, such as INCB24360 or indoximod.

In certain embodiments, the immune checkpoint inhibitor is selected from atezolizumab, avelumab, durvalumab, nivolumab, and pembrolizumab.

Additional biologically active agents

According to another embodiment, the active compounds described herein can be administered in an effective amount to treat abnormal tissue of the male reproductive system, such as prostate or testicular cancer, in combination or alternation with an effective amount of an androgen inhibitor (such as testosterone), including, but not limited to, a selective androgen receptor modulator, a selective androgen receptor degrader, a complete androgen receptor degrader, or another form of partial or complete androgen antagonist. According to certain embodiments, the prostate or testicular cancer is androgen-resistant. Non-limiting examples of antiandrogen compounds are provided in WO 2011/156518 and U.S. Patent Nos. 8,455,534 and 8,299,112. Additional non-limiting examples of antiandrogen compounds include enzalutamide, apalutamide, cyproterone acetate, chlormadinone acetate, spironolactone, canrenone, drospirenone, ketoconazole, topilutamide, abiraterone acetate, and cimetidine.

In certain embodiments, the biologically active agent is an ALK inhibitor. Examples of ALK inhibitors include, but are not limited to, crizotinib, alectinib, ceritinib, TAE684 (NVP-TAE684), GSK1838705A, AZD3463, ASP3026, PF-06463922, entrectinib (RXDX-101), and AP26113.

In certain embodiments, the biologically active agent is an EGFR inhibitor. Examples of EGFR inhibitors include erlotinib (Tarceva), gefitinib (Iressa), afatinib (Gilotrif), rociletinib (CO-1686), osimertinib (Tagrisso), olmutinib (Olita), naquatinib (ASP8273), nazartinib (EGF816), PF-06747775 (Pfizer), icotinib (BPI-2009), neratinib (HKI-272, PB272), avitinib (AC0010), EAI045, tarloxotinib (TH-4000, PR-610), PF-06459988 (Pfizer), tesevatinib (XL647, EXEL-7647, KD-019), transtinib, WZ-3146, WZ8040, CNX-2006 and dacomitinib (PF-00299804, Pfizer).

In certain embodiments, the biologically active agent is a HER-2 inhibitor. Examples of HER-2 inhibitors include trastuzumab, lapatinib, ado-trastuzumab emtansine, and pertuzumab.

In certain embodiments, the biologically active agent is a CD20 inhibitor. Examples of CD20 inhibitors include obinutuzumab, rituximab, fatumumab, ibritumomab, tositumomab, and ocrelizumab.

In certain embodiments, the biologically active agent is a JAK3 inhibitor. Examples of JAK3 inhibitors include tasocitinib.

In certain embodiments, the biologically active agent is a JAK inhibitor, such as ruxolitinib.

In certain embodiments, the biologically active agent is a BCL-2 inhibitor. Examples of BCL-2 inhibitors include venetoclax, ABT-199 (4-[4-[[2-(4-chlorophenyl)-4,4-dimethylcyclohex-1-en-1-yl]methyl]piperazin-1-yl]-N-[[3-nitro-4-[[(tetrahydro-2H-pyran-4-yl)methyl]amino]phenyl]sulfonyl]-2-[(1H-pyrrolo[2,3-b]pyridin-5-yl)oxy]benzamide), ABT-737 (4-[4-[[2-(4-chlorophenyl)phenyl]methyl]piperazin-l-yl]-N-[4-[[(2R)-4-(dimethylamino)-1-phenylsulfanylbutan-2-yl]amino]-3-nitrophenyl]sulfonylbenzamide) (navitoclax), ABT-263 ((R)-4-(4-((4'-chloro-4,4-dimethyl-3,4,5,6-tetrahydro-[1,1'-biphenyl]-2-yl)methyl)piperazin-1-yl)-N-((4-((4-morpholino-1-(phenylthio)butan-2-yl)amino)-3((trifluoromethyl)sulfonyl)phenyl)sulfonyl)benzamide), GX15-070 (obatoclax mesylate, (2Z)-2-[(5Z)-5-[(3,5-dimethyl-1H-pyrrol-2-yl)methylidene]-4-methoxypyrrol-2-ylidene]indole, methanesulfonic acid))), 2-methoxy-antimycin A3, YC137 (4-(4,9-dioxo-4,9-dihydronaphtho[2,3-d]thiazol-2-ylamino)phenyl ester), pogosin, ethyl 2-amino-6-bromo-4-(1-cyano-2-ethoxy-2-oxoethyl)-4H-chromene-3-carboxylate, nilotinib-d3, TW-37 (N-[4-[[2-(1,1-dimethylethyl)phenyl]sulfonyl]phenyl]-2,3,4-trihydroxy-5-[[2-(1-methylethyl)phenyl]methyl]benzamide), apogossypolone (ApoG2), HA14-1, AT101, sabutoclax, gambogic acid, or G3139 (oblimersen).

In certain embodiments, the biologically active agent is venetoclax.

In certain embodiments, the biologically active agent is a MEK inhibitor. MEK inhibitors are well known and include, for example, trametinib/GSK1120212 (N-(3-{3-cyclopropyl-5-[(2-fluoro-4-iodophenyl)amino]-6,8-dimethyl-2,4,7-trioxo-3,4,6,7-tetrahydropyrido[4,3-d]pyrimidin-1(2H-yl}phenyl)acetamide), selumetinib (6-(4-bromo-2-chloroanilino)-7-fluoro-N-(2-hydroxyethoxy)-3-methylbenzimidazole-5-carboxamide), pimasertib/AS703026/MSC1935369 ((S)-N-(2,3-dihydroxypropyl)-3-((2-fluoro-4-iodophenyl)amino)isonicotinamide), XL-518/GDC-0973 (1-({3,4-difluoro-2-[(2-fluoro-4-iodophenyl)amino]phenyl}carbonyl)-3-[(2S)-piperidin-2-yl]azetidin-3-ol), refametinib/BAY869766/RDEAl 19 (N-(3,4-difluoro-2-(2-fluoro-4-iodophenylamino)-6-methoxyphenyl)-1-(2,3-dihydroxypropyl)cyclopropane-1-sulfonamide), PD-0325901 (N-[(2R)-2,3-dihydroxypropoxy]-3,4-difluoro-2-[(2-fluoro-4-iodophenyl)amino]-benzamide), TAK733 ((R)-3-(2,3-dihydroxypropyl)-6-fluoro-5-(2-fluoro-4-iodophenylamino)-8-methylpyrido[2,3-d]pyrimidine-4,7(3H,8H)-dione), MEK162/ARRY438162 (5-[(4-bromo-2-fluorophenyl)amino]-4-fluoro-N-(2-hydroxyethoxy)-1-methyl-1H-benzimidazol-6-carboxamide), R05126766 (3-[[3-fluoro-2-(methylsulfamoylamino)-4-pyridyl]methyl]-4-methyl-7-pyrimidin-2-yloxychromen-2-one), WX-554, R04987655/CH4987655 (3,4-difluoro-2-((2-fluoro-4-iodophenyl)amino)-N-(2-hydroxyethoxy)-5-((3-oxo-1,2-oxazinan-2-yl)methyl)benzamide) or AZD8330 (2-((2-fluoro-4-iodophenyl)amino)-N-(2-hydroxyethoxy)-1,5-dimethyl-6-oxo-1,6-dihydropyridine-3-carboxamide), U0126-EtOH, PD184352 (CI-1040), GDC-0623, BI-847325, cobimetinib, PD98059, BIX 02189, BIX 02188, bimetinib, SL-327, TAK-733, PD318088.

In certain embodiments, the biologically active agent is a Raf inhibitor. Raf inhibitors are known and include, for example, vemurafenib (N-[3-[[5-(4-chlorophenyl)-1H-pyrrolo[2,3-b]pyridin-3-yl]carbonyl]-2,4-difluorophenyl]-1-propanesulfonamide), sorafenib tosylate (4-[4-[[4-chloro-3-(trifluoromethyl)phenyl]carbamoylamino]phenoxy]-N-methylpyridine-2-carboxamide, 4-methylbenzenesulfonate), AZ628 (3-(2-cyanopropan-2-yl)-N-(4-methyl-3-(3-methyl-4-oxo-3,4-dihydroquinazolin-6-ylamino)phenyl)benzamide), NVP-BHG712 (4-methyl-3-(1-methyl-6-(pyridin-3-yl)-1H-pyrazolo[3,4-d]pyrimidin-4-ylamino)-N-(3-(trifluoromethyl)phenyl)benzamide), RAF-265 (1-methyl-5-[2-[5-(trifluoromethyl)-1H-imidazol-2-yl]pyridin-4-yl]oxy-N-[4-(trifluoromethyl)phenyl]benzimidazole-2-amine), 2-bromaldisine (2-bromo-6,7-dihydro-1H,5H-pyrrolo[2,3-c]azepine-4,8-dione), Raf kinase inhibitor IV (2-chloro-5-(2-phenyl-5-(pyridin-4-yl)-1H-imidazol-4-yl)phenol), sorafenib N-oxide (4-[4-[[[[4-chloro-3(trifluoromethyl)phenyl]amino]carbonyl]amino]phenoxy]-N-methyl-2-pyridinecarboxamide 1-oxide), PLX-4720, dabrafenib (GSK2118436), GDC-0879, RAF265, AZ 628, SB590885, ZM336372, GW5074, TAK-632, CEP-32496, LY3009120, and GX818 (encorafenib).

In certain embodiments, the biologically active agent is an AKT inhibitor, including but not limited to MK-2206, GSK690693, perifosine (KRX-0401), GDC-0068, triciribin, AZD5363, honokiol, PF-04691502 and miltefosine, a FLT-3 inhibitor, including but not limited to P406, dovitinib, quizartinib (AC220), amuvatinib (MP-470), tandutinib (MLN518), ENMD-2076 and KW-2449, or a combination thereof.

In certain embodiments, the biologically active agent is an mTOR inhibitor. Examples of mTOR inhibitors include, but are not limited to, rapamycin and its analogs, everolimus (Afinitor), temsirolimus, ridaforolimus, sirolimus, and deforolimus. Examples of MEK inhibitors include, but are not limited to, tametinib/GSK1120212 (N-(3-{3-cyclopropyl-5-[(2-fluoro-4-iodophenyl)amino]-6,8-dimethyl-2,4,7-trioxo-3,4,6,7-tetrahydropyrido[4,3-d]pyrimidin-1(2H-yl}phenyl)acetamide), selumetinob (6-(4-bromo-2-chloroanilino)-7-fluoro-N-(2-hydroxyethoxy)-3-methylbenzimidazole-5-carboxamide), pimasertib/A8703026/MSC1935369 ((S)-N-(2,3-dihydroxypropyl)-3-((2-fluoro-4-iodophenyl)amino)isonicotinamide), XL-518/GDC-0973 (1-({3,4-difluoro-2-[(2-fluoro-4-iodophenyl)amino]phenyl}carbonyl)-3-[(2S)-piperidin-2-yl]azetidin-3-ol) (cobimetinib), refametinib/BAY869766/RDEA119 (N-(3,4-difluoro-2-(2-fluoro-4-iodophenylamino)-6-methoxyphenyl)-1-(2,3-dihydroxypropyl)cyclopropane-1-sulfonamide), PD-0325901 (N-[(2R)-2,3-dihydroxypropoxy]-3,4-difluoro-2-[(2-fluoro-4-iodophenyl)amino]-benzamide), TAK733 ((R)-3-(2,3-dihydroxypropyl)-6-fluoro-5-(2-fluoro-4-iodophenylamino)-8-methylpyrido[2,3d]pyrimidine-4,7(3H,8H)-dione), MEK162/ARRY438162 (5-[(4-bromo-2-fluorophenyl)amino]-4-fluoro-H-(2-hydroxyethoxy)-1-methyl-1H-benzimidazol-6 carboxamide), R05126766 (3-[[3-fluoro-2-(methylsulfamoylamino)-4-pyridyl]methyl]-4-methyl-7-pyrimidin-2-yloxychromen-2-one), WX-554, R04987655/CH4987655 (3,4-difluoro-2-((2-fluoro-4-iodophenyl)amino)-N-(2-hydroxyethoxy)-5-((3-oxo-1,2-oxazinan-2-yl)methyl)benzamide) or AZD8330 (2-((2-fluoro-4-iodophenyl)amino)-N-(2-hydroxyethoxy)-1,5-dimethyl-6-oxo-1,6-dihydropyridine-3-carboxamide).

In certain embodiments, the biologically active agent is a RAS inhibitor. Examples of RAS inhibitors include, but are not limited to, reolysin and siG12D LODER.

In certain embodiments, the biologically active agent is an HSP inhibitor. HSP inhibitors include, but are not limited to, geldanamycin or 17-N-allylamino-17-demethoxygeldanamycin (17AAG) and radicicol.

In certain embodiments, the biologically active agent is a bisphosphonate. Examples of bisphosphonates include, but are not limited to, clodronate, pamidronate, and zoledronic acid.

Additional biologically active compounds include, for example, everolimus, trabectedin, abraxane, TLK 286, AV-299, DN-101, pazopanib, GSK690693, RTA 744, ON 0910.Na, AZD 6244 (ARRY-142886), AMN-107, TKI-258, GSK461364, AZD 1152, enzastaurin, vandetanib, ARQ-197, MK-0457, MLN8054, PHA-739358, R-763, AT-9263, FLT-3 inhibitor, VEGFR inhibitor, aurora kinase inhibitor, PIK-1 modulator, HDAC inhibitor, c-MET inhibitor, PARP inhibitor, Cdk inhibitor, IGFR-TK, anti-HGF antibody, focal adhesion kinase inhibitor, Map kinase inhibitor (mek), VEGF decoy antibody, pemetrexed, panitumumab, amrubicin, oregovomab, Lep-etu, nolatrexed, azd2171, batabulin, ofatumumab, zanolimumab, edotecarin, tetrandrine, rubitecan, tesmilifen, oblimersen, ticilimumab, ipilimumab, gossypol, Bio 111, 131-I-TM-601, ALT-110, BIO 140, CC 8490, cilengitide, gimatecan, IL13-PE38QQR, INO 1001, IPdR ₁ KRX-0402, lucanton, LY317615, Neuradiab, Vitespan, Rta 744, Sdx 102, Talampanel, Atrasentan, Xr 311, Romidepsin, ADS-100380, Sunitinib, 5-Fluorouracil, Vorinostat, Etoposide, Gemcitabine, Doxorubicin, Liposomal Doxorubicin, 5'-Deoxy-5-Fluorouridine, Vincristine, Temozolomide, ZK-304709, Seliciclib, PD0325901, AZD-6244, Capecitabine, L-Glutamic Acid, N-[4-[2-(2-Amino-4,7-dihydro-4-oxo-1H-pyrrolo[2,3-d]pyrimidin-5-yl)ethyl]benzoyl]-, Disodium Salt, heptahydrate, camptothecin, PEG-labeled irinotecan, tamoxifen, toremifene citrate, anastrazole, exemestane, letrozole, BE8(diethylstilbestrol), estradiol, estrogen, conjugated estrogen, bevacizumab, IMC-1C11, CHIR-258), 3-[5-(methylsulfonylpiperadinemethyl)-indolyl-quinolone, vatalanib, AG-013736, AVE-0005, goserelin acetate, leuprolide acetate, triptorelin pamoate, medroxyprogesterone acetate, hydroxyprogesterone caproate, megestrol acetate, raloxifene, bicalutamide, flutamide, nilutamide, megestrol acetate, CP-724714, TAK-165, HKI-272, erlotinib, lapatainibi, canertinib, ABX-EGF antibody, erbitux, EKB-569, PKI-166, GW-572016, ionafarnib, BMS-214662, tipifarnib, amifostine, NVP-LAQ824, suberoylanalide, hydroxamic acid, valproic acid, trichostatin A, FK-228, SU11248, sorafenib, KRN951, aminoglutethimide, arnsacrine, anagrelide, L-asparaginase, bacille Calmette-Guérin (BCG) vaccine, adriamycin, bleomycin, buserelin, busulfan, carboplatin, carmustine, chlorambucil, cisplatin, cladribine, clodronate, cyproterone, cytarabine, dacarbazine, dactinomycin, daunorubicin, diethylstilbestrol, epirubicin, fludarabine, fludrocortisone, fluoxymesterone, flutamide, glevec, gemcitabine, hydroxybicarbine, tinofamide, ifosidarumide leuprolide, levamisole, lomustine, mechlorethamine, melphalan, 6-mercaptopurine, mesna, methotrexate, mitomycin, mitotane, mitoxantrone, nilutamide, octreotide, oxaliplatin, pamidronate, pentostatin, plicamycin, porfimer, procarbazine, raltitrexed, rituximab, streptozocin, teniposide, testosterone thalidomide, Thioguanine, Thiotepa, Tretinoin, Vindesine, 13-cis-retinoic acid, Phenylalanine mustard, Uracil mustard, Estramustine, Altretamine, Floxuridine, 5-Deoxyuridine, Cytosine Arabinoside, 6-Mecaptopurine, Deoxycoformycin, Calcitriol, Valrubicin, Mithramycin, Vinblastine, Vinorelbine, Topotecan, Razoxin, Marimastat, COL-3, Neovastat, BMS-275291, Squalamine, Endostatin, SU56618, SU66618, EMD121974, Interleukin-12, IM862, Angiostatin, Vitaxin, Droloxifene, Idocifene, Spironolactone, Finasteride, Cimetidine, trastuzumab, denileukin diftitox, gefitinib, bortezomib, paclitaxel, paclitaxel without cremophor, docetaxel, epithilon B, BMS-247550, BMS-310705, droloxifene, 4-hydroxytamoxifen, pipendoxifene, ERA-923, arzoxifene, fulvestrant, acolbifene, lasofoxifene, idoxifene, TSE-424, HMR-3339, ZK186619, topotecan, PTK787/ZK 222584, VX-745, PD 184352, rapamycin, 40-O-(2-hydroxyethyl)-rapamycin, temsirolimus, AP-23573, RAD001, ABT-578, BC-210, LY294002, LY292223, LY292696, LY293684, LY293646, wortmannin, ZM336372, L-779,450, PEG-filgrastim darbepoietin, erythropoietin, granulocyte colony stimulating factor, zoledronate, prednisolone, cetuximab, granulocyte macrophage colony stimulating factor, histrelin, pegylated interferon alpha-2a, interferon alpha-2a, pegylated interferon alpha-2b, interferon alpha-2b, azacitidine, PEG-L-asparaginase, lenalidomide, gemtuzumab, hydrocortisone, interleukin-11, dexrazoxane, alemtuzumab, all-trans retinoic acid, ketoconazole, interleukin-2, megestrol, immunoglobulin, nitrogen mustard, methylprednisolone, ibritgumomab tiuxetan, androgens, decitabine, hexamethylmelamine, bexarotene, tositumomab, arsenic trioxide, cortisone, edtronate, mitotane, cyclosporine, liposomal daunorubicin, edvina-asparaginase, strontium 89, casopitant, netupitant, NK-1 receptor antagonist, palonosetron, aprepitant, diphenhydramine, hydroxyzine, metoclopramide, lorazepam, alprazolam, haloperidol, droperidol, dronabinol, dexamethasone, prochlorpernizoline, ondansetron, dolasetron, tropisetron, pegfilgrastim, erythropoietin, epoetin alfa, darbepoetin alfa and their mixtures.

In certain embodiments, the biologically active agent is selected from, but is not limited to, imatinib mesylate (Gleevac®), dasatinib (Sprycel®), nilotinib (Tasigna®), bosutinib (Bosulif®), trastuzumab (Herceptin®), trastuzumab-DM1, pertuzumab (PerjetaTM), lapatinib (Tykerb®), gefitinib (Iressa®), erlotinib (Tarceva®), cetuximab (Erbitux®), panitumumab (Vectibix®), vandetanib (Caprelsa®), vemurafenib (Zelboraf®), vorinostat (Zolinza®), romidepsin (Istodax®), bexarotene (Tagretin®), alitretinoin (Panretin®), tretinoin (Vesanoid®), carfilizomib (KyprolisTM), pralatrexate (Folotyn®), bevacizumab (Avastin®), ziv-aflibercept (Zaltrap®), sorafenib (Nexavar®), sunitinib (Sutent®), pazopanib (Votrient®), regorafenib (Stivarga®), and cabozantinib (CometriqTM).

According to certain aspects, the biologically active agent is an anti-inflammatory agent, a chemotherapeutic agent, a radiotherapeutic agent, an adjunctive therapeutic agent, or an immunosuppressant.

Suitable chemotherapeutic biologically active agents include, but are not limited to, a radioactive molecule, a toxin, also called a cytotoxin or cytotoxic agent, which includes any agent that is detrimental to cell viability, and liposomes or other vesicles containing chemotherapeutic compounds. Common anticancer pharmaceuticals include: vincristine (Oncovin®) or liposomal vincristine (Marqibo®), daunorubicin (daunomycin or Cerubidine®) or doxorubicin (Adriamycin®), cytarabine (cytosine arabinoside, Aga-C or Cytosar®), L-asparaginase (Elspar®) or PEG-L-asparaginase (pegaspargase or Oncaspar®), etoposide (VP-16), teniposide (Vumon®), 6-mercaptopurine (6-MP or Purinethol®), methotrexate, cyclophosphamide (Cytoxan®), prednisone, dexamethasone (Decadron), imatinib (Gleevec®), dasatinib (Sprycel®), nilotinib (Tasigna®), bosutinib (Bosulif®), and ponatinib (Iclusig™). Examples of additional suitable chemotherapeutic agents include, but are not limited to, 1-dehydrotestosterone, 5-fluorouracil decarbazine, 6-mercaptopurine, 6-thioguanine, actinomycin D, adriamycin, aldesleukin, an alkylating agent, allopurinol sodium, altretamine, amifostine, anastrozole, anthramycin (AMC), an antimitotic agent, cis-dichlorodiamine platinum (II) (DDP) cisplatin), diaminodichloroplatin, anthracycline, an antibiotic, an antimetabolite, asparaginase, BCG live (intravesical), betamethasone sodium phosphate and betamethasone acetate, bicalutamide, bleomycin sulfate, busulfan, calcium leucourin, calicheamicin, capecitabine, carboplatin, lomustine (CCNU), carmustine (BSNU), chlorambucil, cisplatin, cladribine, colchicine, conjugated estrogens, cyclophosphamide, cyclophosphamide, cytarabine, cytarabine, cytochalasin B, cytoxan, dacarbazine, dactinomycin, dactinomycin (formerly actinomycin), daunirubicin HCL, daunorubicin citrate, denileukin diftitox, dexrazoxane, dibromomannitol, dihydroxyanthracindione, docetaxel, dolasetron mesylate, doxorubicin HCL, dronabinol, E. coli L-asparaginase, emetine, epoetin-α, Erwinia L-asparaginase, esterified estrogens, estradiol, estramustine sodium phosphate, ethidium bromide, ethinyl estradiol, etidronate, etoposide citronellal factor, etoposide phosphate, filgrastim, floxuridine, fluconazole, fludarabine phosphate, fluorouracil, flutamide, folinic acid, gemcitabine HCL, glucocorticoids, goserelin acetate, gramicidin D, granisetron HCL, hydroxyurea, idarubicin HCL, ifosfamide, interferon α-2b, irinotecan HCL, letrozole, leucovorin calcium, leuprolide acetate, levamisole HCL, lidocaine, lomustine, maytansinoid, mechlorethamine HCL, medroxyprogesterone acetate, megestrol acetate, melphalan HCL, mercaptipurine, mesna, methotrexate, methyltestosterone, mithramycin, mitomycin C, mitotane, mitoxantrone, lutoxantrone, octreotide acetate, ondansetron HCL, paclitaxel, disodium pamidronate, pentostatin, pilocarpine HCL, plymycin, polypheprosan 20 with carmustine implant, sodium porfimer, procaine, procarbazine HCL, propranolol, rituximab, sargramostim, streptozotocin, tamoxifen, taxol, teniposide, tenoposide, testolactone, tetracaine, thioepa chlorambucil, thioguanine, thiotepa, topotecan HCL, toremifene citrate, trastuzumab, tretinoin, valrubicin, vinblastine sulfate, vincristine sulfate and vinorelbine tartrate.

In some embodiments, a compound of the present invention is administered in combination with a chemotherapeutic agent (e.g., a cytotoxic agent or other chemical entity useful for treating cancer). Examples of chemotherapeutic agents include alkylating agents, antimetabolites, folic acid analogs, pyrimidine analogs, purine analogs and related inhibitors, vinca alkaloids, epipodopyillotoxins, antibiotics, L-asparaginase, topoisomerase inhibitors, interferons, platinum coordination complexes, anthracenedione substituted urea, methylhydrazine derivatives, adrenocortical suppressor, adrenocorticosteroids, progestins, estrogens, antiestrogens, androgens, antiandrogens, and a gonadotropin-releasing hormone analog. Also included are 5-fluorouracil (5-FU), leucovorin (LV), irenotecan, oxaliplatin, capecitabine, paclitaxel, and doxetaxel. Non-limiting examples of chemotherapeutic agents include alkylating agents such as thiotepa and cyclophosphamide, alkyl sulfonates such as busulfan, improsulfan and piposulfan, aziridines such as benzodopa, carboquone, meturedopa and uredopa, ethyleneimines and methylamelamines including altretamine, triethylenemelamine, triethylenephosphoramide, triethylenethiophosphoramide and trimethylolomelamine, acetogenins (especially bullatacin and bullatacinone), camptothecin (including the synthetic analog topotecan), bryostatin, callistatin, CC-1065 (including its synthetic analogs adozelesin, carzelesin and bizelesin), cryptophycins (particularly cryptophycin 1 and cryptophycin 8), dolastatin, duocarmycin (including synthetic analogs KW-2189 and CB1-TM1), eleutherobin, pancratistatin, sarcodictin, spongistatin, nitrogen mustards such as chlorambucil, chlornaphazine, cholophosphamide, estramustine, ifosfamide, mechlorethamine, mechlorethamine oxide hydrochloride, melphalan, novembichin, phenesterine, prednimustine, trofosfamide, uracil mustard, nitrosoureas such as carmustine, chlorzotocin, fotemustine, lomustine, nimustine, and ranimnustine, antibiotics such as enediyne antibiotics (e.g., calicheamicin, especially calicheamicin gammaal and calicheamicin omegall (see, e.g., Agnew, Chem. Inti. Ed Engl. 33:183-186 (1994)), dynemicin including dynemicin A, bisphosphonates such as clodronate, esperamicin, and the neocarzinostatin chromophore and related chromoprotein enediyne antibiotic chromophores), aclacinomycins, actinomycin, authramycin, azaserine, bleomycins, cactinomycin, carbine, caminomycin, carzinophylline, chromomycin, dactinomycin, daunorubicin, 6-detorubicin, detorubicin-oxo-L-norleucine, ADRIAMYCIN® (doxorubicin including morpholino-doxorubicin, cyanomorpholino-doxorubicin, 2-pyrrolino-doxorubicin and deoxydoxorubicin), epirubicin, esorubicin, idarubicin, marcellomycin, mitomycins such as mitomycin C, mycophenolic acid, nogalamycin, olivomycins, peplomycin, potfiromycin, puromycin, kelamicin, rodorubicin, streptonigrin, streptozocin, tubercidin, ubenimex, zinostatin, zorubicin, antimetabolites such as methotrexate and 5-fluorouracil (5-FU), folic acid analogues such as denopterin, methotrexate, pteropterin, trimetrexate, purine analogues such as fludarabine, 6-mercaptopurine, thiamiprine, thioguanine, pyrimidine analogues such as ancitabine, azacitidine, 6-azauridine, carmofur, cytarabine, dideoxyuridine, doxifluridine, enocitabine, floxuridine, androgens such as calusterone, dromostanolone propionate, epithiostanol, mepitiostane, testolactone, anti-adrenal agents such as aminoglutethimide, mitotane, trilostane, folate replenisher such as frolinic acid, aceglatone, aldophosphamide glycoside, aminolevulinic acid, eniluracil, amsacrine, bestrabucil, bisantrene, edatraxate, defamine, demecolcine, diaziquone, elfomitin, elliptinium acetate, epothilone, etoglucide, gallium nitrate, hydroxyurea, lentinan, lonidainine, maytansinoids such as maytansine and ansamitocins, mitoguazone, mitoxantrone, mopidanmol, nitraerin, pentostatin, fenamet, pirarubicin, losoxantrone, podophyllic acid, 2-ethylhydrazide, procarbazine, PSK® polysaccharide complex (JHS Natural Products, Eugene, OR), razoxane, rhizoxin, sizofuran, spirogermanium, tenuazonic acid, triaziquone, 2,2',2''- trichlorotriethylamine, trichothepines (especially T-2 toxin, verracurin A, roridin A, and anguidine), urethane, vindesine, dacarbazine, manmustine, mitobronitol, mitolactol, pipobroman, gacytosin, arabinoside ("Ara-C"), cyclophosphamide, thiotepa, taxoids such as TAXOL® (paclitaxel, Bristol-Myers Squibb Oncology, Princeton, NJ), ABRAXANE®, a cremophor-free formulation of paclitaxel engineered with albumin (American Pharmaceutical Partners, Schaumberg, IL) and TAXOTERE® doxetaxel (Rhone-Poulenc Rorer, Antony, France), chloranbucil, GEMZAR® gemcitabine, 6-thioguanine, mercaptopurine, methotrexate, platinum coordination complexes such as cisplatin, oxaliplatin, and carboplatin, vinblastine, platinum, etoposide (VP-16), ifosfamide, mitoxantrone, vincristine, NAVELBINE® vinorelbine, novantrone, teniposide, edatrexate, daunomycin, aminopterin, xeloda, ibandronate, irinotecan (e.g., CPT-11), the topoisomerase inhibitor RFS 2000, difluoromethylornithine (DMFO), retinoids such as retinoic acid, capecitabine, and pharmaceutically acceptable salts, acids or derivatives of any of the foregoing. Two or more chemotherapeutic agents can be used in admixture for administration in combination with a compound of the present invention. Suitable dosing regimens for combination chemotherapy are known in the art. For example, combination dosing regimens are described in Saltz et al., Proc. Am. Soc. Clin. Oncol. 18:233a (1999) and Douillard et al., Lancet 355(9209): 1041-1047 (2000).

Additional therapeutic agents that can be administered in combination with a compound disclosed herein include bevacizumab, sutinib, sorafenib, 2-methoxyestradiol or 2ME2, finasunate, vatalanib, vandetanib, aflibercept, volociximab, etaracizumab (MEDI-522), cilengitide, erlotinib, cetuximab, panitumumab, gefitinib, trastuzumab, dovitinib, figitumumab, atacicept, rituximab, alemtuzumab, aldesleukin, atlizumab, tocilizumab, temsirolimus, everolimus, lucatumumab, dacetuzumab, HLL1, huN901-DM1, atiprimod, natalizumab, bortezomib, carfilzomib, marizomib, tanespimycin, saquinavir mesylate, ritonavir, nelfinavir mesylate, indinavir sulfate, belinostat, panobinostat, mapatumumab, lexatumumab, dulanermin, ABT-737, oblimersen, plitidepsin, talmapimod, P276-00, enzastaurin, tipifarnib, perifosine, imatinib, dasatinib, lenalidomide, thalidomide, simvastatin, celecoxib, bazedoxifene, AZD4547, rilotumumab, oxaliplatin (Eloxatin), PD0332991, ribociclib (LEE011), amebaciclib (LY2835219), HDM201, fulvestrant (Faslodex), exemestane (Aromasin), PIM447, ruxolitinib (INC424), BGJ398, necitumumab, pemetrexed (Alimta), and ramucirumab (IMC-1121B).

In certain embodiments, the adjunctive therapy is a monoclonal antibody (MAb). Some MAbs stimulate an immune response that destroys cancer cells. Like antibodies naturally produced by B cells, these MAbs can “coat” the surface of a cancer cell, causing the immune system to destroy it. For example, bevacizumab targets vascular endothelial growth factor (VEGF), a protein secreted by tumor cells and other cells in the tumor microenvironment that promotes the development of tumor blood vessels. When bound to bevacizumab, VEGF is unable to interact with its cellular receptor, preventing signaling that leads to the growth of new blood vessels. Similarly, cetuximab and panitumumab target the epidermal growth factor receptor (EGFR), and trastuzumab targets human epidermal growth factor receptor 2 (HER-2). MAbs that bind to cell-surface growth factor receptors prevent target receptors from sending their normal growth-promoting signals. They can also trigger apoptosis and activate the immune system to destroy tumor cells.

According to one aspect of the present invention, the biologically active agent is an immunosuppressant. The immunosuppressant may be a calcineurin inhibitor, such as cyclosporine or ascomycin, such as cyclosporine A (NEORAL®), FK506 (tacrolimus), pimecrolimus, an mTOR inhibitor, such as rapamycin or a derivative thereof, such as sirolimus (RAPAMUNE®), everolimus (Certican®), temsirolimus, zotarolimus, biolimus-7, biolimus-9, rapalog, egridaforolimus, azathioprine, campath 1H, an S1P receptor modulator, such as fingolimod or an analogue thereof, an anti-IL-8 antibody, mycophenolic acid or a salt thereof, such as the sodium salt, or a prodrug thereof, such as mycophenolate mofetil (CELLCEPT®), OKT3 (ORTHOCLONE OKT3®), prednisone, ATGAM®, THYMOGLOBULIN®, brequinar sodium, OKT4, T10B9.A-3A, 33B3.1, 15-deoxyspergualin, tresperimus, leflunomide ARAVA®, CTLAI-Ig, amn-CD25, anti-IL2R, basiliximab (SIMULECT®), daclizumab (ZENAPAX®), mizorbin, methotrexate, dexamethasone, ISAtx-247, SDZ ASM 981 (pimecrolimus, Elidel®), CTLA4lg (abatacept), belatacept, LFA3lg, etanercept (brand name Enbrel® by Immunex), adalimumab (Humira®), infliximab (Remicade®), anti-LFA-1 antibody, natalizumab (Antegren®), enlimomab, gavilimomab, antithymocyte immunoglobulin, siplizumab, alefacept, efalizumab, pentase, mesalazine, asacol, codeine phosphate, benorylate, fenbufen, naprosyn, diclofenac, etodolac and indomethacin, aspirin and ibuprofen.

In some embodiments, the biological agent is a therapeutic agent that is a biological agent, such as a cytokine (e.g., an interferon or an interleukin (e.g., IL-2)) used to treat cancer. In some embodiments, the biological agent is an anti-angiogenic agent, such as an anti-VEGF agent, such as bevacizumab (AVASTIN®). In some embodiments, the biological agent is an immunoglobulin-based biological agent, such as a monoclonal antibody (e.g., a humanized antibody, a fully human antibody, an Fc fusion protein, or a functional fragment thereof) that agonizes a target to stimulate an anti-cancer response or counteracts an antigen important for cancer. Such agents include RITUXAN® (rituximab), ZENAPAX® (daclizumab), SIMULECT® (basiliximab), SYNAGIS® (palivizumab), REMICADE® (infliximab), HERCEPTIN® (trastuzumab), MYLOTARG® (gemtuzumab ozogamicin), CAMPATH® (alemtuzumab), ZEVALIN® (ibritutomab tiuxetan), HUMIRA® (adalimumab), XOLAIR® (omalizumab), BEXXAR® (tositumomab-1-131), RAPTIVA® (efalizumab), ERBITUX® (cetuximab), AVASTIN® (bevacizumab), TYSABRI® (natalizumab), ACTEMRA® (tocilizumab), VECTIBIX® (panitumumab), LUCENTIS® (ranibizumab), SOURIS® (eculizumab), CIMZIA® (certolizumab pegol), SIMPONI® (golimumab), ILARIS® (canakinumab), STELARA® (ustekinumab), ARZERRA® (ofatumumab), PROLIA® (denosumab), NUMAX® (motavizumab), ABTHRAX® (raxibacumab), BENLYSTA® (belimumab), YERVOY® (ipilimumab), ADCETRIS® (brentuximab vedotin), PERJETA® (pertuzumab), KADCYLA® (adotrastuzumab emtansine), and GAZYVA® (obinutuzumab). Also included are antibody-drug conjugates.

In certain embodiments, the additional therapy is bendamustine. In certain embodiments, the additional therapy is obinutuzmab. In certain embodiments, the additional therapy is a proteasome inhibitor, such as ixazomib or oprosomib. In certain embodiments, the additional therapy is a histone deacetylase inhibitor, such as ACY241. In certain embodiments, the additional therapy is a BET inhibitor, such as GSK525762A, OTX015, BMS-986158, TEN-010, CPI-0610, INCB54329, BAY1238097, FT-1101, ABBV-075, BI 894999, GS-5829, GSK1210151A (I-BET-151), CPI-203, RVX-208, XD46, MS436, PFI-1, RVX2135, ZEN3365, XD14, ARV-771, MZ-1, PLX5117, 4-[2-(cyclopropylmethoxy)-5-(methanesulfonyl)phenyl]-2-methylisoquinolin-1(2H)-one, EP11313 and EP11336. In certain embodiments, the additional therapy is an MCL-1 inhibitor, such as AZD5991, AMG176, MIK665, S64315, or S63845. In certain embodiments, the additional therapy is an LSD-1 inhibitor, such as ORY-1001, ORY-2001, INCB-59872, IMG-7289, TAK-418, GSK-2879552, 4-[2-(4-amino-piperidin-1-yl)-5-(3-fluoro-4-methoxy-phenyl)-1-methyl-6-oxo-1,6-dihydropyrimidin-4-yl]-2-fluoro-benzonitrile, or a salt thereof. In certain embodiments, the additional therapy is a CS1 antibody, such as elotuzumab. In certain embodiments, the additional therapy is an anti-CD38 antibody, such as daratumumab or isatuximab. In certain embodiments, the additional therapy is a BCMA antibody or antibody conjugate, such as GSK2857916 or BI 836909.

In certain embodiments, the biologically active agent is selinexor. In certain embodiments, Compound 1 is administered in combination with selinexor. In certain embodiments, Compound 1 is administered in combination with aspirin.

In certain embodiments, Compound 2, Compound 3, Compound 4, Compound 5, Compound 6, Compound 7, Compound 8, Compound 9, Compound 10, Compound 11, Compound 12, or Compound 13 are administered in combination with selinexor. In certain embodiments, Compound 2, Compound 3, Compound 4, Compound 5, Compound 6, Compound 7, Compound 8, Compound 9, Compound 10, Compound 11, Compound 12, or Compound 13 are administered in combination with aspirin.

In other embodiments, the compound described herein is administered in combination with a drug selected from selinexor, oxafenamide, belantamab mafodotin, denosumab, zoledronic acid, plerixafor, eltrombopag, ipilimumab, palbociclib, ricolinostat, afuresertib, dinaciclib, filanesib, indatuximab ravtansine, masitinib, sonidegib, sotatercept, ulocuplumab, and urelumab.

In certain embodiments, Compound 1 is administered in combination with a chemokine receptor antagonist, such as plerixafor.

In certain embodiments, Compound 1 is administered in combination with a vaccine, such as PVX-410.

In certain embodiments, Compound 1 is administered in combination with a chemotherapeutic agent, such as bendamustine, busulfan, carmustine, cyclophosphamide, doxorubicin, etoposide, fludarabine, melphalan, or vincristine.

In certain embodiments, Compound 1 is used in combination with a conjugated antibody, such as belantamab mafodotin, CC-99712, HDP-101, or MEDI2228.

In certain embodiments, Compound 1 is used in combination with a phospholipid drug conjugate, such as CLR 131.

In certain embodiments, a compound described herein is used in combination with a chemokine receptor antagonist, such as plerixafor.

In certain embodiments, a compound described herein is administered in combination with a vaccine, such as PVX-410.

In certain embodiments, a compound described herein is used in combination with a chemotherapeutic agent, such as bendamustine, busulfan, carmustine, cyclophosphamide, doxorubicin, etoposide, fludarabine, melphalan, or vincristine.

In certain embodiments, a compound described herein is used in combination with a conjugated antibody, such as belantamab, mafodotin, CC-99712, HDP-101, or MEDI2228.

In certain embodiments, a compound described herein is used in combination with a phospholipid drug conjugate, such as CLR 131.

In certain embodiments, a compound described herein is used in combination with a CHOP (cyclophosphamide, vincristine, prednisolone, and daunorubicin) regimen. In certain embodiments, Compound 1 is used in combination with a CHOP (cyclophosphamide, vincristine, prednisolone, and daunorubicin) regimen. In certain embodiments, Compound 1, rituximab, and CHOP are administered in combination.

In certain embodiments, a compound described herein is used in combination with a CVP (cyclophosphamide, vincristine, and prednisone) regimen. In certain embodiments, Compound 1 is used in combination with a CVP (cyclophosphamide, vincristine, and prednisolone) regimen. In certain embodiments, Compound 1, rituximab, and CVP are administered in combination.

In certain embodiments, a compound described herein is used in combination with romidepsin, belinostat, or brentuximab to treat a cancer, such as T-NHL. In certain embodiments, Compound 1 is used in combination with a romidepsin, belinostat, or brentuximab to treat a cancer, such as T-NHL.

In certain embodiments, a compound described herein is used in combination with polatuzumab, tafastemab, CAR-T, a BTK inhibitor, or a PI3-kinase inhibitor to treat a cancer, such as B-NHL. In certain embodiments, Compound 1 is used in combination with polatuzumab, tafastemab, CAR-T, a BTK inhibitor, or a PI3-kinase inhibitor to treat a cancer, such as B-NHL.

In certain embodiments, Compound 1 is used to treat relapsed and/or refractory mantle cell leukemia. In certain embodiments, Compound 1 and rituximab are used to treat relapsed and/or refractory mantle cell leukemia. In certain embodiments, Compound 1, rituximab, and bendamustine are used to treat relapsed and/or refractory mantle cell leukemia. In certain embodiments, Compound 1, rituximab, and ibrutinib are used to treat relapsed and/or refractory mantle cell leukemia.

In certain embodiments, Compound 1 is used to treat relapsed and/or refractory marginal zone cell leukemia. In certain embodiments, Compound 1 and rituximab are used to treat relapsed and/or refractory marginal zone cell leukemia. In certain embodiments, Compound 1, rituximab, and dexamethasone are used to treat relapsed and/or refractory marginal zone cell leukemia.

In certain embodiments, Compound 1 is used in combination with rituximab and DHAP (dexamethasone, cytarabine, and cisplatin). In certain embodiments, Compound 1 is used in combination with DHAP (dexamethasone, cytarabine, and cisplatin). In certain embodiments, Compound 1 is used in combination with rituximab and ICE (ifosfamide, carboplatin, and etoposide). In certain embodiments, Compound 1 is used in combination with ICE (ifosfamide, carboplatin, and etoposide). In certain embodiments, Compound 1 is used in combination with rituximab and GemOx (gemcitabine and oxaliplatin). In certain embodiments, Compound 1 is used in combination with GemOx (gemcitabine and oxaliplatin). In certain embodiments, Compound 1 is used in combination with polatuzumab and brentuximab. In certain embodiments, Compound 1 is used in combination with polatuzumab. In certain embodiments, Compound 1 is used in combination with tafasitimab. In certain embodiments, Compound 1 is used in combination with tafasitimab and lenolidomide. In certain embodiments, Compound 1 is used in combination with rituximab and lenolidomide. In certain embodiments, Compound 1 is used in combination with an anti-CD19 CART (e.g., axicabtagene, lisocabtagene, tisafenlucleucel, loncastuximab, or tesirine). In certain embodiments, one of the above combinations is used to treat DLBCL or B-NHL.

In certain embodiments, Compound 1 is used in combination with praltrexate. In certain embodiments, Compound 1 is used in combination with bendamustine. In certain embodiments, Compound 1 is used in combination with brexucabtagene. In certain embodiments, Compound 1 is used in combination with bortezomib and rituximab. In certain embodiments, Compound 1 is used in combination with bendamustine and rituximab. In certain embodiments, Compound 1 is used in combination with VR-CAP (rituximab, cyclophosphamide, doxorubicin, prednisolone, and borzomib).

In certain embodiments, Compound 1 is administered in combination with carfilzomib and daratumumab.

In certain embodiments, Compound 1 is used in combination with abecma. In certain embodiments, Compound 1 is used in combination with melflufen. In certain embodiments, Compound 1 is used in combination with ciltacabtagene autoleucel.

In certain embodiments, a compound described herein, such as Compound 1, is used in a therapy according to the table below. In other embodiments, Compound 1 is replaced with another compound described herein.

VI. Pharmaceutical compositions

Any of the compounds disclosed herein may be administered as a pure chemical, but are typically administered as a pharmaceutical composition that contains an effective amount for a host, typically a human, in need of such treatment for any of the disorders described herein. Accordingly, the present invention provides pharmaceutical compositions comprising an effective amount of the compound or a pharmaceutically acceptable salt together with at least one pharmaceutically acceptable carrier for any of the uses described herein. The pharmaceutical composition may comprise the compound or a salt thereof as the sole active agent, or in an alternative embodiment, the compound and at least one additional active agent.

According to certain embodiments, the pharmaceutical composition is in a dosage form that contains from about 0.1 mg to about 2000 mg, from about 10 mg to about 1000 mg, from about 50 mg to about 600 mg, or from about 100 mg to about 400 mg of the active compound. According to another embodiment, the pharmaceutical composition is in a dosage form that contains from about 0.1 mg to about 100 mg, from about 0.5 mg to about 100 mg, from about 1 mg to about 50 mg, or from about 2 mg to about 25 mg of the active compound. According to another embodiment, the pharmaceutical composition is in a dosage form that contains from about 0.1 mg to about 10 mg, from about 0.5 mg to about 8 mg, from about 0.5 mg to about 6 mg, or from about 0.5 mg to about 5 mg of the active compound. Examples include dosage forms containing at least, or in some embodiments, no more than, 1, 5, 10, 25, 50, 100, 200, 250, 300, 400, 500, 600, 700, or 750 mg of the active compound or salt thereof.

According to certain embodiments, the pharmaceutical composition is in a dosage form that contains from about 1 μg to about 2000 μg, from about 10 μg to about 1000 μg, from about 50 μg to about 600 μg, or from about 100 μg to about 400 μg of the active compound. According to another embodiment, the pharmaceutical composition is in a dosage form that contains from about 1 μg to about 400 μg, from about 5 μg to about 400 μg, from about 10 μg to about 250 μg, or from about 25 μg to about 250 μg of the active compound. Examples include dosage forms containing at least, or in some embodiments, no more than, 0.1, 1, 5, 10, 25, 50, 100, 200, 250, 300, 400, 500, 600, 700, or 750 μg of the active compound or salt thereof.

The pharmaceutical composition may also include a molar ratio of the active compound to the additional biologically active agent. For example, the pharmaceutical composition may comprise a molar ratio of about 0.0005:1, about 0.001:1, about 0.01:1, about 0.05:1, or about 0.1:1 of the active compound to the anti-inflammatory agent or immunosuppressant. For example, the pharmaceutical composition may comprise a molar ratio of about 0.5:1, about 1:1, about 2:1, about 3:1, or from about 1.5:1 to about 4:1 of the active compound to the anti-inflammatory agent or immunosuppressant. The compounds disclosed herein may be administered orally, topically, parenterally, by inhalation or spray, sublingually, by implant, including an ocular implant, transdermally, by buccal administration, rectally, as an ophthalmic solution, by injection, including ocular injection, by intravenous route, intra-aortic route, intracranial route, subdermal route, intraperitoneal route, subcutaneous route, transnasal route, sublingual or rectal route, or other routes, in unit dosage formulations containing conventional pharmaceutically acceptable carriers. For ocular delivery, the compound can be administered as needed, for example, by intravitreal, intrastromal, intracameral, sub-Tenon, subretinal, retrobulbar, peribulbar, suprachondrium, conjunctival, subconjunctival, episcleral, periocular, transscleral, retrobulbar, posterior parascleral, pericorneal or lacrimal duct injection, or through a mucosal, mucinous or mucosal barrier, with immediate or controlled release, or through an ocular device.

The pharmaceutical composition may be formulated in any pharmaceutically useful form, such as an aerosol, cream, gel, pill, injection or infusion solution, capsule, tablet, syrup, transdermal patch, subcutaneous patch, dry powder, inhalation formulation, medical device, suppository, buccal or sublingual formulation, parenteral formulation or ophthalmic solution. Some dosage forms, such as tablets and capsules, are divided into appropriately sized unit doses containing appropriate amounts of the active ingredients, such as an amount effective to achieve the desired purpose.

Carriers include excipients and diluents and must be of sufficiently high purity and sufficiently low toxicity to render them suitable for administration to the patient being treated. The carrier may be inert or may have its own pharmaceutical advantages. The amount of carrier used in combination with the compound is sufficient to provide a practical amount of material for administration per unit dose of the compound.

Classes of carriers include, but are not limited to, binders, buffering agents, colorants, diluents, disintegrants, emulsifiers, flavors, glidants, lubricants, preservatives, stabilizers, surfactants, tabletting agents, and wetting agents. Some carriers may be classified into more than one class, for example, a vegetable oil may be used as a lubricant in some formulations and as a diluent in others. Pharmaceutically acceptable carriers are carriers that do not cause any severe adverse reactions in the human body when administered at a dose in the amount that would be used in the corresponding pharmaceutical composition. Examples of pharmaceutically acceptable carriers include sugars, starches, celluloses, powdered tragacanth, malt, gelatin, talc, and vegetable oils. Optional active agents that do not significantly affect the activity of the compound of the present invention may be included in the pharmaceutical composition.

Pharmaceutical compositions/combinations can be formulated for oral administration. These compositions can contain any amount of active compound that achieves the desired result, for example, from 0.1 to 99% by weight (wt. %) of the compound, and usually at least about 5 wt. % of the compound. According to some embodiments, from about 25 wt. % to about 50 wt. % or from about 5 wt. % to about 75 wt. % of the compound is contained.

Formulations suitable for rectal administration are usually presented as unit-dose suppositories. These may be prepared by mixing the active compound with one or more conventional solid carriers, such as cocoa butter, and then molding the resulting mixture.

Formulations suitable for topical application to the skin are preferably in the form of an ointment, cream, lotion, paste, gel, spray, aerosol or oil. Carriers that may be used include petrolatum, lanolin, polyethyleneglycols, alcohols, transdermal enhancers and combinations of two or more of these.

Formulations suitable for transdermal administration may be presented as discrete patches adapted to remain in close contact with the epidermis of the recipient for an extended period of time. Formulations suitable for transdermal administration may also be delivered by iontophoresis (see, for example, Pharmaceutical Research 3 (6): 318 (1986)) and typically take the form of an optionally buffered aqueous solution of the active compound. According to certain embodiments, patches or microneedle devices are provided for delivering drugs through or into biological tissue, particularly the skin. The patches or microneedle devices enable the drug to be delivered at a clinically significant rate through or into the skin or other tissue barriers with minimal or no tissue damage, pain, or irritation.

Formulations suitable for pulmonary administration can be delivered using a wide range of single/multiple dose dry powder inhalers (DPIs), both breath-actuated and power-driven. The devices most commonly used for respiratory delivery include nebulisers, metered dose inhalers and dry powder inhalers. Several types of nebulisers are available, including jet nebulisers, ultrasonic nebulisers and vibrating mesh nebulisers. The choice of an appropriate pulmonary delivery device depends on parameters such as the nature of the drug and its formulation, the site of action and the pathophysiology of the lung.

In certain embodiments, a compound described herein, such as Compound 1, is formulated as described in Table A.

According to certain embodiments, the present invention relates to a pharmaceutical composition comprising Compound 1, a non-aqueous solvent, a preservative, and/or a pH adjusting agent.

In certain embodiments, a compound described herein, e.g., Compound 1, is formulated as described herein, e.g., as described in Table B or Table C.

Non-limiting examples of preservatives include alcohols (e.g., ethanol, benzyl alcohol), aluminum acetate, benzalkonium chloride, benzethonium chloride, benzoic acid and its salts (e.g., potassium benzoate, sodium benzoate), boric acid and its salts (e.g., sodium borate), bronopol, butylene glycol, butylhydroxyanisole, calcium acetate, calcium chloride, cetrimide, chlorhexidine, chlorobutanol, chlorocresol, chloroxylenol, citric acid monohydrate, cresol, dimethyl ether, edetic acid and its salts, gelatin, glycerin, hexetidine, imidurea, lactic acid and its salts (e.g., calcium lactate, sodium lactate), monothioglycerin, parabens (e.g., butylparaben, ethylparaben, methylparaben, propylparaben, sodium propylparaben), pentetic acid and its salts, phenol, phenoxyethanol, phenylethyl alcohol, phenylmercuric acetate, phenylmercuric borate, phenylmercuric nitrate, propyl gallate, potassium metabisulfite, sorbic acid and its salts (e.g. potassium sorbate), propionic acid and its salts (e.g. sodium propionate), propylene glycol, sodium acetate, sodium sulfite, sulfur dioxide, thimerosal and xylitol.

The term pH adjusting agent may be used interchangeably with pH adjusting agent, pH regulator and/or buffering agent. Non-limiting examples of pH adjusting agents include acetic acid, glacial acetic acid, adipic acid, concentrated ammonia solution and its salts (e.g., ammonium carbonate, ammonium chloride, ammonium phosphate), arginine, boric acid and its salts (e.g., sodium borate), calcium carbonate, calcium hydroxide, calcium lactate, tribasic calcium phosphate, citric acid (anhydrous or monohydrate) and its salts (e.g., potassium citrate, sodium citrate), diethanolamine, fumaric acid and its salts, glycine, gluconic acid, hydrochloric acid, dilute hydrochloric acid, alpha-lactalbumin, lactic acid and its salts (e.g., sodium lactate solution), lysine hydrochloride, maleic acid, malic acid, methionine, monoethanolamine, sodium glutamate, meglumine, nitric acid, phosphoric acid and its salts (e.g. dibasic sodium phosphate, monobasic sodium phosphate, potassium phosphate, dibasic potassium hydrogen phosphate, potassium phosphate), dilute phosphoric acid, potassium bicarbonate, potassium hydroxide, monobasic potassium metaphosphate, propionic acid, racemethionine, sodium acetate, sodium bicarbonate, sodium carbonate, sodium hydroxide, succinic acid, sulfuric acid, tartaric acid, trolamine, hydroxide, amine and their salts.

Non-limiting examples of buffering agents include, but are not limited to, adipic acid, ammonia solution, boric acid, calcium carbonate, calcium hydroxide, calcium lactate, calcium phosphate, citric acid, sodium phosphate, diethanolamine, maleic acid, malic acid, methionine, monoethanolamine, monosodium glutamate, phosphoric acid, potassium citrate, sodium acetate, sodium bicarbonate, sodium borate, sodium carbonate, sodium citrate, sodium hydroxide, sodium lactate, and triethanolamine.

According to certain embodiments, the buffering agent is selected from carbonates, citrates, gluconates, lactates, phosphates, tartrates, potassium metaphosphate, monobasic potassium phosphate, sodium acetate, anhydrous and dihydrate sodium citrate.

According to certain embodiments, the pharmaceutical composition comprises water.

According to certain embodiments, the pharmaceutical composition includes a non-aqueous or anhydrous solvent.

Non-limiting examples of non-aqueous solvents include alcohols (e.g., ethanol), PEG 300, PEG 400, propylene glycol, cremophor, caplex 355, CapryolTM 90, LauroglycolTM 90, TranscutolHP, butylated hydroxytoluene, benzyl alcohol, citric acid, triacetin, propylene glycol, fats and oils.

1. According to certain embodiments, Compound 1 is provided as a capsule filled with a liquid containing a solvent, a preservative, and a pH adjusting agent.

2. The pharmaceutical composition of embodiment 1, wherein the capsule is filled with a solution containing about 0.001, 0.005, 0.01, 0.015, 0.02, 0.025, 0.03, 0.033, 0.035, 0.04, 0.045, 0.05, 0.06, 0.07, 0.08, 0.09, 0.1, 0.15, 0.2, 0.25, 0.3, 0.35, 0.4, 0.45, 0.5, 0.55, 0.6, 0.65, 0.7, 0.75, 0.8, 0.85, 0.9, 0.95, 1, 1.5, 2, 2.5, 3, 3.5, 4, 4.5 or 5 wt.% Compound 1.

3. The pharmaceutical composition of embodiment 1, wherein the capsule is filled with a solution containing at least about 0.001, 0.005, 0.01, 0.015, 0.02, 0.025, 0.03, 0.033, 0.035, 0.04, 0.045, 0.05, 0.06, 0.07, 0.08, 0.09, 0.1, 0.15, 0.2, 0.25, 0.3, 0.35, 0.4, 0.45, 0.5, 0.55, 0.6, 0.65, 0.7, 0.75, 0.8, 0.85, 0.9, 0.95, 1, 1.5, 2, 2.5, 3, 3.5, 4, 4.5 or 5 wt.% Compound 1.

4. The pharmaceutical composition according to any one of embodiments 1-3, wherein the solvent is non-aqueous.

5. The pharmaceutical composition according to embodiment 4, wherein the solvent is PEG.

6. The pharmaceutical composition according to embodiment 5, wherein the solvent is PEG 400.

7. The pharmaceutical composition according to any one of embodiments 1-6, wherein the capsule is filled with a solution containing about 95, 95.5, 96, 96.5, 97, 97.5, 98, 98.1, 98.2, 98.3, 98.4, 98.5, 98.6, 98.65, 98.7, 98.75, 98.8, 98.85, 98.9, 99, 99.1, 99.2, 99.3, 99.4, or 99.5 wt. % of a solvent.

8. The pharmaceutical composition according to any one of embodiments 1-6, wherein the capsule is filled with a solution containing at least about 85, 90, 95, 95.5, 96, 96.5, 97, 97.5, 98, 98.1, 98.2, 98.3, 98.4, 98.5, 98.6, 98.65, 98.7, 98.75, 98.8, 98.85, 98.9, 99, 99.1, 99.2, 99.3, 99.4, or 99.5 wt. % of a solvent.

9. The pharmaceutical composition according to any one of embodiments 1-8, wherein the preservative is butylated hydroxytoluene.

10. The pharmaceutical composition according to any one of embodiments 1-9, wherein the capsule is filled with a solution containing at least about 0.01, 0.05, 0.1, 0.15, 0.2, 0.25, 0.3, 0.35, 0.4, 0.45, 0.5, 0.55, 0.6, 0.65, 0.8, 0.85, 0.9, 0.95, or 1 wt. % of a preservative.

11. The pharmaceutical composition according to any one of embodiments 1-9, wherein the capsule is filled with a solution containing about 0.01, 0.05, 0.1, 0.15, 0.2, 0.25, 0.3, 0.35, 0.4, 0.45, 0.5, 0.55, 0.6, 0.65, 0.8, 0.85, 0.9, 0.95, or 1 wt. % of a preservative.

12. The pharmaceutical composition according to any one of embodiments 1-11, wherein the pH adjusting agent is citric acid.

13. The pharmaceutical composition according to any one of embodiments 1-11, wherein the pH adjusting agent is anhydrous citric acid.

14. The pharmaceutical composition according to any one of embodiments 1-13, wherein the capsule is filled with a solution containing at least about 0.01, 0.05, 0.1, 0.15, 0.2, 0.25, 0.3, 0.35, 0.4, 0.45, 0.5, 0.55, 0.6, 0.65, 0.8, 0.85, 0.9, 0.95, 1, 1.1, 1.2, 1.3, 1.4, 1.5, 1.6, 1.7, 1.8, 1.9, 2, 2.25, 2.5, 2.75, or 3 wt. % of a pH adjusting agent.

15. The pharmaceutical composition according to any one of embodiments 1-13, wherein the capsule is filled with a solution containing about 0.01, 0.05, 0.1, 0.15, 0.2, 0.25, 0.3, 0.35, 0.4, 0.45, 0.5, 0.55, 0.6, 0.65, 0.8, 0.85, 0.9, 0.95, 1, 1.1, 1.2, 1.3, 1.4, 1.5, 1.6, 1.7, 1.8, 1.9, 2, 2.25, 2.5, 2.75, or 3 wt. % of a pH adjusting agent.

Additional excipients

In certain embodiments, a compound described herein is administered as a pharmaceutical composition comprising one or more excipients according to the Handbook of Pharmaceutical Excipients ^9th Edition (or earlier edition).

In certain embodiments, the compound described herein is dissolved or dispersed in oil or fat components. When administered orally, the oil or fat component can form a microemulsion in the digestive system, thereby allowing the compound to be absorbed more quickly into the body.

According to certain embodiments, the oil is an edible oil, a medicinal oil, a pharmaceutically acceptable fat and oil, or edible fats and oils. For example, pharmaceutically acceptable fats and oils or edible fats and oils include, but are not limited to, vegetable oil, animal oil, fish oil, or mineral oil.

According to certain embodiments, the edible oil is selected from the group consisting of medium chain triglycerides of fatty acids, amaranth oil, apricot kernel oil, apple oil, argan oil, artichoke oil, avocado oil, almond oil, acai berry extract, peanut oil, buffalo pumpkin oil, borage seed oil, borage oil, babassu oil, coconut oil, corn oil, cottonseed oil (cottonseed oil), cashew oil, carob oil, coriander oil, camellia oil (camellia oil), cauliflower oil, cape chestnut oil, blackcurrant oil, fallow deer oil, evening primrose oil, grape syrup oil Oila (hibiscus oil), grape seed oil, pumpkin oil, hazelnut oil, hemp oil, kapok oil, krill oil, flaxseed oil, walnut oil Macadamia nut, Mongolian oil, Moringa oil, Malula oil, Meadowfoam oil, Mustard oil, Nougat oil, Olive oil, Okra oil, Hibiscus oil, Palm oil, Palm kernel oil, Peanut oil, Pecan oil, Pine oil, Pistachio oil, Pumpkin seed oil, Papaya oil, Perryl oil (perryl oil), Poppy seed oil, Prune oil, Saw palmetto oil, Quinoa oil, Canola oil, Rice germ oil, Rice bran oil, Rice oil, Rarelman oil, Safflower oil (safflower oil), Soybean oil, Sesame oil, Sunflower oil, Milk thistle oil, Tomato oil, Wheat germ oil, Walnut oil, Watermelon oil, Docosahexaenoic acid (DHA), Eicosapentaenoic acid (EPA), and any combination thereof.

According to certain embodiments, the fat-soluble oil is a vitamin, including but not limited to vitamin A oil, vitamin D oil, vitamin E oil, vitamin K oil and derivatives thereof, and glycerophospholipids such as lecithin, and any combination thereof can also be used as fats and oils according to the present invention.

According to certain embodiments, the oil of the present invention may be liquid (e.g., fatty oil) or solid (fat, etc.) at room temperature. According to certain embodiments, the oil of the present invention is liquid at living body temperature (in particular, at stomach temperature of about 37°C). According to certain embodiments, the fat comprises many saturated fatty acids (e.g., palmitic acid, stearic acid), and/or the fatty oils contain many unsaturated fatty acids (e.g., oleic acid, linoleic acid, linolenic acid), and wherein these fatty acids and oils may be esterified.

According to certain embodiments, fatty acids containing 2 to 4 carbon atoms are called short-chain fatty acids (lower fatty acids), fatty acids containing 5 to 12 fatty acids are called medium-chain fatty acids, and fatty acids with 12 or more carbon atoms are called long-chain fatty acids (higher fatty acids). Since fatty acids generally have high hydrophilicity when the number of carbon atoms is small, the oils of the present invention include medium-chain fatty acids (e.g., triglycerides of medium-chain fatty acids such as tri(caprylic/capric acid)glycerol and glyceryl tricaprylate) and long-chain fatty acids.

According to certain embodiments, the fatty acid is used in combination with a saturated polyglycolated glyceride obtained by polyglycolysis of hydrogenated vegetable oil with polyethylene glycol, a mono-, di- or triglyceride, and a mono- or diester of a fatty acid and polyethylene glycol.

According to certain embodiments, the pharmaceutical composition includes a diluent. Non-limiting examples of a diluent used in capsules include, but are not limited to, calcium carbonate, dibasic or tribasic calcium phosphate, kaolin, lactose, lactitol, mannitol, microcrystalline cellulose, powdered cellulose, cellulose acetate, sorbitol, starch, calcium sulfate, dextrates, dextrin, dextrose, maltodextrin, erythritol, glyceryl palmitostearate, isomalt, magnesium carbonate, magnesium oxide, mannitol, sodium chloride, sucrose, sulfobutyl ether b-cyclodextrin, talc, and xylitol.

VII. Metabolites

According to certain aspects of the present invention, a metabolite of Compound 1 is used as either (i) an active agent administered to a patient in need thereof in an effective amount for the treatment of any of the disorders and in any of the methods described herein, or (ii) as an intermediate in a synthetic process. Examples of metabolites of Compound 1 include:

According to certain embodiments, the metabolite of Compound 1 is enantiomerically or diastereomerically enriched:

In certain embodiments, a metabolite of Compound 1 is used for the treatment described herein.

In certain embodiments, a metabolite of Compound 1 is used as an intermediate to produce Compound 1.

VIII. General Synthesis

The compounds described herein can be prepared by methods known to those skilled in the art. According to one non-limiting example, the disclosed compounds can be prepared using the following schemes.

Compounds of the present invention with stereocenters may conveniently be prepared without stereochemistry. It will be apparent to one skilled in the art that pure or enriched enantiomers and diastereomers may be prepared by methods known in the art. Examples of methods for preparing optically active substances include at least the following:

i) physical separation of crystals is a method in which macroscopic crystals of individual enantiomers are separated manually. This technique can be used if crystals of individual enantiomers exist, i.e. the material is a conglomerate and the crystals are visually distinguishable,

ii) simultaneous crystallization is a process in which the individual enantiomers are crystallized separately from a solution of the racemate, which is only possible if the enantiomer is a conglomerate in the solid state,

iii) enzymatic resolution is a process in which partial or complete separation of a racemate occurs through the different rates of reaction of the enantiomers with an enzyme,

iv) enzymatic asymmetric synthesis is a synthetic process in which at least one step of the synthesis uses an enzymatic reaction to obtain an enantiomerically pure or enriched synthetic precursor of the desired enantiomer,

v) chemical asymmetric synthesis is a synthetic method in which the desired enantiomer is synthesized from an achiral precursor under conditions that induce asymmetry (i.e. chirality) in the product, which may be achieved by using chiral catalysts or chiral auxiliaries,

vi) separation of diastereomers is a process in which a racemic compound is reacted with an enantiomerically pure reagent (chiral auxiliary), which converts the individual enantiomers into diastereomers. The resulting diastereomers are then separated by chromatography or crystallization due to their now more distinct structural differences, the chiral auxiliary is later removed to yield the desired enantiomer,

vii) first and second order asymmetric transformations are a process in which diastereomers from a racemate rapidly equilibrate to form a predominant diastereomer of the desired enantiomer in solution, whereby preferential crystallization of the diastereomer from the desired enantiomer upsets the equilibrium so that ultimately, in principle, all of the material is converted to a crystalline diastereomer of the desired enantiomers. The desired enantiomer is then liberated from the diastereomer,

viii) kinetic resolution is a process that refers to achieving partial or complete resolution of a racemate (or further resolution of a partially resolved compound) due to unequal rates of reaction of the enantiomers with a chiral, non-racemic reagent or catalyst under kinetic conditions,

ix) enantiospecific synthesis from non-racemic precursors is a synthetic route in which the desired enantiomer is obtained from non-chiral starting materials and in which the stereochemical integrity is not or only minimally disturbed during the synthesis,

x) Chiral liquid chromatography is a technique in which the enantiomers of a racemate are separated in a liquid mobile phase by virtue of their different interactions with the stationary phase (including chiral vial HPLC). The stationary phase may be prepared from a chiral substance, or the mobile phase may contain an additional chiral substance in order to induce different interactions,

xi) chiral gas chromatography is a technique in which the racemate is volatilized and the enantiomers are separated by their different interactions in a gaseous mobile phase with a column containing a fixed non-racemic chiral adsorbent phase,

xii) chiral solvent extraction is a method of separating enantiomers by preferentially dissolving one enantiomer in a particular chiral solvent,

xiii) Transport across chiral membranes is a mode in which the racemate is in contact with a thin membrane barrier. The barrier typically separates two miscible liquids, one of which contains the racemate, and a driving force such as concentration or pressure difference causes preferential transport across the membrane barrier. The separation occurs as a result of the non-racemic chiral nature of the membrane, which allows only one enantiomer of the racemate to pass,

xiv) According to one embodiment, simulated moving bed chromatography is used. A wide variety of chiral stationary phases are commercially available.

Examples of the present invention

Example 1. Synthesis of 3-(6-(4-(morpholinomethyl)benzyl)-2-oxobenzo[cd]indol-1(2H)-yl)piperidin-2,6-dione (Compound 5) and enantiomer 1 3-(6-(4-(morpholinomethyl)benzyl)-2-oxobenzo[cd]indol-1(2H)-yl)piperidin-2,6-dione (Compound 1)

Step 1. Synthesis of 6-bromo-1H-benzo[cd]indol-2-one (2): A solution of molecular bromine (354.23 g, 2.22 mol, 113.53 mL) in chloroform (500 mL) was added dropwise at 0 °C to a stirred suspension of 1H-benzo[cd]indryl-2-one (1) (250 g, 1.48 mol) in chloroform (2.5 L), and the reaction mixture was stirred at room temperature for 16 h. After completion of the reaction (monitored by TLC), the reaction mass was poured into a saturated solution of sodium thiosulfate in water. The resulting yellow solid was filtered through a sintered funnel, washed with water, washed with pentane and distilled with toluene to give 6-bromo-1H-benzo[cd]indol-2-one (2) (350 g, 90% yield) as a yellow solid, which was stored in a Tarson plastic bottle at ambient temperature. LC MS: ES+ 2 (248.2 and 250.2).

Step 2. Synthesis of 6-(4,4,5,5-tetramethyl-1,3,2-dioxaborolan-2-yl)-1H-benzo[cd]indol-2-one (3): Bis(pinacolato)diborone (30.71 g, 120.93 mmol) followed by well-dried potassium acetate (23.74 g, 241.86 mmol, 15.12 mL) were added to a stirred solution of 6-bromo-1H-benzo[cd]indol-2-one (2) (20 g, 80.62 mmol) in 1,4-dioxane (500 mL). The resulting reaction mixture was well degassed with argon for 15 min. Pd ₂ (dba) ₃ (6.58 g, 8.06 mmol) was then added and the reaction mixture was heated at 100 °C for 16 h. After completion of the reaction (monitored by TLC), the reaction mixture was cooled to room temperature (RT), filtered through a pad of celite and washed with ethyl acetate (1 L). The combined filtrate was then washed with cold water (3×300 mL), dried over anhydrous sodium sulfate and concentrated under reduced pressure to give crude 6-(4,4,5,5-tetramethyl-1,3,2-dioxaborolan-2-yl)-1H-benzo[cd]indol-2-one (3) (23 g, 46.76 mmol, 58.00% yield) as a brown gum, which was stored in a round bottom flask at 5 °C inside a refrigerator. It was then sent without further purification, LC MS: ES+ 295.7.

Step 3. Synthesis of 4-(4-(chloromethyl)benzyl)morpholine (6): To a stirred solution of morpholine (5) (8 g, 91.83 mmol, 8.03 mL) in analytical grade acetone (15 mL) was added anhydrous 99% potassium carbonate (12.69 g, 91.83 mmol, 5.54 mL) at room temperature and the resulting reaction mixture was heated at 50 °C for 20 min. 1,4-Bis(chloromethyl)benzene (4) (16.07 g, 91.83 mmol, 11.32 mL) was then added to the reaction mixture and heating was continued for 3 h. After completion of the reaction (monitored by TLC and LCMS), the volatiles were removed under vacuum and the solid thus obtained was diluted in ethyl acetate (50 mL), washed with water (3 x 25 mL) and brine (2 x 15 mL), dried over anhydrous sodium sulfate and concentrated under reduced pressure. The crude material was purified by column chromatography (silica, gradient: 10-30% ethyl acetate in hexane) to give 4-[[4-(chloromethyl)phenyl]methyl]morpholine (6) (10 g, 44.30 mmol, 48.25% yield) as a colorless sticky solid, which was stored in a round-bottomed flask at 5 °C in a refrigerator, LC MS: ES+ 226.2.

Step 4. Synthesis of 6-(4-(morpholinomethyl)benzyl)benzo[cd]indol-2(1H)-one (7): To a well degassed solution of 4-[[4-(chloromethyl)phenyl]methyl]morpholine (6) (8 g, 35.44 mmol) and 6-(4,4,5,5-tetramethyl-1,3,2-dioxaborolan-2-yl)-1H-benzo[cd]indol-2-one (3) (20.92 g, 70.89 mmol) in a mixture of ethanol (20 mL) and toluene (40 mL) was added anhydrous potassium phosphate tribasic (22.57 g, 106.33 mmol), followed by the addition of tri-o-tolylphosphine (2.16 g, 7.09 mmol) and Pd ₂ (dba) ₃ (3.25 g, 3.54 mmol). The resulting mixture was then heated at 90 °C for 12 h. After completion of the reaction (LCMS monitoring), the reaction mixture was filtered through a pad of celite and washed with ethyl acetate (200 mL). The combined filtrate was then washed with water (3 x 50 mL) and brine (2 x 40 mL), dried over anhydrous sodium sulfate and concentrated under reduced pressure. The crude material was purified by column chromatography (silica, gradient: 0-20% ethyl acetate in DCM) to give 6-[[4-(morpholinomethyl)phenyl]methyl]-1H-benzo[cd]indol-2-one (7) (6.5 g, 17.59 mmol, 49.63% yield) as a yellow solid, which was stored in a Tarson plastic bottle at ambient temperature. LC MS: ES+ 359.3.

Step 5. Synthesis of 3-(6-(4-(morpholinomethyl)benzyl)-2-oxobenzo[cd]indol-1(2H)-yl)piperidine-2,6-dione: To an ice-cooled solution of 6-[[4-(morpholinomethyl)phenyl]methyl]-1H-benzo[cd]indol-2-one (7) (4.8 g, 13.39 mmol) in dry THF (50 mL), sodium hydride (60% dispersion in mineral oil) (3.08 g, 133.92 mmol) was added portionwise while maintaining the temperature at < 5 °C. Immediately after complete addition, the reaction mixture was stirred for 15 min at room temperature. The reaction mixture was then cooled again to 0 °C and 3-bromopiperidine-2,6-dione (8) (12.86 g, 66.96 mmol) was added thereto in portions. After complete addition, the resulting solution was heated at 70 °C for 1 h. Upon completion (apparent based on TLC), the reaction mixture was cooled again to 0 °C and quenched with ice-cold water (40 mL). The aqueous layer was extracted with ethyl acetate (3 x 50 mL). The combined extracts were dried over anhydrous sodium sulfate and concentrated under reduced pressure. The crude material was purified by flash chromatography (silica, gradient: 2.5% MeOH in DCM) to give 3-[6-[[4-(morpholinomethyl)phenyl]methyl]-2-oxo-benzo[cd]indol-1-yl]piperidine-2,6-dione Compound 5 (4 g, 8.36 mmol, 62.44% yield) as a yellow solid, which was stored in a round-bottomed flask at 5 °C in a refrigerator. ¹ H NMR (400 MHz, DMSO-d6) δ 11.12 (s, 1H), 8.31 (d, J=8.2 Hz, 1H), 8.06 (d, J=6.92 Hz, 1H), 7.80 (t, J=7.58 Hz, 1H), 7.39 (d, J=7.24 Hz, 1H), 7.24-7.17 (m, 4H), 7.10 (d, J=7.2 Hz, 1H), 5.44 (dd, J=12.36, 4.76 Hz, 1H), 4.36 (s, 1H), 3.51 (br s, 4H), 3.36 (s, 2H), 2.98-2.90 (m, 1H), 2.79-2.73 (m, 1H), 2.69-2.62 (m, 1H), 2.28 (br s, 4H), 2.10-2.07 (m, 1H), LC MS: ES+ 470.2.

Step 6. Chiral resolution: preparation of R- and S-3-(6-(4-(morpholinomethyl)benzyl)-2-oxobenzo[cd]indol-1(2H)-yl)piperidine-2,6-dione: 3.8 g of 3-(6-(4-(morpholinomethyl)benzyl)-2-oxobenzo[cd]indol-1(2H)-yl)piperidine-2,6-dione Compound 5 was separated into enantiomers by the chiral preparative normal phase HPLC method described below. The fractions were first evaporated separately under reduced pressure to give a solid mass. The solid was then suspended in a mixture of acetonitrile and water (2:3) and then placed in a dry ice/acetone bath until the acetonitrile-water mixture solidified. The frozen mixture was then freeze-dried using a lyophilizer for 20 h to give 3-(6-(4-(morpholinomethyl)benzyl)-2-oxobenzo[cd]indol-1(2H)-yl)piperidine-2,6-dione Compound 1 (first eluted peak, retention time = 9.33 min, 'S' ABS) (1.3 g, %ee 99.9).

Preparative chiral HPLC method:

Column: Chiralpak IC (20 x 250 mm), 5 µm Detection wavelength:258nm Detection wavelength:258nm Injection volume: 500 µl Injection volume: 500 µl Flow rate: 18 ml/minute Flow rate: 18 ml/minute Column temperature: NA Sample temperature: NA Travel time: 25 minutes Travel time: 25 minutes Diluent:Mobile phase Diluent:Mobile phase Flushing the needle: DCM Output flush: NA

Mobile phase: 500 ml DCM and 500 ml isopropyl alcohol were mixed and sonicated for degassing.

Example 2. Synthesis of Compound 2, Compound 7, Compound 9 and Compound 13

To a stirred solution of amine (1.0 equiv.) in THF (mL/mmol) was added triethylamine (2.0 equiv.) at 0 °C. Then, aldehyde (1.0 equiv.), phenylsilane (1.0 equiv.), and dibutyltin dichloride (1.2 equiv.) were added to the reaction mixture. The resulting solution was stirred at 90 °C for 16 h. The reaction mixture was then diluted with ethyl acetate and washed with aqueous NaHCO ₃ , water (×3), and brine. The organic layer was then dried over anhydrous Na ₂ SO ₄ and concentrated under reduced pressure. The crude material was then purified through a CombiFlash ISCO column eluting with 3% methanol in DCM to afford the title compound.

Connection 2:

Yellow solid, 3.8 g, 71.92% yield, 99.49% purity. ^1H NMR (400 MHz, DMSO-d6) δ 11.11 (s, 1H), 8.37 (d, J=8.2 Hz, 1H), 8.08 (d, J=6.96 Hz, 1H), 7.82 (t, J=7.6 Hz, 1H), 7.57 (s, 1H), 7.35 (d, J=7.24 Hz, 1H), 7.28 (s, 1H), 7.06 (d, J=7.24 Hz, 1H), 5.43 (dd, J=12.64, 4.8 Hz, 1H), 4.17 (s, 2H), 3.99-3.96 (m, 1H), 2.94-2.90 (m, 1H), 2.75-2.62 (m, 4H), 2.22 (s, 2H), 2.09-2.02 (m, 3H), 1.99-1.69 (m, 8H), 1.61-1.56 (m, 2H), 1.13 (s, 3H), LC MS: ES+ 526.4.

Compound 7:

Yellow solid, 119.0 mg, 43.26% yield, 96.88% purity. ^1H NMR (400 MHz, DMSO-d6) δ 11.11 (s, 1H), 8.37 (d, J=8.16 Hz, 1H), 8.08 (d, J=6.88 Hz, 1H), 7.83 (t, J=7.64 Hz, 1H), 7.58 (s, 1H), 7.35 (d, J=7.24 Hz, 1H), 7.29 (s, 1H), 7.07 (d, J=7.24 Hz, 1H), 5.44-5.42 (m, 1H), 4.18 (s, 2H), 4.00-3.99 (m, 1H), 3.00-2.91 (m, 3H), 2.80-2.73 (m, 1H), 2.67-2.62 (m, 1H), 2.11-2.10 (m, 3H), 1.91-1.88 (m, 6H), 1.01 (s, 3H), 0.26-0.21 (m, 4H), LC MS: ES+ 512.3.

Compound 9:

Yellow solid, 75.0 mg, 42.55% yield, 97.14% purity. ¹ H NMR (400 MHz, DMSO-d6) δ 11.12 (s, 1H), 8.38 (d, J=8.32 Hz, 1H), 8.08 (d, J=6.76 Hz, 1H), 7.83 (t, J=7.54 Hz, 1H), 7.57 (s, 1H), 7.36 (d, J=7.64 Hz, 1H), 7.29 (s, 1H), 7.07 (d, J=7.24 Hz, 1H), 5.44-5.42 (m, 1H), 4.17 (s, 2H), 3.99-3.97 (m, 1H), 2.99-2.91 (m, 1H), 2.81-2.77 (m, 2H), 2.67-2.62 (m, 1H), 2.33-2.27 (m, 2H), 2.05-2.04 (m, 3H), 1.89-1.83 (m, 4H), 1.60-1.58 (m, 1H), 0.83 (s, 9H), LC MS: ES+ 514.7.

Compound 13:

Yellow solid, 100.0 mg, 59.76% yield, 96.30% purity. ¹ H NMR (400 MHz, DMSO-d6) δ 11.11 (s, 1H), 8.37 (d, J=8.24 Hz, 1H), 8.08 (d, J=6.96 Hz, 1H), 7.83 (t, J=7.6 Hz, 1H), 7.59 (s, 1H), 7.35 (d, J=7.32 Hz, 1H), 7.30 (s, 1H), 7.07 (d, J=7.28 Hz, 1H), 5.43 (dd, J=12.6, 5.0 Hz, 1H), 4.18 (s, 2H), 4.03-4.01 (m, 1H), 3.12-2.90 (m, 3H), 2.80-2.62 (m, 4H), 2.22-2.19 (m, 2H), 2.09-2.07 (m, 1H), 1.90-1.88 (m, 4H), 0.99-0.95 (m, 2H), 0.66-0.65 (m, 2H), LCMS: ES+ 516.3.

Example 3. Synthesis of 3-[6-[[4-[[4-(2-fluorophenyl)piperazin-1-yl]methyl]phenyl]methyl]-2-oxo-benzo[cd]indol-1-yl]piperidin-2,6-dione

(Connection 3)

Step 1. Synthesis of 1-[[4-(chloromethyl)phenyl]methyl]-4-(2-fluorophenyl)piperazine (3): To a stirred solution of 1-(2-fluorophenyl)piperazine (1) (2 g, 11.10 mmol) in dry grade DMF (5 mL), DIPEA (4.30 g, 33.29 mmol, 5.80 mL) was added followed by 1,4-bis(chloromethyl)benzene (2) (3.89 g, 22.19 mmol, 2.74 mL). The resulting reaction mixture was heated at 60 °C for 12 h. After completion of the reaction (apparent based on LC MS), ice-cold water (25 mL) was added to the reaction mixture and extracted with ethyl acetate (3 x 30 mL). The organic portion was separated, dried over anhydrous sodium sulfate and concentrated under reduced pressure. The crude reaction mixture was purified by column chromatography (silica, gradient: 0-40% ethyl acetate in hexane) to give 1-[[4-(chloromethyl)phenyl]methyl]-4-(2-fluorophenyl)piperazine (3) (3 g, 8.47 mmol, 76.31% yield, 90% purity) as a yellow solid, LC MS: ES+ 319.4.

Step 2. Synthesis of 6-[[4-[[4-(2-fluorophenyl)piperazin-1-yl]methyl]phenyl]methyl]-1H-benzo[cd]indol-2-one (5): To a stirred solution of 6-(4,4,5,5-tetramethyl-1,3-dioxolan-2-yl)-1H-benzo[cd]indol-2-one (4) (500 mg, 1.68 mmol) and 1-[[4-(chloromethyl)phenyl]methyl]-4-(2-fluorophenyl)piperazine (3) (536.10 mg, 1.68 mmol) in ethanol (1 mL) in toluene (2 mL) was added tripotassium phosphate (892.33 mg, 4.20 mmol) followed by 0.5 mL of water, and the reaction mixture was degassed under nitrogen atmosphere for 10 minutes. Then tris-o-tolylphosphane (102.36 mg, 336.31 μmol) and (1E,4E)-1,5-diphenylpenta-1,4-dien-3-one palladium (153.98 mg, 168.15 μmol) were added to this reaction mass and the resulting reaction mixture was heated at 90 °C overnight. After completion of the reaction, the reaction mixture was filtered through a pad of celite and washed with ethyl acetate (50 mL). The filtrate was collected and washed with water (2 x 20 mL)/brine (20 mL). The combined organic layers were separated, dried over sodium sulfate and concentrated in vacuo. The crude material was purified by column chromatography (silica, gradient: 0-40% ethyl acetate in hexane) to give 6-[[4-[[4-(2-fluorophenyl)piperazin-1-yl]methyl]phenyl]methyl]-1H-benzo[cd]indol-2-one (5) (280 mg, 539.49 μmol, 32.08% yield, 87% purity) as a yellow solid, LC MS: 452.4.

Step 3. Preparation of 3-[6-[[4-[[4-(2-fluorophenyl)piperazin-1-yl]methyl]phenyl]methyl]-2-oxo-benzo[cd]indol-1-yl]piperidine-2,6-dione: To an ice-cooled solution of 6-[[4-[[4-(2-fluorophenyl)piperazin-1-yl]methyl]phenyl]methyl]-1H-benzo[cd]indol-2-one 5 (80 mg, 177.17 μmol) in dry THF (8 mL), 60% sodium hydride (in oil dispersion) 60% in mineral oil (67.89 mg, 1.77 mmol, 60% purity) was added portionwise, maintaining the temperature at < 5°C. Immediately after the addition was complete, the reaction mixture was stirred for 15 min at room temperature. The reaction mixture was then cooled again to 0 °C and 3-bromopiperidine-2,6-dione 6 (170.10 mg, 885.87 μmol) was added thereto in portions. After the addition was complete, the resulting solution was heated at 70 °C for 1 h. Upon completion (apparent based on TLC), the reaction mixture was cooled again to 0 °C and quenched with ice-cold water (40 mL). The aqueous layer was extracted with ethyl acetate (3 x 50 mL). The combined extracts were dried over anhydrous sodium sulfate and concentrated under reduced pressure. The crude material was purified by flash chromatography (silica, gradient: 3% MeOH in DCM) to afford 3-[6-[[4-[[4-(2-fluorophenyl)piperazin-1-yl]methyl]phenyl]methyl]-2-oxo-benzo[cd]indol-1-yl]piperidine-2,6-dione Compound 3 (20 mg, 34.69 μmol, 19.58% yield, 97.59% purity). ¹ H NMR (400 MHz, DMSO-d6) δ 11.12 (s, 1H), 8.32 (d, J=8.16 Hz, 1H), 8.07 (d, J=6.92 Hz, 1H), 7.81 (t, J=7.92 Hz, 1H), 7.41 (d, J=7.24 Hz, 1H), 7.26-7.20 (m, 4H), 7.11-7.05 (m, 3H), 7.0-6.93 (br m, 2H), 5.43 (dd, J=12.64, 4.76 Hz, 1H), 4.38 (s, 2H), 3.45 (s, 2H), 2.96 (br s, 5H), 2.76-2.73 (m, 1H), 2.66-2.62 (m, 1H), 2.50 (br s, 4H), 2.10-2.09 (m, 1H), LC MS: ES+ 563.5.

Example 4. Synthesis of 4-[4-[[4-[[1-(2,6-dioxo-3-piperidyl)-2-oxo-benzo[cd]indol-yl]methyl]phenyl]methyl]piperazin-1-yl]-3-fluoro-benzonitrile (Compound 4)

Step 1. Synthesis of tert-butyl 4-(4-cyano-2-fluorophenyl)piperazine-1-carboxylate (3): To a stirred solution of 3,4-difluorobenzonitrile (1) (13 g, 93.46 mmol) in DMSO (80 mL), potassium carbonate (19.37 g, 140.18 mmol, 8.46 mL) and tert-butyl piperazine-1-carboxylate (2) (19.15 g, 102.80 mmol) were added and the resulting reaction mixture was heated at 100 °C for 16 h. After completion (monitored by TLC), the reaction mixture was allowed to cool and water (500 mL) was added to it. The resulting solid was then filtered, washed with water and dried under vacuum to give tert-butyl 4-(4-cyano-2-fluoro-phenyl)piperazine-1-carboxylate (3) (20 g, 66% yield) as a white solid. LC MS: ES+ 306.2.

Step 2. Synthesis of 3-fluoro-4-piperazin-1-yl-benzonitrile hydrochloride salt (4): To a stirred solution of tert-butyl 4-(4-cyano-2-fluoro-phenyl)piperazine-1-carboxylate (3) (20 g, 65.50 mmol) in dioxane (15 mL) was added dioxane-HCl (65.50 mmol, 50 mL) and the reaction mixture was stirred at room temperature for 3 h. All volatiles were removed under reduced pressure. The resulting solid was triturated with ether to give 3-fluoro-4-piperazin-1-yl-benzonitrile hydrochloride (4) (17 g, 88% yield) as a white solid. LC MS: ES+ 206.4.

Step 3. Synthesis of 4-[4-[[4-(chloromethyl)phenyl]methyl]piperazin-1-yl]-3-fluorobenzonitrile (6): To a stirred solution of 3-fluoro-4-piperazin-1-ylbenzonitrile hydrochloride (4) (15 g, 62.06 mmol) in DMF (75 mL) was added DIPEA (24.06 g, 186.19 mmol, 32.43 mL) and the reaction mixture was stirred for 5 min. Then 1,4-bis(chloromethyl)benzene (5) (10.86 g, 62.06 mmol, 7.65 mL) was added in one portion and the reaction was heated at 60 °C for 16 h. After completion (TLC monitoring), the reaction mixture was diluted with ethyl acetate and washed with water and brine, dried over sodium sulfate and concentrated. The crude material thus obtained was purified by column chromatography (silica, gradient: 10-30% EtOAc in hexane) to give 4-[4-[[4-(chloromethyl)phenyl]methyl]piperazin-1-yl]-3-fluorobenzonitrile (6) (7 g, 32% yield) as a white solid. LC MS: (Es, ES+2) 344.2, 346.4.

Step 4. Synthesis of 3-fluoro-4-[4-[[4-[(2-oxo-1H-benzo[cd]indol-6-yl)methyl]phenyl]methyl]piperazin-1-yl]benzonitrile (7): To a well degassed solution of 4-[4-[[4-(chloromethyl)phenyl]methyl]piperazin-1-yl]-3-fluorobenzonitrile (6) (7 g, 20.36 mmol) and 6-(4,4,5,5-tetramethyl-1,3-dioxolan-2-yl)-1H-benzo[cd]indol-2-one (3) (9.08 g, 30.54 mmol) in ethanol (30 mL) in toluene (60 mL), anhydrous potassium phosphate tribasic (10.80 g, 50.90 mmol) was added followed by by adding tri-o-tolylphosphine (1.24 g, 4.07 mmol) and Pd2(dba)3 (1.86 g, 2.04 mmol). The resulting mixture was then heated at 100 °C for 16 h. After completion of the reaction (monitored by LCMS), the reaction mixture was filtered through a pad of celite and washed with ethyl acetate. The combined filtrate was washed with water and brine, dried over sodium sulfate and concentrated under reduced pressure. The crude material was purified by column chromatography (silica, gradient: 0-40% ethyl acetate in hexane) to give 3-fluoro-4-[4-[[4-[(2-oxo-1H-benzo[cd]indol-6-yl)methyl]phenyl]methyl]piperazin-1-yl]benzonitrile (7) (5.5 g, 52% yield) as a yellow solid. LC MS: ES+ 477.4.

Step 5. Synthesis of 4-[4-[[4-[[1-(2,6-dioxo-3-piperidyl)-2-oxo-benzo[cd]indol-6-yl]methyl]phenyl]methyl]piperazin-1-yl]-3-fluoro-benzonitrile: To a cooled solution of fluoro-4-[4-[[4-[(2-oxo-1H-benzo[cd]indol-6-yl)methyl]phenyl]methyl]piperazin-1-yl]benzonitrile (7) (5.5 g, 11.54 mmol) in dry THF (30 mL), sodium hydride (60% dispersion in mineral oil) (2.65 g, 115.41 mmol) was added portionwise, maintaining the temperature at < 5 °C. Immediately after complete addition, the reaction mixture was stirred for 15 min at room temperature. The reaction mixture was then cooled again to 0 °C and 3-bromopiperidine-2,6-dione (8) (11.08 g, 57.71 mmol) was added portionwise. After complete addition, the resulting solution was heated at 70 °C for 1 h. Upon completion (apparent based on TLC), the reaction mixture was cooled to 0 °C and quenched by addition of ice-cold water. The aqueous layer was extracted with ethyl acetate (3×100 mL). The combined organic phases were separated, dried over sodium sulfate and concentrated under reduced pressure. The crude material was purified by column chromatography (silica, gradient: 30-100% EtOAc in DCM) to give 4-[4-[[4-[[1-(2,6-dioxo-3-piperidyl)-2-oxo-benzo[cd]indol-6-yl]methyl]phenyl]methyl]piperazin-1-yl]-3-fluoro-benzonitrile Compound 4 (4.4 g, 63% yield) as a yellow solid. ¹ H NMR (400 MHz, DMSO-d6) δ 11.12 (s, 1H), 8.32 (d, J=8.28 Hz, 1H), 8.07 (d, J=6.92 Hz, 1H), 7.80 (t, J=7.66 Hz, 1H), 7.66 (d, J=12.4 Hz, 1H), 7.54 (d, J=8.36 Hz, 1H), 7.40 (d, J=7.28 Hz, 1H), 7.26-7.19 (m, 4H), 7.11-7.05 (m, 2H), 5.44 (dd, J=12.64, 4.84 Hz, 1H), 4.37 (s, 2H), 3.49 (s, 2H), 3.12 (br s, 4H), 2.98-2.90 (m, 1H), 2.79-2.73 (m, 1H), 2.70-2.62 (m, 1H), 2.45 (br s, 4H), 2.10-2.07 (m, 1H), LC MS: ES+ 588.48.

Example 5. Conditions for amide binding

To an equimolar mixture of amine and acid in DMF (6 mL/mmol) were added HATU (1.5 equiv) and DIPEA (5.0 equiv) at 0 °C. The resulting solution was stirred at ambient temperature for 16 h. The reaction mixture was then diluted with ethyl acetate and washed with aqueous NaHCO ₃ , water (×3), and brine. The organic layer was then dried over anhydrous Na ₂ SO ₄ and concentrated under reduced pressure. The crude material was then purified on a CombiFlash ISCO column eluting with 2% methanol in DCM.

Compound 6:

Yellow solid, 110 mg, 48.55% yield, 98.94% purity. ¹ H NMR (400 MHz, DMSO-d6) δ 11.11 (s, 1H), 8.39 (d, J=Hz, 1H), 8.08 (d, J=Hz, 1H), 7.82 (t, J=Hz, 1H), 7.78 (s, 1H), 7.36-7.33 (m, 2H), 7.07 (d, J=Hz, 1H), 5.43 (dd, J=Hz, 1H), 4.20 (s, 2H), 3.69-3.67 (m, 1H), 3.29-3.28 (m, 1H), 3.07-3.05 (m, 2H), 2.99-2.90 (m, 1H), 2.80-2.69(m, 1H), 2.67-2.62 (m, 1H), 2.39-2.28 (m, 4H), 2.09-2.07 (m, 1H), 1.94-1.83 (m, 1H), 1.78-1.74 (m, 4H), 1.62-1.55 (m, 1H), 1.34 (s, 3H), 1.31 (s, 3H), LC MS: ES+ 554.2.

Compound 11:

Yellow solid, 120 mg, 43.48% yield, 99.32% purity. ¹ H NMR (400 MHz, DMSO-d6) δ 11.11 (s, 1H), 8.39 (d, J=8.2 Hz, 1H), 8.08 (d, J=6.92 Hz, 1H), 7.83 (t, J=7.58 Hz, 1H), 7.78 (s, 1H), 7.35-7.33 (m, 1H), 7.07 (d, J=7.08 Hz, 1H), 5.44-5.42 (m, 1H), 4.21 (s, 2H), 3.70-3.68 (m, 2H), 3.25-3.22 (m, 2H), 2.98-2.91 (m, 1H), 2.76-2.72 (m, 1H), 2.67-2.62 (m, 1H), 2.27-2.26 (m, 2H), 2.09-2.08 (m, 1H), 1.76-1.71 (m, 2H), 1.35 (s, 3H), 1.16 (s, 9H), LC MS: ES+ 542.2.

Example 6. Synthesis of Compound 8 and Compound 10

To a stirred solution of amine (1.0 equiv.) in THF (6 mL/mmol) was added triethylamine (2.0 equiv.) at 0 °C. Then, aldehyde (1.0 equiv.), phenylsilane (1.0 equiv.), and dibutyltin dichloride (1.2 equiv.) were added to the reaction mixture. The resulting solution was stirred at 90 °C for 16 h. The reaction mixture was then diluted with ethyl acetate and washed with aqueous NaHCO ₃ , water (×3), and brine. The organic layer was then dried over anhydrous Na ₂ SO ₄ and concentrated under reduced pressure. The crude material was then purified on a CombiFlash ISCO column eluting with 3% methanol in DCM.

Compound 8

Yellow solid, 130.0 mg, 60.22% yield, 98.59% purity. ¹ H NMR (400 MHz, DMSO-d6) δ 11.11 (s, 1H), 8.38 (d, J=8.24 Hz, 1H), 8.08 (d, J=6.88 Hz, 1H), 7.81 (t, J=7.58 Hz, 1H), 7.71 (s, 1H), 7.37-7.31 (m, 2H), 7.07 (d, J=7.24 Hz, 1H), 5.43 (dd, J=12.64, 4.8 Hz, 1H), 4.20 (s, 2H), 2.99-2.90 (m, 1H), 2.80-2.69 (m, 1H), 2.66-2.62 (m, 1H), 2.50-2.49 (m, 3H), 2.34-2.32 (m, 2H), 2.15-1.92 (m, 4H), 1.82-1.78 (m, 2H), 1.29 (s, 3H), 1.00 (s, 3H), LC MS: ES+ 526.6.

Compound 10:

Yellow solid, 130.0 mg, 57% yield, 94.32% purity. ¹ H NMR (400 MHz, DMSO-d6) δ 11.10 (s, 1H), 8.39 (d, J=8.24 Hz, 1H), 8.08 (d, J=6.96 Hz, 1H), 7.82 (t, J=7.62 Hz, 1H), 7.70 (s, 1H), 7.34 (d, J=7.24 Hz, 1H), 7.30 (s, 1H), 7.07 (d, J=7.24 Hz, 1H), 5.43 (dd, J=12.64, 4.88 Hz, 1H), 4.20 (s, 2H), 2.97-2.90 (m, 1H), 2.77-2.72 (m, 1H), 2.67-2.62 LC MS: ES+ 528.5.

Example 7. Synthesis of 3-(6-(4-((1-oxa-8-azaspiro[4.5]decan-8-yl)methyl)benzyl)-2-oxobenzo[cd]indol-1(2H)-yl)piperidine-2,6-dione (Compound 12)

Step 1. Synthesis of 6-bromo-1H-benzo[cd]indol-2-one (2): To a stirred suspension of 1H-benzo[cd]indol-2-one (1) (250 g, 1.48 mol) in chloroform (2.5 L), a solution of molecular bromine (354.23 g, 2.22 mol, 113.53 mL) in chloroform (500 mL) was added dropwise at 0 °C and the reaction mixture was stirred at room temperature for 16 h. After completion of the reaction (monitored by TLC), the reaction mass was poured into a saturated solution of sodium thiosulfate in water. The resulting yellow solid was filtered through a sintered column, washed with water, washed with pentane and then distilled with toluene to give 6-bromo-1H-benzo[cd]indol-2-one (2) (350 g, 90% yield) as a yellow solid. LC MS: ES+ 2 (248.2 and 250.2).

Step 2. Synthesis of 6-(4,4,5,5-tetramethyl-1,3,2-dioxaborolan-2-yl)-1H-benzo[cd]indol-2-one: To a stirred solution of 6-bromo-1H-benzo[cd]indol-2-one (2) (100 g, 403.10 mmol) in 1,4-dioxane (1 L) was added bis(pinacolato)diborone (153.55 g, 604.66 mmol) followed by well-dried potassium acetate (118.68 g, 1.21 mol, 75.60 mL). The resulting reaction mixture was thoroughly degassed with argon for 15 min. PdCl2(dppf).DCM (32.92 g, 40.31 mmol) was added and the reaction mixture was heated at 100 °C for 16 h. After the reaction was complete (TLC monitoring), the reaction mixture was cooled to room temperature, filtered through a pad of celite and washed with ethyl acetate. The combined filtrate was then washed with chilled water, dried over anhydrous sodium sulfate and concentrated under reduced pressure to give crude 6-(4,4,5,5-tetramethyl-1,3,2-dioxaborolan-2-yl)-1H-benzo[cd]indol-2-one (3) (110 g, 64% yield) as a brown gum. It was then used without further purification, LC MS: ES+ 295.7.

Step 3. Synthesis of 8-(4-(chloromethyl)benzyl)-1-oxa-8-azaspiro[4.5]decane (6): To a stirred solution of 1-oxa-8-azaspiro[4.5]decane hydrochloride (5) (5 g, 28.14 mmol) in dry-grade acetone (50 mL) was added DIPEA (3.64 g, 28.14 mmol, 4.90 mL) followed by 99% anhydrous potassium carbonate (11.67 g, 84.43 mmol, 5.10 mL) at room temperature and the resulting reaction mixture was heated at 50 °C for 20 min. 1,4-Bis(chloromethyl)benzene (4) (9.85 g, 56.28 mmol, 6.94 mL) was then added to the reaction mixture and heating was continued for 3 h. After completion of the reaction (monitored by TLC and LCMS), the volatiles were removed under vacuum and the solid thus obtained was diluted in ethyl acetate (20 mL), washed with water and brine (×3), dried over anhydrous sodium sulfate and concentrated under reduced pressure. The crude material was purified by column chromatography (silica, gradient: 0-5% MeOH in DCM) to give 8-[[4-(chloromethyl)phenyl]methyl]-1-oxa-8-azaspiro[4.5]decane (6) (4.68 g, 16.56 mmol, 58.84% yield, 99% purity) as a colorless sticky solid. LC MS: ES+ 280.4.

Step 4. Synthesis of 6-(4-((1-oxa-8-azaspiro[4.5]decan-8-yl)methyl)benzyl)benzo[cd]indol-2(1H)-one (8): To a well degassed solution of 8-[[4-(chloromethyl)phenyl]methyl]-1-oxa-8-azaspiro[4.5]decane (6) (4.68 g, 16.73 mmol) and 6-(4,4,5,5-tetramethyl-1,3,2-dioxaborolan-2-yl)-1H-benzo[cd]indol-2-one (7) (9.87 g, 33.45 mmol) in ethanol (20.0 mL) - toluene (40.0 mL), anhydrous potassium phosphate tribasic (10.65 g, 50.18 mmol) was added followed by tri-o-tolylphosphine (1.02 g, 3.35 mmol) and Pd2(dba)3 (1.53 g, 1.67 mmol). The resulting mixture was then heated at 90 °C for 12 h. After completion of the reaction (monitored by LCMS), the reaction mixture was filtered through a pad of celite, washed with ethyl acetate. The combined filtrate was washed with water and brine, dried over anhydrous sodium sulfate and concentrated under reduced pressure. The crude material was purified by column chromatography (silica, gradient: 0-5% MeOH in DCM) to give 6-[[4-(1-oxa-8-azaspiro[4.5]decan-8-ylmethyl)phenyl]methyl]-1H-benzo[cd]indol-2-one (8) (2.83 g, 6.17 mmol, 36.91% yield, 90% purity) as a yellow solid. LC MS: ES+ 413.0.

Step 5. Synthesis of 3-(6-(4-((1-oxa-8-azaspiro[4.5]decan-8-yl)methyl)benzyl)-2-oxobenzo[cd]indol-1(2H)-yl)piperidine-2,6-dione: To an ice-cooled solution of 6-[[4-(1-oxa-8-azaspiro[4.5]decan-8-ylmethyl)phenyl]methyl]-1H-benzo[cd]indol-2-one (8) (2.83 g, 6.86 mmol) in dry THF (20 mL), sodium hydride (60% dispersion in mineral oil) (2.63 g, 68.60 mmol, 60% purity) was added portionwise. The temperature was maintained at < 5 °C. Immediately after the addition was complete, the reaction mixture was stirred for 15 min at room temperature. Then the reaction mixture was cooled again to 0 °C and 3-bromopiperidine-2,6-dione (9) (6.59 g, 34.30 mmol) was added thereto in portions. After the addition was complete, the resulting solution was heated at 70 °C for 1 h. After completion (apparent by TLC), the reaction mixture was cooled again to 0 °C and quenched with ice-cold water. The aqueous layer was extracted with ethyl acetate (3 x 20 mL). The combined extract was dried over anhydrous sodium sulfate and concentrated under reduced pressure. The crude material was washed with diethyl ether/pentane to give the racemate, which was then chirally resolved to give (S)-3-[6-[[4-(1-oxa-8-azaspiro[4.5]decan-8-ylmethyl)phenyl]methyl]-2-oxo-benzo[cd]indol-1-yl]piperidine-2,6-dione Compound 12 (2.7 g, 5.15 mmol, 75.1% yield) as a yellow solid. LC MS: ES+ 524.3.

Step 6. Chiral resolution: 1.2 g of 3-(6-(4-((1-oxa-8-azaspiro[4.5]decan-8-yl)methyl)benzyl)-2-oxobenzo[cd]indol-1(2H)-yl)piperidine-2,6-dione was separated into enantiomers by a chiral preparative normal phase HPLC method below. The fractions were first evaporated separately under reduced pressure to give a solid mass. The solid was then suspended in a mixture of acetonitrile and water (2:3) and kept in a dry ice/acetone bath until the acetonitrile-water mixture solidified. The frozen mixture was then freeze-dried using a lyophilizer for 20 h to give 3-[6-[[4-(1-oxa-8-azaspiro[4.5]decan-8-ylmethyl)phenyl]methyl]-2-oxo-benzo[cd]indol-1-yl]piperidine-2,6-dione Compound 12 (first eluted peak, RT=6.29 m 'S' ABS) (420 mg, %ee 99.28).

Preparative chiral HPLC method:

Column: Chiralpak IC (20 x 250 mm), 5 µm Detection wavelength:224nm Detection wavelength:224nm Injection volume: 500 µl Injection volume: 500 µl Flow rate: 18 ml/minute Flow rate: 18 ml/minute Column temperature: NA Sample temperature: NA Travel time: 20 minutes Travel time: 20 minutes Diluent:Mobile phase Diluent:Mobile phase Needle flushing: DCM Output flush: NA

Mobile phase: 600 ml DCM and 400 ml isopropyl alcohol were mixed and sonicated for degassing.

General experimental information

The various cell lines described in the examples below were obtained and cultured as described in Table 1.

Example 8. Unique properties of Compound 1

Biomarker assessment in MM and ALCL tumor studies showed profound destruction (>75%) of both IKZF1/3 by Compound 1 within 24 hours. Compound 1 was effective in pomalidomide-resistant animal models of MM. Mice bearing human RPMI-8226 MM tumor xenografts failed to respond to pomalidomide treatment (3000 mcg/kg/day, a clinically relevant dose) for 17 days, and immediately after Compound 1 was administered to these animals at a dose of 100 mcg/day, it resulted in rapid tumor regression.

CB17 SCID mice bearing RPMI-8226 xenografts were treated with pomalidomide (3000 μg/kg/day) daily PO for 3 weeks. Pomalidomide treatment was then discontinued and mice were treated with Compound 1 (100 μg/kg/day) on a daily schedule for 2 weeks. Data are presented as mean tumor volumes ± SEM.

In one study in MM tumors, Compound 1 alone achieved regression, but the combination of dexamethasone (5 mg/kg, QW) with Compound 1 (10 mcg/kg/day) provided longer-lasting regression compared to Compound 1 or dexamethasone alone.

IKZF1/3 are not degraded in mouse cells treated with thalidomide or its IMiD ^® analogues due to a single amino acid difference in mouse cereblon compared to human cereblon that causes steric hindrance and prevents IKZF1/3 recruitment. Ex vivo studies were performed in mouse, rat, dog and monkey peripheral blood cells (PBMCs) to identify suitable IND species that would allow toxicity studies based on the extent of IKZF1 degradation by Compound 1. IKZF1 was not degraded in mouse or rat cells by Compound 1, so although the rat was not considered a pharmacologically relevant species for human risk assessment, it was chosen as the rodent species for toxicology studies because it is widely used in toxicity testing and has an extensive historical toxicology database. Compound 1 was highly effective in disrupting IKZF1/3 in cynomolgus macaque cells but had no effect on IKZF1/3 in canine cells. Therefore, cynomolgus macaques were chosen for toxicology studies in non-rodent animals.

Secondary pharmacodynamic (PD) studies were conducted to evaluate the off-target activity of Compound 1. Global proteomic studies of ALCL tumor xenografts isolated from mice 4 hours after a single dose of Compound 1 (100 μg/kg) showed that Compound 1 is highly selective, degrading only IKZF1/3 out of >7900 proteins detected in the tissue.

Specific studies were conducted to evaluate the ability of Compound 1 to cleave Sal-like protein 4 (SALL4), a protein implicated in teratogenicity induced by the ^IMiD® class of degraders IKZF1/IKZF3. In the KELLY neuroblastoma cell line, Compound 1 promoted the degradation of SALL4 by greater than 85% at 10 nM, suggesting that Compound 1 has teratogenic potential. A small amount of the E3 ligase modulators cereblon has been shown to promote the degradation of the translation termination factor GSPT1. In a targeted cell assay, Compound 1 had no significant effect on G1 to S phase transition 1 (GSPT1) up to 10 μM.

In vitro testing of specific binding or activity of Compound 1 (100 nM) against 87 potential unintended molecular targets revealed no targets inhibited by >50% at 500-fold the free plasma exposure (C _max ) associated with maximal in vivo potency. Viability assays performed in HepG2 liver cancer cells lacking IKZF1 and IKZF3 showed no significant effect of Compound 1 on viability at concentrations up to 3.3 μM.

Example 9. Efficiency and catalytic rate of Compound 1

The potency of Compound 1 was determined using the NCI-H929 viability assay and compared to pomalidomide. NCIH929 cell viability was determined based on ATP quantification using the CellTiter-Glo® 2.0 Luminescent Assay Kit, which signals the presence of metabolically active cells. Briefly, test compound was added to 384-well plates at a maximum concentration of 1 μM in 10-point, semi-logarithmic titrations with replicates. NCIH929 cells were seeded in 384-well plates in RPMI medium containing 10% FBS and 0.05 mM 2-mercaptoethanol at a cell density of 750 cells per well. Cells treated in the absence of test compound served as negative controls, and cells treated in the absence of CellTiter-Glo® 2.0 served as positive controls. Cells treated with test compounds were incubated at 37°C with 5% _CO2 for 96 hours. CellTiter-Glo reagent was then added to the cells and luminescence was recorded using an EnVision™ Multichannel Reader (PerkinElmer, Santa Clara, CA, USA). % Viability was determined by normalizing the signal to positive and DMSO-treated negative controls on the same microtiter plate. The results are shown in Fig. 1A and Table 2. As shown in Table 2, Compound 1 exhibited an _IC50 of 0.071 μM. The data in Fig. 1A are expressed as the mean ± SD of 3 independent experiments, each with n=2. Curve fitting and IC ₅₀ determination were performed using 4-parameter regression analysis. Data in Table 1 represent the geometric mean and 95% confidence interval of the IC ₅₀ values and the linear mean ± SD of the E _max values determined for the indicated number of independent experiments.

The effect of Compound 1 on IKZF1 degradation in NCI-H929 was also determined. Endogenous IKZF1 expression in NCIH929 cells was determined by adding an N-terminal HiBiT tag to the NCIH929 cell line using CRISPR-Cas9. Briefly, test compound was added to 384-well plates at a maximum concentration of 1 μM in a 10-point, semi-logarithmic titration with replicates. NCIH929 cells expressing HiBiT-tagged IKZF1 were seeded in 384-well plates in RPMI medium containing 10% FBS and 0.05 mM 2-mercaptoethanol at a cell density of 15,000 cells per well. Cells treated in the absence of test compound served as negative controls, and wells containing medium alone served as positive controls. Following compound treatment, cells were incubated at 37°C with 5% _CO2 for 1.5 h. HiBiT signal was determined using the Nano-Glo™ HiBiT Lytic Assay System (Promega, N3050) and luminescence was recorded using the EnVision™ Multichannel Reader (PerkinElmer, Santa Clara, CA, USA). The % of IKZF1 remaining was determined by normalizing the signal with positive and negative controls on the same microtiter plate. The results are shown in Fig. 1B and Table 2. As shown in Table 2, Compound 1 exhibited an _IC50 of 0.27 μM. The data in Fig. 1B are presented as the mean ± SD of 2 independent experiments, each with n=2. Curve fitting and IC ₅₀ determination were performed using 4-parameter regression analysis. Data in Table 1 represent the geometric mean and 95% confidence interval of the IC ₅₀ values, as well as the linear mean ± SD determined from the indicated number of independent experiments.

The data in Fig. 29 were generated by measuring the degradation of IKZF1 following addition of an N-terminal HiBiT tag to endogenously expressed IKZF1 in the NCIH929 cell line using CRISPR-Cas9. The test compound was added to 384-well plates at a maximum concentration of 1 μM for compound 1 and 10 μM for pomalidomide in 22-point, 2-fold duplicate titrations. NCIH929 cells expressing HiBiT-tagged IKZF1 were seeded in 384-well plates in RPMI medium containing 10% FBS and 0.05 mM 2-mercaptoethanol at a cell density of 15,000 cells per well. Cells treated in the absence of test compound served as negative controls, and wells containing medium alone served as positive controls. Following compound treatment, cells were incubated at 37°C with 5% _CO2 for 1 or 2 h. HiBiT signal was determined using the Nano-Glo™ HiBiT Lytic Assay System (Promega, N3050) and luminescence was recorded using the En Vision™ Multichannel Reader (PerkinElmer, Santa Clara, CA, USA). % IKZF1 remaining was determined by normalizing the signal with positive and negative controls on the same microtiter plate. Curve fitting and _IC50 determination were performed using 4-parameter regression analysis. Compound 1 induced rapid degradation of IKZF1, resulting in 50% degradation after 1 hour of treatment and >80% maximal degradation after 2 hours of treatment with a DC50 of 21 pM. Pomalidomide degraded less than 25% of IKZF1 at 1 hour, and Compound 1 was >2000-fold more potent than pomalidomide at 2 hours.

Example 10. Selectivity of Compound 1 in ALCL Tumor Xenografts Using Global Proteomics

NOD SCI mice bearing ALCL ALK+ DL40 xenografts were treated orally with 100 μg/kg Compound 1. After 1, 4, 24, and 48 h of treatment, biological replicates of tumor cells were collected, washed twice with PBS, and snap-frozen in liquid nitrogen. Samples were resuspended in lysis buffer [8 M urea, 50 mM EPPS (pH 8.5), 50 mM NaCl, 1× protease inhibitor cocktail] and lysed by sonication. Lysates were centrifuged at maximum speed, reduced for 1 h at room temperature with 5 mM TCEP, and then cysteine residues were alkylated with 15 mM iodoacetamide (room temperature, in the dark, 30 min). The protein content was extracted twice by methanol-chloroform precipitation followed by ice-cold acetone washes. The protein pellets were resuspended in 8 M urea, 50 mM EPPS buffer (pH 8.5) and protein concentrations were measured by the BCA assay. Samples were then diluted to 4 M urea with 50 mM EPPS (pH 8.5) and digested at 37°C for 1 h with endoproteinase Lys-C at an enzyme/protein ratio of 1/100. The mixtures were then diluted to 1 M urea with 50 mM EPPS (pH 8.5) and trypsin was added at an enzyme/protein ratio of 1/50. The reaction was incubated overnight at 37°C and stopped by acidification with formic acid 0.5% (v/v). Peptides were loaded into tC18 SepPak solid phase extraction cartridges and lyophilized.

For peptide labeling, 100 μg peptides per sample were prepared at a concentration of 1 μg/μl in 200 mM EPPS (pH 8.5), ACN was added to a final concentration of 30%, followed by 50 μg of each TMT10 reagent. After 1 h of incubation at room temperature, the reaction mixture was quenched with 0.3% hydroxylamine for 15 min and mixed uniformly. The mixed sample was desalted using tC18 SepPak solid phase extraction cartridges and lyophilized. The dried peptides were resuspended in 5% ACN, 10 mM _NH4HCO3 pH ₈ and fractionated by basic pH reverse phase chromatography using HPLC equipped with an XBridge Peptide BEH C18 3.5 μm column. Ninety-six fractions were collected and pooled into 24, of which only 12 non-contiguous samples were analyzed. Samples were desalted, dried by vacuum centrifugation, and reconstituted in 12 µL 5% formic acid for LC-MS/MS analysis.

One-third of each sample (4 μL) was separated by reversed-phase chromatography using an EASY-Spray C18 column (2 μm particle size, 250 mm length × 75 μm i.d.) mounted in an EASY-nLC 1200 LC pump coupled to an Orbitrap Fusion Lumos Tribid mass spectrometer. Peptides were separated using a 420 min gradient from 2 to 30% acetonitrile at a flow rate of ~300 nL/min. Each analysis was performed using multichannel scanning MS3 (SPS-MS3). For identification and relative quantification, all RAW files obtained from the mass spectrometer were transformed using SEQUEST software. Briefly, mass spectra were compared against the human Uniprot database (February 2016) combined with a database consisting of all protein sequences in reverse order plus known contaminants. In all SEQUEST searches, precursor ion tolerance was set to 25 ppm and product ion tolerance to 0.9 Da, including methionine oxidation (+15.9949 Da) and cysteine carbamidomethylation (+57.0215 Da) as variable modifications. TMT tag (+229.1629 Da) on lysine residues and the N-termini of the peptide were set as static modifications. Peptide spectrum matches (PSM) were performed using linear discriminant analysis and corrected to 2% FDR2. Peptide intensities were quantified by signal-to-noise ratio extraction, and proteins were further degraded to a final FDR of 2% protein level. Protein quantification values were exported to Excel, GraphPad Prism 7, or Perseus 14 for further analysis. The resulting data are shown in Fig. 2A, and the 24-hour experiment is shown in Fig. 2B.

The identified proteins are shown in Table 3 and Table 4 below.

Example 11. Compound 1-induced destruction of Ikaros in Ki-JK (ALK ⁺ ) cells

Flow cytometry was used to assess the degradation characteristics of Ikaros and to evaluate the mechanism of Compound 1. Ki-JK cells (DSMZ, ACC 695) were seeded in 96-well plates pre-coated with Compound 1 in a dose-response relationship (11 points, duplicate, 0.001-100 nM) with or without bortezomib (proteasome inhibitor, 10 μM) or MLN-4924 (neddylation inhibitor, 10 μM). Cells were prepared for staining using FOXP3 Fix/Perm buffer kit (BioLegend, 421403) according to the manufacturer's instructions at 1.5, 3, and 6 h. IKZF1-AF488 (564867) and IgGl-488 (557782) (BD Biosciences) were diluted 1:100 in Perm Buffer. Signal was detected using a Guava® easyCyte™ flow cytometer. As shown in Fig. 3, a strong dose-time response degradation was observed for Compound 1, with maximal degradation observed at 6 hours. No degradation was observed with the combination of Compound 1 and bortezomib or MLN-4924.

Example 12. Antiproliferative activity of Compound 1 in hematological cancer

The antiproliferative activity of Compound 1 in hematological cancer cell lines was assessed and compared with pomalidomide. Cells were treated for 96 hours and the results are shown in Table 5. Compound 1 is more active than pomalidomide in every cell line and is more than 5000-fold more active in several cell lines.

Example 13. Efficacy of Compound 1 in Multiple Myeloma

The efficacy of Compound 1 in two multiple myeloma cell lines (NCI-H929) and (RPMI-8226) was assessed at four similar doses: 3 μg/kg, 10 μg/kg, 30 μg/kg, and 100 μg/kg. Compound 1 was administered QD (every day) orally (PO). The efficacy of Compound 1 was compared with pomalidomide (administered at 3000 μg/kg). The results for the NCI-H929 and RPMI-8826 cell lines are shown in Figs. 4 and 5, respectively. Table 6 describes the statistical significance of the experiment in each cell line for each dose. Compound 1 at all four concentrations reduced tumor volume to a greater extent than pomalidomide administered at a dose of 3000 μg/kg.

For NCI-H929 cells, a xenograft study was performed in female NOD SCID mice bearing NCI-H929 multiple myeloma tumors. Female SCID mice were inoculated subcutaneously in the right flank with 5× ¹⁰⁶ tumor cells in 0.2 ml PBS supplemented with Matrigel (PBS:Matrigel=1:1). Tumor volume was measured twice a week in two dimensions using a caliper, and the volume was calculated using the formula: (w2×1)/2= ^mm3 , assuming that 1 mg is equivalent to 1 ^mm3 of tumor volume. Once tumor volume reached the range of 84-267 ^mm3 (18 days after implantation), animals were randomly divided into groups of 6, stratified to obtain approximately equal mean tumor sizes in each treatment group. Treatment was initiated on day 0 with established subcutaneous NCI-H929 tumors with a mean tumor volume (MTV) of 149-150 mm ³ .

All treatments were administered to NCI-H929 tumor-bearing mice on day 0 and dosed PO daily for 21 days except for the vehicle group, which was dosed for 18 days. Compound 1 was administered at 3, 10, 30, or 100 μg/kg and formulated in PEG400 (30% v/v) + 70% v/v HPMC (1% w/v) in citrate buffer (pH 5). Pomalidomide was administered at 3000 μg/kg and the same formulation was used as for Compound 1. Body weight and MTV were measured on a 2× weekly schedule, and the study endpoint was an MTV of 2460 ^mm3 in the vehicle control group on day 18. Statistical analysis was performed using two-way analysis of variance (ANOVA). Data are expressed as MTV ± SEM.

For the RPMI-8226 MM model, female CB17 SCID mice were subcutaneously inoculated in the right flank with 10× ¹⁰⁶ RPMI-8226 cells in 0.2 ml PBS supplemented with Matrigel (PBS:Matrigel=1:1). Tumor volume was measured twice a week in two dimensions using a caliper, and the volume was calculated using the formula: (w2×1)/2= ^mm3 , assuming that 1 mg is equivalent to 1 ^mm3 of tumor volume. Once the tumor volume reached 109-158 ^mm3 (28 days after implantation), the animals were randomly divided into groups of 6, stratified to obtain approximately equal mean tumor sizes in each treatment group. Treatment was initiated on day 0 with established RPMI-8226 subcutaneous tumors with a mean tumor volume (MTV) of 130 mm ³ .

All agents were administered to RPMI-8226 tumor-bearing mice on day 0 and dosed PO daily for 31 days. Compound 1 was administered at 3, 10, 30, or 100 μg/kg and formulated in PEG400 (30% v/v) + 70% v/v HPMC (1% w/v) in nitrate buffer (pH 5). Body weight and MTV were measured 2× weekly, and the study endpoint was an MTV of 2211 ^mm3 in the vehicle-treated control group on day 31. Statistical analysis was performed using two-way analysis of variance (ANOVA). Data are expressed as MTV ± SEM.

Example 14. Compound 1 demonstrates rapid tumor elimination in a multiple myeloma model

Compound 1 was administered at 300 μg/kg QD orally to female NOD SCID mice bearing disseminated MM1.S multiple myeloma tumors with measurable MM1.S tumor burden on day 0 and administered PO daily for 3 weeks. Compound 1 was formulated in PEG400 (30% v/v) + 70% v/v HPMC (1% w/v) in citrate buffer (pH 5). Body weight and tumor burden were measured 2× weekly and the study endpoint was 20 days post-dose, day 41. Data are expressed as mean BLI±SEM.

A xenograft study was performed. Female SCID animals were irradiated with a 200 rad Co60 source and 24 h later the animals were inoculated by tail vein injection with 1 x 10 ⁷ MM1.S-Luc in 200 μl PBS. Tumor burden was measured twice weekly using image analysis. Mice were injected intraperitoneally with 15 mg/ml D-luciferin and anesthetized with 1-2% isoflurane by inhalation. Ten minutes after luciferin injection, mice were imaged with an IVIS Lumina II (Perkin Elmer) and the total bioluminescence signal (BLI, photons/s) in the region of interest (ROI) was measured. BLI from the ROI was quantified and used as an indicator of tumor burden. At 29 days post tumor cell inoculation, mice were randomized into two groups, with 3 mice per group and a mean BLI of 17× ¹⁰⁶ photons. Figure 6 shows images of Compound 1-treated mice compared to vehicle-treated mice. Images were collected on days 4, 7, 11, 14, and 18. As shown in Figure 8, the total bioluminescent signal of Compound 1-treated mice was significantly lower than that of vehicle-treated mice. These data are also shown in Figure 34.

Example 15. Compound 1 in pomalidomide-resistant or -refractory myeloma models

Compound 1 was tested in a pomalidomide-resistant multiple myeloma cell line (H929 PomR) and a non-responsive multiple myeloma cell line (RPMI-8226). The results are shown in Fig. 8 (H929 PomR cells) and Fig. 9 (RPMI-8226 cells).

Pomalidomide-resistant H929 multiple myeloma cells (H929 PomR) were generated by continued in vitro treatment with pomalidomide until cell proliferation was no longer inhibited. H929 PomR cells were implanted subcutaneously into the right flank of NOD SCID mice at a concentration of 10 × 10 ⁶ tumor cells in 0.2 ml PBS supplemented with Matrigel (PBS:Matrigel = 1:1). Mice were then treated daily with pomalidomide (5000 μg/kg/day) for 14 days until they were divided into groups of 8, stratified to obtain approximately equal mean tumor sizes in each treatment group.

After a 1-day drug washout, mice were treated with Compound 1 (100 μg/kg/day) or pomalidomide (3000 μg/kg/day) on a daily PO schedule. Both compounds were formulated as PEG400 (30% v/v) + 70% v/v HPMC (1% w/v) in citrate buffer (pH 5). Body weight and mean tumor volume (MTV) were measured on a 2× weekly schedule, and the study endpoint was an MTV of 1048 ^mm3 in the pomalidomide-treated control group on day 24. Data are expressed as MTV ± SEM. As shown in Fig. 8, tumors in Compound 1-treated mice were smaller over the course of the study than in pomalidomide-treated mice.

For the RPMI-8226 MM model, female CB17 SCID mice were subcutaneously inoculated in the right flank with 10× ¹⁰⁶ RPMI-8226 cells in 0.2 ml PBS supplemented with Matrigel (PBS:Matrigel=1:1). Tumor volume was measured twice a week in two dimensions using a caliper, and the volume was calculated using the formula: (w2×1)/2= ^mm3 , assuming that 1 mg is equivalent to 1 ^mm3 of tumor volume. Once the tumor volume reached the range of 109-158 ^mm3 (28 days after implantation), the animals were randomly divided into groups of 6, stratified to obtain approximately equal mean tumor sizes in each treatment group. Treatment was initiated on day 0 with established RPMI-8226 subcutaneous tumors with a mean tumor volume (MTV) of 130 mm ³ .

All agents were administered to RPMI-8226 tumor-bearing mice on day 0 and dosed PO. Pomalidomide was administered at 3000 μg/kg for 17 days and formulated in PEG400 (30% v/v) + 70% v/v HPMC (1% w/v) in citrate buffer (pH 5). Vehicle-treated animals were treated for 28 days until tumor MTV reached 2211 mm ³ . Pomalidomide-treated animals were switched to Compound 1 (100 μg/kg/day) beginning on day 18 and dosed for 21 additional days. Data are expressed as MTV ± SEM. As shown in Fig. 9, tumor volume began to decrease when mice were switched from pomalidomide to Compound 1.

For the RPMI-8226 MM model, female CB17 SCID mice were subcutaneously inoculated in the right flank with 10× ¹⁰⁶ RPMI-8226 cells in 0.2 ml PBS supplemented with Matrigel (PBS:Matrigel=1:1). Tumor volume was measured twice a week in two dimensions using a caliper, and the volume was calculated using the formula: (w2×1)/2= ^mm3 , assuming that 1 mg is equivalent to 1 ^mm3 of tumor volume. Once the tumor volume reached 109-158 ^mm3 (28 days after implantation), the animals were randomly divided into groups of 6, stratified to obtain approximately equal mean tumor sizes in each treatment group. Treatment was initiated on day 0 with established RPMI-8226 subcutaneous tumors with a mean tumor volume (MTV) of 130 mm ³ .

All agents were administered to RPMI-8226 tumor-bearing mice on day 0 and dosed PO daily for 31 days. Compound 1 was administered at 3, 10, 30, or 100 μg/kg and formulated in PEG400 (30% v/v) + 70% v/v HPMC (1% w/v) in citrate buffer (pH 5). Body weight and MTV were measured 2× weekly, and the study endpoint was an MTV of 2211 ^mm3 in the vehicle control group on day 31. Statistical analysis was performed using two-way analysis of variance (ANOVA). Data are expressed as MTV ± SEM. The data from this experiment are presented in Fig. 35.

In another experiment, each mouse was inoculated subcutaneously in the right flank with RPMI-8226 tumor cells (10× ¹⁰⁶ ) in 0.2 ml PBS supplemented with Matrigel (PBS:Matrigel=1:1) for tumor development. Tumor growth was monitored throughout the study. Tumor volume was measured in two dimensions using a caliper, and the volume was calculated using the formula: (w2×1)/2= ^mm3 , assuming that 1 mg is equivalent to 1 mm3 of tumor volume. Measurements were taken twice a week. The health of the mice was monitored and noteworthy clinical observations were recorded. Acceptable toxicity was defined as a mean group body weight loss of less than 20% during the study and no more than one treatment-related death among ten treated animals, or 10%.

Once tumor volumes reached the range of 108-158 ^mm3 (28 days after implantation), animals were randomized into groups of 6. Treatment was initiated on day 0 with established subcutaneous RPMI-8226 tumors with an MTV of 130 ^mm3 . Animals were weighed on a twice-weekly schedule. Mice were administered compound (100 μg/kg/day) or pomalidomide (3000 μg/kg/day) on a PO schedule daily. Both compounds were formulated in PEG400 (30% v/v) + 70% v/v HPMC (1% w/v) in citrate buffer (pH 5). Seventeen days after pomalidomide administration, a one-day drug washout was performed and mice in this group were treated with Compound 1 (100 μg/kg/day) beginning on day 18. Body weight and mean tumor volume (MTV) were measured 2× weekly, and the study endpoint was reached when the vehicle control group reached an MTV of 2211 mm ³ . The data from this experiment are presented in Fig. 40.

Example 16. Compound 1 in lymphoma cell lines

Compound 1 was tested in both a mantle cell lymphoma xenograft model (REC1) and a DLBLC tumor xenograft model (TMD8). The results are shown in Fig. 10 (REC1) and Fig. 11 (TMD8). Table 7 describes the statistical significance of the experiment in each model for each dose.

A xenograft study was performed in female balb/c nude mice bearing REC1 mantle cell lymphoma tumors. Female nude mice were inoculated subcutaneously in the right flank with 5× ¹⁰⁶ tumor cells in 0.2 ml PBS supplemented with Matrigel (PBS:Matrigel=1:1). Tumor volume was measured twice a week in two dimensions using a caliper, and the volume was calculated using the formula: (w2×1)/2= ^mm3 , assuming that 1 mg is equivalent to 1 ^mm3 of tumor volume. Once tumor volume reached the range of 102-121 ^mm3 (10 days after implantation), animals were randomly divided into groups of 6, stratified to obtain approximately equal mean tumor sizes in each treatment group. Treatment was initiated on day 0 with established subcutaneous REC1 tumors with a mean tumor volume (MTV) of 112 mm ³ .

All agents were administered to REC1 tumor-bearing mice on day 0 and dosed po daily for 18 days. Compound 1 was administered at 3, 10, 30, or 100 μg/kg and formulated in PEG400 (30% v/v) + 70% v/v HPMC (1% w/v) in nitrate buffer (pH 5). Pomalidomide was administered at 3000 μg/kg and the same formulation was used as for Compound 1. Body weight and MTV were measured 2× weekly and the endpoint of the study was an MTV of 1608 ^mm3 in the vehicle control group on day 18. Statistical analysis was performed using two-way analysis of variance (ANOVA). Data are expressed as MTV ± SEM. As shown in Fig. 10, tumor volume was decreased in mice administered 10, 30, or 100 μg/kg Compound 1, but increased in mice administered pomalidomide.

A xenograft study was also performed in female NOD SCID mice bearing DLBCL TMD8 tumors. Female SCID mice were inoculated subcutaneously in the right flank with 1× ¹⁰⁷ tumor cells in 0.1 ml PBS supplemented with Matrigel (PBS:Matrigel=1:1). Tumor volume was measured twice a week in two dimensions using calipers, and the volume was calculated using tumor: (w2×1)/2= ^mm3 , assuming that 1 mg is equivalent to 1 ^mm3 of tumor volume. Once tumor volume reached 94-238 ^mm3 (12 days after implantation), animals were randomly divided into groups of 8, stratified to obtain approximately equal mean tumor sizes in each treatment group. Treatment was initiated on day 0 with established subcutaneous TMD8 tumors with a mean tumor volume (MTV) of 187 mm ³ .

All agents were administered to TMD8 tumor-bearing mice on day 0 and daily po for 18 days. Compound 1 was administered at 100 μg/kg and pomalidomide was administered at 3000 μg/kg. Both compounds were formulated in PEG400 (30% v/v) + 70% v/v HPMC (1% w/v) in citrate buffer (pH 5). Body weight and MTV were measured 2× weekly, and the study endpoint was an MTV of 2120 mm ³ in the vehicle-treated control group on day 18. Statistical analysis was performed using two-way analysis of variance (ANOVA). Data are expressed as MTV ± SEM. As shown in Fig. 11, Tumor volume was decreased in Compound 1-treated mice but increased in pomalidomide-treated mice.

Example 17. Compound 1 induces profound and long-lasting loss of IZKF1 and IRF4 in ALCL KI-JK tumors

A xenograft study was performed in female NOD SCID mice bearing KI-JK ALCL tumors. Female SCID mice were inoculated subcutaneously in the right flank with 10× ¹⁰⁶ tumor cells in 0.2 ml PBS supplemented with Matrigel (PBS:Matrigel=1:1). Tumor volume was measured twice a week in two dimensions using a caliper, and the volume was calculated using the formula: (w2×1)/2= ^mm3 , assuming that 1 mg is equivalent to 1 ^mm3 of tumor volume. Once tumor volume reached the range of 117-272 ^mm3 (27 days after implantation), animals were randomly divided into groups of 3, stratified to obtain approximately equal mean tumor sizes in each treatment group. Treatment was initiated on day 0 with established subcutaneous KI-JK tumors with a mean tumor volume (MTV) of 195–196 mm ³ .

All agents were administered to KI-JK tumor-bearing mice on day 0 and dosed PO daily for 21 days. Compound 1 was administered at 30 or 100 μg/kg and was formulated in PEG400 (30% v/v) + 70% v/v HPMC (1% w/v) in citrate buffer (pH 5). Pomalidomide was administered at 3000 μg/kg and the same formulation was used as for Compound 1. Body weight and MTV were measured 2× weekly and the study endpoint was an MTV of 1339 ^mm3 in the vehicle-treated control group on day 21. Data are expressed as MTV ± SEM. In Fig. 12 is a graph of tumor volume in mice administered Compound 1 or pomalidomide over a 21-day period. As shown in Fig. 12, tumor volume was reduced in mice administered Compound 1.

A pharmacodynamic study was also performed in female NOD SCID mice bearing KI-JK ALCL tumors. Female SCID mice were inoculated subcutaneously in the right flank with 10× ¹⁰⁶ tumor cells in 0.2 ml PBS supplemented with Matrigel (PBS:Matrigel=1:1). Tumor volume was measured twice a week in two dimensions using a caliper, and the volume was calculated using the formula: (w2×1)/2= ^mm3 , assuming that 1 mg is equivalent to 1 ^mm3 of tumor volume. Once the tumors reached a volume of 450 ^mm3 , they were divided into 3 treatment groups consisting of 4 mice each.

A single dose of vehicle, pomalidomide (3000 μg/kg), or Compound 1 (30 or 100 μg/kg) was administered to mice bearing KI-JK tumors. All compounds were formulated in PEG400 (30% v/v) + 70% v/v HPMC (1% w/v) in citrate buffer (pH 5). Mice were sacrificed and tumors were harvested at 6 and 24 h after a single dose. Tumors were mechanically homogenized and protein was extracted using RIPA buffer (Sigma Aldrich). Protein concentration was quantified using the Pierce™ BCA Protein Assay Kit, samples were reduced, and then equal amounts of protein were loaded onto a Western blot gel for analysis. Samples were analyzed for the expression of IKZF1 (Invitrogen, PA5-23728) and IRF-4 (CST, 15106). IKZF1 expression is shown in Fig. 13A and IRF-4 expression is shown in Fig. 13B. Data are expressed as percentages of IKZF1 or IRF-4 compared to vehicle control and normalized to GAPDH levels to control for protein loading. Error bars represent values ± SEM. As shown in Figs. 13A and 13B, the percentage of IKZF1 and IRF-4 was lower in Compound 1-treated mice compared to pomalidomide-treated mice at both 6 and 24 hours.

Example 18. Compound 1 induces profound and long-lasting loss of IZKF1 and IRF4 in ALCL DL-40 tumors

A xenograft study was performed in female NOD SCID mice bearing DL-40 ALCL tumors. Female SCID mice were inoculated subcutaneously in the right flank with 10× ¹⁰⁶ tumor cells in 0.2 ml PBS supplemented with Matrigel (PBS:Matrigel=1:1). Tumor volume was measured twice a week in two dimensions using a caliper, and the volume was calculated using the formula: (w2×1)/2= ^mm3 , assuming that 1 mg is equivalent to 1 ^mm3 of tumor volume. Once tumor volume reached the range of 187-335 ^mm3 (31 days after implantation), animals were randomly divided into groups of 3, stratified to obtain approximately equal mean tumor sizes in each treatment group. Treatment was initiated on day 0 with established DL-40 subcutaneous tumors with a mean tumor volume (MTV) of 241–247 mm ³ .

All agents were administered to DL-40 tumor-bearing mice on day 0 and administered PO daily for 21 days. Compound 1 was administered at 3, 10, 30, 100, or 300 μg/kg and was formulated in PEG400 (30% v/v) + 70% v/v HPMC (1% w/v) in citrate buffer (pH 5). Pomalidomide was administered at 3000 μg/kg and used in the same formulation as Compound 1. Body weight and MTV were measured 2× weekly, and the endpoint of the study was an MTV of 1110 ^mm3 in the vehicle-treated control group on day 21. Data are expressed as MTV ± SEM. In Fig. 14 shows a graph of tumor volume in mice over a 21-day period, and as shown in Fig. 14, Compound 1 was more effective in reducing tumor volume than pomalidomide.

A pharmacodynamic study was performed in female NOD SCID mice bearing DL-40 ALCL tumors. Female SCID mice were inoculated subcutaneously in the right flank with 10× ¹⁰⁶ tumor cells in 0.2 ml PBS supplemented with Matrigel (PBS:Matrigel=1:1). Tumor volume was measured twice a week in two dimensions using a caliper, and the volume was calculated using the formula: (w2×1)/2= ^mm3 , assuming that 1 mg is equivalent to 1 ^mm3 of tumor volume. Once the tumors reached a volume of 324 ^mm3 , they were divided into 3 treatment groups.

A single dose of vehicle, pomalidomide (3000 μg/kg), or Compound 1 (30 or 100 μg/kg) was administered to mice bearing DL-40 tumors. All compounds were formulated in PEG400 (30% v/v) + 70% v/v HPMC (1% w/v) in citrate buffer (pH 5). In the vehicle control group, three mice were sacrificed and tumors were collected 24 h post-dose. Pomalidomide-treated mice were sacrificed at 1, 4, and 24 h post-dose, with two mice at each time point. Mice in the Compound 1-treated groups were sampled at 1, 4, 24, and 48 h post-dose, with four tumors collected at each time point. Tumors were then mechanically homogenized and protein was extracted using RIPA buffer (Sigma Aldrich). Protein concentration was quantified using the Pierce™ BCA Protein Assay Kit, samples were concentrated, and equal amounts of protein were then loaded onto a Western blot gel for analysis as shown in Fig. 15B. Samples were analyzed for IKZF1 expression (Invitrogen, PA5-23728) as shown in Fig. 15A. Data are expressed as percentages of IKZF1 compared to vehicle control and normalized to GAPDH levels for protein loading control. Error bars represent values ± SEM.

Example 19. Efficacy of Compound 1 in DL-40 Tumor-Bearing Mice

A xenograft study was performed in female NOD SCID mice bearing DL-40 ALCL tumors. Female SCID mice were inoculated subcutaneously in the right flank with 10× ¹⁰⁶ tumor cells in 0.2 ml PBS supplemented with Matrigel (PBS:Matrigel=1:1). Tumor volume was measured twice a week in two dimensions using a caliper, and the volume was calculated using the formula: (w2×1)/2= ^mm3 , assuming that 1 mg is equivalent to 1 ^mm3 of tumor volume. Once tumor volume reached the range of 135-372 ^mm3 (32 days after implantation), animals were randomly divided into groups of 6, stratified to obtain approximately equal mean tumor sizes in each treatment group. Treatment was initiated on day 0 with established DL-40 subcutaneous tumors with a mean tumor volume (MTV) of 238-240 ^mm3 . Table 8 describes the statistical significance of each dose of Compound 1 compared to vehicle.

All agents were administered to DL-40 tumor-bearing mice on day 0 and administered daily PO for 21 days. Compound 1 was dosed at 3, 10, 30, 100, or 300 μg/kg and was formulated in PEG400 (30% v/v) + 70% v/v HPMC (1% w/v) in citrate buffer (pH 5). Pomalidomide was administered at 3000 μg/kg and the same formulation was used as for Compound 1. Body weight and MTV were measured according to a 2× weekly schedule, and the study endpoint was MTV 2007 mm ³ in the vehicle control group on day 21. Statistical analysis was performed using a two-way analysis of variance (ANOVA). Data are expressed as MTV ± SEM. In Fig. 16 is a graph of tumor volume in mice treated with Compound 1 or pomalidomide for 21 days, showing that Compound 1 at doses of 30, 100, and 300 μg/kg was able to reduce tumor volume. Fig. 17 is a graph of body weight change in mice over the 21-day study period.

Example 20. Compound 1 promotes robust loss of IRF4 and upregulation of caspase 3 (MoA) and IRF-1 (BmX) in an ALCL model

A xenograft study was performed in female NOD SCID mice bearing ALCL KI-JK tumors. Female SCID mice were inoculated subcutaneously in the right flank with 10× ¹⁰⁶ tumor cells in 0.2 ml PBS supplemented with Matrigel (PBS:Matrigel=1:1). Tumor volume was measured twice a week in two dimensions using a caliper, and the volume was calculated using the formula: (w2×1)/2= ^mm3 , assuming that 1 mg is equivalent to 1 ^mm3 of tumor volume. Once tumors reached the volume range of 147-365 ^mm3 (39 days after implantation), animals were randomly divided into groups of 5, stratified to obtain approximately equal mean tumor sizes in each treatment group. Treatment was initiated on day 0 with established subcutaneous KI-JK tumors with a mean tumor volume (MTV) of 257–269 mm ³ .

Compound 1 (100 μg/kg) and vehicle control were administered to KI-JK tumor-bearing mice on day 0 and administered PO daily for 21 days. Compound 1 was formulated in PEG400 (30% v/v) + 70% v/v HPMC (1% w/v) in citrate buffer (pH 5), which was also used as vehicle control. Body weight and MTV were measured 2× weekly and the endpoint of the study was MTV 1885 ^mm3 in the vehicle control group on day 24. Data are expressed as MTV ± SEM. Figure 18 shows a graph of tumor volume over the course of the study and shows that Compound 1 reduced tumor volume.

The second xenograft study was performed in female NOD SCID mice bearing KI-JK ALCL tumors. Female SCID mice were inoculated subcutaneously in the right flank with 10× ¹⁰⁶ tumor cells in 0.2 ml PBS supplemented with Matrigel (PBS:Matrigel=1:1). Tumor volume was measured twice weekly in two dimensions using calipers, and the volume was calculated using the formula: (w2×1)/2= ^mm3 , assuming that 1 mg is equivalent to 1 ^mm3 of tumor volume. Once the tumors reached a mean tumor volume (MTV) of 450 ^mm3 , they were divided into two groups.

Compound 1 (100 μg/kg) or vehicle control were administered PO to KI-JK tumor-bearing mice. Compound 1 was formulated in PEG400 (30% v/v) + 70% v/v HPMC (1% w/v) in citrate buffer (pH 5). In the vehicle control group, 2 mice were sacrificed and tumors were collected 24 h after a single dose. Mice in the Compound 1 group were sampled 4 and 24 h after a single dose, with two tumors collected at each time point. Additional mice were dosed daily for 5 or 7 days with tumors collected 24 h after the last dose, with samples collected from 2 mice at each time point. Tumors were then mechanically homogenized and protein was extracted using RTPA buffer (Sigma Aldrich). Protein concentration was quantified using the Pierce™ BCA Protein Assay Kit, samples were concentrated, and then equal amounts of protein were loaded onto a Western blot gel for analysis. Tumors were analyzed for the expression of IKZF1 (Invitrogen, PA5-23728), IKZF3 (CST, 15103), IRF-1 (CST, 8478), IFR-4 (CST, 15106), and caspase-3 (CST, 9662). As shown in Fig. 19, the percentage of IKZF1, IKZF3, and IRF-4 decreased, while the concentration of caspase-3 and IRF-1 increased. Data are expressed as the percentage of each target compared to the target level measured in the medium control and normalized to the GAPDH levels for the protein loading control. Error bars represent values ± SEM.

Example 21. Combination of Compound 1 and Ibrutinib

A xenograft study was performed in female CB17 SCID mice bearing DLBCL TMD8 tumors. Female SCID mice were inoculated subcutaneously in the right flank with 5× ¹⁰⁶ tumor cells in 0.2 ml PBS supplemented with Matrigel (PBS:Matrigel=1:1). Tumor volume was measured twice a week in two dimensions using a caliper, and the volume was calculated using the formula: (w2×1)/2= ^mm3 , assuming that 1 mg is equivalent to 1 ^mm3 of tumor volume. Once tumor volume reached 82-130 ^mm3 (13 days after implantation), animals were randomly divided into groups of 6, stratified such that each treatment group had approximately the same mean tumor size. Treatment was initiated on day 0 with established subcutaneous TMD8 tumors with a mean tumor volume of 107-108 ^mm3 .

All agents were administered to mice bearing TMD8 tumors on day 0 and dosed PO daily until individual tumor volumes reached or exceeded 1500 ^mm3 , at which point dosing was stopped and the individual mouse was excluded from the study. Compound 1 was dosed at 50 μg/kg and was formulated in PEG400 (30% v/v) + 70% v/v HPMC (1% w/v) in citrate buffer (pH 5). Ibrutinib was dosed at 12.5 mg/kg and was formulated in 0.5% MC + 0.4% Cremophor EL + 0.1% sodium lauryl sulfate. Compound 1 was also administered in combination with ibrutinib at the corresponding doses of the individual agents. Body weight and MTV were measured 2× weekly, and the study endpoint was when all mice were sacrificed on day 54. Statistical analysis was performed using survival analysis and the Logrank test (Mantel-Cox test). Data are expressed as MTV ± SEM. Figure 20 shows a graph of tumor volume in mice treated with vehicle, Compound 1, Ibrutinib, or the combination of Compound 1 and Ibrutinib, and shows that the combination was the most effective in reducing tumor volume. Figure 21 shows a graph of the percentage of mice surviving during the study. The highest percentage of surviving mice was observed with the combination. Table 9 describes the statistical significance of vehicle versus vehicle, and Table 10 describes the statistical significance of the combination versus ibrutinib and Compound 1.

Example 22. Combination of Compound 1 and Dexamethesone

A xenograft study was performed in female CB17 SCID mice bearing RPMI-8226 multiple myeloma tumors. Female SCID mice were inoculated subcutaneously in the right flank with 10× ¹⁰⁶ tumor cells in 0.2 ml PBS supplemented with Matrigel (PBS:Matrigel=1:1). Tumor volume was measured twice a week in two dimensions using a caliper, and the volume was calculated using the formula: (w2×1)/2= ^mm3 , assuming that 1 mg is equivalent to 1 ^mm3 of tumor volume. Once tumor volume reached the range of 108-158 ^mm3 (28 days after implantation), animals were randomly divided into groups of 6, stratified to obtain approximately equal mean tumor sizes in each treatment group. Treatment was initiated on day 0 with established RPMI-8226 subcutaneous tumors with a mean tumor volume (MTV) of 130 mm ³ .

All agents were administered to mice bearing RPMI-8226 tumors on day 0 and dosed until individual mouse tumor volumes reached or exceeded 1000 ^mm3 , at which point mice were removed from the study and dosing was stopped. Compound 1 was dosed PO daily at 10 mcg/kg and was formulated in PEG400 (30% v/v) + 70% v/v HPMC (1% w/v) in citrate buffer (pH 5). Dexamethasone was dosed IV once weekly at 5000 mcg/kg and was formulated in saline. Compound 1 was also dosed in combination with Dexamethasone at their respective dose levels and schedules. Body weight and MTV were measured according to a 2× weekly schedule. The study endpoint was a TV greater than 1000 mm ³ , after which any mouse with a TV reaching this TV threshold was removed from the study. The last mouse was removed from the study on day 52. The Logrank (Mantel-Cox) test was used as a survival analysis. Data are expressed as MTV ± SEM. Figure 22 shows a plot of tumor volume in mice treated with vehicle, Compound 1, dexamethasone, or a combination of Compound 1 and dexamethasone, and shows that the combination was most effective in reducing tumor volume. The highest percentage of surviving mice was observed with the combination (Figure 23). Table 11 describes the statistical significance of vehicle versus vehicle, and Table 12 describes the statistical significance of the combination versus ibrutinib and Compound 1. These data are also shown in Figure 41 and Figure 42.

Example 23. Concentration of IKZF1 and IKZF3 in Compound 1-treated monkeys

One male cynomolgus monkey was administered Compound 1 formulated in 1% sodium carboxymethylcellulose + 20% PEG400 in citrate buffer at 30 μg/kg via oral gavage in the fasted state. At the indicated time points, blood was collected from the peripheral vessel of the restrained, unsedated monkey. 50 μl of whole blood was added to a flow tube containing 1 ml of PBS, centrifuged for 5 min at 400×g and the supernatant discarded. Blood was lysed in 1 ml of lysis buffer for 10-15 min at room temperature, centrifuged for 5 min at 400× and the supernatant discarded. PBS (1 ml) was added to each tube, centrifuged for 5 min at 400×g and the supernatant discarded. PBS (1 ml) was added along with 1 μl of Zombie NIR dye to the leukocytes and incubated for 20 min at room temperature. The cells were centrifuged for 5 min at 400×g and the supernatant was removed. Fixation buffer (1 ml) was added and incubated for 20 min at room temperature. The cells were washed with permeabilization buffer (1 ml) and centrifuged for 5 min at 400×g and the supernatant was removed. Permeabilization buffer (1 ml) was added, incubated for 20 min at room temperature, the cells were centrifuged for 5 min at 400×g and the supernatant was removed. Fc block (1 ml) was added, incubated for 15 min at room temperature. IKZF1-AF488 and IKZF3-AF647 or control IgG were added and incubated for 60 min at room temperature with shaking (200 rpm). Cells were washed twice with PBS (1 ml), resuspended in PBS and analyzed on a BD LSR Fortessa Flow Cytometer. Figure 24 shows a plot of the percent mean fluorescence intensity of IKZF1 and IZKF3 at 4 h and 24 h post-dose compared to the pre-dose sample. At both 4 h and 24 h, IKZF1 and IKZF3 levels were less than pre-dose concentrations.

Example 24. Interconversion of Compound 1 in rats and monkeys

Compound 1 was administered to rats at 30 mg/kg PO and to monkeys at either 60 or 100 μg/kg PO. At the indicated time points shown in Fig. 25A and Fig. 25B, plasma samples were collected and the C _max , half-life, and AUC _0-inf were determined for both Compound 1 and the Compound 1 metabolite, Compound 14. The percentage of Compound 14 was also determined (Table 13). PK values were normalized to 1 mg/kg. Fig. 25A is a graph of the concentration of Compound 1 or Compound 14 measured over 24 hours in monkeys administered 100 μg/kg or 60 μg/kg Compound 1. Fig. 25B shows a graph of the concentration of Compound 1 or Compound 14 measured over 24 hours in a rat administered 30 mg/kg Compound 1.

Example 25. Efficacy of selected compounds against multiple myeloma

A xenograft study was performed in female NOD SCID mice bearing NCI-H929 multiple myeloma tumors. Female SCID mice were inoculated subcutaneously in the right flank with 5× ¹⁰⁶ tumor cells in 0.2 ml PBS supplemented with Matrigel (PBS:Matrigel=1:1). Tumor volume was measured twice a week in two dimensions using a caliper, and the volume was calculated using the formula: (w2×1)/2= ^mm3 , assuming that 1 mg is equivalent to 1 ^mm3 of tumor volume. Once tumor volume reached the range of 112-194 ^mm3 (14 days after implantation), animals were randomly divided into groups of 2, stratified to obtain approximately equal mean tumor sizes in each treatment group. Treatment was initiated on day 0 with established NCI-H929 subcutaneous tumors with a median tumor volume (MTV) of 146-154 mm ³ .

Compound 1, Compound 11, and Compound 13 were administered to NCI-H929-bearing mice on day 0 and dosed PO daily for 5 days. Compounds were administered at 100 μg/kg and were formulated in PEG400 (30% v/v) + 70% v/v HPMC (1% w/v) in citrate buffer (pH 5). Body weight and MTV were measured 2× weekly, and the study endpoint was after 5 days of dosing. Figs. 26A–26K and Fig. 26M show plots of tumor volume measurements in mice over the course of the study. Data are expressed as MTV ± SEM.

The second xenograft study was performed in female NOD SCID mice bearing ALCL DL-40 tumors. Female SCID mice were inoculated subcutaneously in the right flank with 10 × 10 ⁶ tumor cells in 0.2 ml PBS supplemented with Matrigel (PBS:Matrigel = 1:1). Tumor volume was measured twice a week in two dimensions using calipers, and the volume was calculated using the formula: (w2 × 1)/2 = mm ³ , assuming that 1 mg is equivalent to 1 mm ³ of tumor volume. Once tumor volume reached the range of 187-335 mm ³ (31 days after implantation), animals were randomly divided into groups of 2, stratified to obtain approximately equal mean tumor sizes in each treatment group. Treatment was initiated on day 0 with established DL-40 subcutaneous tumors with a mean tumor volume (MTV) of 242–244 mm ³ .

Compound 12 and vehicle control were administered to DL-40-bearing mice on day 0 and dosed PO for 5 days. Compound 12 was formulated in PEG400 (30% v/v) + 70% v/v HPMC (1% w/v) in citrate buffer (pH 5). Body weight and MTV were measured 2× weekly and the study endpoint was after 7 days of dosing. Figure 26L shows a graph of tumor volumes measured in mice over the 5-day study. Data are expressed as MTV ± SEM.

Example 26: Fluorescence Polarization

Compounds were dispensed from serially diluted DMSO stock solution into low dead volume plates in black 384-well FP-compatible plates using acoustic technology to 1% of the total reaction volume. Compounds were stacked vertically in rows A through P. Concentration series were stacked horizontally: columns 1–11, followed by duplicates in columns 12–22. Columns 23 and 24 were reserved for 0% (5 nM probe) and 100% controls (high protein with 5 nM probe), respectively.

20 μl of a mixture containing 10 nM cereblon-DDBl and 5 nM probe dye in 50 mM HEPES, pH 7.4, 200 mM NaCl, 1 mM TCEP, 0.1% BSA, and 0.05% pluronic acid F-127 were added to the wells containing compound and incubated at room temperature for 1.5 h. Control wells with 100% probe bound contained 100 nM cereblon. To correct for background fluorescence, corresponding control plates without cereblon-DDBl were used. Plates were read on an Envision plate reader with the appropriate FP filter sets. The results of testing Compound 1 and pomalidomide in this assay are shown in Fig. 27.

Example 27. Competitive analysis of cereblon in a cell

Cell permeability and binding affinity of Compound 1 and pomalidomide to cellular cereblon (CRBN) were determined by competitive displacement of NanoBRET™ tracer reversibly linked to cereblon-NanoLuc ^® fusion protein in 293T cells. 293T cells were modified by lentiviral transfection to express the cereblon-NanoLuc ^® luciferase fusion. The modified cereblon-NanoLuc 293T cell line was co-treated with varying concentrations of test compound and probe conjugated to the NanoBRET fluorescent tracer at its specified K _D concentration (300 nM) and incubated for 2 h at 37°C to reach equilibrium. Affinity of the test compound was determined by the shift in the NanoBRET indicator signal after addition of NanoBRET reagents (Promega). Forty microliters of Cereblon-NanoLuc 293T cells suspended in OptiMEM at 2 x ¹⁰⁵ cells/mL (8000 cells/well) were dispensed using a Multidrop Combi Reagent Dispenser (Thermo Fisher) into each well of 384-well TC-treated white microplates (Corning 3570). A 10 mM stock solution of the test compound in DMSO was serially diluted (half-log) with DMSO to generate an 11-point dose series in an acoustically prepared low dead volume 384-well microplate (Labcyte). Using an Echo 550 Acoustic Liquid Handler (Labcyte), 40 nL of serially diluted compound solutions were dispensed in duplicate into each 384-well TC-treated white microplate containing cereblon-NanoLuc 293T cells. All control wells were loaded with 40 nL DMSO. 40 nL NanoBRET indicator was loaded into all wells in columns 1–23 at its predetermined _KD concentration (300 nM). An additional 40 nL DMSO was loaded into column 24. The final DMSO concentration was 0.2% for all samples. Plates were briefly rotated and cells were incubated at 37°C, 5% CO2 for 2 h. 20 μl of NanoBRET TE assay reagents were added to each well and the NanoBRET signal was recorded using an En Vision Multilabel Reader (PerkinElmer). Donor emission of cereblon-NanoLuc was detected at 450 nm using a NanoLuc 460/50 filter and acceptor fluorescence of the NanoBRET tracer-pomalidomide (618 nm) was detected using a 600 nm long-pass NanoBRET filter. The acceptor signal/donor signal ratio was calculated for each well. Column 24 (cells without added NanoBRET tracer-pomalidomide) was used as a positive control (P). The percent response of the compound-treated samples (T) was calculated by normalizing the acceptor/donor ratio for each well to the DMSO-treated negative (N) controls on the same microtiter plate after subtracting the background (i.e., positive control) signal: % response = 100 × (Signal(T) - Mean(P))/(Mean(N) - Mean(P)). The results of testing Compound 1 and pomalidomide in this assay are shown in Fig. 28.

Example 28. Destruction of IKZF1/IKZF3

Western blotting was used to assess the Compound 1 degradation characteristics of Aiolos (IKZF3) and Ikaros (IKZF1). NCI-H929 cells (ATCC, CRL-9068) were seeded in 6-well plates pre-coated with Compound 1 or pomalidomide according to a dose-response relationship (5 points, 0.01-100 nM). After 4 h incubation with compounds, pellets were washed with PBS and frozen at -80°C. Cell pellets were lysed in lysis buffer [RIPA (Thermo, Ref 89901), 1× Halt protease and phosphatase inhibitor mix (Thermo, Pro#1361281), Benzonase (Sigma, E1014-5JU)] for 10 min on ice. Insoluble proteins were removed from the lysates by centrifugation (21.2×g, 10 min).

Protein concentrations were measured using the BSA Protein Assay Kit (Thermo, 23228). The BSA protein standard curve and protein concentration of the samples were read using an Envision Multichannel Reader (PerkinElmer). Lysate concentrations were normalized with Laemmli 6X Lysis Buffer, SDS Sample Buffer, Reducing Buffer (Boston BioProducts, Inc. Part #BP-111R-50 mL). Normalized samples and Chameleon® Duo Prestained Protein Marker Kit (LI-COR, 928-60000) were loaded onto a Criterion™ Tris-HCl Protein Gel containing 4-15% (Bio-Rad, #3450028). Gels were run at 120 V for 1.5 hours. Protein transfer was completed using the Trans-Blot Turbo Transfer System (Bio-Rad, 1704150EDU) at 25 V for 7 min using the Trans-Blot Turbo RTA Midi 0.2 µm Nitrocellulose Transfer Kit (Bio-Rad, cat# 1704271) according to the manufacturer's recommendations.

Membranes were blocked with rocking for one hour in Intercept® Blocking Buffer (TBS) (LI-COR, catalog #927-50000). Primary antibodies were diluted in Intercept® T20 (TBS) protein-free antibody diluent (LI-COR, catalog #927-85001) and incubated with rocking at 4°C overnight.

Membranes were washed 3 times for 5 minutes in TBS-T with rocking. Secondary antibodies were diluted in Intercept® T20 (TBS) in protein-free antibody diluent (LI-COR, Cat# 927-85001) and incubated on membranes for 1 hour with rocking at room temperature. Membranes were washed as described previously and visualized on an Odyssey CLx. Data were expressed as percentage of Aiolos or Ikaros compared to medium control and normalized to the β-actin loading control. The resulting Western blot is shown in Fig. 30.

Example 29. Apoptosis induction assay

Induction of apoptosis in NCIH929 multiple myeloma cells was determined by measuring caspase 3/7 activity after 72 h of treatment with Compound 1 using the Caspase 3/7 Assay System (Promega, G8091). Briefly, test compound was added to white 384-well plates treated with TC at a maximum concentration of 100 nM for Compound 1 and 10 μM for pomalidomide in a 10-point, semi-logarithmic titration with replicates. 30 μl of NCIH929 cells were seeded into 384-well plates in RPMI medium containing 10% FBS and 0.05 mM 2-mercaptoethanol at a cell density of 1000 cells/well. Cells treated in the absence of test compound served as negative controls. Following compound treatment, cells were incubated at 37°C with 5% _CO2 for 72 h. 30 μl of reconstituted caspase 3/7 detection assay reagent was added to each well and luminescence was recorded using an En Vision™ Multichannel Plate Reader (PerkinElmer, Santa Clara, CA, USA). Maximum caspase 3/7 activity >2-fold was observed at 72 h with an EC50 of 0.58 nM for Compound 1 and 407 nM for pomalidomide. Data from this assay are shown in Fig. 31.

Example 30. Analysis of antiproliferative activity

Growth of eight multiple myeloma cell lines was determined following 96 hours of treatment with Compound 1 or pomalidomide based on ATP quantification using the CellTiter-Glo® 2.0 Luminescent Assay Kit, which signals the presence of metabolically active cells. Briefly, test compound was added to 384-well plates at a maximum concentration of 100 nM for Compound 1 and 10 μM for pomalidomide in 10-point, semi-logarithmic titrations in duplicate. 50 µl of cells suspended in growth medium at the cell density indicated for each cell line in Table 14 were dispensed using a multi-drop combination reagent dispenser (Thermo Fisher) into 384-well black microplates treated with TC containing test compounds at duplicate concentration ranges and DMSO controls. Cells treated in the absence of test compound served as negative controls and cells treated in the absence of CellTiter-Glo® 2.0 served as positive controls. Cells were incubated at 37°C with 5% _CO2 for 96 h. CellTiter-Glo reagent was then added to the cells and luminescence was recorded on an EnVision™ Multichannel Reader (PerkinElmer, Santa Clara, CA, USA). % viability was determined by normalizing the signal with positive and DMSO-treated negative controls on the same microtiter plate. Compound 1 inhibited the growth of all eight multiple myeloma cell lines with a maximal inhibition of greater than 50% and an IC50 of less than 1 nM and was 1000-fold more potent than pomalidomide. The data are shown in Fig. 32.

Example 31. Efficacy of Compound 1 in Multiple Myeloma

The efficacy of Compound 1 on a multiple myeloma cell line (NCI-H929) was assessed at four different concentrations: 3 μg/kg, 10 μg/kg, 30 μg/kg, and 100 μg/kg. Compound 1 was administered QD (every day) orally (PO). The efficacy of Compound 1 was compared to pomalidomide (administered at 3000 μg/kg). The results for NCI-H929 are shown in Fig. 7. Table 15 describes the statistical significance of the experiment for each dose. Treatment with Compound 1 at doses of 10, 30, and 100 μg/kg resulted in a greater reduction in tumor volume than pomalidomide administered at a dose of 3000 μg/kg.

For NCI-H929 cells, a xenograft study was performed in female NOD SCID mice bearing NCI-H929 multiple myeloma tumors. Female SCID mice were inoculated subcutaneously in the right flank with 5× ¹⁰⁶ tumor cells in 0.2 ml PBS supplemented with Matrigel (PBS:Matrigel=1:1). Tumor volume was measured twice a week in two dimensions using a caliper, and the volume was calculated using the formula: (w2×1)/2= ^mm3 , assuming that 1 mg is equivalent to 1 ^mm3 of tumor volume. Once tumor volume reached the range of 84-267 ^mm3 (18 days after implantation), animals were randomly divided into groups of 6, stratified to obtain approximately equal mean tumor sizes in each treatment group. Treatment was initiated on day 0 with established subcutaneous NCI-H929 tumors with a mean tumor volume (MTV) of 149-150 mm ³ .

All agents were administered to NCI-H929 tumor-bearing mice on day 0 and dosed PO daily for 21 days except for the vehicle group, which was dosed for 18 days. Following the dosing period, tumor growth was monitored for 45 days. Compound 1 was administered at 3, 10, 30, or 100 μg/kg and was formulated in PEG400 (30% v/v) + 70% v/v HPMC (1% w/v) in citrate buffer (pH 5). Pomalidomide was dosed at 3000 μg/kg and administered in the same formulation as Compound 1. Body weight and MTV were measured on a 2× weekly schedule. After the first 21 days of dosing, all Compound 1-treated groups were monitored for tumor growth. The 10 μg/kg group was discontinued on day 39. Tumors in the 30 μg/kg group began to grow, and daily PO dosing was resumed beginning on day 40 for an additional 23 days. Statistical analysis was performed using two-way analysis of variance (ANOVA). Data are expressed as MTV ± SEM. The resulting data are shown in Fig. 33.

Example 32. Destruction of IKZF3 depending on dose

A xenograft study was performed in female CB17 SCID mice bearing RPMI- multiple myeloma tumors. Female SCID mice were inoculated subcutaneously in the right flank with 10× ¹⁰⁶ tumor cells in 0.2 ml PBS supplemented with Matrigel (PBS:Matrigel=1:1). Tumor volume was measured twice weekly in two dimensions using a caliper, and the volume was calculated using the formula: (w2×1)/2= ^mm3 , assuming that 1 mg is equivalent to 1 ^mm3 of tumor volume. Once tumors reached a mean volume of 265 ^mm3 for the multiple-dose study or 380 ^mm3 for the single-dose study, they were randomized into groups of 3.

Compound 1 (100 μg/kg) and vehicle control were administered to RPMI-8226-bearing mice on day 0 and dosed PO daily for 5 days. Compound 1 was formulated in PEG400 (30% v/v) + 70% v/v HPMC (1% w/v) in citrate buffer (pH 5), which was also used as vehicle control. Body weight and MTV were measured on a 2× weekly schedule. In the vehicle control group, 3 mice were sacrificed and tumors were collected 24 h after a single dose. Mice in the Compound 1 group were sampled 4 and 24 h after a single dose, with 3 tumors collected at each time point. Additional mice were dosed daily for 3 or 5 days, with tumors harvested 24 hours after the last dose from 3 mice at each time point. Tumors were then mechanically homogenized and protein was extracted using RIPA buffer (Sigma Aldrich). Protein concentration was quantified using the Pierce™ BCA Protein Assay Kit, samples were concentrated, and equal amounts of protein were then loaded onto a Western blot gel for analysis. Tumors were analyzed for expression of IKZF1 (Invitrogen, PA5-23728), IKZF3 (CST, 15103), or IRF-4 (CST, 15106). Individual band intensities were measured for data analysis using Image Studio NIR software. Protein expression was quantified relative to the reference protein, GAPDH, to control for total protein concentration. The data were then normalized to the amount of target in Compound 1-treated samples compared to the vehicle control samples. Data are presented as the percentage of target present in the vehicle control and are normalized to total protein. Error bars represent values ± SEM. This study was used to generate the data shown in Fig. 36 and Fig. 37.

Example 33. Decreased expression of cereblon with prolonged administration of IMiD dose

Cells were cultured in 10 μM lenalidomide for two months. A subset of cells were then passaged in 1 μM pomalidomide or continued in parallel in lenalidomide for an additional two months. Cells were lysed in RIPA medium with protease inhibitors, 30 μg of protein were loaded into each well of a 4-12% T TGX gel and exposed to 90 minutes at 120 volts. Protein was transferred to a nitrocellulose membrane and blotted for the indicated protein (cereblon: Sigma Cat# HPA045910, IKZF1 Cat# CST 14859, IKZF3 Cat# CST 15103, vinculin EMD Cat# MAB3574) for 1 hour at room temperature. Membranes were washed three times and incubated with anti-rabbit 800 or anti-mouse 700 (LiCor) and visualized on an Odyssey CLx. This experiment yielded the data shown in Fig. 38.

Example 34. Compound 1 retains activity in IMiD-resistant H929 MM cells

The human multiple myeloma cell line NCIH929 was continuously cultured in medium containing 10 μM lenalidomide for 8 weeks followed by 1 μM pomalidomide for 4 weeks to develop IMiD resistance. The growth of IMiD-resistant NCIH929 cells was determined after 96 hours of treatment with Compound 1 or pomalidomide based on ATP quantification using the CellTiter-Glo 2.0 Luminescent Assay Kit, which is proportional to the number of metabolically active cells. Briefly, test compound was added to 384-well plates at a maximum concentration of 10 μM in 14-point, semi-logarithmic titration with replicates. NCIH929 cells were seeded in 384-well plates in RPMI medium containing 10% FBS and 0.05 mM 2-mercaptoethanol at a cell density of 750 cells per well. Cells treated in the absence of test compound served as negative controls, and cells treated in the absence of CellTiter-Glo® 2.0 served as positive controls. Cells treated with test compounds were incubated at 37°C with 5% _CO2 for 96 h. CellTiter-Glo reagent was then added to the cells and luminescence was recorded on an EnVision™ Multichannel Reader (PerkinElmer, Santa Clara, CA, USA). % viability was determined by normalizing the signal with positive and DMSO-treated negative controls on the same microtiter plate. Compound 1 inhibited the proliferation of IMiD-resistant NCIH929 cells with an IC _{50 of} 2.3 nM and a maximal inhibition of 70%, while pomalidomide induced growth inhibition of less than 50% at concentrations up to 10 μM. This experiment yielded the data shown in Fig. 39.

Example 35 Compound 1 is active in REC1 cells

Compound 1 (30 μg/kg) and vehicle control were administered to REC1-bearing mice on day 0 and dosed PO daily for 3 days. Compound 1 was formulated in PEG400 (30% v/v) + 70% v/v HPMC (1% w/v) in citrate buffer (pH 5), which was also used as vehicle control. Body weight and MTV were measured on a 2× weekly schedule. In the vehicle control group, 3 mice were sacrificed and tumors were collected 24 h after a single dose. Mice in the Compound 1 group were sampled 4 and 24 h after a single dose, with 3 tumors collected at each time point. Additional mice were dosed daily for 3 days, with tumors harvested 24 hours after the last dose, and 3 mice were sampled at each time point. Tumors were then mechanically homogenized and protein was extracted using RIPA buffer (Sigma Aldrich). Protein concentration was quantified using the Pierce™ BCA Protein Assay Kit, samples were concentrated, and then equal amounts of protein were loaded onto a Western blot gel for analysis. Tumors were analyzed for the expression of IKZF1 (bivitrogen, PA5-23728), IKZF3 (CST, 15103), IRF-4 (CST, 15106), cyclin D1 (CST, 2922), and E2F1 (CST, 3742). Individual band intensities were measured for data analysis using Image Studio NIR software. Protein expression was quantified relative to a reference protein, GAPDH, to control for total protein concentration. Data were then normalized to the amount of target in Compound 1-treated samples compared to medium controls. Data are presented as the percentage of target present in the medium control and normalized to total protein. Error bars represent values ± SEM. This analysis was used to generate the data shown in Figs. 43 and 44.

Example 36. Cell viability analysis

Materials

RPMI 1640 medium, fetal bovine serum (FBS), and 2-mercaptoethanol were purchased from Gibco (Grand Island, NY, USA). CellTiter-Glo® 2.0 assay was purchased from Promega (Madison, WI, USA). NCIH929.1 cell line was purchased from ATCC (Manassas, VA, USA). Cell culture flasks and 384-well microplates were purchased from VWR (Radnor, PA, USA).

Cell viability analysis

NCIH929.1 cell viability was determined based on ATP quantification using the CellTiter-Glo® 2.0 Luminescent Assay Kit, which signals the presence of metabolically active cells. Briefly, test compound was added to 384-well plates at a maximum concentration of 1 μM in 10-point, semi-logarithmic titrations with replicates. NCIH929.1 cells were seeded in 384-well plates in RPMI medium containing 10% FBS and 0.05 mM 2-mercaptoethanol at a cell density of 750 cells per well. Cells treated in the absence of test compound served as a negative control, and cells treated in the absence of CellTiter-Glo® 2.0 served as a positive control. On the same day of compound treatment, CellTiter-Glo® 2.0 was added to the plate with cells treated in the absence of test compound to establish a cytostatic control value (C _T0 ). Cells treated with test compound were incubated at 37°C with 5% CO2 for 96 hours. CellTiter-Glo reagent was then added to the cells and luminescence was recorded on an En Vision™ Multichannel Reader (PerkinElmer, Santa Clara, CA, USA).

Using the above analysis, GI ₅₀ data were determined for the illustrative compounds listed in Table 16 below.

Example 37. NHL cell line growth inhibition assay

Growth of NHL cell lines was determined following 96 h of treatment with Compound 1, pomalidomide, or CC-92480 based on ATP quantification using the CellTiter-Glo® 2.0 Luminescent Assay Kit, which is indicative of the presence of metabolically active cells. Test compound was added to 384-well plates at a maximum concentration of 100 nM or 10 μM for Compound 1 and 10 μM for pomalidomide and CC-92480 in a 10-point, semilogarithmic titration with replicates. 50 μl of cells suspended in growth medium at the cell density indicated for each cell line in Table 17 were dispensed using a multi-drop combination reagent dispenser (Thermo Fisher) into 384-well black microplates treated with TC containing a range of test compounds in duplicate and DMSO controls. Cells treated in the absence of test compound served as negative controls and cells treated in the absence of CellTiter-Glo® 2.0 served as positive controls. Cells were incubated at 37°C with 5% CO2 for 96 h. CellTiter-Glo reagent was then added to the cells and luminescence was recorded on an En Vision™ Multichannel Reader (PerkinElmer, Santa Clara, CA, USA). % viability was determined by normalizing the signal with positive and DMSO-treated negative controls on the same microtiter plate. The resulting data are shown in Fig. 45A, Fig. 45B, and Fig. 45C.

Example 38. ALCL KI-JK xenograft study

A xenograft study was performed in female CB17 SCID mice bearing ALCL KI-JK tumors. Female SCID mice were inoculated subcutaneously in the right flank with 10× ¹⁰⁶ tumor cells in 0.2 ml PBS supplemented with Matrigel (PBS:Matrigel=1:1). Tumor volume was measured twice a week in two dimensions using a caliper, and the volume was calculated using the formula: (w2×1)/2= ^mm3 , assuming that 1 mg is equivalent to 1 ^mm3 of tumor volume. Once the tumors reached a mean volume of 313 ^mm3 , mice were randomized into groups of 4.

Compound 1 (100 μg/kg) and vehicle control were administered to KI-JK-bearing mice on day 0 and dosed PO daily for 5 days. Compound 1 was formulated in PEG400 (30% v/v) + 70% v/v HPMC (1% w/v) in citrate buffer (pH 5), which was also used as vehicle control. Body weight and MTV were measured on a 2× weekly schedule. In the vehicle control group, 3 mice were sacrificed and tumors were collected 24 h after a single dose. Mice in the Compound 1 group were sampled 4 and 24 h after a single dose, with 3 tumors collected at each time point. Additional mice were dosed daily for 5 days with tumors harvested 24 hours after the last dose, with samples collected from 3 mice at each time point. Tumors were then mechanically homogenized and protein was extracted using RIPA buffer (Sigma Aldrich). Protein concentration was quantified using the Pierce™ BCA Protein Assay Kit, samples were concentrated, and equal amounts of protein were then loaded onto a Western blot gel for analysis. Tumors were analyzed for IKZF3 (CST, 15103) or IRF-4 (CST, 15106) expression. Individual band intensities were measured for data analysis using Image Studio NIR software. Protein expression was quantified relative to the reference protein, GAPDH, to control for total protein concentration. The data were then normalized to the amount of target in Compound 1-treated samples compared to the vehicle control samples. Data are presented as the percentage of target present in the vehicle control and are normalized to total protein. Error bars represent values ± SEM. The resulting data are shown in Fig. 46.

Example 39. Mino xenograft study

A xenograft study was performed in female CB17 SCID mice bearing Mino mantle cell lymphoma tumors. Female SCID mice were inoculated subcutaneously in the right flank with 10× ¹⁰⁶ tumor cells in 0.2 ml PBS supplemented with Matrigel (PBS:Matrigel=1:1). Tumor volume was measured twice weekly in two dimensions using calipers, and the volume was calculated using the formula: (w2×1)/2= ^mm3 , assuming that 1 mg is equivalent to 1 ^mm3 of tumor volume. Once the tumors reached a mean tumor volume of 129 ^mm3 (21 days after implantation), animals were randomly divided into groups of 4, stratified to obtain approximately equal mean tumor sizes in each treatment group. Treatment was initiated on day 0 with established subcutaneous Mino tumors with a mean tumor volume (MTV) of 134 mm ³ .

All agents were administered to Mino tumor-bearing mice on day 0 and dosed for 34 days. Compound 1 was dosed PO daily at 30 μg/kg and was formulated in PEG400 (30% v/v) + 70% v/v HPMC (1% w/v) in citrate buffer (pH 5). Rituximab was dosed IV once weekly at 10 mg/kg and was formulated in saline. Compound 1 was also administered in combination with Rituximab at their respective dose levels and schedules. Body weight and MTV were measured according to a 2× weekly schedule. The study endpoint was day 35, when vehicle control reached an MTV of 2320 mm ³ . Statistical analysis was performed using the paired t-test. Data are expressed as MTV ± SEM. The resulting data are shown in Fig. 1. 47.

Example 40. NCI-H929 Xenograft Study

A xenograft study was performed in female NOD SCID mice bearing MM NCI-H929 tumors. Female SCID mice were inoculated subcutaneously in the right flank with 10× ¹⁰⁶ tumor cells in 0.2 ml PBS supplemented with Matrigel (PBS:Matrigel=1:1). Tumor volume was measured twice a week in two dimensions using a caliper, and the volume was calculated using the formula: (w2×1)/2= ^mm3 , assuming that 1 mg is equivalent to 1 ^mm3 of tumor volume. To evaluate the efficacy of this study, once the tumors reached a mean tumor volume of 419 ^mm3 , the mice were randomly divided into 4 groups of 4 mice each. For the pharmacodynamic and pharmacokinetic portion of the study, once the tumors reached a mean tumor volume of 620 ^mm3 , the mice were randomly divided into 4 groups of 12 mice each.

For the efficacy portion of the study, Compound 1 (100 μg/kg), CC-92480 (1000 μg/kg), pomalidomide (3000 μg/kg), and vehicle control were administered to NCI-H929-bearing mice on day 0 and dosed PO daily for 18 days. Compound 1, CC-92480, and pomalidomide were formulated in PEG400 (30% v/v) + 70% v/v HPMC (1% w/v) in citrate buffer (pH 5), which also served as vehicle control. Body weight and MTV were measured on a twice-weekly schedule. The study endpoint was day 18, when vehicle control reached an MTV of 2460 mm ³ . Data are expressed as MTV ± SEM.

For the pharmacodynamic and pharmacokinetic portions of this study, 3 mice were sacrificed at each time point with tumors and plasma collected, with mice in the vehicle control group sacrificed 24 hours after the single dose. For mice in the Compound 1 (100 μg/kg), CC-92480 (1000 μg/kg), and pomalidomide (3000 μg/kg) treated groups, samples were collected at 1, 4, 24, and 48 hours after the single dose from 3 mice collected per time point. Tumors were then mechanically homogenized and protein was extracted using RIPA buffer (Sigma Aldrich). Protein concentration was quantified using the Pierce™ BCA Protein Assay Kit, samples were concentrated, and then equal amounts of protein were loaded onto a Western blot gel for analysis. Tumors were analyzed for IKZF3 expression (CST, 15103). The intensity of individual bands was measured for data analysis using Image Studio NIR software. Protein expression was quantified relative to the reference protein, GAPDH, to control for total protein concentration. Data were then normalized to the amount of target in Compound 1-treated samples compared to vehicle control samples. Data are presented as the percentage of target present in the vehicle control and normalized to total protein. For pharmacokinetic analysis of plasma and tumor samples, tumor samples were mechanically homogenized using homogenization solution (MeOH/15 mM PBS (1:2, v:v)) and plasma proteins were precipitated. Samples were quenched and compared to a standard curve by LC-MS/MS analysis. Error bars represent values ± SEM. The resulting data are shown in Fig. 48, Fig. 49 and Fig. 50.

Example 41 Induction of Apoptosis in TMD8 Cells

Induction of apoptosis in TMD8 cells was determined by measuring caspase 3/7 activity after 48 h treatment with Compound 1 or pomalidomide using the caspase 3/7 assay system (Promega, G8091). Test compound was added to white 384-well plates treated with TC at a maximum concentration of 10 μM for Compound 1 and pomalidomide with 10-14 point, semi-logarithmic titration with replicates. 30 μl of TMD8 cells were seeded in 384-well plates in RPMI medium containing 10% FBS at a cell density of 2000 cells/well. Cells treated in the absence of test compound served as negative controls. Following compound treatment, cells were incubated at 37°C with 5% _CO2 for 48 h. A total of 30 μl of reconstituted caspase 3/7 detection assay reagent was added to each well and luminescence was recorded using an En Vision™ Multichannel Reader (PerkinElmer, Santa Clara, CA, USA). Maximum caspase 3/7 activity >8-fold was observed at 48 h with an EC50 of 2.7 nM for Compound 1 and 1.2 μM for pomalidomide. % caspase 3/7 activity was determined by normalizing the signal with DMSO-treated controls on the same microtiter plate. The resulting data are shown in Fig. 51.

Example 42. Cell viability in TMD8 cells

TMD8 cell viability was determined based on ATP quantification using the CellTiter-Glo® 2.0 Luminescent Assay Kit, which is indicative of the presence of metabolically active cells. Test compound was added to 384-well plates at a maximum concentration of 100 nM for Compound 1 or 10 μM for pomalidomide in an 11-point, semi-logarithmic titration with replicates. TMD8 cells were seeded in 384-well plates in RPMI medium containing 10% FBS at a cell density of 6,000 cells per well. Cells treated in the absence of test compound served as a negative control normalized to 100% viability, and cells treated in the absence of CellTiter-Glo® 2.0 served as a positive control normalized to 0% viability. The cells were incubated at 37°C with 5% CO2 for 96 h. CellTiter-Glo reagent was then added to the cells and luminescence was recorded using an En Vision™ multichannel plate reader (PerkinElmer, Santa Clara, CA, USA). The data obtained are shown in Fig. 52.

Example 43. Gene ontology research

ALCL DL-40 xenograft tumors were isolated from mice 48 h after oral administration of 100 μg/kg Compound 1 or vehicle and quantitative global proteomic analysis was performed as described in Example 10. Gene ontology analysis was performed using BINGO, a Java-based tool implemented as an extension to Cytoscape. Default parameters were used to assess overexpression of GO categories and interactions between altered proteins were determined using the STRING database with a cutoff of 700. Interferon (IFN) signaling was the most prominent network affected by Compound 1. Genes with significant changes in expression are listed in Table 18. The study indicates that the primary effect of Compound 1 is on interferon response genes.

Example 44. Study of the CB17 SCID mantle cell lymphoma Mino xenograft

A xenograft study was performed in female CB17 SCID mice bearing Mino multiple mantle cell lymphoma tumors. Female SCID mice were inoculated subcutaneously in the right flank with 10× ¹⁰⁶ tumor cells in 0.2 ml PBS supplemented with Matrigel (PBS:Matrigel=1:1). Tumor volume was measured twice weekly in two dimensions using a caliper, and the volume was calculated using the formula: (w2×1)/2= ^mm3 , assuming that 1 mg is equivalent to 1 ^mm3 of tumor volume. Once the tumors reached a mean tumor volume of 129 ^mm3 (21 days after implantation), animals were randomly divided into groups of 5, stratified to obtain approximately equal mean tumor sizes in each treatment group. Treatment was initiated on day 0 with established subcutaneous Mino tumors with a mean tumor volume (MTV) of 134 mm ³ .

All agents were administered to Mino tumor-bearing mice on day 0 and dosed for 34 days. Compound 1 was dosed PO daily at 30 μg/kg and was formulated in PEG400 (30% v/v) + 70% v/v HPMC (1% w/v) in citrate buffer (pH 5). Ibrutinib was dosed PO daily at 25 mg/kg and was formulated in 0.5% MC + 0.4% EL + 0.1% SDS. Compound 1 was also administered in combination with Ibrutinib at their respective dose levels and schedules. Body weight and MTV were measured according to a 2× weekly schedule. The study endpoint was day 35, when vehicle control reached an MTV of 2320 mm ³ . Statistical analysis was performed using a paired t-test. Data are expressed as MTV ± SEM.

The obtained data are shown in Fig. 53.

Example 45. CB17 SCID RPMI-8226 xenograft study

A xenograft study was performed in female CB17 SCID mice bearing RPMI-8226 multiple myeloma tumors. Female SCID mice were inoculated subcutaneously in the right flank with 10× ¹⁰⁶ tumor cells in 0.2 ml PBS supplemented with Matrigel (PBS:Matrigel=1:1). Tumor volume was measured twice weekly in two dimensions using a caliper, and the volume was calculated using the formula: (w2×1)/2= ^mm3 , assuming that 1 mg is equivalent to 1 ^mm3 of tumor volume. Once tumors reached a mean volume of 101 ^mm3 for the efficacy study or 342 ^mm3 for the multiple-dose pharmacodynamic study, mice were randomized into groups of 4 or 3, respectively.

For the efficacy portion of the study, Compound 1 (100 μg/kg), pomalidomide (3000 μg/kg), CC-92480 (1000 μg/kg), and vehicle control were administered to RPMI-8226 tumor-bearing mice on day 0 and administered PO daily for 19 days. Compounds were formulated in PEG400 (30% v/v) + 70% v/v HPMC (1% w/v) in citrate buffer (pH 5), which also served as vehicle control. Body weight and MTV were measured as per the twice weekly schedule. The study endpoint was day 19, when vehicle control reached an MTV of 1789 mm ³ . Statistical analysis was performed using the paired t-test. Data are expressed as MTV ± SEM.

For the pharmacodynamic portion of the study, mice were dosed daily. Compound 1 (100 μg/kg), pomalidomide (3000 μg/kg), CC-92480 (1000 μg/kg), and vehicle control were administered to mice bearing RPMI-8226 tumors starting on day 0. Mice were dosed orally for 7 days, with tumors harvested at 4 and 24 h after a single dose or 24 h after 3, 5, or 7 daily doses. Tumors were then mechanically homogenized and protein was extracted using RIPA buffer (Sigma Aldrich). Protein concentration was quantified using the Pierce™ BCA Protein Assay Kit, samples were concentrated, and then equal amounts of protein were loaded onto a Western blot gel for analysis. Tumors were analyzed for IKZF3 expression (CST, 15103). Individual band intensities were measured for data analysis using Image Studio NIR software. Protein expression was quantified relative to a reference protein, GAPDH, to control for total protein concentration. Data were then normalized to the amount of target in Compound 1-treated samples compared to vehicle control samples. Data are presented as the percentage of target present in the vehicle control and normalized to total protein. Error bars represent values ± SEM.

The obtained data are shown in Fig. 54 and Fig. 55.

Example 46. Cytokine profile

Human CD4 ⁺ /CD8 ⁺ T cells were isolated from Leukopak using a magnetic negative selection enrichment kit (StemCell T cell enrichment catalog #17851). T cells were plated on anti-COD (clone OKT3, 10 μg/mL)-coated cells at 500,000 cells/mL in RPMI1640 + 10% fetal bovine serum and treated with a seven-point dilution series of pomalidomide, CC-92480, Compound 1, Compound 15, or Compound 2 in duplicate. After six days, supernatants were collected, centrifuged to clear debris, and snap frozen until assayed. A Procarta 45-plex panel was used to determine cytokine concentrations using the manufacturer's protocol (Thermo catalog #EPX450-12171-901). Briefly, antibody-coated beads were incubated with supernatants, washed, and biotinylated detection antibodies, which are detected by the addition of streptavidin-RPE, were added. Absolute values of each cytokine were extrapolated from standard curves using nonlinear regression. Cytokine levels were normalized and fold change levels were determined compared to DMSO-treated controls.

Compound 15 is

The results are shown graphically in Figs. 58–64 and are listed in Table 19.

Table 19 shows the effect of Compound 1, Compound 2, Compound 15, CC-92480, or pomalidomide on the levels of cytokines secreted by anti-CD3-stimulated T cells after six days of incubation. Values represent fold changes compared to DMSO-treated control wells.

All publications and patent applications cited in this specification are herein incorporated by reference to the same extent as if each individual publication or patent application were specifically and individually indicated to be incorporated by reference.

Although the present invention has been described in some detail above by way of illustration and example for purposes of clarity of understanding, it will be apparent to one skilled in the art that, in light of the teachings of the present invention, certain changes and modifications can be made without departing from the spirit or scope of the present invention as defined in the claims.

Claims (129)

1. Administration of a compound at a dose of no more than about 500 micrograms (mcg) once daily (QD) or twice daily (BID), wherein the compound is selected from the group consisting of: And or a pharmaceutically acceptable salt thereof, for the treatment of Ikaros and/or Aiolos mediated cancer. 2. The use according to claim 1, wherein the compound is intended for use in a dose of from about 500 mcg to 1 mcg. 3. Use according to claim 1 or 2, wherein the compound is intended for use over several days with drug holidays between subsequent treatment cycles. 4. Use according to claim 3, wherein the compound is intended for use once or twice a day for at least 13, 14, 15, 16, 17, 18, 19, 20, 21, 22, 23, 24, 25, 26 or 27 consecutive days, followed by a drug holiday until the next cycle of 28 days. 5. Use according to item 3, wherein the compound is intended for use once or twice a day for 21 days, followed by a 7-day rest period. 6. Use according to any of paragraphs. 1-5, wherein the dose is less than or equal to about 400 mcg. 7. Use according to any of paragraphs. 1-5, wherein the dose is less than or equal to about 300 mcg. 8. Use according to any of paragraphs. 1-5, wherein the dose is less than or equal to about 200 mcg. 9. Use according to any of paragraphs. 1-5, wherein the dose is less than or equal to about 100 mcg. 10. Use according to any of paragraphs. 1-5, wherein the dose is less than or equal to about 50 mcg. 11. Use according to any of paragraphs. 1-5, wherein the dose is less than or equal to about 25 mcg. 12. Use according to any of paragraphs. 1-5, wherein the dose is less than or equal to about 10 mcg. 13. Use according to any of paragraphs. 1-5, wherein the dose is less than or equal to about 5 mcg. 14. Use according to any of paragraphs. 1-5, wherein the dose is about 50 mcg. 15. Use according to any of paragraphs. 1-5, wherein the dose is about 25 mcg. 16. Use according to any of paragraphs. 1-5, wherein the dose is about 10 mcg. 17. Use according to any of paragraphs. 1-5, wherein the dose is about 5 mcg. 18. The use according to any one of claims 1-17, wherein the cancer is diffuse large B-cell lymphoma. 19. The use according to claim 18, wherein the diffuse large B-cell lymphoma is activated B-cell lymphoma. 20. The use according to claim 18, wherein the diffuse large B-cell lymphoma is germinal center B-cell lymphoma. 21. The use according to any one of claims 1-17, wherein the cancer is anaplastic large cell lymphoma. 22. The use according to any one of claims 1-17, wherein the cancer is cutaneous T-cell lymphoma. 23. The use according to any one of claims 1-17, wherein the cancer is mantle cell lymphoma. 24. The use according to any one of claims 1-17, wherein the cancer is multiple myeloma. 25. The use according to any one of claims 1-24, wherein the cancer is resistant to treatment with first generation IMiD drugs. 26. The use according to claim 25, wherein the cancer is resistant to treatment with thalidomide. 27. The use according to claim 25, wherein the cancer is resistant to treatment with pomalidomide. 28. The use according to claim 25, wherein the cancer is resistant to treatment with lenalidomide. 29. The use according to claim 25, wherein the cancer is resistant to treatment with iberdomide. 30. Use of a compound selected from the group consisting of: (Compound 1) and (Compound 5) or a pharmaceutically acceptable salt thereof, for the treatment of Ikaros (IKZF1) and/or Aiolos (IKZF3) mediated cancer in a patient, wherein a blood or tissue sample taken from the patient has a statistically lower concentration of one or more biomarkers selected from IRF-1, caspase-3, IL-2 and IFN-γ than in a healthy individual. 31. The use according to claim 30, wherein the statistically lower concentration of the biomarker(s) is 5% lower than the average for a healthy individual. 32. The use according to claim 30, wherein the statistically lower concentration of the biomarker(s) is 20% lower than the average for a healthy individual. 33. Use of a compound selected from the group consisting of: (Compound 1) and (Connection 5) or a pharmaceutically acceptable salt thereof, for the treatment of Ikaros (IKZF1) and/or Aiolos (IKZF3) mediated cancer in a patient, wherein a blood or tissue sample taken from the patient has a statistically higher concentration of one or more biomarkers selected from cyclin D1, E2F1, ZFP91, SALL4, IRF-4, BLIMP1 and MYC than in a healthy individual. 34. The use according to claim 33, wherein the statistically higher concentration of the biomarker(s) is 5% higher than the average for a healthy individual. 35. The use according to claim 33, wherein the statistically higher concentration of the biomarker(s) is 20% higher than the average for a healthy individual. 36. Use of a compound selected from the group consisting of: (Compound 1) and (Connection 5) or a pharmaceutically acceptable salt thereof, for the treatment of Ikaros and/or Aiolos-mediated cancer, wherein the Ikaros or Aiolos-mediated cancer is activated diffuse large B-cell lymphoma or germinal center large B-cell lymphoma. 37. Use of a compound selected from the group consisting of: (Compound 1) and (Connection 5) or a pharmaceutically acceptable salt thereof, for the treatment of Ikaros and/or Aiolos-mediated cancer, in combination with a BTK inhibitor selected from acalabrutinib, spebrutinib, zanubrutinib, LOXO-305, evobrutinib, TG-1701, tolebrutinib, BIIB091, DZD-9008, HZ-A-018, orelabrutinib, AC0058TA, SN1011, rilzabrutinib, ARQ 531, DTRMWXHS-12, JNJ-64264681, branebrutinib and fenebrutinib. 38. Use of a compound selected from the group consisting of: (Compound 1) and (Connection 5) or a pharmaceutically acceptable salt thereof, for the treatment of Ikaros and/or Aiolos mediated cancer, in combination with a proteasome inhibitor selected from ixazomib citrate, oprozomib, delanozomib, lactacystin, epoxomicin, MG132, MG-262, CEP-18770, NEOSH101, TQB3602 and KZR-616. 39. The use according to any one of paragraphs 1-38, wherein the compound is (Compound 1) or a pharmaceutically acceptable salt thereof. 40. The use according to any one of paragraphs 1-29, wherein the compound is (Connection 2) or a pharmaceutically acceptable salt thereof. 41. The use according to any one of paragraphs 1-29, wherein the compound is (Connection 3) or a pharmaceutically acceptable salt thereof. 42. The use according to any one of paragraphs 1-29, wherein the compound is (Connection 4) or a pharmaceutically acceptable salt thereof. 43. The use according to any one of paragraphs 1-38, wherein the compound is (Connection 5) or a pharmaceutically acceptable salt thereof. 44. The use according to any one of paragraphs 1-29, wherein the compound is (Compound 6) or a pharmaceutically acceptable salt thereof. 45. The use according to any one of paragraphs 1-29, wherein the compound is (Compound 7) or a pharmaceutically acceptable salt thereof. 46. The use according to any one of paragraphs 1-29, wherein the compound is (Compound 8) or a pharmaceutically acceptable salt thereof. 47. The use according to any one of paragraphs 1-29, wherein the compound is (Compound 9) or a pharmaceutically acceptable salt thereof. 48. The use according to any one of paragraphs 1-29, wherein the compound is (Compound 10) or a pharmaceutically acceptable salt thereof. 49. The use according to any one of paragraphs 1-29, wherein the compound is (Compound 11) or a pharmaceutically acceptable salt thereof. 50. The use according to any one of paragraphs 1-29, wherein the compound is (Compound 12) or a pharmaceutically acceptable salt thereof. 51. The use according to any one of paragraphs 1-29, wherein the compound is (Compound 13) or a pharmaceutically acceptable salt thereof. 52. The use according to any one of claims 1-51, further comprising the use of a Bruton tyrosine kinase inhibitor. 53. The use according to claim 52, wherein the Bruton tyrosine kinase inhibitor is ibrutinib. 54. Use according to any one of paragraphs 1-53, further comprising the use of a corticosteroid. 55. Use according to claim 54, wherein the corticosteroid is dexamethasone. 56. The use according to any one of claims 1-55, further comprising the use of CAR T-cell therapy. 57. The use according to any one of claims 1-55, further comprising the use of a conjugate of an antibody and a drug. 58. The use according to any one of paragraphs 1-55, further comprising the use of BiTE therapy. 59. The use according to any one of claims 1-55, further comprising the use of a bispecific antibody. 60. The use according to any one of claims 1-55, further comprising the use of a monoclonal antibody. 61. The use according to any one of claims 1 to 55, wherein a BTK inhibitor selected from acalabrutinib, spebrutinib, zanubrutinib, LOXO-305, evobrutinib, TG-1701, tolebrutinib, BIIB091, DZD-9008, HZ-A-018, orelabrutinib, AC0058TA, SN1011, rilzabrutinib, ARQ 531, DTRMWXHS-12, JNJ-64264681, branebrutinib, ibrutinib and fenebrutinib is also used. 62. The use according to any one of claims 1-55, wherein an anti-CD38 antibody selected from felzartamab, daratumumab, GBR 1342, TAK-573, CID-103, OKT10, STI-6129, SGX301, TAK-079 and mesagitamab is also used. 63. The use according to any one of claims 1-55, wherein a proteasome inhibitor selected from ixazomib citrate, oprozomib, delanozomib, lactacystin, bortezomib, carfilzomib, VLX1570, epoxomicin, MG132, MG-262, CEP-18770, NEOSH101, TQB3602 and KZR-616 is also used. 64. Use according to any one of claims 1-55, wherein an IMiD selected from pomalidomide, lenalidomide, thalidomide, iberdomide, CC-92480, CC-90009 and CC-99282 is also used. 65. The use according to any one of claims 1 to 55, wherein an HDAC inhibitor selected from trapoxin B, sodium phenylbutyrate, tacedinalin, mocetinostat, BRD73954, BG45, domatinostat, cay10603, HPOB, TMP269, nexturastat A, santacruzamate A, splitomycin, LMK-235, sodium butyrate, pivaloyloxymethylbutyrate, pyroxamide, abexinostat, resminostat, givinostat, quisinostat, psammapline A, KD5170, 1-alanine chlamidocin, depudecin and CUDC-101 is also used. 66. The use according to any one of claims 1 to 55, wherein a compound selected from selinexor, oxafenamide, belantamab mafodotin, denosumab, zoledronic acid, plerixafor, eltrombopag, ipilimumab, palbociclib, ricolinostat, afuresertib, dinaciclib, filanesib, indatuximab ravtansine, masitinib, sonidegib, sotatercept, ulocuplumab and urelumab is also used. 67. Use according to any one of claims 1 to 66, wherein the compound selected from Compound 1, Compound 2, Compound 3, Compound 4, Compound 5, Compound 6, Compound 7, Compound 8, Compound 9, Compound 10, Compound 11, Compound 12 and Compound 13 or a pharmaceutically acceptable salt thereof, is administered once a day. 68. Use according to any one of claims 1 to 66, wherein the compound selected from Compound 1, Compound 2, Compound 3, Compound 4, Compound 5, Compound 6, Compound 7, Compound 8, Compound 9, Compound 10, Compound 11, Compound 12 and Compound 13 or a pharmaceutically acceptable salt thereof, is administered twice a day. 69. The use according to any one of claims 1-68, wherein the cancer is non-Hodgkin's lymphoma. 70. The use according to any one of claims 1-68, wherein the cancer is multiple myeloma. 71. Use according to any one of claims 1-70, wherein the cancer is recurrent. 72. The use according to any one of claims 1-70, wherein the cancer is refractory to a standard treatment regimen. 73. The use according to any one of claims 1-70, wherein the cancer is recurrent and refractory to a standard treatment regimen. 74. A method for treating cancer mediated by Ikaros (IKZF1) and/or Aiolos (IKZF3), wherein the patient is administered an effective dose of a compound selected from (Compound 1) and (Connection 5) or a pharmaceutically acceptable salt thereof, and then a blood or tissue sample is taken from the patient and the concentration of one or more biomarkers selected from IRF-1, caspase-3, IL-2 and IFN-γ is determined, wherein if the concentration of the biomarker(s) is not increased by at least 1.25 times, then the dose of the compound is increased. 75. The method of claim 74, wherein the dose of the compound is increased if the concentration of the biomarker(s) is not increased by at least 100%. 76. The method of claim 74, wherein the dose of the compound is increased if the concentration of the biomarker(s) is not increased by at least 200%. 77. A method for treating cancer mediated by Ikaros (IKZF1) and/or Aiolos (IKZF3), wherein the patient is administered an effective dose of a compound selected from (Compound 1) and (Connection 5) or a pharmaceutically acceptable salt thereof, and then a blood or tissue sample is taken from the patient and the concentration of one or more biomarkers selected from cyclin D1, E2F1, ZFP91, SALL4, IRF-4, BLIMP1 and MYC is determined, wherein if the concentration of the biomarker(s) is not reduced by at least 1.25 times, then the dose of the compound is increased. 78. The method of claim 77, wherein the dose of the compound is increased if the concentration of the biomarker(s) is not reduced by at least 50%.