Modulators of BCL6 proteolysis and related methods of use thereof

Abstract

The present disclosure relates to bifunctional compounds, their preparation, and the use of these bifunctional compounds in the treatment of a disease or condition caused by the aggregation or accumulation of B cell lymphoma 6 protein in a subject in need thereof.

Description

[0001] Cross - Reference to Related Applications

[0002] This application claims the benefit and priority of U.S. Provisional Application No. 63 / 417,657, filed on October 19, 2022, and U.S. Provisional Application No. 63 / 417,628, filed on October 19, 2022, the contents of each of which are hereby incorporated by reference in their entirety for all purposes. Technical Field

[0003] The present disclosure provides imide - based compounds and related methods of use, the imide - based compounds including bifunctional compounds that comprise imide - based compounds. The bifunctional compounds can be used as regulators of targeted ubiquitination (especially with respect to various polypeptides and other proteins), which regulators are degraded and / or otherwise inhibited by the bifunctional compounds according to the present disclosure. Background Art

[0004] Most small - molecule drugs bind to enzymes or receptors in a tight and well - defined pocket. On the other hand, protein - protein interactions are notoriously difficult to target with small molecules due to the large contact surfaces and shallow grooves or flat interfaces involved in protein - protein interactions. E3 ubiquitin ligases, of which hundreds are known in the human body, confer substrate specificity for ubiquitination and thus represent attractive therapeutic targets. The development of ligands for E3 ligases has proven challenging, in part due to the fact that the ligand must disrupt a protein - protein interaction. However, recent developments have provided specific ligands that bind to these ligases.

[0005] One E3 ubiquitin ligase with therapeutic potential is cereblon. Cereblon is a protein encoded by the CRBN gene in the human body. Thalidomide and its analogs, such as pomalidomide and lenalidomide, are known to bind to cereblon. These agents bind to cereblon, altering the specificity of the complex and thus inducing the ubiquitination and degradation of transcription factors essential for multiple myeloma growth. In fact, higher expression of cereblon is associated with increased efficacy of imide drugs in treating multiple myeloma.

[0006] However, non - specific effects and the inability to fully target and regulate certain classes of proteins, such as transcription factors, remain obstacles in the development of effective anti - cancer agents. Thus, small - molecule therapeutic agents that utilize or enhance the substrate specificity of cereblon and are at the same time "tunable" to specifically target and regulate a wide range of protein classes would be very useful as therapeutic agents. Summary of the Invention

[0007] The present disclosure describes bifunctional compounds for recruiting endogenous proteins to an E3 ubiquitin ligase for degradation and methods of using the same. In particular, the present disclosure provides bifunctional or proteolysis-targeting chimeric compounds that are suitable for use as targeted ubiquitination modulators of a variety of polypeptides and other proteins, which are then degraded and / or inhibited by the bifunctional compounds described herein. Additionally, the specification provides methods of treating or ameliorating a disease condition using an effective amount of a compound described herein, the disease condition such as breast cancer, ovarian cancer, leukemia, lymphoma, benign lymphoma, malignant lymphoma, Burkitt's lymphoma, non-Hodgkin's lymphoma, B-cell non-Hodgkin's lymphoma, sarcoma, Ewing's sarcoma, angiosarcoma, Kaposi's sarcoma, liposarcoma, rhabdomyosarcoma, synovial sarcoma, meningosarcoma, carcinosarcoma, acute myeloid leukemia (AML), acute lymphoblastic leukemia (ALL), T-lineage acute lymphoblastic leukemia (T-ALL), T-lineage lymphoblastic lymphoma (T-LL), peripheral T-cell lymphoma, adult T-cell leukemia, precursor B acute lymphoblastic leukemia, precursor B lymphoma, B-cell lymphoma, large B-cell lymphoma, diffuse large B-cell lymphoma, B-cell acute lymphoblastic leukemia (ALL), Philadelphia chromosome-positive acute lymphoblastic leukemia (ALL), Philadelphia chromosome-positive chronic myeloid leukemia (CML), follicular lymphoma, intravascular large B-cell lymphoma, angioimmunoblastic T-cell lymphoma (AITL), T-cell lymphoma, B-cell leukemia, chronic myeloid leukemia, non-small cell lung cancer, systemic lupus erythematosus (SLE), brain tumor or central nervous system cancer.

[0008] In one aspect, the disease or disorder is a cancer associated with abnormal BCL6 expression or activity.

[0009] In one aspect, the disease or disorder is associated with BCL6 accumulation and aggregation.

[0010] In one aspect, the disease or disorder is a cancer associated with BCL6 accumulation and aggregation.

[0011] In another aspect, the specification provides a method for using a compound according to the present disclosure to identify the effect of degradation of a protein of interest in a biological system.

[0012] In one aspect, the present application relates to a bifunctional compound of formula (I):

[0013]

[0014] or a pharmaceutically acceptable salt, enantiomer, stereoisomer or isotopic derivative thereof, wherein:

[0015] R 1 is H or C1-C6 alkyl;

[0016] Q is

[0017] X is N or CH;

[0018] Y1, Y2 and Y3 are each independently N or CR 3 ;

[0019] Z1 and Z2 are each independently N or CH;

[0020] R 2 is H or C1-C6 alkyl;

[0021] Each R 3 is independently H, halogen, C1-C6 alkyl, C1-C6 haloalkyl, -O-(C1-C6 alkyl), -O-(C1-C6 haloalkyl); and

[0022] wherein the of Q indicates the point of attachment to X or the glutarimide.

[0023] On the one hand, the present application relates to a bifunctional compound of formula (II):

[0024]

[0025] or a pharmaceutically acceptable salt, enantiomer, stereoisomer or isotopic derivative thereof, wherein:

[0026] R 1a is H or halogen;

[0027] R 2a is H or C1-C3 alkyl;

[0028] X 3a is CHR 3a or C(O);

[0029] R 3a is H or C1-C3 alkyl;

[0030] X 4a and X 6a are each independently CH or N; and

[0031] R 5a is H, C1-C3 alkyl or halogen.

[0032] On the one hand, the present application relates to a bifunctional compound of formula (III):

[0033]

[0034] or a pharmaceutically acceptable salt, enantiomer, stereoisomer or isotopic derivative thereof, wherein L is

[0035] X 6b is CHR 6b or C(O);

[0036] R 6b is H or C1-C3 alkyl;

[0037] R 1b 、R 2b 、R 3b and R 4b are each independently H or halogen, wherein at least one of R 1b 、R 2b 、R 3b 、R 4b is halogen;

[0038] R 5b is H or halogen;

[0039] R 6b is H or C1-C3 alkyl;

[0040] X 1b and X 2b are each independently CH or N, wherein at least one of X 1b and X 2b is N; and

[0041] wherein each of L indicates a point of attachment.

[0042] On the one hand, the present application relates to a bifunctional compound of any one of formulas (I)-(III) or a pharmaceutically acceptable salt thereof.

[0043] On the one hand, the present application relates to a bifunctional compound of any one of formulas (I)-(III).

[0044] On the one hand, the present application relates to a bifunctional compound of any one of formulas (I)-(III), wherein the compound is as shown in Table 1, or a pharmaceutically acceptable salt, enantiomer, stereoisomer or isotopic derivative thereof.

[0045] On the one hand, the present application relates to a bifunctional compound of any one of formulas (I)-(III), wherein the compound is as shown in Table 1, or a pharmaceutically acceptable salt thereof.

[0046] On the one hand, the present application relates to a bifunctional compound of any one of formulas (I)-(III), wherein the compound is as shown in Table 1.

[0047] In one embodiment, the present application provides a pharmaceutical composition comprising the bifunctional compound described herein, and one or more pharmaceutically acceptable excipients.

[0048] In one embodiment, the composition is formulated as a tablet and comprises one or more of the following: an emulsifier; a surfactant; a binder; a disintegrant; a glidant; and a lubricant.

[0049] In one embodiment, the composition further comprises an effective amount of at least one additional anti-cancer agent.

[0050] In one embodiment, the present application provides a method for treating cancer in a subject, the method comprising administering to a subject in need thereof a therapeutically effective amount of the bifunctional compound described herein, or a therapeutically effective amount of the pharmaceutical composition described herein.

[0051] In one embodiment, the therapeutically effective amount of the bifunctional compound is orally administered to the subject.

[0052] In one embodiment, the therapeutically effective amount of the bifunctional compound is administered to the subject once a day, twice a day, three times a day, or four times a day.

[0053] In one embodiment, the therapeutically effective amount of the bifunctional compound is administered to the subject once a day.

[0054] In one embodiment, the therapeutically effective amount of the bifunctional compound is administered to the subject all at once, or in two, three, or four divided doses.

[0055] In one embodiment, the therapeutically effective amount of the bifunctional compound is about 1 mg to about 1000 mg.

[0056] In one embodiment, the therapeutically effective amount of the bifunctional compound is about 5 mg to about 750 mg.

[0057] In one embodiment, the therapeutically effective amount of the bifunctional compound is about 10 mg to about 500 mg.

[0058] In one embodiment, the therapeutically effective amount of the bifunctional compound is about 20 mg to about 250 mg.

[0059] In one embodiment, the subject is in a fed state at the time of administration.

[0060] In one embodiment, the subject is in a fasting state at the time of administration.

[0061] In one embodiment, the method further comprises administering to the subject in need thereof an effective amount of at least one additional anti-cancer agent. BRIEF DESCRIPTION OF THE DRAWINGS

[0062] The drawings incorporated in and constituting a part of this specification illustrate several embodiments of the present disclosure and, together with the description, serve to explain the principles of the present disclosure. The drawings are only for the purpose of illustrating embodiments of the present disclosure and should not be construed as limiting the present disclosure. Additional objects, features, and advantages of the present disclosure will become apparent from the following detailed description in conjunction with the drawings, which show illustrative embodiments of the present disclosure, wherein:

[0063] Figure 1A and 1B is a diagram of the general principle of the function of a proteolysis-targeting chimeric compound. Figure 1A represents an exemplary proteolysis-targeting chimeric compound that comprises a protein targeting moiety (PTM; rectangle), a human cereblon E3 ubiquitin ligase binding moiety (CLM; triangle), and a linker moiety (black line) that couples or tethers the PTM to the CLM. Figure 1B Shows the functional use of the proteolysis-targeting chimeric compound as described herein. Briefly, the CLM recognizes and binds to the human cereblon, an E3 ubiquitin ligase, and the PTM binds and recruits an intracellular target protein, bringing it into close proximity to the human cereblon E3 ubiquitin ligase. Typically, the human cereblon E3 ubiquitin ligase complexes with an E2 ubiquitin-conjugated protein and, alone or through the E2 protein, catalyzes the ligation of ubiquitin (dark circles) to lysine on the target protein via an isopeptide bond. The targeted polyubiquitinated protein (far right) is then targeted for degradation by the cell's proteasome machinery. DETAILED DESCRIPTION

[0064] DEFINITIONS

[0065] The term "ubiquitin ligase" refers to a family of proteins that facilitate the transfer of ubiquitin to specific substrate proteins, thereby targeting the substrate proteins for degradation. For example, cerebellin is an E3 ubiquitin ligase protein that, alone or in combination with an E2 ubiquitin-conjugating enzyme, causes ubiquitin to attach to lysine on a target protein and subsequently targets specific protein substrates for degradation by the proteasome. Thus, an E3 ubiquitin ligase, alone or complexed with an E2 ubiquitin-conjugating enzyme, is responsible for transferring ubiquitin to a target protein. Generally, ubiquitin ligases are involved in polyubiquitination such that a second ubiquitin attaches to the first ubiquitin; a third ubiquitin attaches to the second ubiquitin, and so on. Polyubiquitination marks the protein for degradation by the proteasome. However, there are some ubiquitination events that are limited to monoubiquitination, in which the ubiquitin ligase adds only a single ubiquitin to the substrate molecule. Monoubiquitinated proteins are not targeted to the proteasome for degradation but can, for example, alter their cellular location or function by binding to other proteins that have a domain capable of binding ubiquitin. Adding to the complexity, an E3 can target different lysines on ubiquitin to make chains. The most common lysine is Lys48 on the ubiquitin chain. This is the lysine that is recognized by the proteasome for ubiquitin preparation.

[0066] As used herein, the term "compound", "bifunctional compound", or "compound of the present disclosure" refers to the compounds disclosed by the structures in the following tables and examples.

[0067] "Halogen" or "halo group" refers to fluorine (F), chlorine (Cl), bromine (Br), or iodine (I).

[0068] "C1-C6 alkyl" refers to a straight-chain or branched-chain saturated hydrocarbon containing 1-6 carbon atoms. Examples of C1-C6 alkyl include, but are not limited to, methyl, ethyl, propyl, butyl, pentyl, hexyl, isopropyl, isobutyl, sec-butyl, tert-butyl, isopentyl, neopentyl, and isohexyl.

[0069] "C1-C6 haloalkyl" refers to a straight-chain or branched-chain saturated hydrocarbon containing 1-6 carbon atoms that is substituted with one or more halogens. Examples of C1-C6 haloalkyl include, but are not limited to, trifluoromethyl, difluoromethyl, and fluoromethyl.

[0070] As used herein, "pharmaceutically acceptable salts" of the compounds of the present disclosure mean salt forms of the compounds of the present disclosure and hydrates of salt forms in which one or more water molecules are present. Such salt and hydrate forms retain the biological activity of the compounds of the present disclosure and exhibit minimal (if any) toxicological effects in non-biological or other undesirable aspects. Representative "pharmaceutically acceptable salts" include, for example, water-soluble and water-insoluble salts such as acetate, amsonate (4,4-diaminostilbene-2,2-disulfonate), benzenesulfonate, benzoate, bicarbonate, bisulfate, bitartrate, borate, bromide, butyrate, calcium, calcium edetate, camphorsulfonate, carbonate, chloride, citrate, clavulariate, dihydrochloride, edetate, ethanedisulfonate, dodecyl sulfate propionate, ethanesulfonate, fumarate, glucoheptonate, gluconate, glutamate, glycolylarsanilate, hexafluorophosphate, hexylresorcinolate, hydrabamine, hydrobromide, hydrochloride, hydroxynaphthoate, iodide, isothiocyanate, lactate, lactobionate, laurate, magnesium, malate, maleate, mandelate, mesylate, methyl bromide, methyl nitrate, methyl sulfate, mucate, naphthalenesulfonate, nitrate, N-methylglucamine ammonium salt, 3-hydroxy-2-naphthoate, oleate, oxalate, palmitate, pamoate (1,1'-methylene-bis-2-hydroxy-3-naphthoate, einbonate), pantothenate, phosphate / diphosphate, picrate, polygalacturonate, propionate, p-toluenesulfonate, salicylate, stearate, basic acetate, succinate, sulfate, sulfosalicylate, suramate, tannate, tartrate, theoclate, toluenesulfonate, triethyl iodide, and valerate.

[0071] The term "isomers" refers to salts and / or compounds having the same composition and molecular weight but different in physical and / or chemical properties. The structural differences can lie in the composition (geometric isomers) or the ability to rotate the plane of polarized light (stereoisomers). With respect to stereoisomers, the salts of the compounds of the present disclosure can have one or more asymmetric carbon atoms and can exist as racemates, racemic mixtures, and as individual enantiomers or diastereoisomers.

[0072] The compounds of the present disclosure can exist in non-solvated as well as solvated forms such as hydrates.

[0073] "Solvate" means a solvate addition form containing a stoichiometric or non-stoichiometric amount of a solvent. Non-limiting examples of suitable solvates include ethanolates, methanolates, and the like. Some compounds have a tendency to trap a fixed molar ratio of solvent molecules in the crystalline solid state, thereby forming a solvate. If the solvent is water, the solvate formed is a hydrate, and when the solvent is an alcohol, the solvate formed is an alcoholate. Hydrates are formed by the combination of one or more water molecules with one of the substances in which water retains its molecular state as H2O, and such combinations are capable of forming one or more hydrates. In hydrates, the water molecules are connected by intermolecular forces, especially hydrogen bridges, through secondary valences. Solid hydrates contain water as so-called water of crystallization in a stoichiometric ratio, where the water molecules do not have to be equivalent to their bound state. Examples of hydrates are sesquihydrates, monohydrates, dihydrates, or trihydrates. Also suitable are hydrates of salts of the compounds of the present disclosure.

[0074] As mentioned herein, "isotope derivative" relates to a compound of the present disclosure that is isotopically enriched or labeled (with respect to one or more atoms of the compound) with one or more stable isotopes. Thus, in the present application, the compounds of the present disclosure include, for example, compounds that are isotopically enriched or labeled with one or more atoms such as deuterium.

[0075] As used herein, "treatment" describes the management and care of a subject for the purpose of combating a disease, condition, or disorder, and includes reducing or alleviating symptoms or complications, or eliminating the disease, condition, or disorder.

[0076] As used herein, "prevention" describes the stopping of the onset of symptoms or complications of a disease, condition, or disorder.

[0077] "Administer" means introducing an agent, such as a compound of the present disclosure, into a subject. The related terms "administering" and "administration of..." (and grammatical equivalents) both refer to direct administration, which can be administered to a subject by a medical professional or self-administered by the subject; and / or indirect administration, which can be the act of prescribing a drug. For example, a doctor who instructs a patient to self-administer a drug and / or provides a prescription for a drug to the patient administers the drug to the patient.

[0078] The terms "co-administration" and "co-administering" or "combination therapy" refer to simultaneous administration (administering two or more therapeutic agents simultaneously) and time-variable administration (administering one or more therapeutic agents at a different time than one or more additional therapeutic agents), provided that the therapeutic agents are present in the patient to some extent, preferably in an effective amount. In certain preferred aspects, one or more of the compounds of the invention described herein are co-administered in combination with at least one additional bioactive agent (bioactive agents particularly including anti-cancer agents). In particularly preferred aspects, the co-administration of the compounds results in synergistic activity and / or therapy, including anti-cancer activity.

[0079] As used herein, "therapeutically effective amount" means an amount of the free base of a compound of the present disclosure sufficient to treat, ameliorate or prevent a designated disease (e.g., lymphoma), disease symptom, disorder or condition or exhibit a detectable therapeutic or inhibitory effect. The effect can be detected by any assay known in the art. The effective amount for a particular subject may depend on the subject's weight, size and health; the nature and extent of the condition; and whether additional therapeutic agents are to be administered to the subject. The therapeutically effective amount for a given situation can be determined by routine experimentation within the skill and judgment of the clinician.

[0080] As used herein, "C max " refers to the maximum (peak) plasma concentration of a particular compound in a subject observed after administration of a single dose of the particular compound to the subject.

[0081] As used herein, "AUC" refers to the total area under the plasma concentration-time curve, which is a measure of the exposure of the target compound and is the integral of the concentration-time curve after a single dose or at steady state. AUC is expressed in units of ng*H / mL (ng x H / mL), where "H" refers to hours.

[0082] As used herein, "AUC tau " refers to the AUC from 0 hours to the end of the dosing interval.

[0083] "AUC 0-24 " means the AUC from 0 hours to 24 hours after administration of a single dose.

[0084] As used herein, "controlled release" or "CR" with respect to an oral dosage form refers to a compound of the present disclosure released from the dosage form according to a predetermined profile, which may include when and where release occurs after oral administration and / or a specified release rate over a specified time period.

[0085] As used herein, a "controlled release agent" with respect to the oral dosage forms of the present disclosure refers to one or more substances or materials that regulate the release of the compounds of the present disclosure from the dosage form. The controlled release agent can be an organic or inorganic, naturally occurring or synthetic material, such as a polymeric material, a triglyceride, a derivative of a triglyceride, a fatty acid and a fatty acid salt, talc, boric acid, colloidal silica, and combinations thereof.

[0086] As used herein, an "enteric coating" with respect to the dosage forms of the present disclosure refers to a pH-dependent material that surrounds a core containing the compounds of the present disclosure and remains substantially intact in the acidic environment of the stomach but dissolves in the pH environment of the intestine.

[0087] As applied to the CR oral dosage forms described herein, "gastric tolerance" or "GR" means that the release of the compounds of the present disclosure in the stomach of a subject should not exceed 5%, 2.5%, 1%, or 0.5% of the total amount of the compounds of the present disclosure in the dosage form.

[0088] As used herein, an "oral dosage form" refers to a finished pharmaceutical product containing a specific amount (dose) of a compound of the present disclosure or a pharmaceutically acceptable salt and / or solvate thereof as an active ingredient, as well as inactive components (excipients), and the finished product is formulated into a specific configuration suitable for oral administration, such as an oral tablet, a liquid, or a capsule. In one embodiment, the composition is in the form of a tablet that can be scored.

[0089] As used in the present disclosure, the term "carrier" encompasses pharmaceutically acceptable excipients and diluents and refers to a material, composition, or vehicle, such as a liquid or solid filler, diluent, excipient, solvent, or encapsulating material, which is involved in carrying or transporting a pharmaceutical agent from one organ or part of the body to another organ or another part of the body of a subject.

[0090] The term "about" as part of a quantitative expression such as "about X" includes any value that is 10% higher or lower than X, and also includes any numerical value that falls between X - 10% and X + 10%. Thus, for example, a weight of about 40 g includes weights from 36 g to 44 g.

[0091] As used in connection with a particular dosage form, composition, use, method or process described or claimed herein, "comprising" means that the dosage form, composition, use, method or process includes all of the elements recited in the specific description or claim, but does not exclude other elements. "Consists essentially of" and "consisting essentially of" mean that the composition, dosage form, method, use or process described or claimed does not exclude other materials or steps that do not materially affect the physical, pharmacological, pharmacokinetic properties or therapeutic action of the composition, dosage form, method, use or process. "Consists of" and "consisting of" mean the exclusion of other ingredients and substantial method or process steps in excess of trace elements.

[0092] As used herein in connection with a subject, a "fasted condition" or "fasted state" means that the subject has not eaten for at least 4 hours prior to a target time point (e.g., the time of administration of a compound of the present disclosure). In one embodiment, a subject in a fasted state has not eaten for any one of at least 6 hours, 8 hours, 10 hours or 12 hours prior to administration of a compound of the present disclosure.

[0093] As used herein in connection with a subject, a "fed condition" or "fed state" means that the subject has eaten within less than 4 hours prior to a target time point (e.g., the time of administration of a compound of the present disclosure). In one embodiment, a subject in a fed state has eaten within any one of at least 3 hours, 2 hours, 1 hour or 0.5 hours prior to administration of a compound of the present disclosure.

[0094] As used herein, the term "anticancer agent" is used to describe an anticancer agent or a therapeutic agent co-administered with an anticancer agent (e.g., palonosetron), which can be co-administered and / or co-formulated with the compounds of the present disclosure to treat cancer and side effects associated with cancer treatment. These agents include, for example, everolimus, venetoclax, palbociclib, tazemetostat, apelisib, olaparib, MK2206, ibrutinib, acalabrutinib, bendamustine, prednisone, cyclophosphamide, gemcitabine, polatuzumab, upadacitinib, abrocitinib, panobinostat, trabectedin, abraxane, TLK 286, AV-299, DN-101, pazopanib, GSK690693, RTA744, 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, EGFR TK inhibitor, aurora kinase inhibitor, PIK-1 regulator, Bcl-2 inhibitor, HDAC inhibitor, c-MET inhibitor, PARP inhibitor, Cdk inhibitor, EGFRTK inhibitors, IGFR-TK inhibitors, anti-HGF antibodies, PI3 kinase inhibitors, AKT inhibitors, mTORC1 / 2 inhibitors, JAK / STAT inhibitors, checkpoint-1 or 2 inhibitors, focal adhesion kinase inhibitors, Map kinase (mek) inhibitors, VEGF trap antibodies, pemetrexed, erlotinib, dasatinib, nilotinib, dacomitinib, panitumumab, amrubicin, ogavuzumab, Lep-etu, lolatrexed, azd2171, batabulin, ofatumumab, zanolimumab, edotecarin, inverted arch, rubitecan, tesmilifen, oblimumab, ticilimumab, ipilimumab, cottonpol, Bio 111, 131-I-TM-601, ALT-110, BIO 140, CC8490, cilengitide, gimatecan, IL13-PE38QQR, INO 1001, IPdR1KRX-0402, lucanthone, 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, celecoxib (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, anastrozole, exemestane, letrozole, DES (diethylstilbestrol), estradiol, estrogen, conjugated estrogens, bevacizumab, IMC-1 C11, CHIR-258); 3-[5-(methylsulfonylpiperidinylmethyl)-indolyl-quinolone, vatalanibine, AG-013736, AVE-0005, dapoxetine acetate, leuprolide acetate, triptorelin pamoate, medroxyprogesterone acetate, hydroxyprogesterone caproate, megestrol acetate, raloxifene, bicalutamide, flutamide, nilutamide, megestrol acetate, CP-7247 14; TAK-165, HKI-272, erlotinib, lapatinib, canertinib, ABX-EGF antibody, Erbitux, EKB-569, PKI-166, GW-572016, Ionafarnib, BMS-214662, tipifarnib; amifostine, NVP-LAQ824, suberoyl analide hydroxamic acidhydroxamic acid), valproic acid, trichostatin A, FK-228, SU11248, sorafenib, KRN951, aminoglutethimide, arnsacrine, anagrelide, L-asparaginase, Bacillus Calmette-Guérin (BCG) vaccine, doxorubicin, bleomycin, buserelin, busulfan, carboplatin, carmustine, chlorambucil, cisplatin, cladribine, clodronate, cyproterone, cytarabine, dacarbazine, actinomycin, daunorubicin, diethylstilbestrol, epirubicin, fludarabine, fludrocortisone, fluoxymesterone, flutamide, Gleevec, gemcitabine, hydroxyurea, idarubicin, ifosfamide, imatinib, leuprorelin, levamisole, lomustine, mechlorethamine, melphalan, 6-mercaptopurine, mesna, methotrexate, mitomycin, mitotane, mitoxantrone, nilutamide, octreotide, oxaliplatin, pamidronate disodium, pentostatin, plicamycin, porfimer sodium, procarbazine, raltitrexed, rituximab, streptozocin, teniposide, testosterone, thalidomide, thioguanine, thiotepa, tretinoin, vindesine, 13-cis-retinoic acid, phenylalanine mustard, uracil mustard, estramustine, altretamine, floxuridine, 5-deoxyuridine, cytarabine, 6-mercaptopurine, deoxycoformycin, calcitriol, valrubicin, mithramycin, vinblastine, vinorelbine, topotecan, razoxane, marimastat, COL-3, canertinib, BMS-275291, squalamine, endostatin, SU5416, SU6668, EMD121974, interleukin-12, IM862, endostatin, vitaxin, droloxifene, idoxyfene, spironolactone, finasteride, cimetidine, trastuzumab, denileukin diftitox, gefitinib, bortezimib, paclitaxel, Cremophor-free paclitaxel, docetaxel, epothilone B, BMS-247550, BMS-310705, droloxifene, 4-hydroxytamoxifen, pipendoxifene, ERA-923, azoxifene, fulvestrant, acolbifene, lasofoxifene, idoxyfene, TSE-424, HMR-3339, ZK186619, topotecan, PTK787 / ZK 222584, VX-745, PD184352, 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, darbepoetin, erythropoietin, granulocyte colony-stimulating factor, zolendronate, prednisone, cetuximab, granulocyte macrophage colony-stimulating factor, histrelin, pegylated interferon α-2a, interferon α-2a, pegylated interferon α-2b, interferon α-2b, azacitidine, PEG-L-asparaginase, lenalidomide, gemtuzumab, hydrocortisone, interleukin-11, dexrazoxane, alemtuzumab, all-trans retinoic acid, ketoconazole, interleukin-2, megestrol acetate, immunoglobulin, mechlorethamine, methylprednisolone, ibritumomab tiuxetan, androgen, decitabine, altretamine, bexarotene, tositumomab, arsenic trioxide, cortisone, editronate, mitotane, cyclosporine, liposomal daunorubicin, Edwina-asparaginase, strontium 89, casopitant, netupitant, NK-1 receptor antagonist, palonosetron, aprepitant, diphenhydramine, hydroxyzine, metoclopramide, lorazepam, alprazolam, haloperidol, droperidol, dronabinol, dexamethasone, methylprednisolone, prochlorperazine, granisetron, ondansetron, dolasetron, tropisetron, pegylated filgrastim, erythropoietin, darbepoetin alfa, darbepoetin α, and mixtures thereof.

[0095] In one embodiment, the anti-cancer agent is selected from the group consisting of temozolomide, capecitabine, irinotecan, tamoxifen, anastrozole, exemestane, letrozole, DES, estradiol, estrogen, bevacizumab, goserelin acetate, leuprorelin acetate, triptorelin pamoate, medroxyprogesterone acetate, hydroxyprogesterone caproate, raloxifene, megestrol acetate, carboplatin, cisplatin, dacarbazine, methotrexate, vinblastine, vinorelbine, topotecan, finasteride, asoxifene, fulvestrant, prednisone, abiraterone, enzalutamide, apalutamide, darolutamide, sipuleucel-T, pembrolizumab, nivolumab, cemiplimab, atezolizumab (Tecentriq), avelumab (Bavencio), durvalumab (Imfinzi), docetaxel (Taxotere), cabazitaxel (Jevtana), mitoxantrone (Novantrone), estramustine (Emcyt), docetaxel, ketoconazole, histrelin, triptorelin, buserelin, cyproterone, flutamide, bicalutamide, nilutamide, pamidronate, and zolendronate.

[0096] As used in this disclosure, the articles "a" or "an" refer to one or more than one (i.e., at least one) of the grammatical objects of the article. By way of example, "an element" means one element or more than one element.

[0097] Unless otherwise indicated, in this disclosure, the term "and / or" is used to mean "and" or "or".

[0098] The terms "patient" and "subject" are used interchangeably herein and refer to a mammal, such as a human, mouse, rat, guinea pig, dog, cat, horse, cow, pig, or non-human primate, such as a monkey, chimpanzee, baboon, or rhesus monkey.

[0099] In one embodiment, the subject is a human.

[0100] In one embodiment, the subject is a human who has been diagnosed with multiple myeloma.

[0101] In one embodiment, the subject is a human who has been diagnosed with lymphoma.

[0102] In one embodiment, the subject is a human who has been diagnosed with any of the following: B-cell non-Hodgkin lymphoma, large B-cell lymphoma, Burkitt lymphoma, follicular lymphoma, intravascular large B-cell lymphoma, B-cell leukemia, B-cell acute lymphoblastic leukemia, chronic myeloid leukemia, or non-small cell lung cancer.

[0103] The compounds of the present disclosure

[0104] In one aspect, the present invention relates to bifunctional or multifunctional compounds for modulating protein activity by inducing the degradation of a target protein. In some embodiments, the bifunctional compound comprises an E3 ubiquitin ligase-binding moiety and a protein targeting moiety, preferably linked by a linker moiety, as further described herein, wherein the E3 ubiquitin ligase-binding moiety is conjugated to the protein targeting moiety, and wherein the E3 ubiquitin ligase-binding moiety recognizes a ubiquitin pathway protein (e.g., a ubiquitin ligase, preferably an E3 ubiquitin ligase) and the protein targeting moiety recognizes a target protein such that when the target protein is in the vicinity of the ubiquitin ligase, the target protein will be degraded, thereby resulting in the degradation / inhibition of the action of the target protein and the control of protein levels. In certain embodiments, the bifunctional compound comprises a CLM and a PTM conjugated (e.g., covalently, directly, or indirectly) to a chemical linker L, and the bifunctional compound can be described as:

[0105] PTM-L-CLM

[0106] CLM recognizes and binds to the E3 ubiquitin ligase human cerebellin protein. The PTM is the small molecule protein-binding moiety that binds and recruits an intracellular target protein or polypeptide, bringing it closer to CLM for degradation of the target protein, thereby causing ubiquitination of the target protein. In certain embodiments, the PTM is the B cell lymphoma 6 protein (BCL6) targeting moiety.

[0107] On the one hand, for the compounds described herein, the PTM comprises the following chemical structure:

[0108]

[0109] wherein the of the PTM indicates the point of attachment to the L.

[0110] In some embodiments, for the compounds described herein, the L comprises the following chemical structure:

[0111]

[0112]

[0113] wherein the of the L indicates the point of attachment to the PTM or CLM.

[0114] In some embodiments of the compounds described herein, the CLM comprises the following chemical structure:

[0115] wherein the of the CLM indicates the point of attachment to the L.

[0116] On the one hand, the present application relates to a bifunctional compound of formula (I):

[0117]

[0118] or a pharmaceutically acceptable salt, enantiomer, stereoisomer or isotopic derivative thereof, wherein:

[0119] R 1 is H or C1-C6 alkyl;

[0120] Q is

[0121] X is N or CH;

[0122] Y1, Y2 and Y3 are each independently N or CR 3 ;

[0123] Z1 and Z2 are each independently N or CH;

[0124] R 2 is H or a C1-C6 alkyl group;

[0125] Each R 3 is independently H, halogen, a C1-C6 alkyl group, a C1-C6 haloalkyl group, -O-(C1-C6 alkyl), -O-(C1-C6 haloalkyl); and

[0126] wherein the indicates the point of attachment to X or the glutarimide.

[0127] In one aspect, the present application relates to a bifunctional compound of formula (I):

[0128]

[0129] or a pharmaceutically acceptable salt thereof, wherein:

[0130] R 1 is H or a C1-C6 alkyl group;

[0131] Q is

[0132] X is N or CH;

[0133] Y1, Y2 and Y3 are each independently N or CR 3 ;

[0134] Z1 and Z2 are each independently N or CH;

[0135] R 2 is H or a C1-C6 alkyl group;

[0136] Each R 3 is independently H, halogen, a C1-C6 alkyl group, a C1-C6 haloalkyl group, -O-(C1-C6 alkyl), -O-(C1-C6 haloalkyl); and

[0137] wherein the indicates the point of attachment to X or the glutarimide.

[0138] In one aspect, the present application relates to a bifunctional compound of formula (I):

[0139]

[0140] wherein:

[0141] R 1 is H or a C1-C6 alkyl group;

[0142] Q is

[0143] X is N or CH;

[0144] Y1, Y2, and Y3 are each independently N or CR 3 ;

[0145] Z1 and Z2 are each independently N or CH;

[0146] R 2 is H or C1-C6 alkyl;

[0147] Each R 3 is independently H, halogen, C1-C6 alkyl, C1-C6 haloalkyl, -O-(C1-C6 alkyl), -O-(C1-C6 haloalkyl); and

[0148] wherein the designation of Q indicates the point of attachment to X or the glutarimide.

[0149] In some embodiments, R 1 is hydrogen. In some embodiments, R 1 is methyl.

[0150] In some embodiments, Q is

[0151] In some embodiments, Q is

[0152] In some embodiments, Q is

[0153] In some embodiments, Q is

[0154] In some embodiments, Q is

[0155] In some embodiments, Q is

[0156] In some embodiments, Q is

[0157] In some embodiments, each of Y1, Y2, and Y3 is CH.

[0158] In some embodiments, one of Y1, Y2, and Y3 is N, and the other two of Y1, Y2, or Y3 are CH. In some embodiments, Y1 is N, and Y2 and Y3 are CH. In some embodiments, Y2 is N, and Y1 and Y3 are CH. In some embodiments, Y3 is N, and Y1 and Y2 are CH.

[0159] In some embodiments, two of Y1, Y2, and Y3 are N, and the other one of Y1, Y2, or Y3 is CH. In some embodiments, Y1 and Y2 are N, and Y3 is CH. In some embodiments, Y1 and Y3 are N, and Y2 is CH. In some embodiments, Y2 and Y3 are N, and Y1 is CH.

[0160] In some embodiments, each of Z1 and Z2 is N.

[0161] In some embodiments, each of Z1 and Z2 is CH.

[0162] In some embodiments, Z1 is N, and Z2 is CH.

[0163] In some embodiments, Z1 is CH, and Z2 is N.

[0164] In some embodiments, X is N. In some embodiments, X is CH.

[0165] In some embodiments, R 2 is H, methyl, ethyl, or isopropyl.

[0166] In some embodiments, R 2 is H.

[0167] In some embodiments, R 2 is methyl, ethyl, or isopropyl.

[0168] In some embodiments, R 2 is methyl.

[0169] In some embodiments, R 2 is ethyl.

[0170] In some embodiments, R 2 is isopropyl.

[0171] In some embodiments, each R 3 is independently hydrogen, methyl, fluoro, or methoxy.

[0172] In one aspect, the present application relates to a bifunctional compound of formula (II):

[0173]

[0174] or a pharmaceutically acceptable salt, enantiomer, stereoisomer, or isotopic derivative thereof, wherein:

[0175] R 1a is H or halogen; R 2a is H or C1-C3 alkyl;

[0176] X 3ais CHR 3a or C(O);

[0177] R 3a is H or C1-C3 alkyl;

[0178] X 4a and X 6a are each independently CH or N; and

[0179] R 5a is H, C1-C3 alkyl or halogen.

[0180] On the one hand, the present application relates to a bifunctional compound of formula (II):

[0181]

[0182] or a pharmaceutically acceptable salt thereof, wherein:

[0183] R 1a is H or halogen;

[0184] R 2a is H or C1-C3 alkyl;

[0185] X 3a is CHR 3a or C(O);

[0186] R 3a is H or C1-C3 alkyl;

[0187] X 4a and X 6a are each independently CH or N; and

[0188] R 5a is H, C1-C3 alkyl or halogen.

[0189] On the one hand, the present application relates to a bifunctional compound of formula (II):

[0190]

[0191] wherein:

[0192] R 1a is H or halogen;

[0193] R 2a is H or C1-C3 alkyl;

[0194] X 3a is CHR 3a or C(O);

[0195] R 3a is H or C1-C3 alkyl;

[0196] X 4a and X 6a are each independently CH or N; and

[0197] R 5a is H, C1-C3 alkyl or halogen.

[0198] In some embodiments, the compound of formula (II) is a compound of formula (II-a):

[0199]

[0200] or a pharmaceutically acceptable salt thereof.

[0201] In some embodiments, the compound of formula (II) is a compound of formula (II-a).

[0202] In some embodiments, the compound of formula (II) is a compound of formula (II-b), formula (II-c), formula (II-d), formula (II-e), formula (II-f), formula (II-g), formula (II-h), formula (II-i) or formula (II-j):

[0203]

[0204]

[0205]

[0206]

[0207] or a pharmaceutically acceptable salt thereof.

[0208] In some embodiments, the compound of formula (II) is a compound of formula (II-b), formula (II-c), formula (II-d), formula (II-e), formula (II-f), formula (II-g), formula (II-h), formula (II-i) or formula (II-j).

[0209] In some embodiments, R 1a is H. In some embodiments, R 1a is halogen.

[0210] In some embodiments, R 1a is fluorine. In some embodiments, R 1a is Cl.

[0211] In some embodiments, R 2a is H. In some embodiments, R 2a is C1-C3 alkyl. In some embodiments, R 2ais H or CH3. In some embodiments, R 2a is CH3.

[0212] In some embodiments, X 3a is CHR 3a . In some embodiments, X 3a is C(O). In some embodiments, X 3a is CH2 or C(O). In some embodiments, X 3a is CH2. In some embodiments, X 3a is CH(CH3).

[0213] In some embodiments, R 3a is H. In some embodiments, R 3a is C1-C3 alkyl. In some embodiments, R 3a is CH3.

[0214] In some embodiments, R 1a is F, X 4a and X 6a are each N, and R 3a and R 5a are each CH3.

[0215] In some embodiments, R 1a is Cl, X 4a and X 6a are each N, and R 3a and R 5a are each CH3.

[0216] In some embodiments, at least one of X 4a and X 6a is N. In some embodiments, X 4a is N. In some embodiments, X 4a is C. In some embodiments, X 6a is N. In some embodiments, X 6a is C.

[0217] In some embodiments, X 4a and X 6a are each N, and R 2a is CH3.

[0218] In some embodiments, R 5a is H. In some embodiments, R 5a is H, CH3 or halogen. In some embodiments, R 5a is CH3 or halogen. In some embodiments, R 5a is H, CH3 or F. In some embodiments, R 5ais CH3 or F. In some embodiments, R 5a is CH3. In some embodiments, R 5a is a halogen. In some embodiments, R 5a is F.

[0219] On the one hand, the present application relates to a bifunctional compound of formula (III):

[0220]

[0221] or a pharmaceutically acceptable salt, enantiomer, stereoisomer or isotopic derivative thereof, wherein:

[0222] L is

[0223] X 6b is CHR 6b or C(O);

[0224] R 6b is H or C1-C3 alkyl;

[0225] R 1b 、R 2b 、R 3b and R 4b are each independently H or a halogen, wherein at least one of R 1b 、R 2b 、R 3b 、R 4b is a halogen;

[0226] R 5b is H or a halogen;

[0227] R 6b is H or C1-C3 alkyl;

[0228] X 1b and X 2b are each independently CH or N, wherein at least one of X 1b and X 2b is N; and

[0229] wherein each of L indicates a point connection.

[0230] On the one hand, the present application relates to a bifunctional compound of formula (III):

[0231]

[0232] or a pharmaceutically acceptable salt thereof, wherein:

[0233] L is

[0234] X 6b is CHR 6b or C(O);

[0235] R 6b is H or C1-C3 alkyl;

[0236] R 1b 、R 2b 、R 3b and R 4b are each independently H or halogen, where at least one of R 1b 、R 2b 、R 3b 、R 4b is halogen;

[0237] R 5b is H or halogen;

[0238] R 6b is H or C1-C3 alkyl;

[0239] X 1b and X 2b are each independently CH or N, where at least one of X 1b and X 2b is N; and

[0240] where each of L's indicates a point connection.

[0241] On the one hand, the present application relates to a bifunctional compound of formula (III):

[0242]

[0243] wherein:

[0244] L is

[0245] X 6b is CHR 6b or C(O);

[0246] R 6b is H or C1-C3 alkyl;

[0247] R 1b 、R 2b 、R 3b and R 4b are each independently H or halogen, where at least one of R 1b 、R 2b 、R 3b 、R 4b is halogen;

[0248] R 5bis H or a halogen;

[0249] R 6b is H or a C1-C3 alkyl;

[0250] X 1b and X 2b are each independently CH or N, where at least one of X 1b and X 2b is N; and

[0251] where each of L indicates a point connection.

[0252] In some embodiments, R 5b is H. In some embodiments, R 5b is a halogen. In some embodiments, R 5b is F.

[0253] In some embodiments, L is In some embodiments, L is In some embodiments, L is In some embodiments, L is In some embodiments, L is In some embodiments, L is In some embodiments, L is In some embodiments, L is In some embodiments, L is In some embodiments, L is

[0254] In some embodiments, L is

[0255] In some embodiments, X 6b is CHR 3a . In some embodiments, X 6b is C(O). In some embodiments, X 6b is CH2 or C(O). In some embodiments, X 6b is CH2. In some embodiments, X 6b is CH(CH3).

[0256] In some embodiments, R 5b is F, and X 6b is CH.

[0257] In some embodiments, at least two of R 1b 、R 2b 、R 3b 、R 4b are halogens. In some embodiments, R 1b, R 2b , R 3b , R 4b Exactly two of them are halogens. In some embodiments, R 1b , R 2b , R 3b , R 4b At least two of them are halogens. In some embodiments, R 1b , R 2b , R 3b , R 4b Exactly one of them is a halogen.

[0258] In some embodiments, R 1b , R 2b , R 3b , R 4b At least two of them are F. In some embodiments, R 1b , R 2b , R 3b , R 4b Exactly two of them are F. In some embodiments, R 1b , R 2b , R 3b , R 4b At least two of them are F. In some embodiments, R 1b , R 2b , R 3b , R 4b Exactly two of them are F.

[0259] In some embodiments, R 6b is H. In some embodiments, R 6b is a C1-C3 alkyl group. In some embodiments, R 6b is CH3.

[0260] In some embodiments, X 1b and X 2b At least one of them is N. In some embodiments, X 1b is N. In some embodiments, X 1b is CH. In some embodiments, X 2b is N. In some embodiments, X 2b is CH.

[0261] In some embodiments, L is

[0262] In some embodiments, L is In some embodiments, L is In some embodiments, L is

[0263] In some embodiments, L is

[0264] In some embodiments, L is

[0265] In some embodiments, L is

[0266] In some embodiments, L is

[0267] In some embodiments, L is

[0268] On the other hand, the present application relates to a compound, or a pharmaceutically acceptable salt, enantiomer, stereoisomer, solvate or isotopic derivative thereof, wherein the compound is as shown in Table 1.

[0269]

[0270]

[0271]

[0272]

[0273]

[0274]

[0275]

[0276]

[0277]

[0278]

[0279]

[0280]

[0281]

[0282]

[0283]

[0284]

[0285]

[0286]

[0287]

[0288] On the other hand, the present application relates to a compound, or a pharmaceutically acceptable salt thereof, wherein the compound is as shown in Table 1.

[0289] On the other hand, the present application relates to a compound, wherein the compound is as shown in Table 1.

[0290] On the other hand, the present application relates to a compound, or a pharmaceutically acceptable salt, enantiomer, stereoisomer, solvate or isotopic derivative thereof, wherein the compound is as shown in Table 2.

[0291]

[0292]

[0293]

[0294]

[0295]

[0296]

[0297]

[0298]

[0299]

[0300]

[0301]

[0302]

[0303]

[0304]

[0305] On the other hand, the present application relates to a compound, or a pharmaceutically acceptable salt thereof, wherein the compound is as shown in Table 2.

[0306] On the other hand, the present application relates to a compound, wherein the compound is as shown in Table 2.

[0307] On the other hand, the present application relates to a compound, or a pharmaceutically acceptable salt, enantiomer, stereoisomer, solvate or isotopic derivative thereof, wherein the compound is as shown in Table 3.

[0308]

[0309]

[0310]

[0311]

[0312]

[0313]

[0314] On the other hand, the present application relates to a compound, or a pharmaceutically acceptable salt thereof, wherein the compound is as shown in Table 3.

[0315] On the other hand, the present application relates to a compound, wherein the compound is as shown in Table 3.

[0316] The compounds of the present disclosure can be synthesized using standard synthetic methods and procedures for the preparation of organic molecules and the transformation and manipulation of functional groups, including the use of protecting groups, as can be obtained from relevant scientific literature or standard reference textbooks in the art according to the present disclosure. Although not limited to any one or more sources, recognized reference textbooks on organic synthesis include: Smith, M.B.; March, *March's Advanced Organic Chemistry: Reactions, Mechanisms, and Structure*, 5th Edition; John Wiley & Sons: New York, 2001; and Greene, T.W.; Wuts, P.G.M. *Protective Groups in Organic Synthesis*, 3rd Edition; John Wiley & Sons: New York, 1999. The synthetic methods described in International Applications No. PCT / US2020 / 056145 and No. PCT / US2022 / 025041 are incorporated herein by reference in their entirety.

[0317] In one embodiment, the compounds of the present disclosure can be prepared according to the procedures and methods disclosed herein. Other bifunctional compounds of the present disclosure can be prepared from common intermediates or their derivatives using similar methods.

[0318] General synthetic routes for preparing the compounds of the present disclosure

[0319] Scheme 1

[0320]

[0321] The compound of formula I' (commercially available or easily preparable) can react with the compound of formula II' (also commercially available or easily preparable) in a solvent such as DMSO or DMF with a base such as triethylamine or DIEA and be heated to produce the compound of formula III'. In this case, X on compound II' can be a leaving group such as a halogen, and Q6 and Q7 are such that the selective substitution shown here is favorable. Non-limiting examples are those where X = Cl and both Q6 and Q7 are N. The compound of formula III' can be heated in the presence of a base (such as DIEA) in a solvent (such as DMSO) and react with the compound of formula IV to produce the PROTAC of formula V TM . The compound of formula IV' is an advanced building block, where a part of ULM, the linker, and PTM form a complete subunit. Where represents a 4-8 membered ring amine or spirocyclic amine of any 2-ring combination selected from 4,4, 4,5, 4,6, 5,4, 5,5, 5,6, 6,4, 6,5, and 6,6, and optionally includes a second nitrogen atom (N) if there are more than 2 carbons between them. L' can be a bond, a linker, or a part of a linker

[0322] Scheme 2

[0323]

[0324] The compound of formula I' in Scheme 1 can be prepared using the procedures found and / or adapted in Kerres et al., 2017, Cell Reports 20, 2860 - 2875, and as shown in Scheme 2. When G1 is NO2, the compound of formula VI' can be dissolved in a solvent (such as DMF), treated with a base (such as but not limited to K2CO3), and alkylated with R PTM1 -X' alkyl. In this case, X' can be a leaving group such as but not limited to iodine or bromine. Generally, R PTM1 -X' is commercially available or easily preparable. Alternatively, R PTM1The boronic acid analogues can be linked to the compound of formula VI' using the Chan-Lam coupling reaction (Chen et al., 2020, *Advanced Synthesis and Catalysis* 62(16), 3311 - 3331), where the boronic acid and the compound of formula VI are combined with a copper salt (such as Cu(OAc)2), a base (such as Na2CO3) in a solvent (such as DCE), and heated. In this case, it is preferred that G1 = H, and nitration as shown in the third step of Scheme 2 is carried out using KNO3 under acidic conditions. Those skilled in the art will realize that when alkylating 5-nitroisatin (VI', where G1 = NO2) with R PTM1 -X', the nitration step is skipped as the compound of formula VIII' is directly produced. The compound of formula VIII can react with TMS-diazomethane under basic conditions (see Duplantier et al., 2009, *J. Med. Chem.* 52, 3576–3585 and references cited therein) to obtain the ring-expanded compound of formula IX'. The hydroxyl group of the compound of formula X' can be deprotected by treating the compound of formula IX' with BBR3. The compound of formula I can be obtained in 2 additional steps by alkylation of the hydroxyl group of X' with 2-haloacetamide, followed by reduction of the nitro group. Those skilled in the art can use various methods to achieve nitro reduction.

[0325] Method for ubiquitinating / degrading a target protein in a cell

[0326] The present disclosure provides a method for ubiquitinating / degrading a target protein in a cell. The method comprises administering a bifunctional composition comprising an E3 ubiquitin ligase binding moiety and a protein targeting moiety, preferably linked by a linker moiety, as further described herein, wherein the E3 ubiquitin ligase binding moiety is conjugated to the protein targeting moiety, and wherein the E3 ubiquitin ligase binding moiety recognizes a ubiquitin pathway protein (e.g., a ubiquitin ligase, preferably an E3 ubiquitin ligase) and the protein targeting moiety recognizes the target protein such that when the target protein is in the vicinity of the ubiquitin ligase, the target protein will be degraded, thereby resulting in the effect of degrading / inhibiting the target protein and controlling the protein level. The control of the protein level provided by the present disclosure provides a treatment for a disease state or condition that is regulated by the target protein by reducing the level of the protein in the patient's cells.

[0327] In one embodiment, the present disclosure relates to a method of treating a patient having a disease state or condition that is regulated by a protein, wherein degradation of the protein would provide a therapeutic benefit to the patient, the method comprising administering to the patient in need thereof an effective amount of a compound of any one of formulas (I-III) disclosed herein or a pharmaceutically acceptable salt, enantiomer, stereoisomer, solvate or isotopic derivative thereof, optionally in combination with another anti-cancer agent. The disease state or condition can be a disease caused by a microbial agent or other exogenous agent (such as a virus, bacterium, fungus, protozoan or other microbe), or can be a disease state caused by overexpression of a protein, which results in the disease state and / or condition.

[0328] Treatment method

[0329] In one aspect, the present application relates to a method of treating and / or preventing cancer, the method comprising administering to a subject in need thereof a therapeutically effective amount of a compound of the present disclosure, or a pharmaceutically acceptable salt, enantiomer, stereoisomer, solvate or isotopic derivative thereof.

[0330] In one aspect, the present application relates to a method of treating and / or preventing cancer, the method comprising administering to a subject in need thereof a therapeutically effective amount of a combination of a compound of the present disclosure or a pharmaceutically acceptable salt, enantiomer, stereoisomer, solvate, polymorph or isotopic derivative thereof with one or more additional anti-cancer agents.

[0331] In one aspect, the cancer to be treated or prevented is breast cancer, ovarian cancer, leukemia, lymphoma, benign lymphoma, malignant lymphoma, Burkitt's lymphoma, non-Hodgkin's lymphoma, B-cell non-Hodgkin's lymphoma, sarcoma, Ewing's sarcoma, angiosarcoma, Kaposi's sarcoma, liposarcoma, rhabdomyosarcoma, synovial sarcoma, meningeal sarcoma, carcinosarcoma, acute myeloid leukemia (AML), acute lymphoblastic leukemia (ALL), T-lineage acute lymphoblastic leukemia (T-ALL), T-lineage lymphoblastic lymphoma (T-LL), peripheral T-cell lymphoma, adult T-cell leukemia, precursor B acute lymphoblastic leukemia, precursor B lymphoma, B-cell lymphoma, large B-cell lymphoma, diffuse large B-cell lymphoma, B-cell acute lymphoblastic leukemia (ALL), Philadelphia chromosome-positive acute lymphoblastic leukemia (ALL), Philadelphia chromosome-positive chronic myeloid leukemia (CML), follicular lymphoma, intravascular large B-cell lymphoma, angioimmunoblastic T-cell lymphoma (AITL), T-cell lymphoma, B-cell leukemia, chronic myeloid leukemia, non-small cell lung cancer, systemic lupus erythematosus (SLE), brain tumor or central nervous system cancer.

[0332] On the one hand, the cancer to be treated or prevented is large B-cell lymphoma, diffuse large B-cell lymphoma, Burkitt lymphoma, follicular lymphoma, or angioimmunoblastic T-cell lymphoma (AITL).

[0333] The methods for treating cancer described herein result in a reduction in tumor size. Alternatively or additionally, the cancer is metastatic cancer, and the treatment method includes inhibiting the invasion of metastatic cancer cells.

[0334] On the one hand, the present application relates to a method for treating cancer, the method comprising administering to a subject in need thereof a therapeutically effective amount of a bifunctional compound of the present disclosure, i.e., a compound of any of formulas (I)-(III) or Tables 1-3 as defined herein, or a pharmaceutically acceptable salt, enantiomer, stereoisomer, solvate, or isotopic derivative thereof.

[0335] On the one hand, the present application relates to a method for treating cancer, the method comprising administering to a subject in need thereof a therapeutically effective amount of a bifunctional compound of the present disclosure, i.e., a compound of any of formulas (I)-(III) or Tables 1-3 as defined herein, or a pharmaceutically acceptable salt thereof.

[0336] On the one hand, the present application relates to a method for treating cancer, the method comprising administering to a subject in need thereof a therapeutically effective amount of a bifunctional compound of the present disclosure, i.e., a compound of any of formulas (I)-(III) or Tables 1-3 as defined herein.

[0337] On the one hand, the present application relates to a method for treating cancer, the method comprising administering to a subject in need thereof a therapeutically effective amount of a bifunctional compound of the present disclosure, i.e., any compound of formula (I) as defined herein, or a pharmaceutically acceptable salt, enantiomer, stereoisomer, solvate, or isotopic derivative thereof.

[0338] On the one hand, the present application relates to a method for treating cancer, the method comprising administering to a subject in need thereof a therapeutically effective amount of a bifunctional compound of the present disclosure, i.e., any compound of formula (I) as defined herein, or a pharmaceutically acceptable salt thereof.

[0339] On the one hand, the present application relates to a method for treating cancer, the method comprising administering to a subject in need thereof a therapeutically effective amount of a bifunctional compound of the present disclosure, i.e., any compound of formula (I) as defined herein.

[0340] On the one hand, the present application relates to a method for treating cancer, the method comprising administering to a subject in need thereof a therapeutically effective amount of a bifunctional compound of the present disclosure, namely, any compound of formula (II) as defined herein, or a pharmaceutically acceptable salt, enantiomer, stereoisomer, solvate or isotopic derivative thereof.

[0341] On the one hand, the present application relates to a method for treating cancer, the method comprising administering to a subject in need thereof a therapeutically effective amount of a bifunctional compound of the present disclosure, namely, any compound of formula (II) as defined herein, or a pharmaceutically acceptable salt thereof.

[0342] On the one hand, the present application relates to a method for treating cancer, the method comprising administering to a subject in need thereof a therapeutically effective amount of a bifunctional compound of the present disclosure, namely, any compound of formula (II) as defined herein.

[0343] On the one hand, the present application relates to a method for treating cancer, the method comprising administering to a subject in need thereof a therapeutically effective amount of a bifunctional compound of the present disclosure, namely, any compound of formula (III) as defined herein, or a pharmaceutically acceptable salt, enantiomer, stereoisomer, solvate or isotopic derivative thereof.

[0344] On the one hand, the present application relates to a method for treating cancer, the method comprising administering to a subject in need thereof a therapeutically effective amount of a bifunctional compound of the present disclosure, namely, any compound of formula (III) as defined herein, or a pharmaceutically acceptable salt thereof.

[0345] On the one hand, the present application relates to a method for treating cancer, the method comprising administering to a subject in need thereof a therapeutically effective amount of a bifunctional compound of the present disclosure, namely, any compound of formula (III) as defined herein.

[0346] On the one hand, the present application relates to a method for treating cancer, the method comprising administering to a subject in need thereof a therapeutically effective amount of a bifunctional compound selected from any one of Tables 1-3 or a pharmaceutically acceptable salt, enantiomer, stereoisomer, solvate or isotopic derivative thereof.

[0347] On the one hand, the present application relates to a method for treating cancer, the method comprising administering to a subject in need thereof a therapeutically effective amount of a bifunctional compound selected from any one of Tables 1-3 or a pharmaceutically acceptable salt thereof.

[0348] On the one hand, the present application relates to a method for treating cancer, the method comprising administering to a subject in need thereof a therapeutically effective amount of a bifunctional compound selected from any one of Tables 1-3.

[0349] On the one hand, the present application relates to a method for treating cancer, the method comprising administering to a subject in need thereof a therapeutically effective amount of a bifunctional compound selected from Table 1 or a pharmaceutically acceptable salt, enantiomer, stereoisomer, solvate or isotopic derivative thereof.

[0350] On the one hand, the present application relates to a method for treating cancer, the method comprising administering to a subject in need thereof a therapeutically effective amount of a bifunctional compound selected from Table 1 or a pharmaceutically acceptable salt thereof.

[0351] On the one hand, the present application relates to a method for treating cancer, the method comprising administering to a subject in need thereof a therapeutically effective amount of a bifunctional compound selected from Table 1.

[0352] On the one hand, the present application relates to a method for treating cancer, the method comprising administering to a subject in need thereof a therapeutically effective amount of a bifunctional compound selected from Table 2 or a pharmaceutically acceptable salt, enantiomer, stereoisomer, solvate or isotopic derivative thereof.

[0353] On the one hand, the present application relates to a method for treating cancer, the method comprising administering to a subject in need thereof a therapeutically effective amount of a bifunctional compound selected from Table 2 or a pharmaceutically acceptable salt thereof.

[0354] On the one hand, the present application relates to a method for treating cancer, the method comprising administering to a subject in need thereof a therapeutically effective amount of a bifunctional compound selected from Table 2.

[0355] On the one hand, the present application relates to a method for treating cancer, the method comprising administering to a subject in need thereof a therapeutically effective amount of a bifunctional compound selected from Table 3 or a pharmaceutically acceptable salt, enantiomer, stereoisomer, solvate or isotopic derivative thereof.

[0356] On the one hand, the present application relates to a method for treating cancer, the method comprising administering to a subject in need thereof a therapeutically effective amount of a bifunctional compound selected from Table 3 or a pharmaceutically acceptable salt thereof.

[0357] On the one hand, the present application relates to a method for treating cancer, the method comprising administering to a subject in need thereof a therapeutically effective amount of a bifunctional compound selected from Table 3.

[0358] On the one hand, the present application relates to treating cancer with a combination of a compound of the present disclosure and another anti-cancer agent. In one embodiment, the cancer treated with the combination of a compound of the present disclosure and another anti-cancer agent is lymphoma.

[0359] In one embodiment, the cancer treated with the combination of a compound of the present disclosure and another anti-cancer agent is multiple myeloma. In one aspect, treating the cancer results in a reduction in tumor size. A reduction in tumor size may also be referred to as "tumor regression". Preferably, after treatment, the tumor size is reduced by 5% or more relative to its size before treatment; more preferably, the tumor size is reduced by 10% or more; more preferably, by 20% or more; more preferably, by 30% or more; more preferably, by 40% or more; even more preferably, by 50% or more; and most preferably, by more than 75% or more. The tumor size can be measured by any reproducible measurement means. In a preferred aspect, the tumor size can be measured as the diameter of the tumor.

[0360] In another aspect, treating the cancer results in a reduction in tumor volume. Preferably, after treatment, the tumor volume is reduced by 5% or more relative to its volume before treatment; more preferably, the tumor volume is reduced by 10% or more; more preferably, by 20% or more; more preferably, by 30% or more; more preferably, by 40% or more; even more preferably, by 50% or more; and most preferably, by more than 75% or more. The tumor volume can be measured by any reproducible measurement means.

[0361] In another aspect, treating the cancer results in a reduction in the number of tumors. Preferably, after treatment, the number of tumors is reduced by 5% or more relative to the number before treatment; more preferably, the number of tumors is reduced by 10% or more; more preferably, by 20% or more; more preferably, by 30% or more; more preferably, by 40% or more; even more preferably, by 50% or more; and most preferably, by more than 75%. The number of tumors can be measured by any reproducible measurement means. In a preferred aspect, the number of tumors can be measured by counting the tumors visible to the naked eye or visible at a specified magnification. In a preferred aspect, the specified magnification is 2x, 3x, 4x, 5x, 10x or 50x.

[0362] In another aspect, treating the cancer results in a reduction in the number of metastatic lesions in other tissues or organs away from the primary tumor site. Preferably, after treatment, the number of metastatic lesions is reduced by 5% or more relative to the number before treatment; more preferably, the number of metastatic lesions is reduced by 10% or more; more preferably, by 20% or more; more preferably, by 30% or more; more preferably, by 40% or more; even more preferably, by 50% or more; and most preferably, by more than 75%. The number of metastatic lesions can be measured by any reproducible measurement means. In a preferred aspect, the number of metastatic lesions can be measured by counting the metastatic lesions visible to the naked eye or visible at a specified magnification. In a preferred aspect, the specified magnification is 2x, 3x, 4x, 5x, 10x or 50x.

[0363] On the other hand, treating cancer increases the average survival time of the treated subject population compared to the population receiving only the vehicle. Preferably, the average survival time is increased by more than 30 days; more preferably, by more than 60 days; more preferably, by more than 90 days; and most preferably, by more than 120 days. The increase in the average survival time of the population can be measured by any reproducible means. In a preferred aspect, the increase in the average survival time of the population can be measured, for example, by calculating the average survival length of the population after starting treatment with the active agent or compound of the present disclosure. In another preferred aspect, the increase in the average survival time of the population can also be measured, for example, by calculating the average survival length of the population after completing the first round of treatment with the active agent or compound of the present disclosure.

[0364] On the other hand, treating cancer results in an increase in the average survival time of the treated subject population compared to the population of untreated subjects. Preferably, the average survival time is increased by more than 30 days; more preferably, by more than 60 days; more preferably, by more than 90 days; and most preferably, by more than 120 days. The increase in the average survival time of the population can be measured by any reproducible means. In a preferred aspect, the increase in the average survival time of the population can be measured by calculating the average survival length of the population after starting treatment with the active agent or compound of the present disclosure. In another preferred aspect, the increase in the average survival time of the population can be measured by calculating the average survival length of the population after completing the first round of treatment with the compound of the present disclosure.

[0365] On the other hand, treating cancer reduces the tumor growth rate. Preferably, after treatment, the tumor growth rate is reduced by at least 5% relative to the growth rate before treatment; more preferably, the tumor growth rate is reduced by at least 10%; more preferably, by at least 20%; more preferably, by at least 30%; more preferably, by at least 40%; more preferably, by at least 50%; even more preferably, by at least 50%; and most preferably, by at least 75%. The tumor growth rate can be measured by any reproducible measurement means. In a preferred aspect, the tumor growth rate is measured based on the change in tumor diameter per unit time.

[0366] On the other hand, treating cancer results in a reduction of tumor regrowth. Preferably, after treatment, the tumor regrowth is less than 5%; more preferably, less than 10%; more preferably, less than 20%; more preferably, less than 30%; more preferably, less than 40%; more preferably, less than 50%; even more preferably, less than 50%; and most preferably, less than 75%. Tumor regrowth can be measured by any reproducible measurement means. In a preferred aspect, tumor regrowth is measured by measuring the increase in the diameter of the tumor after the previous tumor shrinkage after treatment. In another preferred aspect, the reduction of tumor regrowth is indicated by the failure of the tumor to recur after treatment is stopped.

[0367] The dosage of the disclosed compounds for any of the methods and uses described herein will vary depending upon the agent, the age, weight and clinical condition of the recipient subject, as well as the experience and judgment of the clinician or practitioner administering the therapy, and other factors that influence the chosen dosage.

[0368] A therapeutically effective amount of the disclosed compounds can be administered one or more times a day for up to 30 days or more, followed by one or more days without administration of the disclosed compounds. This type of treatment schedule, i.e., administering the disclosed compounds for several consecutive days followed by several consecutive days without administering the compounds, can be referred to as one treatment cycle. The treatment cycle can be repeated as many times as needed to achieve the desired effect.

[0369] In one embodiment, a therapeutically effective amount of the compounds of the present disclosure is 0.01, 0.05, 0.1, 0.2, 0.3, 0.4, 0.5, 0.6, 0.7, 0.8, 0.9, 1, 1.5, 2, 2.5, 3, 3.5, 4, 4.5, 5, 5.5, 6, 6.5, 7, 7.5, 8, 8.5, 9, 9.5, 10, 11, 12, 13, 14, 15, 16, 17, 18, 19, 20, 21, 22, 23, 24, 25, 26, 27, 28, 29, 30, 31, 32, 33, 34, 35, 36, 37, 38, 39, 40, 41, 42, 43, 44, 45, 46, 47, 48, 49, 50, 51, 52, 53, 54, 55, 56, 57, 58, 59, 60, 61, 62, 63, 64, 65, 66, 67, 68, 69, 70, 71, 72, 73, 74, 75, 76, 77, 78, 79, 80, 81, 82, 83, 84, 85, 86, 87, 88, 89, 90, 91, 92, 93, 94, 95, 96, 97, 98, 99, 100, 105, 110, 115, 120, 125, 130, 135, 140, 145, 150, 155, 160, 165, 170, 175, 180, 185, 190, 195, 200, 205, 210, 215, 220, 225, 230, 235, 240, 245, 250, 255, 260, 265, 270, 275, 280, 285, 290, 295, 300, 305, 310, 315, 320, 325, 330, 335, 340, 345, 350, 355, 360, 365, 370, 375, 380, 385, 390, 395, 400, 405, 410, 415, 420, 425, 430, 435, 440, 445, 450, 455, 460, 465, 470, 475, 480, 485, 490, 495, 500, 505, 510, 515, 520, 525, 530, 535, 540, 545, 550, 555, 560, 565, 570, 575, 580, 585, 590, 595, 600, 605, 610, 615, 620, 625, 630, 635, 640, 645, 650, 655, 660, 665, 670, 675, 680, 685, 690, 695, 700, 705, 710, 715, 720, 725, 730, 735, 740, 745, 750, 755, 760, 765, 770, 775, 780, 785, 790, 795, 800, 805, 810, 815, 820, 825, 830, 835, 840, 845, 850, 855,860, 865, 870, 875, 880, 885, 890, 895, 900, 905, 910, 915, 920, 925, 930, 935, 940, 945, 950, 955, 960, 965, 970, 975, 980, 985, 990, 995 or 1,000 mg, administered once, twice, three times, four times or more times per day, or once, twice, three times, four times or more times per day in the form of a single or divided dose, for one day, two days, three days, four days, five days, six days, seven days, eight days, nine days, ten days, eleven days, twelve days, thirteen days, fourteen days, fifteen days, twenty days, twenty-five days, thirty days continuously, or for 2 months, 3 months, 4 months, 5 months, 6 months or longer.

[0370] In one embodiment, a therapeutically effective amount of the compounds of the present disclosure is about 10 to about 40 mg, about 20 to about 50 mg, about 30 to about 60 mg, about 40 to about 70 mg, about 50 to about 80 mg, about 60 to about 90 mg, about 70 to about 100 mg, about 80 to about 110 mg, about 90 to about 120 mg, about 100 to about 130 mg, about 110 to about 140 mg, about 120 to about 150 mg, about 130 to about 160 mg, about 140 to about 170 mg, about 150 to about 180 mg, about 160 to about 190 mg, about 170 to about 200 mg, about 180 to about 210 mg, about 190 to about 220 mg, about 200 to about 230 mg, about 210 to about 240 mg, about 220 to about 250 mg, about 230 to about 260 mg, about 240 to about 270 mg, about 250 to about 280 mg, about 260 to about 290 mg, about 270 to about 300 mg, about 280 to about 310 mg, about 290 to about 320 mg, about 300 to about 330 mg, about 310 to about 340 mg, about 320 to about 350 mg, about 330 to about 360 mg, about 340 to about 370 mg, about 350 to about 380 mg, about 360 to about 390 mg, about 370 to about 400 mg, about 380 to about 410 mg, about 390 to about 420 mg, about 400 to about 430 mg, about 410 to about 440 mg, about 420 to about 450 mg, about 430 to about 460 mg, about 440 to about 470 mg, about 450 to about 480 mg, about 460 to about 490 mg, about 470 to about 500 mg, about 480 to about 510 mg, about 490 to about 520 mg, about 500 to about 530 mg, about 510 to about 540 mg, about 520 to about 550 mg, about 530 to about 560 mg, about 540 to about 570 mg, about 550 to about 580 mg, about 560 to about 590 mg, about 570 to about 600 mg, about 580 to about 610 mg, about 590 to about 620 mg, about 600 to about 630 mg, about 610 to about 640 mg, about 620 to about 650 mg, about 630 to about 660 mg, about 640 to about 670 mg, about 650 to about 680 mg, about 660 to about 690 mg, about 670 to about 700 mg, about 680 to about 710 mg, about 690 to about 720 mg, about 700 to about 730 mg, about 710 to about 740 mg, about 720 to about 750 mg, about 730 to about 760 mg, about 740 to about 770 mg, about 750 to about 780 mg, about 760 to about 790 mg, about 770 to about 800 mg, about 780 to about 810 mg, about 790 to about 820 mg, about 800 to about 830 mg, about 810 to about 840 mg, about 820 to about 850 mg,from about 830 to about 860 mg, from about 840 to about 870 mg, from about 850 to about 880 mg, from about 860 to about 890 mg, from about 870 to about 900 mg, from about 880 to about 910 mg, from about 890 to about 920 mg, from about 900 to about 930 mg, from about 910 to about 940 mg, from about 920 to about 950 mg, from about 930 to about 960 mg, from about 940 to about 970 mg, from about 950 to about 980 mg, from about 960 to about 990 mg or from about 970 to about 1,000 mg, administered once, twice, three times, four times or more times per day in a single dose or divided doses (the dose can be adjusted according to the patient's body weight (kg), body surface area (m, 2 ) and / or age (years)).

[0371] In one embodiment, a therapeutically effective amount of the compound of the present disclosure is from about 70 mg to about 1,000 mg, administered once, twice, three times, four times or more times per day in a single dose or divided doses (the dose can be adjusted according to the patient's body weight (kg), body surface area (m 2 ) and / or age (years)).

[0372] A therapeutically effective amount of the compound of the present disclosure can also be in the range of about 0.01 mg / kg / day to about 100 mg / kg / day. In one aspect, a therapeutically effective amount of the compound of the present disclosure can be in the range of about 0.05 mg / kg / day to about 10 mg / kg / day. In one aspect, a therapeutically effective amount of the compound of the present disclosure can be in the range of about 0.075 mg / kg / day to about 5 mg / kg / day. In one aspect, a therapeutically effective amount of the compound of the present disclosure can be in the range of about 0.10 mg / kg / day to about 1 mg / kg / day. In one aspect, a therapeutically effective amount of the compound of the present disclosure can be in the range of about 0.20 mg / kg / day to about 0.70 mg / kg / day.

[0373] In one embodiment, a therapeutically effective amount of the compound of the present disclosure is about 0.10 mg / kg / day, about 0.15 mg / kg / day, about 0.20 mg / kg / day, about 0.25 mg / kg / day, about 0.30 mg / kg / day, about 0.35 mg / kg / day, about 0.40 mg / kg / day, about 0.45 mg / kg / day, about 0.50 mg / kg / day, about 0.55 mg / kg / day, about 0.60 mg / kg / day, about 0.65 mg / kg / day, about 0.70 mg / kg / day, about 0.75 mg / kg / day, about 0.80 mg / kg / day, about 0.85 mg / kg / day, about 0.90 mg / kg / day, about 0.95 mg / kg / day or about 1.00 mg / kg / day.

[0374] In one embodiment, a therapeutically effective amount of the compounds of the present disclosure is about 1.05 mg / kg / day, about 1.10 mg / kg / day, about 1.15 mg / kg / day, about 1.20 mg / kg / day, about 1.25 mg / kg / day, about 1.30 mg / kg / day, about 1.35 mg / kg / day, about 1.40 mg / kg / day, about 1.45 mg / kg / day, about 1.50 mg / kg / day, about 1.55 mg / kg / day, about 1.60 mg / kg / day, about 1.65 mg / kg / day, about 1.70 mg / kg / day, about 1.75 mg / kg / day, about 1.80 mg / kg / day, about 1.85 mg / kg / day, about 1.90 mg / kg / day, about 1.95 mg / kg / day or about 2.00 mg / kg / day.

[0375] In one embodiment, a therapeutically effective amount of the compounds of the present disclosure is about 2 mg / kg / day, about 2.5 mg / kg / day, about 3 mg / kg / day, about 3.5 mg / kg / day, about 4 mg / kg / day, about 4.5 mg / kg / day, about 5 mg / kg / day, about 5.5 mg / kg / day, about 6 mg / kg / day, about 6.5 mg / kg / day, about 7 mg / kg / day, about 7.5 mg / kg / day, about 8.0 mg / kg / day, about 8.5 mg / kg / day, about 9.0 mg / kg / day, about 9.5 mg / kg / day or about 10 mg / kg / day.

[0376] In one embodiment, a therapeutically effective amount of the compounds of the present disclosure is administered to a subject once a day. In one embodiment, the daily dose of the compounds of the present disclosure is administered to the subject in its entirety as a single dose. In one embodiment, the daily dose of the compounds of the present disclosure is administered to the subject in two parts (i.e., divided doses). In one embodiment, the daily dose of the compounds of the present disclosure is administered to the subject in three divided doses. In one embodiment, the daily dose of the compounds of the present disclosure is administered to the subject in four divided doses. In one embodiment, the daily dose of the compounds of the present disclosure is administered to the subject in five or more divided doses. In one embodiment, these parts or divided doses are administered to the subject at regular intervals throughout the day, such as every 12 hours, every 8 hours, every 6 hours, every 5 hours, every 4 hours, etc.

[0377] A therapeutically effective amount of the compounds of the present disclosure can be initially estimated in cell culture assays or in animal models, typically in rats, mice, rabbits, dogs or pigs. Animal models can also be used to determine appropriate concentration ranges and routes of administration. Then, such information can be used to determine the doses and routes useful for administration in humans. Therapeutic / preventive efficacy and toxicity can be determined by standard pharmaceutical procedures in cell cultures or experimental animals, e.g., ED50 (The dose effective for treatment in 50% of the population) and LD 50 (The dose lethal to 50% of the population). The dose ratio between toxicity and therapeutic effect is the therapeutic index, and it can be expressed as the ratio LD 50 / ED 50 . Pharmaceutical compositions exhibiting a large therapeutic index are preferred. The dose can vary within this range depending on the dosage form employed, the sensitivity of the patient, and the route of administration.

[0378] Adjust the dose and administration to provide an adequate level of the compounds of the present disclosure or to maintain the desired effect. Factors to be considered include the severity of the disease state, the general health of the subject, the age, weight, and sex of the subject, diet, the time and frequency of administration, drug combinations, response sensitivity, and tolerance / response to the therapy. Long-acting pharmaceutical compositions can be administered every 3 to 4 days, weekly, biweekly, or monthly, depending on the half-life and clearance rate of the particular formulation.

[0379] In one embodiment, for a method of treating cancer with a combination of a compound of the present disclosure and a therapeutically effective amount of at least one additional anti-cancer agent, the therapeutically effective amount of the compound of the present disclosure is described herein, and the therapeutically effective amount of at least one additional anti-cancer agent is 0.01, 0.05, 0.1, 0.2, 0.3, 0.4, 0.5, 0.6, 0.7, 0.8, 0.9, 1, 1.5, 2, 2.5, 3, 3.5, 4, 4.5, 5, 5.5, 6, 6.5, 7, 7.5, 8, 8.5, 9, 9.5, 10, 11, 12, 13, 14, 15, 16, 17, 18, 19, 20, 21, 22, 23, 24, 25, 26, 27, 28, 29, 30, 31, 32, 33, 34, 35, 36, 37, 38, 39, 40, 41, 42, 43, 44, 45, 46, 47, 48, 49, 50, 51, 52, 53, 54, 55, 56, 57, 58, 59, 60, 61, 62, 63, 64, 65, 66, 67, 68, 69, 70, 71, 72, 73, 74, 75, 76, 77, 78, 79, 80, 81, 82, 83, 84, 85, 86, 87, 88, 89, 90, 91, 92, 93, 94, 95, 96, 97, 98, 99, 100, 105, 110, 115, 120, 125, 130, 135, 140, 145, 150, 155, 160, 165, 170, 175, 180, 185, 190, 195, 200, 205, 210, 215, 220, 225, 230, 235, 240, 245, 250, 255, 260, 265, 270, 275, 280, 285, 290, 295, 300, 305, 310, 315, 320, 325, 330, 335, 340, 345, 350, 355, 360, 365, 370, 375, 380, 385, 390, 395, 400, 405, 410, 415, 420, 425, 430, 435, 440, 445, 450, 455, 460, 465, 470, 475, 480, 485, 490, 495, 500, 505, 510, 515, 520, 525, 530, 535, 540, 545, 550, 555, 560, 565, 570, 575, 580, 585, 590, 595, 600, 605, 610, 615, 620, 625, 630, 635, 640, 645, 650, 655, 660, 665, 670, 675, 680, 685, 690, 695, 700, 705, 710, 715, 720, 725, 730, 735, 740, 745, 750, 755, 760, 765, 770, 775, 780,785, 790, 795, 800, 805, 810, 815, 820, 825, 830, 835, 840, 845, 850, 855, 860, 865, 870, 875, 880, 885, 890, 895, 900, 905, 910, 915, 920, 925, 930, 935, 940, 945, 950, 955, 960, 965, 970, 975, 980, 985, 990, 995 or 1,000 mg, administered once, twice, three times, four times or more times per day for one, two, three, four, five, six, seven, eight, nine, ten, eleven, twelve, thirteen, fourteen, fifteen or thirty days in a row, or administered once, twice, three times, four times or more times per day in the form of a single or divided dose, for 2, 3, 4, 5, 6 months or longer.

[0380] In one embodiment, for a method of treating cancer with a combination of a compound of the present disclosure and at least one additional anti-cancer agent, a therapeutically effective amount of the at least one additional anti-cancer agent is orally administered once per day in a single or divided dose for one, two, three, four, five, six, seven, eight, nine, ten, eleven, twelve, thirteen, fourteen, fifteen, twenty, twenty-five, thirty days or more in a row. In one embodiment, the combination of a compound of the present disclosure and at least one additional anti-cancer agent is administered to a subject in need thereof in a fasting state. In one embodiment, the subject does not eat for at least two hours before and at least one hour after administration of the combination of a compound of the present disclosure and at least one additional anti-cancer agent.

[0381] In one embodiment, the compound of the present disclosure and at least one additional anti-cancer agent are administered to the subject simultaneously. In one embodiment, the compound of the present disclosure and at least one additional anti-cancer agent are administered to the subject sequentially.

[0382] In one embodiment, the compound of the present disclosure and at least one additional anti-cancer agent are administered to the subject in temporal proximity.

[0383] In some embodiments, "temporal proximity" means that the administration of the compound of the present disclosure occurs within a period of time before or after the administration of the at least one additional anti-cancer agent such that the therapeutic effect of the compound of the present disclosure overlaps with the therapeutic effect of the at least one additional anti-cancer agent. In some embodiments, the therapeutic effect of the compound of the present disclosure completely overlaps with the therapeutic effect of the at least one additional anti-cancer agent. In some embodiments, "temporal proximity" means that the administration of the compound of the present disclosure occurs within a period of time before or after the administration of the at least one additional anti-cancer agent such that there is a synergistic effect between the compound of the present disclosure and the at least one additional anti-cancer agent.

[0384] "Time proximity" can vary according to various factors, including but not limited to the age, sex, weight, genetic background, medical condition, disease history, and treatment history of the subject to whom the therapeutic agent will be administered; the disease or condition to be treated or improved; the therapeutic outcome to be achieved; the dose, dosing frequency, and duration of administration of the therapeutic agent; the pharmacokinetics and pharmacodynamics of the therapeutic agent; and the route of administration of the therapeutic agent. In some embodiments, "time proximity" means within 15 minutes, within 30 minutes, within one hour, within two hours, within four hours, within six hours, within eight hours, within 12 hours, within 18 hours, within 24 hours, within 36 hours, within two days, within three days, within four days, within five days, within six days, within one week, within two weeks, within three weeks, within four weeks, within six weeks, or within eight weeks. In some embodiments, multiple administrations of one therapeutic agent can occur in time proximity to a single administration of another therapeutic agent. In some embodiments, time proximity can change during a treatment cycle or within a dosing regimen.

[0385] Pharmaceutical composition

[0386] In one embodiment, the compounds of the present disclosure are formulated for oral administration. For example, in one embodiment, the compounds of the present disclosure are formulated as tablets, which comprise zero, one, two, or more of each of the following: an emulsifier; a surfactant; a binder; a disintegrant; a glidant; and a lubricant.

[0387] In one embodiment, the emulsifier is hydroxypropyl methylcellulose.

[0388] In one embodiment, the surfactant is vitamin E polyethylene glycol succinate.

[0389] In one embodiment, the binder (also referred to herein as a filler) is selected from the group consisting of microcrystalline cellulose, lactose monohydrate, sucrose, glucose, and sorbitol.

[0390] In one embodiment, the disintegrant is sodium carboxymethylcellulose cross-linked.

[0391] In one embodiment, a glidant is a substance used to promote powder flow by reducing interparticle adhesion. In one embodiment, in the dosage forms of the present disclosure, glidants are selected from the group consisting of silica, anhydrous colloidal silica, starch, and talc.

[0392] In one embodiment, a lubricant is a substance that prevents ingredients from adhering and / or aggregating together in the machines used to prepare the dosage forms of the present disclosure. In one embodiment, in the dosage forms of the present disclosure, lubricants are selected from the group consisting of magnesium stearate, sodium stearyl fumarate, stearic acid, and vegetable stearin.

[0393] A pharmaceutical composition containing the compound of the present disclosure can be prepared in a generally known manner, for example, by means of conventional mixing, dissolving, granulating, sugar coating, milling, emulsifying, encapsulating, entrapping or lyophilization processes. The pharmaceutical composition can be formulated in a conventional manner using one or more pharmaceutically acceptable carriers, which include excipients and / or auxiliaries that contribute to processing the compound of the present disclosure into a pharmaceutically usable formulation. Of course, the appropriate formulation depends on the selected route of administration.

[0394] Pharmaceutical compositions suitable for injectable use include sterile aqueous solutions (in the case of water solubility) or dispersions and sterile powders for the extemporaneous preparation of sterile injectable solutions or dispersions. For intravenous administration, suitable carriers include physiological saline, bacteriostatic water, polyoxyethylene castor oil (Cremophor EL) (BASF, Parsippany, N.J.) TM (BASF, Parsippany, N.J.) or phosphate buffered saline (PBS). In all cases, the composition must be sterile and should be fluid to the extent that easy injectability exists. It must be stable under the conditions of manufacture and storage and must be protected against the contaminating action of microorganisms such as bacteria and fungi. The carrier can be a solvent or a dispersion medium containing, for example, water, ethanol, polyols (e.g., glycerol, propylene glycol, and liquid polyethylene glycol, etc.) and suitable mixtures thereof. Appropriate fluidity can be maintained, for example, by using coatings such as lecithin, by maintaining the required particle size in the case of dispersions, and by using surfactants. Prevention of the action of microorganisms can be achieved by various antibacterial and antifungal agents, for example, parabens, chlorobutanol, phenol, ascorbic acid, thimerosal, etc. In many cases, it will be preferable to include isotonic agents in the composition, such as sugars, polyols (e.g., mannitol, sorbitol), sodium chloride. Prolonged absorption of injectable compositions can be achieved by including agents that delay absorption, such as aluminum monostearate and gelatin.

[0395] A sterile injectable solution can be prepared by incorporating the compound of the present disclosure in an appropriate solvent in the required amount, together with one or a combination of the ingredients listed above, and, if necessary, subsequently filtering for sterilization. Generally, dispersions are prepared by incorporating the active agent or compound into a sterile vehicle that contains a basic dispersion medium and the required other ingredients from those listed above. In the case of sterile powders for the preparation of sterile injectable solutions, the methods of preparation are vacuum drying and freeze drying, which yield a powder of the active ingredient plus any additional desired ingredients from its previously sterile filtered solution.

[0396] Oral compositions generally include an inert diluent or a pharmaceutically acceptable edible carrier. They can be enclosed in gelatin capsules or compressed into tablets. For purposes of oral therapeutic administration, the compounds of the present disclosure can be incorporated with excipients and used in the form of tablets, lozenges, or capsules. Oral compositions can also be prepared with a liquid carrier for use as a mouthwash, where the medicament or compound in the liquid carrier is orally administered and swished and spat out or swallowed. Pharmaceutically compatible binders and / or adjuvant materials can be included as part of the composition. Tablets, pills, capsules, lozenges, etc. can contain any of the following ingredients or compounds of similar nature: binders such as microcrystalline cellulose, acacia, or gelatin; excipients such as starch or lactose; disintegrants such as alginic acid, Primogel, or corn starch; lubricants such as magnesium stearate; glidants such as colloidal silicon dioxide; sweetening agents such as sucrose or saccharin; or flavoring agents such as peppermint, methyl salicylate, or orange flavoring.

[0397] For administration by inhalation, the medicament or compound is delivered in the form of an aerosol spray from a pressurized container or dispenser containing a suitable propellant (e.g., a gas such as carbon dioxide, or a nebulizer).

[0398] Systemic administration can also be effected by transmucosal or transdermal means. For transmucosal or transdermal administration, permeants suitable for the barrier to be permeated are used in the formulation. Such permeants are generally known in the art and include, for example, detergents, bile salts, and fusidic acid derivatives for transmucosal administration. Transmucosal administration can be achieved by using a nasal spray or a suppository. For transdermal administration, the active medicament or compound is formulated as an ointment, paste, gel, or cream as is commonly known in the art.

[0399] In one aspect, the compounds of the present disclosure are prepared with a pharmaceutically acceptable carrier that will protect the medicament or compound from rapid elimination from the body, such as a controlled-release formulation, including implants and microencapsulation delivery systems. Biodegradable, biocompatible polymers such as ethylene vinyl acetate, polyanhydrides, polyglycolic acid, collagen, polyorthoesters, and polylactic acid can be used. Methods for preparing such formulations will be apparent to those skilled in the art. Such materials are also commercially available from Alza Corporation and Nova Pharmaceuticals, Inc. Liposome suspensions (including liposomes targeted to infected cells with monoclonal antibodies against viral antigens) can also be used as pharmaceutically acceptable carriers. These can be prepared according to methods known to those skilled in the art, for example, as described in U.S. Patent No. 4,522,811.

[0400] Particularly advantageous is the formulation of oral or parenteral compositions in unit dosage form for ease of administration and uniform dosing. As used herein, a unit dosage form refers to a physically discrete unit suitable as a unit dose for a subject to be treated; each unit contains a predetermined quantity of the active agent or compound, the predetermined quantity being calculated to produce the desired therapeutic effect associated with the required pharmaceutical carrier. The specifications of the unit dosage forms of the present application are indicated by the unique properties of the compounds of the present disclosure and the particular therapeutic effect to be achieved and depend directly on said properties and the particular therapeutic effect to be achieved.

[0401] The pharmaceutical composition may be included in a container, package or dispenser together with instructions for administration.

[0402] Exemplary modes of administration of the compounds of the present disclosure include systemic or topical administration, such as oral, nasal, parenteral, transdermal, subcutaneous, vaginal, buccal, rectal or topical modes of administration. In one embodiment, the compounds of the present disclosure are administered orally. In one embodiment, the compounds of the present disclosure are administered as tablets, capsules, caplets, solutions, suspensions, syrups, granules, beads, powders or pellets.

[0403] Exemplary pharmaceutical compositions are tablets and gelatin capsules which contain a salt of a compound of the present disclosure and a pharmaceutically acceptable carrier, such as a) diluents, for example, purified water, triglyceride oils such as hydrogenated or partially hydrogenated vegetable oils, or mixtures thereof, corn oil, olive oil, sunflower oil, safflower oil, fish oils such as EPA or DHA, or esters or triglycerides thereof or mixtures thereof, omega-3 fatty acids or derivatives thereof, lactose, glucose, sucrose, mannitol, sorbitol, cellulose, sodium, saccharin, glucose and / or glycine; b) lubricants, for example, silica, talc, stearic acid, its magnesium or calcium salts, sodium oleate, sodium stearate, magnesium stearate, sodium benzoate, sodium acetate, sodium chloride and / or polyethylene glycol; also applicable to tablets; c) binders, for example, magnesium aluminum silicate, starch paste, gelatin, tragacanth, methylcellulose, sodium carboxymethylcellulose, magnesium carbonate, natural sugars such as glucose or beta-lactose, corn sweeteners, natural and synthetic gums such as gum arabic, tragacanth or sodium alginate, waxes and / or polyvinylpyrrolidone, if desired; d) disintegrants, for example, starch, agar, methylcellulose, bentonite, xanthan gum, alginic acid or its sodium salt, or effervescent mixtures; e) absorbents, colorants, flavorants and sweeteners; f) emulsifiers or dispersants, such as Tween 80, Labrasol, HPMC, DOSS, caproyl909, labrafac, labrafil, peceol, transcutol, capmul MCM, capmulPG-12, captex 355, gelucire, vitamin E TGPS or other acceptable emulsifiers; and / or g) reagents for enhancing salt absorption, such as cyclodextrin, hydroxypropyl-cyclodextrin, PEG400 and / or PEG200.

[0404] To prepare pharmaceutical compositions from the compounds or salts or hydrates of the present disclosure, the inert pharmaceutically acceptable carrier can be solid or liquid. Solid form preparations include powders, tablets, dispersible granules, capsules, cachets and suppositories. Powders and tablets may contain from about 5 to about 95% of the active ingredient. Suitable solid carriers are known in the art, for example, magnesium carbonate, magnesium stearate, talc, sugar or lactose. Tablets, powders, cachets and capsules can be used as solid dosage forms suitable for oral administration. Examples of pharmaceutically acceptable carriers and methods of preparation for various compositions can be found in: A. Gennaro (editor), Remington's Pharmaceutical Sciences, 18th edition, (1990), Mack Publishing Co., Easton, Pa.

[0405] Solid form preparations include solutions, suspensions and emulsions. For example, water or water-propylene glycol solutions for parenteral injection or for adding sweeteners and opacifiers to oral solutions, suspensions and emulsions. Liquid form preparations may also include solutions for intranasal administration.

[0406] Liquid (specifically, injectable) compositions can be prepared, for example, by dissolution, dispersion, etc. For example, the disclosed salts are dissolved in a pharmaceutically acceptable solvent (such as, for example, water, saline, aqueous dextrose solution, glycerol, ethanol, etc.) or mixed with the pharmaceutically acceptable solvent to form an injectable isotonic solution or suspension. Proteins (such as albumin, cryomicro particles or serum proteins) can be used to dissolve the disclosed compounds.

[0407] Parenteral injectable administration is commonly used for subcutaneous, intramuscular or intravenous injection and infusion. Injectables can be prepared in conventional form as a liquid solution or suspension or in solid form suitable for dissolution in a liquid before injection.

[0408] Aerosol preparations suitable for inhalation can include solutions and solids in powder form, which can be combined with a pharmaceutically acceptable carrier, such as an inert compressed gas, for example nitrogen.

[0409] Also included are solid form preparations that are intended to be converted to liquid form preparations shortly before use for oral or parenteral administration. Such liquid forms include solutions, suspensions and emulsions.

[0410] Depending on the intended mode of administration, the disclosed compositions can be in solid, semi-solid or liquid dosage forms, such as, for example, injectables, tablets, suppositories, pills, sustained release capsules, elixirs, tinctures, emulsions, syrups, powders, liquids, suspensions, etc., sometimes in unit dosage form and in accordance with conventional pharmaceutical practice. Similarly, they can also be administered intravenously (bolus and infusion), intraperitoneally, intrathecally, subcutaneously or intramuscularly, all in forms well known to those skilled in the pharmaceutical art.

[0411] The pharmaceutical compositions can be prepared respectively according to conventional mixing, granulating or coating methods, and by weight or volume, the pharmaceutical compositions of the present invention can contain about 0.1% to about 99%, about 5% to about 90%, or about 1% to about 20% of the disclosed free base or salt.

[0412] The pharmaceutical compositions containing the compounds disclosed herein can further comprise one or more additional anti-cancer agents, including any of those disclosed herein.

[0413] Unless otherwise indicated, all amounts of any component of the oral dosage forms (such as tablets) described herein indicated based on w / w% refer to the total weight of the oral dosage form.

[0414] Example

[0415] The present disclosure is further illustrated by the following examples, which should not be construed as limiting the scope or spirit of the present disclosure to the specific procedures described herein. It should be understood that the examples are provided to illustrate certain embodiments and are not intended to limit the scope of the present disclosure thereby. It should be further understood that various other embodiments, modifications, and their equivalents that can be contemplated by those skilled in the art can be resorted to without departing from the spirit and / or scope of the appended claims.

[0416] Abbreviations:

[0417] ACN: Acetonitrile

[0418] ADDP: 1,1'-(Azodicarbonyl)dipiperidine

[0419] BAST: N,N-Bis(2-methoxyethyl)aminosulfur trifluoride

[0420] Binap: 2,2'-Bis(diphenylphosphino)-1,1'-binaphthalene

[0421] Boc: tert-Butyloxycarbonyl

[0422] BPO: Benzoyl peroxide

[0423] Cbz: Carbobenzyloxy

[0424] DAST: Diethylaminosulfur trifluoride

[0425] DBE: 1,2-Dibromoethane

[0426] DCE: 1,2-Dichloroethane

[0427] DCM: Dichloromethane

[0428] DEAD: Diethyl azodicarboxylate

[0429] DIAD: Diisopropyl azodicarboxylate

[0430] DIBAL: Diisobutylaluminum hydride

[0431] DIEA or DIPEA: Diisopropylethylamine

[0432] DMA: N,N-Dimethylacetamide

[0433] DMF: N,N-Dimethylformamide

[0434] DMP: Dess-Martin periodinane

[0435] DMSO: Dimethyl sulfoxide

[0436] dtbpf: 1,1'-Bis(di-tert-butylphosphino)ferrocene

[0437] EA: Ethyl acetate

[0438] EDCI: 1-Ethyl-3-(3-dimethylaminopropyl)carbodiimide

[0439] EtOAc: Ethyl acetate

[0440] HATU: 1-[Bis(dimethylamino)methylene]-1H-1,2,3-triazolo[4,5-b]pyridinium 3-oxide hexafluorophosphate

[0441] HBTU: N,N,N'N'-Tetramethyl-O-(1H-benzotriazol-1-yl)uronium hexafluorophosphate

[0442] HMDS: Bis(trimethylsilyl)amine

[0443] HOBt: Hydroxybenzotriazole

[0444] HPLC: High performance liquid chromatography

[0445] HMPA: Hexamethylphosphoramide

[0446] LDA: Lithium diisopropylamide

[0447] LCMS: Liquid chromatography - mass spectrometry

[0448] MCPBA: Meta-chloroperoxybenzoic acid

[0449] MeCN: Acetonitrile

[0450] MsCl: Methanesulfonyl chloride

[0451] M.W: Microwave

[0452] NBS: N-Bromosuccinimide

[0453] NMM: N-Methylmorpholine

[0454] NMP: N-Methylpyrrolidone

[0455] Pd-PEPPSI-IPent: Dichloro[1,3-bis(2,6-di-3-pentylphenyl)imidazol-2-ylidene](3-chloropyridyl)palladium(II), [1,3-bis(2,6-di-3-pentylphenyl)imidazol-2-ylidene](3-chloropyridyl)dichloropalladium(II), [1,3-bis(2,6-di-3-pentylphenyl)imidazol-2-ylidene](3-chloropyridyl)dichloropalladium(II)

[0456] PCC: Pyridinium chlorochromate

[0457] Pd-118 or Pd(dtpf)Cl2: 1,1'-Bis(di-tert-butylphosphino)ferrocene dichloropalladium

[0458] Pd(dppf)Cl2: 1,1'-Bis(diphenylphosphino)ferrocene dichloropalladium

[0459] Pd(dba)2: Bis(dibenzylideneacetone)palladium

[0460] Pd2(dba)3: Tris(dibenzylideneacetone)dipalladium

[0461] PPTS: Pyridinium p-toluenesulfonate

[0462] PTSA: p-Toluenesulfonic acid

[0463] RuPhos-Pd-G3: XPhos-Pd-G3: [(2-Dicyclohexylphosphino-2',6'-diisopropoxy-1,1'-biphenyl)-2-(2'-amino-1,1'-biphenyl)]palladium(II) methanesulfonate

[0464] RuPhos-Pd-G2: Chloro[(2-dicyclohexylphosphino-2',6'-diisopropoxy-1,1'-biphenyl)-2-(2'-amino-1,1'-biphenyl)]palladium(II) methanesulfonate

[0465] SEM-Cl: 2-(Trimethylsilyl)ethoxymethyl chloride

[0466] SFC: Supercritical fluid chromatography

[0467] STAB: Sodium triacetoxyborohydride

[0468] t-BuXPhos-Pd-G3: [(2-Di-tert-butylphosphino-2',4',6'-triisopropyl-1,1'-biphenyl)-2-(2'-amino-1,1'-biphenyl)]palladium(II) methanesulfonate

[0469] TEA: Triethylamine

[0470] THF: Tetrahydrofuran

[0471] TFA: Trifluoroacetic acid

[0472] TLC: Thin layer chromatography

[0473] TMP: 2,2,6,6-Tetramethylpiperidine

[0474] TEMPO: 2,2,6,6-Tetramethylpiperidine-N-oxide

[0475] TosCl or TsCl: p-Toluenesulfonyl chloride

[0476] TsOH: p-Toluenesulfonic acid

[0477] XantPhos: 4,5-Bis(diphenylphosphino)-9,9-dimethylxanthene

[0478] XPhos: 2-Dicyclohexylphosphino-2',4',6'-triisopropylbiphenyl

[0479] XPhos-Pd-G3: [(2-Dicyclohexylphosphino-2',4',6'-triisopropyl-1,1'-biphenyl)-2-(2'-amino-1,1'-biphenyl)]palladium(II) methanesulfonate

[0480] 12354-85-7: Bis(pentamethylcyclopentadienyl)rhodium dichloride

[0481] Example 1 - Synthesis of Compounds 30 and 31

[0482] Step 1: 4-Bromo-2-ethyl-3-fluorobenzoic acid

[0483]

[0484] A solution of 2,2,6,6-tetramethylpiperidine (13.48 g, 95 mmol) in tetrahydrofuran (100 mL) was treated with n-BuLi (6.11 g, 95 mmol) at -20 °C for 30 minutes under a nitrogen atmosphere, and then 4-bromo-3-fluorobenzoic acid (9.5 g, 43 mmol) was added dropwise at -50 °C. The resulting mixture was stirred at -50 °C for 2 hours under a nitrogen atmosphere. At -50 °C, iodoethane (33.83 g, 217 mmol) was added dropwise to the above mixture within 5 minutes. The resulting mixture was stirred at room temperature for 2 hours and then acidified to pH 4 with concentrated hydrochloric acid. The aqueous layer was extracted with ethyl acetate. The resulting mixture was concentrated in vacuo to give 4-bromo-2-ethyl-3-fluorobenzoic acid as a colorless solid (10 g, 93%). MS (ESI): m / z 244.90 [M+H] + .

[0485] Step 2: Methyl 4-bromo-2-ethyl-3-fluorobenzoate

[0486]

[0487] Sulfuric acid (10.0 mL, 187 mmol) was added to a stirred solution of 4-bromo-2-ethyl-3-fluorobenzoic acid (12 g, 49 mmol) in methanol (100 mL). The resulting mixture was stirred overnight at 60 °C under a nitrogen atmosphere and then concentrated in vacuo. The residue was extracted with ethyl acetate and concentrated. The residue was purified by silica gel column chromatography, eluting with petroleum ether / ethyl acetate (8:1) to afford methyl 4-bromo-2-ethyl-3-fluorobenzoate (7.6 g, 60%) as an off-white solid.

[0488] Step 3: Methyl 4-bromo-2-(1-bromoethyl)-3-fluorobenzoate

[0489]

[0490] Azobisisobutyronitrile (0.94 g, 6 mmol) was added to a stirred solution of methyl 4-bromo-2-ethyl-3-fluorobenzoate (7.5 g, 29 mmol) and N-bromosuccinimide (6.14 g, 34 mmol) in dichloroethane (100 mL). The resulting mixture was stirred overnight at 70 °C under a nitrogen atmosphere. It was diluted with saturated ammonium chloride solution and extracted with dichloromethane. The combined organic layers were dried over anhydrous sodium sulfate, filtered and concentrated in vacuo to afford methyl 4-bromo-2-(1-bromoethyl)-3-fluorobenzoate (9 g, 92%) as a brown oil.

[0491] Step 4: 2-[2,6-Bis(benzyloxy)pyridin-3-yl]-5-bromo-4-fluoro-3-methyl-3H-isoindol-1-one

[0492]

[0493] N,N-Diisopropylethylamine (5.1 mL, 29 mmol) was added to a stirred solution of methyl 4-bromo-2-(1-bromoethyl)-3-fluorobenzoate (5.0 g, 15 mmol) and 2,6-bis(benzyloxy)pyridin-3-amine (5.4 g, 18 mmol) in acetonitrile (100 mL). The resulting mixture was stirred overnight at 70 °C under a nitrogen atmosphere and then concentrated in vacuo. The resulting mixture was extracted with dichloromethane. The combined organic layers were dried over anhydrous sodium sulfate, filtered and concentrated. The residue was purified by reverse-phase flash chromatography (column: C18 silica gel; mobile phase: aqueous acetonitrile (10 mmol / L NH4HCO3), 10% to 100% in 30 minutes) to afford 2-[2,6-bis(benzyloxy)pyridin-3-yl]-5-bromo-4-fluoro-3-methyl-3H-isoindol-1-one (7.8 g, 99%) as a brown solid. MS (ESI): m / z 535.15 [M+H] + 。

[0494] Step 5: tert-Butyl 4-[(1r,3r)-3-[(3R)-4-{2-[2,6-bis(benzyloxy)pyridin-3-yl]-4-fluoro-3-methyl-1-oxo-3H-isoindol-5-yl}-3-methylpiperazin-1-yl]cyclobutoxy]piperidine-1-carboxylate

[0495]

[0496] To a degassed solution of 2-[2,6-bis(benzyloxy)pyridin-3-yl]-5-bromo-4-fluoro-3-methyl-3H-isoindol-1-one (1.2 g, 2 mmol) in dioxane (20 mL) at room temperature was added tert-butyl 4-[(1r,3r)-3-[(3R)-3-methylpiperazin-1-yl]cyclobutoxy]piperidine-1-carboxylate (0.95 g, 2.7 mmol), then Cs2CO3 (2.20 g, 7 mmol) and dichloro[1,3-bis(2,6-di-3-pentylphenyl)imidazol-2-ylidene](3-chloropyridyl)palladium(II) (3.15 g, 3.8 mmol). The reaction mixture was stirred at 100 °C for 6 hours. The resulting mixture was filtered and the filter cake was washed with ethyl acetate. The filtrate was concentrated under reduced pressure. The residue was purified by reverse phase flash chromatography (column: C18 silica gel; mobile phase: aqueous acetonitrile (10 mmol / L NH4HCO3), 10% to 70% gradient in 30 minutes) to give tert-butyl 4-[(1r,3r)-3-[(3R)-4-{2-[2,6-bis(benzyloxy)pyridin-3-yl]-4-fluoro-3-methyl-1-oxo-3H-isoindol-5-yl}-3-methylpiperazin-1-yl]cyclobutoxy]piperidine-1-carboxylate as a brown solid (1.1 g, 61%).

[0497] Step 6: tert-Butyl 4-((1R,3r)-3-((R)-4-((S)-2-(2,6-bis(benzyloxy)pyridin-3-yl)-4-fluoro-3-methyl-1-oxoisoindolin-5-yl)-3-methylpiperazin-1-yl)cyclobutoxy)piperidine-1-carboxylate and tert-Butyl 4-((1R,3r)-3-((R)-4-((R)-2-(2,6-bis(benzyloxy)pyridin-3-yl)-4-fluoro-3-methyl-1-oxoisoindolin-5-yl)-3-methylpiperazin-1-yl)cyclobutoxy)piperidine-1-carboxylate

[0498]

[0499] The racemic product (1.1 g) was purified by preparative HPLC (column: (S,S)Whelk-O1 4.6 x 50 mm, 3.5 μm; co-solvent: methanol (0.1% diethylamine); 4 mL / min; gradient (B%): from 10% to 50% in 2.0 min). The first peak (1.635 min) was collected to afford tert-butyl 4-[(1r,3r)-3-[(3R)-4-[(3R)-2-[2,6-bis(benzyloxy)pyridin-3-yl]-4-fluoro-3-methyl-1-oxo-3H-isoindol-5-yl]-3-methylpiperazin-1-yl]cyclobutoxy]piperidine-1-carboxylate (tentative assignment, 610 mg, 55%) as a brown solid. The second peak (1.999 min) was collected to afford tert-butyl 4-[(1r,3r)-3-[(3R)-4-[(3S)-2-[2,6-bis(benzyloxy)pyridin-3-yl]-4-fluoro-3-methyl-1-oxo-3H-isoindol-5-yl]-3-methylpiperazin-1-yl]cyclobutoxy]piperidine-1-carboxylate (tentative assignment, 450 mg, 41%) as a brown solid.

[0500] Step 7: tert-butyl 4-[(1r,3r)-3-[(3R)-4-[(3R)-2-(2,6-dioxopiperidin-3-yl)-4-fluoro-3-methyl-1-oxo-3H-isoindol-5-yl]-3-methylpiperazin-1-yl]cyclobutoxy]piperidine-1-carboxylate

[0501]

[0502] Under a nitrogen atmosphere, 10% Pd / C (200 mg, 1.9 mmol) was added to a solution of tert-butyl 4-[(1r,3r)-3-[(3R)-4-[(3R)-2-[2,6-bis(benzyloxy)pyridin-3-yl]-4-fluoro-3-methyl-1-oxo-3H-isoindol-5-yl]-3-methylpiperazin-1-yl]cyclobutoxy]piperidine-1-carboxylate (450 mg, 0.6 mmol) in ethanol (8 mL). The mixture was degassed and purged with hydrogen several times, then stirred overnight at room temperature under a hydrogen atmosphere using a hydrogen balloon. The mixture was filtered through a pad of diatomaceous earth and concentrated under reduced pressure to afford tert-butyl 4-[(1r,3r)-3-[(3R)-4-[(3R)-2-(2,6-dioxopiperidin-3-yl)-4-fluoro-3-methyl-1-oxo-3H-isoindol-5-yl]-3-methylpiperazin-1-yl]cyclobutoxy]piperidine-1-carboxylate (130 mg, 37%) as a brown solid.

[0503] Step 8: 3-[(3R)-4-Fluoro-3-methyl-5-[(2R)-2-methyl-4-[(1r,3r)-3-(piperidin-4-yloxy)cyclobutyl]piperazin-1-yl]-1-oxo-3H-isoindol-2-yl]piperidine-2,6-dione

[0504]

[0505] To a stirred solution of tert-butyl 4-[(1r,3r)-3-[(3R)-4-[(3R)-2-(2,6-dioxopiperidin-3-yl)-4-fluoro-3-methyl-1-oxo-3H-isoindol-5-yl]-3-methylpiperazin-1-yl]cyclobutoxy]piperidine-1-carboxylate (130 mg, 0.2 mmol) in dioxane (3 mL) was added hydrochloric acid (3 mL). The resulting mixture was stirred at room temperature for 2 h and then concentrated under reduced pressure to afford 3-[(3R)-4-fluoro-3-methyl-5-[(2R)-2-methyl-4-[(1r,3r)-3-(piperidin-4-yloxy)cyclobutyl]piperazin-1-yl]-1-oxo-3H-isoindol-2-yl]piperidine-2,6-dione hydrochloride (109 mg, 99%) as a white solid. MS (ESI), m / z 528.40 [M+H] + 。

[0506] Step 9: 1-Isopropyl-5-nitro-indoline-2,3-dione

[0507]

[0508] To a mixture of 5-nitroindoline-2,3-dione (5.00 g, 26 mmol) in N,N-dimethylformamide (50 mL) was added potassium carbonate (7.19 g, 52 mmol) and 2-iodopropane (3.9 mL, 39 mmol). The mixture was stirred at 25 °C for 48 h. The reaction mixture was poured into water (300 mL) and extracted with ethyl acetate (50 mL×3). The organic layer was washed with brine (100 mL), dried over anhydrous sodium sulfate, filtered and concentrated under reduced pressure to afford 1-isopropyl-5-nitro-indoline-2,3-dione (4.00 g, 66%) as a yellow solid, which was used directly in the next step. 1 H NMR (400 MHz, DMSO) δ 8.46 (dd, J = 8.8, 2.4 Hz, 1H), 8.21 (d, J = 2.4 Hz, 1H), 7.52 (d, J = 8.8 Hz, 1H), 4.60 - 4.45 (m, 1H), 1.46 (d, J = 6.8 Hz, 6H).

[0509] Step 10: 1-Isopropyl-3-methoxy-6-nitroquinolin-2-one

[0510]

[0511] At 25 °C, triethylamine (33 mL) was added to a stirred solution of 1-isopropyl-5-nitroindoline-2,3-dione (25.0 g, 107 mmol) in ethanol (400 mL), followed by addition of hexane solution containing trimethylsilyldiazomethane (2 M, 117 mL). After stirring at 25 °C for 12 h, the reaction mixture was poured into water (1500 mL) and extracted with dichloromethane (500 mL×3). The organic layers were combined and concentrated under reduced pressure. The residue was stirred in a mixture of ethyl acetate (50 mL) and petroleum ether (500 mL) at 25 °C for 2 h and then filtered. The filter cake was dried under reduced pressure to give 1-isopropyl-3-methoxy-6-nitroquinolin-2-one (45 g, crude product) as a yellow solid. MS(ESI) m / z: 263.1 [M+H] + ; 1 H NMR (400 MHz, DMSO) δ 8.59 (d, J = 2.8 Hz, 1H), 8.17 (dd, J = 9.6, 2.8 Hz, 1H), 7.52 (d, J = 9.6 Hz, 1H), 7.49 (s, 1H), 5.45 - 5.28 (m, 1H), 3.84 (s, 3H), 1.55 (d, J = 6.8 Hz, 6H).

[0512] Step 11: 3-Hydroxy-1-isopropyl-6-nitroquinolin-2-one

[0513]

[0514] At 0 °C, a solution of boron tribromide (4.5 mL, 46 mmol) in dichloromethane (40 mL) was added dropwise to a mixture of 1-isopropyl-3-methoxy-6-nitroquinolin-2-one (11 g, 42 mmol) in dichloromethane (400 mL). After stirring at 0 °C for 2 h, the mixture was poured into saturated sodium bicarbonate (1000 mL) and extracted with dichloromethane (500 mL×3). The organic layers were combined, washed with brine (100 mL), dried over anhydrous sodium sulfate, filtered, and concentrated under reduced pressure. The residue was triturated in a mixture of ethyl acetate (50 mL), petroleum ether (500 mL) and acetonitrile (50 mL) at 25 °C for 12 h and then filtered. The filtrate was concentrated under reduced pressure to give 3-hydroxy-1-isopropyl-6-bromoquinolin-2-one (28 g, 90%) as a brown solid. MS(ESI) m / z: 280.2 [M+Na] + ; 11H NMR (400 MHz, DMSO) δ 9.95 (s, 1H), 8.54 (d, J = 2.8 Hz, 1H), 8.14 (dd, J = 9.2, 2.4 Hz, 1H), 7.92 (d, J = 9.2 Hz, 1H), 7.33 (s, 1H), 5.58 - 5.14 (m, 1H), 1.59 (d, J = 6.8 Hz, 6H).

[0515] Step 12: 2 - [(1 - Isopropyl - 6 - nitro - 2 - oxo - 3 - quinolinyl)oxy] - N - methyl - acetamide

[0516]

[0517] To a solution of 3 - hydroxy - 1 - isopropyl - 6 - nitroquinolin - 2 - one (15 g, 60 mmol) and potassium carbonate (16.5 g, 120 mmol) in DMF (500 mL) was added 2 - bromo - N - methylacetamide (9.1 g, 60 mmol). The resulting mixture was stirred at room temperature for 2 h. The mixture was suspended in ice water, filtered and dried to give 2 - [(1 - isopropyl - 6 - nitro - 2 - oxoquinolin - 3 - yl)oxy] - N - methylacetamide (13 g, 67%) as a yellow solid. MS (ESI) m / z: 320.1 [M + 1] + ; 1 1H NMR (400 MHz, DMSO) δ 8.60 (d, J = 2.8 Hz, 1H), 8.21 (dd, J = 9.6, 2.8 Hz, 1H), 8.01 - 7.88 (m, 2H), 7.48 (s, 1H), 5.70 - 5.15 (m, 1H), 4.57 (s, 2H), 2.68 (d, J = 4.8 Hz, 3H), 1.58 (d, J = 7.2 Hz, 6H).

[0518] Step 13: 2 - [(6 - Amino - 1 - isopropyl - 2 - oxoquinolin - 3 - yl)oxy] - N - methylacetamide

[0519]

[0520] 2 - [(1 - Isopropyl - 6 - nitro - 2 - oxoquinolin - 3 - yl)oxy] - N - methylacetamide (13 g) and 10% Pd / C (2 g) were combined under a nitrogen atmosphere. The mixture was degassed and purged with hydrogen three times, then stirred under hydrogen using a hydrogen balloon at room temperature overnight. The mixture was filtered through a pad of diatomaceous earth and concentrated under reduced pressure to give 2 - [(6 - amino - 1 - isopropyl - 2 - oxoquinolin - 3 - yl)oxy] - N - methylacetamide (10 g, 84%) as a brown solid.

[0521] Step 14: 2-[(6-Amino-1-isopropyl-2-oxoquinolin-3-yl)oxy]-N-methylacetamide

[0522]

[0523] At room temperature, N,N-diisopropylethylamine (18.1 mL, 104 mmol) was added to a stirred solution / mixture of 2-[(6-amino-1-isopropyl-2-oxoquinolin-3-yl)oxy]-N-methylacetamide (10 g, 34 mmol) and 5-chloro-2,4-difluoropyrimidine (5.2 g, 34 mmol) in N,N-dimethylformamide. The resulting mixture was stirred at room temperature under a nitrogen atmosphere for 2 hours. The mixture was suspended in water, filtered and washed with water (2 × 20 mL). The resulting solid was dried under infrared light to give 2-({6-[(5-chloro-2-fluoropyrimidin-4-yl)amino]-1-isopropyl-2-oxoquinolin-3-yl}oxy)-N-methylacetamide as a yellow solid (11 g, 76%). MS (ESI): m / z 420.05 [M+H] + 。

[0524] Step 15: 2-({6-[(5-chloro-2-{4-[(1r,3r)-3-[(3R)-4-[(3R)-2-(2,6-dioxopiperidin-3-yl)-4-fluoro-3-methyl-1-oxo-3H-isoindol-5-yl]-3-methylpiperazin-1-yl]cyclobutoxy]piperidin-1-yl}pyrimidin-4-yl)amino]-1-isopropyl-2-oxoquinolin-3-yl}hydroxy)-N-methylacetamide

[0525]

[0526] To a stirred solution of 3-[(3R)-4-fluoro-3-methyl-5-[(2R)-2-methyl-4-[(1r,3r)-3-(piperidin-4-yloxy)cyclobutyl]piperazin-1-yl]-1-oxo-3H-isoindol-2-yl]piperidine-2,6-dione (80 mg, 0.2 mmol) and 2-({6-[(5-chloro-2-fluoropyrimidin-4-yl)amino]-1-isopropyl-2-oxoquinolin-3-yl}oxy)-N-methylacetamide (58 mg, 0.1 mmol) in dimethyl sulfoxide (5 mL) was added N,N-diisopropylethylamine (0.5 mL). The resulting mixture was stirred at 50 °C for 2 h under a nitrogen atmosphere. The residue was purified by reverse phase flash chromatography (column: C18 silica gel; mobile phase: aqueous acetonitrile (10 mmol / L NH4HCO3), 10% to 50% gradient in 30 min) to afford 2-({6-[(5-chloro-2-{4-[(1r,3r)-3-[(3R)-4-[(3R)-2-(2,6-dioxopiperidin-3-yl)-4-fluoro-3-methyl-1-oxo-3H-isoindol-5-yl]-3-methylpiperazin-1-yl]cyclobutoxy]piperidin-1-yl}pyrimidin-4-yl)amino]-1-isopropyl-2-oxoquinolin-3-yl}oxy)-N-methylacetamide (100 mg, 78%) as a brown solid. MS (ESI) m / z 927.55 [M+H] + 。

[0527] Step 16: 2-({6-[(5-chloro-2-{4-[(1r,3r)-3-[(3R)-4-[(3R)-2-[(3S)-2,6-dioxopiperidin-3-yl]-4-fluoro-3-methyl-1-oxo-2,3-dihydro-1H-isoindol-5-yl]-3-methylpiperazin-1-yl]cyclobutoxy]piperidin-1-yl}pyrimidin-4-yl)amino]-2-oxo-1-(propan-2-yl)-1,2-dihydroquinolin-3-yl}oxy)-N-methylacetamide and 2-({6-[(5-chloro-2-{4-[(1r,3r)-3-[(3R)-4-[(3R)-2-[(3R)-2,6-dioxopiperidin-3-yl]-4-fluoro-3-methyl-1-oxo-3H-isoindol-5-yl]-3-methylpiperazin-1-yl]cyclobutoxy]piperidin-1-yl}pyrimidin-4-yl)amino]-1-isopropyl-2-oxoquinolin-3-yl}oxy)-N-methylacetamide

[0528]

[0529] The crude product (50 mg) was purified by preparative HPLC (column: CHIRALPAK ID-3, column size: 4.6 x 50 mm, 3 μm, mobile phase: (hexane:dichloromethane = 1:1, 0.1% diethylamine):ethanol = 60:40, 1.0 mL / min). The first peak (3.421 min) was collected to give 2-({6-[(5-chloro-2-{4-[(1r,3r)-3-[(3R)-4-[(3R)-2-[(3S)-2,6-dioxopiperidin-3-yl]-4-fluoro-3-methyl-1-oxo-3H-isoindol-5-yl]-3-methylpiperazin-1-yl]cyclobutoxy]piperidin-1-yl}pyrimidin-4-yl)amino]-1-isopropyl-2-oxoquinolin-3-yl}oxy)-N-methylacetamide (assumed) (19.5 mg, 18%) as an off-white solid. MS (ESI): m / z 925.65 [M+H] + ; 1 H NMR (400 MHz, DMSO) δ 10.71 (s, 1H), 8.83 (s, 1H), 8.04 (s, 1H), 7.96 - 7.95 (m, 2H), 7.69 (s, 2H), 7.43 - 7.41 (m, 1H), 7.20 - 7.17 (m, 1H), 7.03 (s, 1H), 5.42 - 5.53 (m, 1H), 4.90 - 4.88 (m, 1H), 4.67 - 4.65 (m, 1H), 4.53 (s, 2H), 4.20–4.10 (m, 2H), 3.53 - 3.51 (m, 1H), 3.33 - 3.32 (m, 1H), 3.27 - 3.26 (m, 2H), 2.99 - 2.80 (m, 2H), 2.69 - 2.67 (m, 2H), 2.59 - 2.50 (m, 5H), 2.39 - 2.38 (m, 2H), 2.27 - 2.22 (m, 3H), 2.21 - 2.01 (m, 3H), 1.99 - 1.82 (m, 2H), 1.58 - 1.56 (m, 6H), 1.48 - 1.46 (m, 3H), 1.37 - 1.35 (m, 2H), 1.00 - 0.98 (m, 3H). The second peak (5.15 min) was collected to give 2-({6-[(5-chloro-2-{4-[(1r,3r)-3-[(3R)-4-[(3R)-2-[(3R)-2,6-dioxopiperidin-3-yl]-4-fluoro-3-methyl-1-oxo-3H-isoindol-5-yl]-3-methylpiperazin-1-yl]cyclobutoxy]piperidin-1-yl}pyrimidin-4-yl)amino]-1-isopropyl-2-oxoquinolin-3-yl}oxy)-N-methylacetamide (assumed) (21.5 mg, 43%) as an off-white solid. MS (ESI): m / z 925.65 [M+H] +; 1 H NMR (400 MHz, DMSO) δ 10.71 (s, 1H), 8.83 (s, 1H), 8.04 (s, 1H), 7.96 - 7.95 (m, 2H), 7.69 (s, 2H), 7.43 - 7.41 (m, 1H), 7.20 - 7.17 (m, 1H), 7.03 (s, 1H), 5.42 - 5.53 (m, 1H), 4.90 - 4.88 (m, 1H), 4.67 - 4.65 (m, 1H), 4.53 (s, 2H), 4.20–4.10 (m, 2H), 3.53 - 3.51 (m, 1H), 3.33 - 3.32 (m, 1H), 3.27 - 3.26 (m, 2H), 2.99 - 2.80 (m, 2H), 2.69 - 2.67 (m, 2H), 2.59 - 2.50 (m, 5H), 2.39 - 2.38 (m, 2H), 2.27 - 2.22 (m, 3H), 2.21 - 2.01 (m, 3H), 1.99 - 1.82 (m, 2H), 1.58 - 1.56 (m, 6H), 1.48 - 1.46 (m, 3H), 1.37 - 1.35 (m, 2H), 1.00 - 0.98 (m, 3H).

[0530] Example 2 - Synthesis of Compounds 32 and 33

[0531] Step 1: 4 - Bromo - 2 - ethylbenzoic acid

[0532]

[0533] At - 40 °C under a nitrogen atmosphere, 2,2,6,6 - tetramethylpiperidine (16.23 g, 115 mmol) was added dropwise to a stirred solution of butyllithium (2.5 M, 46 mL, 115 mmol) in tetrahydrofuran (200 mL). The mixture was stirred at - 40 °C for 0.5 h. At - 40 °C, 4 - bromobenzoic acid (10.5 g, 52 mmol) dissolved in THF (20 mL) was added dropwise to the above mixture over 0.5 h. The resulting mixture was stirred at - 40 °C for an additional 4 h. At - 40 °C, iodoethane (32.6 g, 209 mmol) was added dropwise to the above mixture over 0.5 h, and then the mixture was stirred at room temperature for an additional 2 h. Concentrated hydrochloric acid was added until the pH was 6 - 7. The resulting mixture was extracted with ethyl acetate (3 × 100 mL). The combined organic layers were washed with brine (3 × 100 mL), dried over anhydrous sodium sulfate, filtered, and concentrated under reduced pressure. The residue was purified by silica gel column chromatography (petroleum ether / ethyl acetate = 1:1) to give 4 - bromo - 2 - ethylbenzoic acid as a pink solid (11.5 g, 90%). MS (ESI): m / z 228.25 [M + H]+ 。

[0534] Step 2: Methyl 4-bromo-2-ethylbenzoate

[0535]

[0536] A solution of 4-bromo-2-ethylbenzoic acid (11.5 g, 50 mmol) in methanol (100 mL) was treated dropwise with sulfuric acid (1.5 mL, 28 mmol) at 0 °C under a nitrogen atmosphere. The resulting mixture was stirred overnight at 60 °C and then concentrated in vacuo. The residue was suspended in water, filtered and washed with water (3 × 10 mL). The resulting solid was dried in vacuo to give methyl 4-bromo-2-ethylbenzoate (12 g) as an off-white solid. MS (ESI): m / z 243.50 [M+H] + 。

[0537] Step 3: Methyl 4-bromo-2-(1-bromoethyl)benzoate

[0538]

[0539] To a stirred solution of methyl 4-bromo-2-ethylbenzoate (4.1 g, 16 mmol) and N-bromosuccinimide (3.60 g, 20 mmol) in ethyl acetate (50 mL) was added BPO (0.86 g, 3.4 mmol). The resulting mixture was stirred at 70 °C for 5 h under a nitrogen atmosphere. The reaction was quenched with saturated aqueous ammonium chloride solution (100 mL). The aqueous layer was extracted with ethyl acetate (100 mL × 3). The combined organic layers were concentrated under reduced pressure to give methyl 4-bromo-2-(1-bromoethyl)benzoate (5.4 g) as a white solid. MS (ESI): m / z 321.50 [M+H] + 。

[0540] Step 4: 2-[2,6-Bis(benzyloxy)pyridin-3-yl]-5-bromo-3-methyl-3H-isoindol-1-one

[0541]

[0542] To a stirred solution of methyl 4-bromo-2-(1-bromoethyl)benzoate (5.4 g, 17 mmol) and 2,6-bis(benzyloxy)pyridin-3-amine (7.71 g, 25 mmol) in acetonitrile (50 mL) was added N,N-diisopropylethylamine (6.50 g, 50 mmol). Acetic acid (0.5 mL) was added to the above mixture, and then the mixture was stirred overnight at room temperature. The mixture was concentrated under reduced pressure, diluted with water (50 mL), and extracted with dichloromethane (50 mL × 3). The combined organic layers were washed with brine (20 mL × 3), dried over anhydrous sodium sulfate, filtered, and concentrated under reduced pressure. The residue was purified by reverse-phase flash chromatography (column, C18 silica gel; mobile phase: aqueous acetonitrile (10 mmol / L NH4HCO3), 10% to 80% gradient in 30 minutes) to give 2-[2,6-bis(benzyloxy)pyridin-3-yl]-5-bromo-3-methyl-3H-isoindol-1-one as a yellow solid (6.57 g, 76%). MS (ESI): m / z 515.15 [M+H] + 。

[0543] Step 5: tert-Butyl 4-[(1r,3r)-3-[(3R)-4-[(3S)-2-[6-(benzyloxy)-2-oxo-1H-pyridin-3-yl]-3-methyl-1-oxo-3H-isoindol-5-yl]-3-methylpiperazin-1-yl]cyclobutoxy]piperidine-1-carboxylate and tert-Butyl 4-[(1r,3r)-3-[(3R)-4-[(3R)-2-[6-(benzyloxy)-2-oxo-1H-pyridin-3-yl]-3-methyl-1-oxo-3H-isoindol-5-yl]-3-methylpiperazin-1-yl]cyclobutoxy]piperidine-1-carboxylate

[0544]

[0545] To a stirred solution of 2-[2,6-bis(benzyloxy)pyridin-3-yl]-5-bromo-3-methyl-3H-isoindol-1-one (5.95 g, 11 mmol) and tert-butyl 4-[(1r,3r)-3-[(3R)-3-methylpiperazin-1-yl]cyclobutoxy]piperidine-1-carboxylate (4.36 g, 10 mmol) in dioxane (100 mL) was added dichloro[1,3-bis(2,6-di-3-pentylphenyl)imidazol-2-ylidene](3-chloropyridyl)palladium(II) (0.81 g, 1 mmol) and sodium tert-butoxide (2.77 g, 29 mmol). The resulting mixture was stirred at 100 °C for 2 h under a nitrogen atmosphere. The reaction solution was diluted with water (100 mL) and extracted with ethyl acetate (100 mL × 3). The combined organic layers were washed with brine, dried over anhydrous sodium sulfate, filtered and concentrated under reduced pressure. The residue was purified by reverse-phase flash chromatography (column, C18 silica gel; mobile phase, aqueous acetonitrile (10 mmol / L NH4HCO3), 10% to 80% gradient in 30 min) to give tert-butyl 4-[(1r,3r)-3-[(3R)-4-{2-[6-(benzyloxy)-2-oxo-1H-pyridin-3-yl]-3-methyl-1-oxo-3H-isoindol-5-yl}-3-methylpiperazin-1-yl]cyclobutoxy]piperidine-1-carboxylate (3.1 g, 46%) as a yellow solid. The racemic product was purified by preparative HPLC (column: CHIRAL ART Cellulose-SB 3.0x100 mm, 3 μm; isopropanol (0.1% diethylamine); 220 nm). The first peak (0.942 min) was collected to give tert-butyl 4-[(1r,3r)-3-[(3R)-4-[(3R)-2-[6-(benzyloxy)-2-oxo-1H-pyridin-3-yl]-3-methyl-1-oxo-3H-isoindol-5-yl]-3-methylpiperazin-1-yl]cyclobutoxy]piperidine-1-carboxylate (tentative assignment, 1.2 g, 39%) as a yellow solid. The second peak (0.372 min) was collected to give tert-butyl 4-[(1r,3r)-3-[(3R)-4-[(3S)-2-[6-(benzyloxy)-2-oxo-1H-pyridin-3-yl]-3-methyl-1-oxo-3H-isoindol-5-yl]-3-methylpiperazin-1-yl]cyclobutoxy]piperidine-1-carboxylate (tentative assignment, 1.17 g, 38%) as a yellow solid. MS(ES+): m / z 698.65 [M+H] + .

[0546] Step 6: tert-Butyl 4-[(1r,3r)-3-[(3R)-4-[(3R)-2-(2,6-dioxopiperidin-3-yl)-3-methyl-1-oxo-3H-isoindol-5-yl]-3-methylpiperazin-1-yl]cyclobutoxy]piperidine-1-carboxylate

[0547]

[0548] A mixture of tert-butyl 4-[(1r,3r)-3-[(3R)-4-[(3R)-2-[6-(benzyloxy)-2-oxo-1H-pyridin-3-yl]-3-methyl-1-oxo-3H-isoindol-5-yl]-3-methylpiperazin-1-yl]cyclobutoxy]piperidine-1-carboxylate (1.2 g, 1.7 mmol) and Pd / C (1 g) in ethanol (20 mL) was degassed and purged with hydrogen, and then stirred at 30 °C for 2 days under a hydrogen atmosphere. The resulting mixture was filtered through celite and washed with dichloromethane (10 mL × 2). The filtrate was concentrated under reduced pressure to give tert-butyl 4-[(1r,3r)-3-[(3R)-4-[(3R)-2-(2,6-dioxopiperidin-3-yl)-3-methyl-1-oxo-3H-isoindol-5-yl]-3-methylpiperazin-1-yl]cyclobutoxy]piperidine-1-carboxylate (820 mg, 78%) as a colorless oil. MS (ESI): m / z 610.35 [M+H] + 。

[0549] Step 7: tert-Butyl 4-[(1r,3r)-3-[(3R)-4-[(3R)-4-chloro-2-(2,6-dioxopiperidin-3-yl)-3-methyl-1-oxo-3H-isoindol-5-yl]-3-methylpiperazin-1-yl]cyclobutoxy]piperidine-1-carboxylate

[0550]

[0551] To a stirred solution of tert-butyl 4-[(1r,3r)-3-[(3R)-4-[(3R)-2-(2,6-dioxopiperidin-3-yl)-3-methyl-1-oxo-3H-isoindol-5-yl]-3-methylpiperazin-1-yl]cyclobutoxy]piperidine-1-carboxylate (720 mg, 1.2 mmol) and trifluoroacetic acid (296.20 mg, 2.6 mmol) in dichloromethane (1 mL) was added methanol (1 mL) and N-chlorosuccinimide (204.9 mg, 1.5 mmol). The resulting mixture was stirred overnight at 40 °C under a nitrogen atmosphere. The mixture was basified to pH 8 with N,N-diisopropylethylamine. The residue was purified by reverse-phase flash chromatography (column, C18 silica gel; mobile phase, aqueous acetonitrile (10 mmol / L NH4HCO3), 10% to 80% gradient over 30 minutes) to afford tert-butyl 4-[(1r,3r)-3-[(3R)-4-[(3R)-4-chloro-2-(2,6-dioxopiperidin-3-yl)-3-methyl-1-oxo-3H-isoindol-5-yl]-3-methylpiperazin-1-yl]cyclobutoxy]piperidine-1-carboxylate (504 mg, 66%) as a white solid. MS (ESI): m / z 644.40 [M+H] + .

[0552] Steps 8-9: 2-({6-[(5-chloro-2-{4-[(1r,3r)-3-[(3R)-4-[(3R)-4-chloro-2-[(3S)-2,6-dioxopiperidin-3-yl]-3-methyl-1-oxo-2,3-dihydro-1H-isoindol-5-yl]-3-methylpiperazin-1-yl]cyclobutoxy]piperidin-1-yl}pyrimidin-4-yl)amino]-2-oxo-1-(propan-2-yl)-1,2-dihydroquinolin-3-yl}oxy)-N-methylacetamide and 2-({6-[(5-chloro-2-{4-[(1r,3r)-3-[(3R)-4-[(3R)-4-chloro-2-[(3R)-2,6-dioxopiperidin-3-yl]-3-methyl-1-oxo-2,3-dihydro-1H-isoindol-5-yl]-3-methylpiperazin-1-yl]cyclobutoxy]piperidin-1-yl}pyrimidin-4-yl)amino]-2-oxo-1-(propan-2-yl)-1,2-dihydroquinolin-3-yl}oxy)-N-methylacetamide

[0553]

[0554] The preparation of the title compound is similar to that of Compounds 30 and 31. The two diastereomers of 2-({6-[(5-chloro-2-{4-[(1r,3r)-3-[(3R)-4-[(3R)-4-chloro-2-(2,6-dioxopiperidin-3-yl)-3-methyl-1-oxo-3H-isoindol-5-yl]-3-methylpiperazin-1-yl]cyclobutoxy]piperidin-1-yl}pyrimidin-4-yl)amino]-1-isopropyl-2-oxoquinolin-3-yl}hydroxy)-N-methylacetamide (248 mg) were separated by chiral HPLC (column: CHIRALPAK IF, 2 x 25 cm, 5 μm; mobile phase A: methyl tert-butyl ether (0.1% diethylamine), mobile phase B: ethanol:dichloromethane = 1:1; 12 mL / min; gradient: from 40% B to 40% B in 35 minutes). The first peak (5.658 minutes) was collected to give 2-({6-[(5-chloro-2-{4-[(1r,3r)-3-[(3R)-4-[(3R)-4-chloro-2-[(3S)-2,6-dioxopiperidin-3-yl]-3-methyl-1-oxo-3H-isoindol-5-yl]-3-methylpiperazin-1-yl]cyclobutoxy]piperidin-1-yl}pyrimidin-4-yl)amino]-1-isopropyl-2-oxoquinolin-3-yl}oxy)-N-methylacetamide as a white solid (tentatively assigned, 84 mg). 1 H NMR (400 MHz, DMSO) δ 10.97 (s, 1H), 8.84 (s, 1H), 8.05 (s, 1H), 7.96 (m, 2H), 7.69 (m, 2H), 7.63 (m, 1H), 7.37 (m, 1H), 7.03 (s, 1H), 4.77 - 4.69 (m, 2H), 4.55 (s, 2H), 4.20 - 4.13 (m, 3H), 3.54 (s, 2H), 3.40 (s, 3H), 2.83 (s, 2H), 2.68 (m, 6H), 2.70 (m, 2H), 2.60 (m, 1H), 2.20 (s, 2H), 2.00 (s, 4H), 1.83 (s, 2H), 1.58 (m, 6H), 1.51 (m, 3H), 1.39 (m, 2H), 0.87 (m, 4H); MS (ESI): m / z 945.05 [M+H] +Collect the second peak (7.125 minutes) to obtain 2-({6-[(5-chloro-2-{4-[(1r,3r)-3-[(3R)-4-[(3R)-4-chloro-2-[(3R)-2,6-dioxopiperidin-3-yl]-3-methyl-1-oxo-3H-isoindol-5-yl]-3-methylpiperazin-1-yl]cyclobutoxy]piperidin-1-yl}pyrimidin-4-yl)amino]-1-isopropyl-2-oxoquinolin-3-yl}oxy)-N-methylacetamide as a white solid (tentative assignment, 77 mg). 1 H NMR (400 MHz, DMSO) δ 10.93 (s, 1H), 8.84 (s, 1H), 8.05 (s, 1H), 7.96 (m, 2H), 7.69 (m, 2H), 7.61 (m, 1H), 7.36 (m, 1H), 7.03 (s, 1H), 4.82 (m, 1H), 4.69 (d m, 1H), 4.55 (s, 2H), 4.21 - 4.13 (m, 3H), 3.53 (s, 2H), 3.26 - 3.21 (m, 3H), 2.82 (s, 1H), 2.68 (m, 6H), 2.63 - 2.53 (m, 3H), 2.20 (s, 1H), 2.16 (s, 2H), 2.01 (s, 4H), 1.83 (s, 2H), 1.55 (m, 6H), 1.45 (m, 3H), 1.39 (m, 2H), 0.87 (m, 4H); MS (ESI): m / z 945.10 [M+H] + 。

[0555] Example 3 - Synthesis of Compounds 34 and 35

[0556]

[0557] Similar to Compounds 32 and 33, the title compound was prepared starting from tert-butyl 4-[(1r,3r)-3-[(3R)-4-[(3S)-2-[6-(benzyloxy)-2-oxo-1H-pyridin-3-yl]-3-methyl-1-oxo-3H-isoindol-5-yl]-3-methylpiperazin-1-yl]cyclobutoxy]piperidine-1-carboxylate. The two diastereoisomers of 2-({6-[(5-chloro-2-{4-[(1r,3r)-3-[(3R)-4-[(3S)-4-chloro-2-(2,6-dioxopiperidin-3-yl)-3-methyl-1-oxo-3H-isoindol-5-yl]-3-methylpiperazin-1-yl]cyclobutoxy]piperidin-1-yl}pyrimidin-4-yl)amino]-1-isopropyl-2-oxoquinolin-3-yl}hydroxy)-N-methylacetamide (220 mg) were separated by chiral HPLC (column: CHIRALPAK IF, 2 x 25 cm, 5 μm; mobile phase A: methyl tert-butyl ether (0.1% diethylamine), mobile phase B: ethanol:dichloromethane = 1:1; flow rate: 12 mL / min; gradient: from 40% B to 40% B over 50 min). The first peak (3.676 min) was collected to afford 2-({6-[(5-chloro-2-{4-[(1r,3r)-3-[(3R)-4-[(3R)-4-chloro-2-[(3S)-2,6-dioxopiperidin-3-yl]-3-methyl-1-oxo-3H-isoindol-5-yl]-3-methylpiperazin-1-yl]cyclobutoxy]piperidin-1-yl}pyrimidin-4-yl)amino]-1-isopropyl-2-oxoquinolin-3-yl}oxy)-N-methylacetamide (hypothetical) (56 mg, 25%) as a white solid. 1 H NMR (300 MHz, DMSO) δ 10.94 (s, 1H), 8.83 (s, 1H), 8.04 (s, 1H), 7.95 (s, 2H), 7.69 - 7.61 (m, 3H), 7.38 (d, J = 8.2 Hz, 1H), 7.03 (s, 1H), 5.70 - 5.05 (m, 1H), 4.71 (d, J = 21.9 Hz, 2H), 4.53 (s, 2H), 4.21 - 4.06 (m, 3H), 3.55 - 3.46 (m, 2H), 3.20 (d, J = 10.9 Hz, 3H), 2.85 - 2.75 (m, 2H), 2.71 - 2.53 (m, 7H), 2.32 (s, 1H), 2.22 - 2.02 (m, 7H), 1.82 (d, J = 12.0 Hz, 2H), 1.53 (d, J = 15.0 Hz, 9H), 1.37 (d, J = 9.0 Hz, 2H), 0.87 (d, J = 6.3 Hz, 3H); MS (ESI): m / z 943.55 [M+H] +Collect the second peak (5.243 minutes) to obtain 2-({6-[(5-chloro-2-{4-[(1r,3r)-3-[(3R)-4-[(3R)-4-chloro-2-[(3S)-2,6-dioxopiperidin-3-yl]-3-methyl-1-oxo-3H-isoindol-5-yl]-3-methylpiperazin-1-yl]cyclobutoxy]piperidin-1-yl}pyrimidin-4-yl)amino]-1-isopropyl-2-oxoquinolin-3-yl}oxy)-N-methylacetamide as a white solid (56.5 mg, 25%). MS (ESI): m / z 943.55 [M+H] + ; 1 H NMR (300 MHz, DMSO) δ 10.92 (s, 1H), 8.83 (s, 1H), 8.04 (s, 1H), 7.97 (s, 2H), 7.68 (d, J = 1.5 Hz, 2H), 7.59 (d, J = 8.1 Hz, 1H), 7.36 (d, J = 8.1 Hz, 1H), 7.01 (s, 1H), 5.50 - 5.00 (m, 1H), 4.80 (d, J = 6.3 Hz, 1H), 4.66 (d, J = 12.4 Hz, 1H), 4.53 (s, 2H), 4.22 - 4.06 (m, 3H), 3.60 (s, 2H), 3.28 - 3.15 (m, 3H), 2.90 - 2.67 (m, 9H), 2.33 (s, 1H), 2.17 (s, 3H), 1.99 (s, 4H), 1.82 (d, J = 12.0 Hz, 2H), 1.54 (d, J = 9.6 Hz, 9H), 1.37 (d, J = 9.3 Hz, 2H), 0.87 (d, J = 6.0 Hz, 3H).

[0558] Example 4 - Synthesis of Compound 36

[0559] Step 1: 5-Bromo-7-methyl-3H-2-benzofuran-1-one

[0560]

[0561] To a stirred solution of 4-bromo-2,6-dimethylbenzoic acid (10 g, 44 mmol) and N-bromosuccinimide (19.0 g, 109 mmol) in chlorobenzene (100 mL) was added benzoyl peroxide (1.12 g, 4.4 mmol). The resulting mixture was stirred overnight at 80 °C under a nitrogen atmosphere. It was diluted with 40% w / w aqueous sodium bisulfite solution (100 mL), and the phases were separated. The organic phase was washed with saturated aqueous sodium bicarbonate solution (30 mL). The chlorobenzene phase was concentrated by vacuum distillation (30 mL), and dimethylacetamide (25 mL) was added, and then this solution was added to a mixture of sodium borohydride (2.5 g, 1.6 mol) in methyl tert-butyl ether (40 mL) and dimethylacetamide (25 mL). The mixture was quenched with 36% w / w hydrochloric acid (25 mL) dissolved in water (35 mL). The solvent was removed by vacuum distillation to give 5-bromo-7-methyl-3H-2-benzofuran-1-one as a white crystalline solid (4 g, 40%). MS (ESI): m / z 227.0 [M+H] + 。

[0562] Step 2: 5-Bromo-3-hydroxy-7-methyl-3H-2-benzofuran-1-one

[0563]

[0564] To a stirred solution of 5-bromo-7-methyl-3H-2-benzofuran-1-one (4.0 g, 18 mmol) in methanol (100 mL) was added potassium hydroxide (1.48 g, 26 mmol). The resulting mixture was stirred at 60 °C for 2 hours under a nitrogen atmosphere. The mixture was acidified to pH 4 with potassium bisulfate (4.80 g, 35 mmol). The aqueous layer was extracted with ethyl acetate and concentrated. The residue was dissolved in dichloromethane (50 mL), manganese dioxide (15.32 g, 176 mmol) was added and stirred overnight at room temperature. The resulting mixture was filtered, and the filter cake was washed with dichloromethane. The filtrate was concentrated under reduced pressure. This gave 5-bromo-3-hydroxy-7-methyl-3H-2-benzofuran-1-one as an off-white solid (0.8 g, 19%). MS (ESI): m / z 242.90 [M+H] + 。

[0565] Step 3: Methyl 4-bromo-2-formyl-6-methylbenzoate

[0566]

[0567] At room temperature, methyl iodide (0.38 mL, 6 mmol) was added dropwise to a stirred solution of 5-bromo-3-hydroxy-7-methyl-3H-isobenzofuran-1-one (1 g, 4 mmol) and potassium carbonate (1.14 g, 8 mmol) in acetone (20 mL). The resulting mixture was stirred at room temperature for 4 h. The reaction solution was quenched with water (30 mL), extracted with dichloromethane (3 × 10 mL), the combined organic layers were dried over anhydrous sodium sulfate, filtered, and concentrated under reduced pressure. The residue was purified by reverse-phase flash chromatography (column: C18 silica gel; mobile phase: aqueous acetonitrile solution (10 mmol / L NH4HCO3), 10% to 50% gradient within 30 min) to obtain methyl 4-bromo-2-formyl-6-methylbenzoate (0.7 g, 66%) as a colorless oil.

[0568] Step 4: 3-(5-Bromo-7-methyl-1-oxo-3H-isoindol-2-yl)piperidine-2,6-dione

[0569]

[0570] 3-(Chloroamino)piperidine-2,6-dione (668 mg, 4.1 mmol) was added to a stirred solution of methyl 4-bromo-2-formyl-6-methylbenzoate (700 mg, 2.7 mmol) in dichloroethane (10 mL) and methanol (3 mL). Then acetic acid (0.1 mL, 2 mmol) was added to adjust the pH to 8. The resulting mixture was stirred at room temperature overnight. Sodium cyanoborohydride (342 mg, 5.4 mmol) was added to the above mixture. The resulting mixture was stirred at 50 °C for an additional 2 h. The mixture was filtered and the filter cake was washed with dichloromethane. The filtrate was concentrated under reduced pressure. The residue was purified by reverse-phase flash chromatography (column, C18 silica gel; mobile phase: aqueous acetonitrile solution (10 mmol / L NH4HCO3), 10% to 50% gradient within 30 min) to obtain 3-(5-bromo-7-methyl-1-oxo-3H-isoindol-2-yl)piperidine-2,6-dione (420 mg, 46%) as a brown solid.

[0571] Step 4: tert-Butyl 4-[(1r,3r)-3-{4-[2-(2,6-dioxopiperidin-3-yl)-7-methyl-1-oxo-3H-isoindol-5-yl]piperazin-1-yl}cyclobutoxy]piperidine-1-carboxylate

[0572]

[0573] To a solution of 3-(5-bromo-7-methyl-1-oxo-3H-isoindol-2-yl)piperidine-2,6-dione (180 mg, 0.5 mmol) and tert-butyl 4-[(1r,3r)-3-(piperazin-1-yl)cyclobutoxy]piperidine-1-carboxylate (181 mg, 0.5 mmol) in N,N-dimethylformamide (10 mL) was added cesium carbonate (522 mg, 1.6 mmol) and [1,1'-bis(diphenylphosphino)ferrocene]dichloropalladium(II) (44.91 mg, 0.05 mmol). After stirring for 2 h at 100 °C under a nitrogen atmosphere, the mixture was diluted with ethyl acetate. The resulting mixture was filtered and the filter cake was washed with ethyl acetate. The mixture was acidified to pH 6 with saturated ammonium chloride solution. The mixture was extracted with ethyl acetate. The combined organic layers were dried over anhydrous sodium sulfate, filtered, and concentrated. The residue was purified by reverse-phase flash chromatography (column, C18 silica gel; mobile phase, aqueous acetonitrile (10 mmol / L NH4HCO3), 10% to 50% gradient in 30 min) to give tert-butyl 4-[(1r,3r)-3-{4-[2-(2,6-dioxopiperidin-3-yl)-7-methyl-1-oxo-3H-isoindol-5-yl]piperazin-1-yl}cyclobutoxy]piperidine-1-carboxylate (140 mg, 44%) as a brown solid. MS (ESI): m / z 596.40 [M+H] + 。

[0574] Step 5: 3-(7-Methyl-1-oxo-5-{4-[(1r,3r)-3-(piperidin-4-yloxy)cyclobutyl]piperazin-1-yl}-3H-isoindol-2-yl)piperidine-2,6-dione

[0575]

[0576] To a stirred solution of tert-butyl 4-[(1r,3r)-3-{4-[2-(2,6-dioxopiperidin-3-yl)-7-methyl-1-oxo-3H-isoindol-5-yl]piperazin-1-yl}cyclobutoxy]piperidine-1-carboxylate (140 mg, 0.2 mmol) in dioxane (2.0 mL) was added dropwise hydrochloric acid dissolved in dioxane (2 mL). The resulting mixture was stirred at room temperature for 2 h and then concentrated under reduced pressure to give 3-(7-methyl-1-oxo-5-{4-[(1r,3r)-3-(piperidin-4-yloxy)cyclobutyl]piperazin-1-yl}-3H-isoindol-2-yl)piperidine-2,6-dione hydrochloride (116 mg, 99%) as a white solid. MS (ESI): m / z 496.45 [M+H] + 。

[0577] Step 6: 2-({6-[(5-Chloro-2-{4-[(1r,3r)-3-{4-[2-(2,6-dioxopiperidin-3-yl)-7-methyl-1-oxo-3H-isoindol-5-yl]piperazin-1-yl}cyclobutoxy]piperidin-1-yl}pyrimidin-4-yl)amino]-1-isopropyl-2-oxoquinolin-3-yl}oxy)-N-methylacetamide

[0578]

[0579] To a stirred solution of 3-(7-methyl-1-oxo-5-{4-[(1r,3r)-3-(piperidin-4-yloxy)cyclobutyl]piperazin-1-yl}-3H-isoindol-2-yl)piperidine-2,6-dione hydrochloride (110 mg, 0.2 mmol) and 2-({6-[(5-chloro-2-fluoropyrimidin-4-yl)amino]-1-isopropyl-2-oxoquinolin-3-yl}oxy)-N-methylacetamide (71.7 mg, 0.171 mmol) in dimethyl sulfoxide (5 mL) was added dropwise N,N-diisopropylethylamine (1 mL). The resulting mixture was stirred at 50 °C for 2 h. The residue was purified by reverse-phase flash chromatography (column, C18 silica gel; mobile phase, acetonitrile / water (10 mmol / L NH4HCO3), 10% to 50% gradient in 10 min) to afford 2-({6-[(5-chloro-2-{4-[(1r,3r)-3-{4-[2-(2,6-dioxopiperidin-3-yl)-7-methyl-1-oxo-3H-isoindol-5-yl]piperazin-1-yl}cyclobutoxy]piperidin-1-yl}pyrimidin-4-yl)amino]-1-isopropyl-2-oxoquinolin-3-yl}oxy)-N-methylacetamide as a white solid (52.7 mg, 34%). 11H NMR (400 MHz, DMSO-d6) δ 11.32 (s, 1H), 10.93 (s, 1H), 8.84 (s, 1H), 8.05 (s, 1H), 7.97 (d, J = 8.3 Hz, 2H), 7.69 (s, 2H), 7.32 (s, 1H), 7.20 (s, 1H), 7.05 (d, J = 16.9 Hz, 2H), 6.93 (s, 1H), 6.86 (s, 1H), 5.01 (dd, J = 13.3, 5.1 Hz, 1H), 4.55 (s, 2H), 4.28 (d, J = 16.9 Hz, 1H), 4.17 (d, J = 17.1 Hz, 2H), 4.11 (s, 1H), 3.56 (s, 1H), 3.24 (t, J = 11.3 Hz, 3H), 2.97–2.83 (m, 1H), 2.68 (d, J = 4.6 Hz, 3H), 2.59 (d, J = 16.0 Hz, 1H), 2.55 (s, 3H), 2.44–2.32 (m, 1H), 2.20 (s, 2H), 1.95 (d, J = 12.3 Hz, 1H), 1.87–1.80 (m, 2H), 1.58 (d, J = 6.8 Hz, 6H), 1.39 (d, J = 9.8 Hz, 3H). MS (ESI): m / z 843.25 [M+H] + 。

[0580] Example 5 - Synthesis of Compound 37

[0581] Step 1: (1s,3s)-3-(Benzyloxy)cyclobutan-1-ol

[0582]

[0583] At 0 °C, sodium borohydride (13.5 g, 357 mmol) was added portionwise to a solution of 3-(benzyloxy)cyclobutan-1-one (100 g, 567 mmol) in methanol (500 mL). The resulting mixture was stirred overnight at room temperature. After filtration, the filtrate was concentrated under reduced pressure. The residue was diluted with water at 0 °C and extracted with ethyl acetate (3 × 400 mL). The combined organic layers were washed with brine (3 × 200 mL), dried over anhydrous sodium sulfate to afford (1s,3s)-3-(benzyloxy)cyclobutan-1-ol (99 g, 98%) as a colorless oil.

[0584] Step 2: Trimethyl[(1s,3s)-3-(benzyloxy)cyclobutoxy]silane

[0585]

[0586] At 0 °C under a nitrogen atmosphere, triethylamine (134.0 g, 1.3 mol) was added to a solution of (1s,3s)-3-(benzyloxy)cyclobutan-1-ol (79.0 g, 443 mmol) in dichloromethane (700 mL), and then chlorotrimethylsilane (53.0 g, 488 mmol) was added dropwise at 0 °C. The resulting mixture was stirred at room temperature under a nitrogen atmosphere for 2 hours, then diluted with water (500 mL) and extracted with ethyl acetate (500 mL × 2). The combined organic phases were washed with saturated brine (300 mL), dried over anhydrous sodium sulfate, filtered, and concentrated in vacuo to give trimethyl[(1s,3s)-3-(benzyloxy)cyclobutoxy]silane (110 g, 99%) as an off-white oil.

[0587] Step 3: Benzyl 4-[(1s,3s)-3-(benzyloxy)cyclobutoxy]piperidine-1-carboxylate

[0588]

[0589] At -78 °C under a nitrogen atmosphere, triethylsilane (34.5 g, 297 mmol) was added dropwise to a solution of trimethyl[(1s,3s)-3-(benzyloxy)cyclobutoxy]silane (67.6 g, 270 mmol) and benzyl 4-oxopiperidine-1-carboxylate (69.3 g, 297 mmol) in dichloromethane (600 mL). The resulting mixture was stirred at -78 °C for 5 minutes, and then trimethylsilyl trifluoromethanesulfonate (30.0 g, 135 mmol) dissolved in dichloromethane (50 mL) was added dropwise. The reaction mixture was stirred at -78 °C for 10 minutes, slowly warmed to 0 °C and stirred for an additional 2 hours. The mixture was diluted with water (500 mL) and extracted with ethyl acetate (500 mL × 3). The combined organic layers were washed with brine (200 mL × 3), dried over anhydrous sodium sulfate, filtered and concentrated under reduced pressure. The residue was purified by silica gel column chromatography (petroleum ether / ethyl acetate = 10 / 1 to 4 / 1) to give benzyl 4-[(1s,3s)-3-(benzyloxy)cyclobutoxy]piperidine-1-carboxylate (81 g, 76%) as a yellow oil.

[0590] Step 4: tert-Butyl 4-[(1s,3s)-3-hydroxycyclobutoxy]piperidine-1-carboxylate

[0591]

[0592] Under a nitrogen atmosphere, Pd / C (10 g) and Pd(OH)2 / C (10 g) were added to a solution of benzyl 4-[(1s,3s)-3-(benzyloxy)cyclobutoxy]piperidine-1-carboxylate (20 g, 51 mmol) and di-tert-butyl dicarbonate (16.6 g, 76 mmol) in ethanol (200 mL) and tetrahydrofuran (150 mL). The mixture was degassed and purged with hydrogen, and then stirred at 55 °C under hydrogen (15 psi) for 48 hours. The reaction mixture was then cooled to room temperature, filtered through diatomaceous earth, and concentrated. The residue was purified by reverse-phase flash chromatography (column, C18 silica gel; mobile phase: acetonitrile / water (10 mmol / L NH4HCO3), 5% to 40% gradient over 30 minutes) to give tert-butyl 4-[(1s,3s)-3-hydroxycyclobutoxy]piperidine-1-carboxylate (28 g, 82%) as an off-white solid.

[0593] Step 5: tert-Butyl 4-[(1s,3s)-3-(trifluoromethanesulfonyloxy)cyclobutoxy]piperidine-1-carboxylate

[0594]

[0595] Under a nitrogen atmosphere at room temperature, triethylamine (11.2 g, 111 mmol) was added to a solution of tert-butyl 4-[(1s,3s)-3-hydroxycyclobutoxy]piperidine-1-carboxylate (10.0 g, 37 mmol) in dichloromethane (150 mL), and then trifluoromethanesulfonic anhydride (16.6 g, 59 mmol) was added dropwise over a 15-minute period at -40 °C under a nitrogen atmosphere. The mixture was stirred at -40 °C for 2 hours, diluted with dichloromethane (200 mL), washed with brine (100 mL × 3), dried over anhydrous sodium sulfate, filtered, and concentrated in vacuo. The residue was purified by silica gel column chromatography (petroleum ether / ethyl acetate = 9:1) to give tert-butyl 4-[(1s,3s)-3-(trifluoromethanesulfonyloxy)cyclobutoxy]piperidine-1-carboxylate (9.6 g, 65%) as a yellow solid.

[0596] Step 6: tert-Butyl 4-[(1r,3r)-3-[4-(4,4,5,5-tetramethyl-1,3,2-dioxaborolan-2-yl)-3,6-dihydro-2H-pyridin-1-yl]cyclobutoxy]piperidine-1-carboxylate

[0597]

[0598] To a stirred mixture of 4-(4,4,5,5-tetramethyl-1,3,2-dioxaborolan-2-yl)-1,2,3,6-tetrahydropyridine (500 mg, 2.4 mmol) and tert-butyl 4-[(1S,3S)-3-(trifluoromethanesulfonyloxy)cyclobutoxy]piperidine-1-carboxylate (964.6 mg, 2.4 mmol) in acetonitrile (30 mL) was added N,N-diisopropylethylamine (2 mL). The resulting mixture was stirred at room temperature under a nitrogen atmosphere for 5 h and then concentrated. The residue was diluted with water (10 mL) and extracted with EtOAc (3 × 10 mL). The combined organic layers were washed with brine (3 × 10 mL), dried over anhydrous sodium sulfate, filtered, and concentrated under reduced pressure to afford tert-butyl 4-[(1R,3R)-3-[4-(4,4,5,5-tetramethyl-1,3,2-dioxaborolan-2-yl)-3,6-dihydro-2H-pyridin-1-yl]cyclobutoxy]piperidine-1-carboxylate (1 g, 90%) as a yellow oil. MS (ESI): m / z 418.21 [M+H] + .

[0599] Step 7: tert-butyl 4-[(1R,3R)-3-{4-[2-(2,6-dioxopiperidin-3-yl)-7-methyl-1-oxo-3H-isoindol-5-yl]-3,6-dihydro-2H-pyridin-1-yl}cyclobutoxy]piperidine-1-carboxylate

[0600]

[0601] To a solution of tert-butyl 4-[(1R,3R)-3-[4-(4,4,5,5-tetramethyl-1,3,2-dioxaborolan-2-yl)-3,6-dihydro-2H-pyridin-1-yl]cyclobutoxy]piperidine-1-carboxylate (170 mg, 0.4 mmol) and 3-(5-bromo-7-methyl-1-oxo-3H-isoindol-2-yl)piperidine-2,6-dione (247 mg, 0.7 mmol) in dioxane (4 mL) and water (0.5 mL) were added cesium fluoride (111 mg, 0.7 mmol) and Pd(dtbpf)Cl2 (24 mg, 0.04 mmol). After stirring at 90 °C for 2 h under a nitrogen atmosphere, the resulting mixture was concentrated under reduced pressure. The residue was purified by silica gel column chromatography (petroleum ether / ethyl acetate = 1:1) to afford tert-butyl 4-[(1R,3R)-3-{4-[2-(2,6-dioxopiperidin-3-yl)-7-methyl-1-oxo-3H-isoindol-5-yl]-3,6-dihydro-2H-pyridin-1-yl}cyclobutoxy]piperidine-1-carboxylate (140 mg, 64%) as a brown solid. MS (ESI): m / z 593.35 [M+H]+ .

[0602] Step 8: tert-Butyl 4-[(1r,3r)-3-{4-[2-(2,6-dioxopiperidin-3-yl)-7-methyl-1-oxo-3H-isoindol-5-yl]piperidin-1-yl}cyclobutoxy]piperidine-1-carboxylate

[0603]

[0604] Under a nitrogen atmosphere, 10% Pd / C (0.1 g) was added to a solution of tert-butyl 4-[(1r,3r)-3-{4-[2-(2,6-dioxopiperidin-3-yl)-7-methyl-1-oxo-3H-isoindol-5-yl]-3,6-dihydro-2H-pyridin-1-yl}cyclobutoxy]piperidine-1-carboxylate (140 mg, 0.2 mmol) in tetrahydrofuran (5 mL) and isopropanol (5 mL). The mixture was degassed and purged with hydrogen three times, and then stirred overnight at room temperature under a hydrogen atmosphere using a hydrogen balloon. The mixture was filtered through a pad of diatomaceous earth and washed with tetrahydrofuran, and concentrated under reduced pressure to give tert-butyl 4-[(1r,3r)-3-{4-[2-(2,6-dioxopiperidin-3-yl)-7-methyl-1-oxo-3H-isoindol-5-yl]piperidin-1-yl}cyclobutoxy]piperidine-1-carboxylate (140 mg, 99%) as a brown oil. MS (ESI): m / z 595.35 [M+H] + .

[0605] Step 9: 3-(7-Methyl-1-oxo-5-{1-[(1r,3r)-3-(piperidin-4-yloxy)cyclobutyl]piperidin-4-yl}-3H-isoindol-2-yl)piperidine-2,6-dione

[0606]

[0607] HCl dissolved in 1,4-dioxane (2 mL) was added to a stirred solution of tert-butyl 4-[(1r,3r)-3-{4-[2-(2,6-dioxopiperidin-3-yl)-7-methyl-1-oxo-3H-isoindol-5-yl]piperidin-1-yl}cyclobutoxy]piperidine-1-carboxylate (140 mg, 0.2 mmol) in dioxane (2 mL). The resulting mixture was stirred at room temperature for 2 hours and then concentrated in vacuo to give 3-(7-methyl-1-oxo-5-{1-[(1r,3r)-3-(piperidin-4-yloxy)cyclobutyl]piperidin-4-yl}-3H-isoindol-2-yl)piperidine-2,6-dione hydrochloride (100 mg, 85%) as a brown solid. MS (ESI): m / z 495.35 [M+H] + .

[0608] Step 10: 2-({6-[(5-chloro-2-{4-[(1r,3r)-3-{4-[2-(2,6-dioxopiperidin-3-yl)-7-methyl-1-oxo-3H-isoindol-5-yl]piperidin-1-yl}cyclobutoxy]piperidin-1-yl}pyrimidin-4-yl)amino]-1-isopropyl-2-oxoquinolin-3-yl}oxy)-N-methylacetamide

[0609]

[0610] To a stirred solution of 3-(7-methyl-1-oxo-5-{1-[(1r,3r)-3-(piperidin-4-yloxy)cyclobutyl]piperidin-4-yl}-3H-isoindol-2-yl)piperidine-2,6-dione hydrochloride (100 mg, 0.2 mmol) and 2-({6-[(5-chloro-2-fluoropyrimidin-4-yl)amino]-1-isopropyl-2-oxoquinolin-3-yl}oxy)-N-methylacetamide (65 mg, 0.2 mmol) in DMSO (5 mL) was added dropwise N,N-diisopropylethylamine (2 mL). The resulting mixture was stirred at 50 °C for 2 h. The crude material was purified by reverse-phase flash chromatography (column: C18 silica gel; mobile phase: aqueous acetonitrile (10 mmol / L NH4HCO3), 10% to 50% gradient in 30 min) to afford 2-({6-[(5-chloro-2-{4-[(1r,3r)-3-{4-[2-(2,6-dioxopiperidin-3-yl)-7-methyl-1-oxo-3H-isoindol-5-yl]piperidin-1-yl}cyclobutoxy]piperidin-1-yl}pyrimidin-4-yl)amino]-1-isopropyl-2-oxoquinolin-3-yl}oxy)-N-methylacetamide as an off-white solid (45.2 mg, 30%). MS (ESI): m / z 894.30 [M+H] + ; 11H NMR (300 MHz, DMSO) δ 10.97 (s, 1H), 8.83 (s, 1H), 8.05 (s, 1H), 7.96 (m, 2H), 7.69 (m, 2H), 7.27 (s, 1H), 7.15 (s, 1H), 7.03 (s, 1H), 5.34 (s, 1H), 5.06 (m, 1H), 4.55 (s, 2H), 4.35 (m, 1H), 4.22 (m, 1H), 4.15 (s, 2H), 4.10 (s, 1H), 3.53 (s, 1H), 2.99 (m, 2H), 2.82 (s, 2H), 2.68 (m, 3H), 2.59 (s, 3H), 2.55 (s, 1H), 2.44–2.29 (m, 1H), 2.20–2.10 (m, 1H), 1.80 (s, 0H), 1.76 (s, 7H), 1.58 (m, 7H), 1.38 (m, 1H).

[0611] Example 6 - Synthesis of Compounds 38 and 39

[0612] Step 1: Methyl 4-bromo-5-fluoro-2-methylbenzoate

[0613]

[0614] A solution of 4-bromo-5-fluoro-2-methylbenzoic acid (30.0 g, 128 mmol) and sulfuric acid (30 mL) in methanol (300 mL) was stirred overnight at 50 °C and then concentrated in vacuo. The residue was diluted with water and extracted with ethyl acetate (100 mL). The combined organic layers were washed with brine (3 × 10 mL), dried over anhydrous sodium sulfate, filtered and concentrated under reduced pressure to give methyl 4-bromo-5-fluoro-2-methylbenzoate as a yellow oil (30 g, 94%). MS (ESI): m / z 247.2 [M+H] + .

[0615] Step 2: Methyl 5-fluoro-4-(3-fluoropyridin-4-yl)-2-methylbenzoate

[0616]

[0617] To a stirred solution of methyl 4-bromo-5-fluoro-2-methylbenzoate (22 g, 87 mmol) and 3-fluoro-4-(4,4,5,5-tetramethyl-1,3,2-dioxaborolan-2-yl)pyridine (19.5 g, 87 mmol) in dioxane (200 mL) and water (20 mL) was added [1,1'-bis(diphenylphosphino)ferrocene]dichloropalladium(II) (7.1 g, 8.7 mmol) and sodium carbonate (18.4 g, 175 mmol). The reaction mixture was stirred at 80 °C overnight, diluted with water, and extracted with ethyl acetate (100 mL × 3). The combined organic layers were washed with brine (3 × 50 mL), dried over anhydrous sodium sulfate, filtered, and concentrated under reduced pressure. The residue was purified by silica gel column chromatography (petroleum ether / ethyl acetate = 5:1) to give methyl 5-fluoro-4-(3-fluoropyridin-4-yl)-2-methylbenzoate (12 g, 54%) as a yellow solid. MS (ESI): m / z 264.3 [M+H] + 。

[0618] Step 3: Methyl 5-fluoro-4-(3-fluoropiperidin-4-yl)-2-methylbenzoate

[0619]

[0620] At room temperature, Pd(OH)2 / C (1 g) was added to a stirred solution of methyl 5-fluoro-4-(3-fluoropyridin-4-yl)-2-methylbenzoate (12 g, 46 mmol) and sulfuric acid (10 mL) in methanol (200 mL). The resulting mixture was degassed and purged with hydrogen, and then stirred overnight under a hydrogen atmosphere. The resulting mixture was filtered, and the filter cake was washed with methanol (3 × 20 mL). The filtrate was concentrated under reduced pressure to give methyl 5-fluoro-4-(3-fluoropiperidin-4-yl)-2-methylbenzoate (11 g, 80%) as a white oil. MS (ESI): m / z 270.30 [M+H] + 。

[0621] Step 4: tert-Butyl 3-fluoro-4-[2-fluoro-4-(methoxycarbonyl)-5-methylphenyl]piperidine-1-carboxylate

[0622]

[0623] To a stirred solution of methyl 5-fluoro-4-(3-fluoropiperidin-4-yl)-2-methylbenzoate (8.0 g, 29 mmol) and sodium carbonate (601 mg, 58 mmol) in tetrahydrofuran (400 mL) and water (400 mL) was added di-tert-butyl dicarbonate (12.6 g, 58 mmol). The resulting mixture was stirred at room temperature for 2 h, then diluted with water and extracted with ethyl acetate (100 mL×3). The combined organic layers were washed with brine (3×20 mL), dried over anhydrous sodium sulfate, filtered and concentrated under reduced pressure to give tert-butyl 3-fluoro-4-[2-fluoro-4-(methoxycarbonyl)-5-methylphenyl]piperidine-1-carboxylate (5 g, 50%) as a white solid. MS(ESI): m / z 370.2 [M+H] + .

[0624] Step 5: 4-[1-(tert-Butoxycarbonyl)-3-fluoropiperidin-4-yl]-5-fluoro-2-methylbenzoic acid

[0625]

[0626] To a stirred solution of tert-butyl 3-fluoro-4-[2-fluoro-4-(methoxycarbonyl)-5-methylphenyl]piperidine-1-carboxylate (5.0 g, 13 mmol) in tetrahydrofuran (200 mL) and water (200 mL) was added sodium hydroxide (2.71 g, 67 mmol). The resulting mixture was stirred at 50 °C overnight. The mixture was acidified to pH 6 with hydrochloric acid (50 mL). The resulting mixture was extracted with ethyl acetate (100 mL×3). The combined organic layers were washed with brine (3×50 mL), dried over anhydrous sodium sulfate, filtered and concentrated under reduced pressure to give 4-[1-(tert-butoxycarbonyl)-3-fluoropiperidin-4-yl]-5-fluoro-2-methylbenzoic acid (4.5 g, 94%) as a white solid. MS(ESI): m / z 356.3 [M+H] + .

[0627] Step 6: tert-Butyl 3-fluoro-4-(4-fluoro-3-hydroxy-7-methyl-1-oxo-3H-2-benzofuran-5-yl)piperidine-1-carboxylate

[0628]

[0629] At -78 °C, 1.3 M tert-butyllithium (48 mL, 63 mmol) was added dropwise over 5 minutes to a mixture of 4-[1-(tert-butoxycarbonyl)-3-fluoropiperidin-4-yl]-5-fluoro-2-methylbenzoic acid (4.5 g, 13 mmol) and tetrahydrofuran (200 mL) at -78 °C. The resulting mixture was stirred at -78 °C for 3 hours. Then N,N-dimethylformamide (1.8 mL, 25 mmol) was added dropwise over 5 minutes at -78 °C. The resulting mixture was stirred at room temperature for 3 hours. The mixture was acidified to pH 6 with hydrochloric acid (50 mL) at 0 °C and extracted with ethyl acetate (100 mL × 3). The combined organic layers were washed with brine (3 × 50 mL), dried over anhydrous sodium sulfate, filtered and concentrated under reduced pressure. The residue was purified by silica gel column chromatography (petroleum ether / ethyl acetate = 1:1) to afford tert-butyl 3-fluoro-4-(4-fluoro-3-hydroxy-7-methyl-1-oxo-3H-2-benzofuran-5-yl)piperidine-1-carboxylate (4.5 g, 92%) as a white solid. MS (ESI): m / z 384.30 [M+H] + 。

[0630] Step 7: tert-Butyl 3-fluoro-4-[2-fluoro-3-formyl-4-(methoxycarbonyl)-5-methylphenyl]piperidine-1-carboxylate

[0631]

[0632] Potassium carbonate (4.87 g, 35 mmol) was added to a stirred solution of tert-butyl 3-fluoro-4-(4-fluoro-3-hydroxy-7-methyl-1-oxo-3H-2-benzofuran-5-yl)piperidine-1-carboxylate (4.5 g, 12 mmol) and iodomethane (2.50 g, 18 mmol) in N,N-dimethylformamide (50 mL). The resulting mixture was stirred at room temperature overnight, then diluted with water and extracted with ethyl acetate (100 mL × 3). The combined organic layers were washed with brine (3 × 50 mL), dried over anhydrous sodium sulfate, filtered and concentrated under reduced pressure. The residue was purified by silica gel column chromatography (petroleum ether / ethyl acetate = 1:1) to afford tert-butyl 3-fluoro-4-[2-fluoro-3-formyl-4-(methoxycarbonyl)-5-methylphenyl]piperidine-1-carboxylate (4.3 g, 92%) as a white solid. MS (ESI): m / z 398.2 [M+H] + 。

[0633] Step 8: tert-Butyl (3R,4S)-4-{2-[(1S)-4-(tert-butoxy)-1-carbamoyl-4-oxobutyl]-4-fluoro-7-methyl-1-oxo-3H-isoindol-5-yl}-3-fluoropiperidine-1-carboxylate and tert-Butyl (3S,4R)-4-{2-[(1S)-4-(tert-butoxy)-1-carbamoyl-4-oxobutyl]-4-fluoro-7-methyl-1-oxo-3H-isoindol-5-yl}-3-fluoropiperidine-1-carboxylate

[0634]

[0635] To a solution of tert-butyl 3-fluoro-4-[2-fluoro-3-formyl-4-(methoxycarbonyl)-5-methylphenyl]piperidine-1-carboxylate (5.8 g, 15 mmol) in dichloroethane (50 mL) was added (4S)-tert-butyl 4-amino-4-carbamoylbutanoate hydrochloride (3.48 g, 14.5 mmol), and the mixture was stirred overnight at 40 °C under a nitrogen atmosphere. Then sodium cyanoborohydride (2.75 g, 44 mmol) was added, and the resulting mixture was stirred overnight at 40 °C. The reaction solution was quenched with water (50 mL) at room temperature, and the resulting mixture was extracted with dichloromethane (100 mL × 3). The combined organic layers were washed with brine (50 mL × 3), dried over anhydrous sodium sulfate, filtered and concentrated under reduced pressure. The residue was purified by reverse-phase flash chromatography (column, C18 silica gel; mobile phase, acetonitrile / water (10 mmol / L NH4HCO3), 5% to 70% gradient over 50 minutes), and then purified by SFC (column, (S,S)Whelk-O1 4.6x50 mm, 3.5 μm; mobile phase, isopropanol / hexane = 1:2 (0.1% diethylamine), from 10% to 50% in 2.0 minutes). The first peak (2.178 minutes) was collected to give tert-Butyl (3R,4S)-4-{2-[(1S)-4-(tert-butoxy)-1-carbamoyl-4-oxobutyl]-4-fluoro-7-methyl-1-oxo-3H-isoindol-5-yl}-3-fluoropiperidine-1-carboxylate (1.2 g) as a yellow solid. The second peak (2.308 minutes) was collected to give tert-Butyl (3S,4R)-4-{2-[(1S)-4-(tert-butoxy)-1-carbamoyl-4-oxobutyl]-4-fluoro-7-methyl-1-oxo-3H-isoindol-5-yl}-3-fluoropiperidine-1-carboxylate (1.3 g) as a yellow solid. MS(ESI): m / z 356.3 [M+H] + 。

[0636] Step 9: tert-Butyl (4S)-4-carbamoyl-4-{4-fluoro-5-[(3R,4S)-3-fluoropiperidin-4-yl]-7-methyl-1-oxo-3H-isoindol-2-yl}butanoate

[0637]

[0638] A solution of tert-butyl (3R,4S)-4-{2-[(1S)-4-(tert-butoxy)-1-carbamoyl-4-oxobutyl]-4-fluoro-7-methyl-1-oxo-3H-isoindol-5-yl}-3-fluoropiperidine-1-carboxylate (1.2 g, 2.1 mmol) and trimethylchlorosilane (2.36 g, 22 mmol) in 2-propanol (20 mL) was stirred overnight at room temperature under a nitrogen atmosphere. The resulting mixture was diluted with water and extracted with dichloromethane (20 mL × 3). The combined organic layers were washed with brine (3 × 10 mL), dried over anhydrous sodium sulfate, filtered and concentrated under reduced pressure to give tert-butyl (4S)-4-carbamoyl-4-{4-fluoro-5-[(3R,4S)-3-fluoropiperidin-4-yl]-7-methyl-1-oxo-3H-isoindol-2-yl}butanoate (860 mg) as a yellow solid. MS (ESI): m / z 451.30 [M+H] + 。

[0639] Step 10: tert-Butyl 4-[(1r,3r)-3-[(3R,4S)-4-{2-[(1S)-4-(tert-butoxy)-1-carbamoyl-4-oxobutyl]-4-fluoro-7-methyl-1-oxo-3H-isoindol-5-yl}-3-fluoropiperidin-1-yl]cyclobutoxy]piperidine-1-carboxylate

[0640]

[0641] To a stirred solution of tert-butyl (4S)-4-carbamoyl-4-{4-fluoro-5-[(3R,4S)-3-fluoropiperidin-4-yl]-7-methyl-1-oxo-3H-isoindol-2-yl}butanoate (300 mg, 0.7 mmol) and tert-butyl 4-[(1S,3S)-3-(trifluoromethanesulfonyloxy)cyclobutoxy]piperidine-1-carboxylate (670 mg, 1.7 mmol) in acetonitrile (2 mL) was added N,N-diisopropylethylamine (0.5 mL). The resulting mixture was stirred at room temperature under a nitrogen atmosphere for 2 h, then diluted with water and extracted with ethyl acetate (10 mL × 3). The combined organic layers were washed with brine (3 × 10 mL), dried over anhydrous sodium sulfate, filtered and concentrated under reduced pressure. The residue was purified by silica gel column chromatography (petroleum ether / ethyl acetate = 5:1) to afford tert-butyl 4-[(1R,3R)-3-[(3R,4S)-4-{2-[(1S)-4-(tert-butoxy)-1-carbamoyl-4-oxobutyl]-4-fluoro-7-methyl-1-oxo-3H-isoindol-5-yl}-3-fluoropiperidin-1-yl]cyclobutoxy]piperidine-1-carboxylate (300 mg, 38%) as a yellow solid. MS (ESI): m / z 705.2 [M+H] + 。

[0642] Step 11: (3S)-3-{4-Fluoro-5-[(3R,4S)-3-fluoro-1-[(1R,3R)-3-(piperidin-4-yloxy)cyclobutyl]piperidin-4-yl]-7-methyl-1-oxo-3H-isoindol-2-yl}piperidine-2,6-dione

[0643]

[0644] To a stirred solution of tert-butyl 4-[(1R,3R)-3-[(3R,4S)-4-{2-[(1S)-4-(tert-butoxy)-1-carbamoyl-4-oxobutyl]-4-fluoro-7-methyl-1-oxo-3H-isoindol-5-yl}-3-fluoropiperidin-1-yl]cyclobutoxy]piperidine-1-carboxylate (300 mg, 0.4 mmol) in acetonitrile (10 mL) was added (1R)-(-)-10-camphorsulfonic acid (19.7 mg, 1.3 mmol). The resulting mixture was stirred at 80 °C under a nitrogen atmosphere for 3 h, then concentrated in vacuo to afford (3S)-3-{4-fluoro-5-[(3R,4S)-3-fluoro-1-[(1R,3R)-3-(piperidin-4-yloxy)cyclobutyl]piperidin-4-yl]-7-methyl-1-oxo-3H-isoindol-2-yl}piperidine-2,6-dione (200 mg, 89%) as a white solid. MS (ESI): m / z 428.3 [M+H] + 。

[0645] Step 12: 2-({6-[(5-chloro-2-{4-[(1r,3r)-3-[(3R,4S)-4-{2-[(3S)-2,6-dioxopiperidin-3-yl]-4-fluoro-7-methyl-1-oxo-3H-isoindol-5-yl}-3-fluoropiperidin-1-yl]cyclobutoxy]piperidin-1-yl}pyrimidin-4-yl)amino]-1-isopropyl-2-oxoquinolin-3-yl}oxy)-N-methylacetamide

[0646]

[0647] To a stirred solution of (3S)-3-{4-fluoro-5-[(3R,4S)-3-fluoro-1-[(1r,3r)-3-(piperidin-4-yloxy)cyclobutyl]piperidin-4-yl]-7-methyl-1-oxo-3H-isoindol-2-yl}piperidine-2,6-dione (182 mg, 0.3 mmol) and 2-({6-[(5-chloro-2-fluoropyrimidin-4-yl)amino]-1-isopropyl-2-oxoquinolin-3-yl}oxy)-N-methylacetamide (120 mg, 0.3 mmol) in dimethyl sulfoxide (4 mL) was added N,N-diisopropylethylamine (0.5 mL). The resulting mixture was stirred at 50 °C for 2 h and then purified by reverse-phase flash chromatography (column, C18 silica gel; mobile phase, acetonitrile / water (10 mmol / L NH4HCO3), 10% to 50% gradient in 10 min) to afford 2-({6-[(5-chloro-2-{4-[(1r,3r)-3-[(3R,4S)-4-{2-[(3S)-2,6-dioxopiperidin-3-yl]-4-fluoro-7-methyl-1-oxo-3H-isoindol-5-yl}-3-fluoropiperidin-1-yl]cyclobutoxy]piperidin-1-yl}pyrimidin-4-yl)amino]-1-isopropyl-2-oxoquinolin-3-yl}oxy)-N-methylacetamide as an off-white solid (72.0 mg, 27%). 11H NMR (300 MHz, DMSO) δ 11.01 (s, 1H), 8.85 (s, 1H), 8.05 (s, 1H), 7.97 (d, J = 9.6 Hz, 2H), 7.70 (s, 2H), 7.28 (d, J = 6.1 Hz, 1H), 7.03 (s, 1H), 5.35 (s, 1H), 5.08 (dd, J = 13.2, 5.1 Hz, 1H), 4.89 (s, 1H), 4.50 (d, J = 30.0 Hz, 3H), 4.31 (d, J = 17.2 Hz, 1H), 4.13 (d, J = 13.5 Hz, 3H), 3.54 (s, 2H), 3.22 (d, J = 10.8 Hz, 4H), 3.04 (d, J = 10.4 Hz, 1H), 2.89 (d, J = 10.6 Hz, 2H), 2.68 (d, J = 4.6 Hz, 3H), 2.62 (s, 1H), 2.58 (s, 3H), 2.55 (s, 2H), 2.24–2.09 (m, 3H), 1.99 (s, 5H), 1.84 (d, J = 12.1 Hz, 2H), 1.66 (s, 2H), 1.57 (d, J = 6.8 Hz, 5H), 1.38 (d, J = 9.4 Hz, 2H), 1.24 (s, 2H), 0.85 (d, J = 6.7 Hz, 1H); MS (ESI): m / z 930.39 [M+H] + 。

[0648] Step 13: 2-({6-[(5-chloro-2-{4-[(1r,3r)-3-[(3S,4R)-4-{2-[(3S)-2,6-dioxopiperidin-3-yl]-4-fluoro-7-methyl-1-oxo-3H-isoindol-5-yl}-3-fluoropiperidin-1-yl]cyclobutoxy]piperidin-1-yl}pyrimidin-4-yl)amino]-1-isopropyl-2-oxoquinolin-3-yl}oxy)-N-methylacetamide

[0649]

[0650] Similar to Steps 9-12 in this example, the title compound was prepared starting from tert-butyl (3S,4R)-4-{2-[(1S)-4-(tert-butoxy)-1-carbamoyl-4-oxobutyl]-4-fluoro-7-methyl-1-oxo-3H-isoindol-5-yl}-3-fluoropiperidine-1-carboxylate. 11H NMR (400 MHz, DMSO) δ 11.00 (s, 1H), 8.83 (s, 1H), 8.04 (s, 1H), 7.96 (d, J = 4.8 Hz, 2H), 7.69 (d, J = 4.0 Hz, 2H), 7.27 (d, J = 6.0 Hz, 1H), 7.03 (s, 1H), 5.50 - 5.08 (m, 2H), 4.94 - 4.72 (m, 1H), 4.53 - 4.45 (m, 3H), 4.40 (s, 1H), 4.31 - 4.12 (m, 3H), 3.32 (d, J = 9.4 Hz, 1H), 3.31 (s, 2H), 3.29 - 3.18 (m, 3H), 3.03 (d, J = 10.4 Hz, 1H), 2.98 - 2.85 (m, 2H), 2.68 (d, J = 4.6 Hz, 3H), 2.62 (d, J = 3.7 Hz, 4H), 2.58 (s, 1H), 2.47 (s, 1H), 2.24 - 2.12 (m, 3H), 2.02 - 1.95 (m, 4H), 1.83 (d, J = 11.6 Hz, 2H), 1.65 (d, J = 12.3 Hz, 1H), 1.57 (d, J = 6.8 Hz, 6H), 1.38 (d, J = 9.2 Hz, 2H); MS (ESI): m / z 930.45 [M + H] + 。

[0651] Example 7 - Synthesis of Compound 40

[0652] Step 1: Benzyl 4-{2-[1-(tert - butoxycarbonyl)-4 - hydroxypiperidin - 4 - yl]acetyl}piperazine - 1 - carboxylate

[0653]

[0654] At room temperature, N,N-diisopropylethylamine (2.02 mL, 12 mmol) was added to a stirred mixture of [1-(tert-butoxycarbonyl)-4-hydroxypiperidin-4-yl]acetic acid (600 mg, 2.3 mmol) and benzyl piperazine-1-carboxylate (509.7 mg, 2.3 mmol) in N,N-dimethylformamide (10 mL). The resulting mixture was stirred at room temperature for 5 minutes. Then propanephosphonic anhydride (2.94 g, 9.3 mmol) was added to maintain the pH above 9. The resulting mixture was stirred at room temperature for 1 hour, then diluted with water and extracted with ethyl acetate (2 × 300 mL). The combined organic layers were washed with brine (2 × 100 mL), dried over anhydrous sodium sulfate, filtered and concentrated under reduced pressure. The residue was purified by silica gel column chromatography (dichloromethane / ethyl acetate = 1:1) to give benzyl 4-{2-[1-(tert-butoxycarbonyl)-4-hydroxypiperidin-4-yl]acetyl}piperazine-1-carboxylate (930 mg, 87%) as a pale yellow solid. MS (ESI): m / z 462.25 [M+H] + 。

[0655] Step 2: Benzyl 4-[2-(4-hydroxypiperidin-4-yl)acetyl]piperazine-1-carboxylate

[0656]

[0657] At room temperature, hydrochloric acid dissolved in 1,4-dioxane (8 mL) was added dropwise to a stirred solution of benzyl 4-{2-[1-(tert-butoxycarbonyl)-4-hydroxypiperidin-4-yl]acetyl}piperazine-1-carboxylate (500 mg, 1.1 mmol) in dioxane (4 mL). The resulting mixture was stirred at room temperature for 1 hour, then concentrated in vacuo to give benzyl 4-[2-(4-hydroxypiperidin-4-yl)acetyl]piperazine-1-carboxylate hydrochloride (390 mg) as a pale yellow solid. MS (ESI): m / z 362.15 [M+H] + 。

[0658] Step 3: Benzyl 4-(2-{1-[2-(2,6-dioxopiperidin-3-yl)-4-fluoro-1-oxo-3H-isoindol-5-yl]-4-hydroxypiperidin-4-yl}acetyl)piperazine-1-carboxylate

[0659]

[0660] At room temperature, dichloro[1,3-bis(2,6-di-3-pentylphenyl)imidazol-2-ylidene](3-chloropyridyl)palladium(II) (4.93 mg, 0.006 mmol) and cesium carbonate (573 mg, 1.8 mmol) were added to a stirred mixture of benzyl 4-[2-(4-hydroxypiperidin-4-yl)acetyl]piperazine-1-carboxylate (212 mg, 0.6 mmol) and 3-(5-bromo-4-fluoro-1-oxo-3H-isoindol-2-yl)piperidine-2,6-dione (200 mg, 0.6 mmol) in N,N-dimethylformamide (10 mL). The resulting mixture was stirred at 100 °C for 2 h under a nitrogen atmosphere. The reaction solution was quenched with saturated ammonium chloride solution (300 mL) at room temperature. The resulting mixture was extracted with ethyl acetate (2 × 300 mL). The combined organic layers were washed with brine (2 × 50 mL), dried over anhydrous sodium sulfate, filtered and concentrated under reduced pressure. The residue was purified by reverse-phase flash chromatography (column, C18 silica gel; mobile phase, aqueous acetonitrile solution (10 mmol / L NH4HCO3), 0% to 50% gradient in 40 min) to give benzyl 4-(2-{1-[2-(2,6-dioxopiperidin-3-yl)-4-fluoro-1-oxo-3H-isoindol-5-yl]-4-hydroxypiperidin-4-yl}acetyl)piperazine-1-carboxylate (190 mg, 52%) as a pale yellow solid. MS(ESI): m / z 622.25 [M+H] + 。

[0661] Step 4: 3-(4-Fluoro-5-{4-hydroxy-4-[2-oxo-2-(piperazin-1-yl)ethyl]piperidin-1-yl}-1-oxo-3H-isoindol-2-yl)piperidine-2,6-dione

[0662]

[0663] Trifluoroacetic acid (5.0 mL, 67 mmol) was added to benzyl 4-(2-{1-[2-(2,6-dioxopiperidin-3-yl)-4-fluoro-1-oxo-3H-isoindol-5-yl]-4-hydroxypiperidin-4-yl}acetyl)piperazine-1-carboxylate (180 mg, 0.3 mmol) at room temperature. The resulting mixture was stirred at 60 °C for 2 h and then concentrated in vacuo to give 3-(4-fluoro-5-{4-hydroxy-4-[2-oxo-2-(piperazin-1-yl)ethyl]piperidin-1-yl}-1-oxo-3H-isoindol-2-yl)piperidine-2,6-dione (140 mg, 99%) as a brownish yellow oil. MS(ESI): m / z 488.25 [M+H] + 。

[0664] Step 5: 2-{[6-({5-chloro-2-[4-(2-{1-[2-(2,6-dioxopiperidin-3-yl)-4-fluoro-1-oxo-3H-isoindol-5-yl]-4-hydroxypiperidin-4-yl}acetyl)piperazin-1-yl]pyrimidin-4-yl}amino)-1-isopropyl-2-oxoquinolin-3-yl]oxy}-N-methylacetamide

[0665]

[0666] At room temperature, N,N-diisopropylethylamine (2 mL) was added to a stirred mixture of 3-(4-fluoro-5-{4-hydroxy-4-[2-oxo-2-(piperazin-1-yl)ethyl]piperidin-1-yl}-1-oxo-3H-isoindol-2-yl)piperidine-2,6-dione (104.51 mg, 0.2 mmol) and 2-({6-[(5-chloro-2-fluoropyrimidin-4-yl)amino]-1-isopropyl-2-oxoquinolin-3-yl}oxy)-N-methylacetamide (60 mg, 0.1 mmol) in dimethyl sulfoxide (4 mL). The reaction mixture was stirred at 50 °C for 6 hours. The crude material was purified by reverse-phase flash chromatography (column, C18 silica gel; mobile phase, acetonitrile / water (10 mmol / L NH4HCO3), 0% to 40% gradient over 40 minutes) to give 2-{[6-({5-chloro-2-[4-(2-{1-[2-(2,6-dioxopiperidin-3-yl)-4-fluoro-1-oxo-3H-isoindol-5-yl]-4-hydroxypiperidin-4-yl}acetyl)piperazin-1-yl]pyrimidin-4-yl}amino)-1-isopropyl-2-oxoquinolin-3-yl]oxy}-N-methylacetamide (53.3 mg, 40%) as an off-white solid. MS (ESI): m / z 887.40 [M+H] + ; 11H NMR (300 MHz, DMSO-d6) δ 10.98 (s, 1H), 8.90 (s, 1H), 8.09 (s, 1H), 8.01–7.90 (m, 2H), 7.72 (d, J = 3.0 Hz, 2H), 7.46 (d, J = 8.1 Hz, 1H), 7.18 (t, J = 7.9 Hz, 1H), 7.08 (s, 1H), 5.32 (s, 1H), 5.07 (dd, J = 13.2, 5.1 Hz, 1H), 4.96 (s, 1H), 4.56 (s, 2H), 4.48 (d, J = 17.0 Hz, 1H), 4.30 (d, J = 16.9 Hz, 1H), 3.64 (t, J = 13.5 Hz, 8H), 3.13 (t, J = 10.7 Hz, 2H), 3.00–2.82 (m, 1H), 2.71–2.51 (m, 6H), 2.47–2.32 (m, 1H), 1.97 (d, J = 11.9 Hz, 1H), 1.87–1.66 (m, 4H), 1.58 (d, J = 6.8 Hz, 6H), 1.23 (s, 1H).

[0667] Example 8 - Synthesis of Compound 41

[0668]

[0669] The title compound can be prepared analogously to Compound 40(11) by first removing the Cbz protecting group from the intermediate prepared in Step 1. 11H NMR (400 MHz, DMSO) δ 11.00 (s, 1H), 8.83 (s, 1H), 8.05 (s, 1H), 7.98 (d, J = 4.8 Hz, 1H), 7.92 (d, J = 2.3 Hz, 1H), 7.77–7.66 (m, 2H), 7.49 (d, J = 8.1 Hz, 1H), 7.17 (t, J = 7.9 Hz, 1H), 7.02 (s, 1H), 5.34 (s, 1H), 5.09 (dd, J = 13.3, 5.1 Hz, 1H), 5.00 (s, 1H), 4.51 (d, J = 23.6 Hz, 3H), 4.33 (d, J = 17.0 Hz, 1H), 4.15 (s, 2H), 3.69 (d, J = 16.6 Hz, 4H), 3.34–3.26 (m, 1H), 3.12 (d, J = 15.6 Hz, 4H), 2.92 (ddd, J = 17.5, 13.6, 5.4 Hz, 1H), 2.68 (d, J = 4.7 Hz, 3H), 2.61 (d, J = 3.6 Hz, 1H), 2.56 (s, 3H), 2.42 (qd, J = 13.1, 4.5 Hz, 1H), 1.98 (d, J = 12.6 Hz, 1H), 1.63–1.53 (m, 9H); MS (ESI): m / z 887.40 [M + H] + 。

[0670] Example 9 - Synthesis of Compound 42

[0671] Step 1: Benzyl 4-[1-(tert-butoxycarbonyl)-3,3-difluoro-2,6-dihydropyridin-4-yl]piperazine-1-carboxylate

[0672]

[0673] Under a nitrogen atmosphere, a solution of benzyl piperazine-1-carboxylate (1 g, 4.9 mmol) in toluene (1.5 ml) and acetonitrile (15 mL) was treated with sodium acetate (1.0 g, 12 mmol) at room temperature for 15 minutes, and then tert-butyl 3,3-difluoro-4-oxopiperidine-1-carboxylate (1.8 g, 7.5 mmol) was added portionwise at room temperature. The resulting mixture was stirred at 100 °C for 16 hours under a nitrogen atmosphere. The mixture was cooled to room temperature and filtered. The filter cake was washed with dichloromethane (3 × 100 mL), and the filtrate was concentrated under reduced pressure to give benzyl 4-[1-(tert-butoxycarbonyl)-3,3-difluoro-2,6-dihydropyridin-4-yl]piperazine-1-carboxylate (1.1 g, 50%) as a pale yellow oil. MS (ESI): m / z 438.25 [M + H] + 。

[0674] Step 2: Benzyl 4-[1-(tert-butoxycarbonyl)-3,3-difluoropiperidin-4-yl]piperazine-1-carboxylate

[0675]

[0676] A mixture of benzyl 4-[1-(tert-butoxycarbonyl)-3,3-difluoro-2,6-dihydropyridin-4-yl]piperazine-1-carboxylate (1.1 g, 2.5 mmol), acetic acid (1.5 mL), and sodium cyanoborohydride (0.5 g, 7.4 mmol) in dichloroethane (15 mL) was stirred overnight at room temperature. The mixture was diluted with water and extracted with dichloromethane (3 × 80 mL). The combined organic layers were washed with brine (3 × 30 mL), dried over anhydrous sodium sulfate, filtered, and concentrated under reduced pressure. The residue was purified by reverse-phase flash chromatography (column: C18 silica gel; mobile phase: aqueous acetonitrile (10 mmol / L NH4HCO3), 5% to 60% gradient in 30 minutes) to afford benzyl 4-[1-(tert-butoxycarbonyl)-3,3-difluoropiperidin-4-yl]piperazine-1-carboxylate as a pale yellow oil (1.5 g, 71%). MS (ESI): m / z 440.30 [M+H] + 。

[0677] Step 3: tert-Butyl 3,3-difluoro-4-(piperazin-1-yl)piperidine-1-carboxylate

[0678]

[0679] Under a nitrogen atmosphere, 10% Pd(OH)2 / C (300 mg) was added to a solution of benzyl 4-[1-(tert-butoxycarbonyl)-3,3-difluoropiperidin-4-yl]piperazine-1-carboxylate (1.5 g, 3.4 mmol) in isopropanol (30 mL). The mixture was then degassed and purged with hydrogen. The reaction mixture was stirred at 30 °C for 2 hours under a hydrogen atmosphere using a hydrogen balloon, then filtered through a Celite pad and concentrated under reduced pressure to afford tert-Butyl 3,3-difluoro-4-(piperazin-1-yl)piperidine-1-carboxylate as a colorless oil (1.05 g, 92%).

[0680] Step 4: tert-Butyl 4-{4-[2-(2,6-dioxopiperidin-3-yl)-4-fluoro-1-oxo-3H-isoindol-5-yl]piperazin-1-yl}-3,3-difluoropiperidine-1-carboxylate

[0681]

[0682] To a stirred solution of tert-butyl 3,3-difluoro-4-(piperazin-1-yl)piperidine-1-carboxylate (200 mg, 0.7 mmol) and 3-(5-bromo-4-fluoro-1-oxo-3H-isoindol-2-yl)piperidine-2,6-dione (223 mg, 0.7 mmol) in N,N-dimethylformamide (5 mL) was added {1,3-bis[2,6-bis(pentan-3-yl)phenyl]-4,5-dichloro-2,3-dihydro-1H-imidazol-2-yl}dichloro(2-methyl-1λ4-pyridin-1-yl)palladium (55 mg, 0.1 mmol) and cesium carbonate (640 mg, 2.0 mmol). The resulting mixture was stirred at 100 °C for 2 h under a nitrogen atmosphere. The residue was diluted with water (400 mL) and extracted with ethyl acetate (2 × 100 mL). The combined organic layers were washed with brine (1 × 100 mL), dried over anhydrous sodium sulfate, filtered, and concentrated under reduced pressure. The residue was purified by reverse-phase flash chromatography (column: C18 silica gel; mobile phase: acetonitrile / water (10 mmol / L NH4HCO3), 5% to 65% gradient in 30 min) to afford tert-butyl 4-{4-[2-(2,6-dioxopiperidin-3-yl)-4-fluoro-1-oxo-3H-isoindol-5-yl]piperazin-1-yl}-3,3-difluoropiperidine-1-carboxylate (110 mg, 30%) as a brown solid. MS (ESI): m / z 566.40 [M+H] + 。

[0683] Step 5: 3-{5-[4-(3,3-Difluoropiperidin-4-yl)piperazin-1-yl]-4-fluoro-1-oxo-3H-isoindol-2-yl}piperidine-2,6-dione

[0684]

[0685] A solution of tert-butyl 4-{4-[2-(2,6-dioxopiperidin-3-yl)-4-fluoro-1-oxo-3H-isoindol-5-yl]piperazin-1-yl}-3,3-difluoropiperidine-1-carboxylate (110.0 mg, 0.2 mmol) and trifluoroacetic acid (2 mL) in dichloromethane (10 mL) was stirred at room temperature for 2 h. The reaction solution was concentrated to afford 3-{5-[4-(3,3-difluoropiperidin-4-yl)piperazin-1-yl]-4-fluoro-1-oxo-3H-isoindol-2-yl}piperidine-2,6-dione (88.2 mg, 97%) as a brown oil. MS (ESI): m / z 466.25 [M+H] + 。

[0686] Step 6: 2-({6-[(5-Chloro-2-{4-[(4-{4-[2-(2,6-dioxopiperidin-3-yl)-4-fluoro-1-oxo-3H-isoindol-5-yl]piperazin-1-yl}-3,3-difluoropiperidin-1-yl)methyl]piperidin-1-yl}pyrimidin-4-yl)amino]-1-isopropyl-2-oxoquinolin-3-yl}oxy)-N-methylacetamide

[0687]

[0688] To a mixture of 3-{5-[4-(3,3-difluoropiperidin-4-yl)piperazin-1-yl]-4-fluoro-1-oxo-3H-isoindol-2-yl}piperidine-2,6-dione (88.2 mg, 0.2 mmol) in dichloroethane (10 mL) and dimethyl sulfoxide (1 mL) was added 2-[(6-{[5-chloro-2-(4-formylpiperidin-1-yl)pyrimidin-4-yl]amino}-1-isopropyl-2-oxoquinolin-3-yl)oxy]-N-methylacetamide (97.0 mg, 0.2 mmol) and N,N-diisopropylethylamine (0.1 mL). The resulting mixture was stirred overnight at room temperature under a nitrogen atmosphere. Then sodium triacetoxyborohydride (120.2 mg, 0.6 mmol) was added and the mixture was stirred for 2 hours under nitrogen. The reaction solution was quenched with water (30 mL) and extracted with dichloromethane (50 mL × 3). The combined organic layers were washed with brine (30 mL), dried over anhydrous sodium sulfate, filtered and concentrated under reduced pressure. The residue was purified by reverse phase flash chromatography (column, C18 silica gel; mobile phase, acetonitrile / water (10 mmol / L NH4HCO3), 5% to 70% gradient in 30 minutes) to give 2-({6-[(5-chloro-2-{4-[(4-{4-[2-(2,6-dioxopiperidin-3-yl)-4-fluoro-1-oxo-3H-isoindol-5-yl]piperazin-1-yl}-3,3-difluoropiperidin-1-yl)methyl]piperidin-1-yl}pyrimidin-4-yl)amino]-1-isopropyl-2-oxoquinolin-3-yl}oxy)-N-methylacetamide (69.3 mg, 36%) as an off-white solid. 11H NMR (300 MHz, DMSO) δ 11.11 - 10.82 (m, 1H), 8.78 (s, 1H), 8.02 - 7.93 (m, 3H), 7.77 - 7.63 (m, 2H), 7.46 (d, J = 8.4 Hz, 1H), 7.14 (d, J = 7.8 Hz, 1H), 7.01 (s, 1H), 5.60 - 5.33 (m, 1H), 5.06 (d, J = 13.2 Hz, 1H), 4.48 (d, J = 22.2 Hz, 5H), 4.30 (d, J = 16.8 Hz, 1H), 3.09 (s, 4H), 2.99 - 2.79 (m, 10H), 2.66 (d, J = 4.6 Hz, 3H), 2.60 (s, 1H), 2.54 (d, J = 5.9 Hz, 1H), 2.39 - 1.98 (m, 5H), 1.74 (s, 5H), 1.55 (d, J = 6.9 Hz, 6H), 1.12 - 0.90 (d, J = 12.3 Hz, 2H); MS (ESI): m / z 962.50 [M + H] + 。

[0689] Example 10 - Synthesis of Compound 43

[0690] Step 1: 2 - {[6 - ({5 - chloro - 2 - [4 - (dimethoxymethyl)piperidin - 1 - yl]pyrimidin - 4 - yl}amino) - 1 - isopropyl - 2 - oxoquinolin - 3 - yl]oxy} - N - methylacetamide

[0691]

[0692] At room temperature, 2 - ({6 - [(5 - chloro - 2 - fluoropyrimidin - 4 - yl)amino] - 1 - isopropyl - 2 - oxoquinolin - 3 - yl}oxy) - N - methylacetamide (500 mg, 1.2 mmol) and DMSO (5 mL) were added to a 100 mL round - bottom flask. The resulting mixture was stirred at 50 °C for 4 hours, suspended in water at 0 °C, and filtered. The resulting solid was dried under infrared light to obtain 2 - {[6 - ({5 - chloro - 2 - [4 - (dimethoxymethyl)piperidin - 1 - yl]pyrimidin - 4 - yl}amino) - 1 - isopropyl - 2 - oxoquinolin - 3 - yl]oxy} - N - methylacetamide as a brown solid (620 mg, 93%). MS (ESI): m / z 559.05 [M + H] + 。

[0693] Step 2: 2 - ((6 - ((5 - chloro - 2 - (4 - formylpiperidin - 1 - yl)pyrimidin - 4 - yl)amino) - 1 - isopropyl - 2 - oxo - 1,2 - dihydroquinolin - 3 - yl)hydroxy) - N - methylacetamide

[0694]

[0695] To a mixture of 2-[[6-([5-chloro-2-[4-(dimethoxymethyl)piperidin-1-yl]pyrimidin-4-yl]amino)-1-isopropyl-2-oxoquinolin-3-yl]oxy]-N-methylacetamide (220 mg) in water (1.0 mL) was added trifluoroacetic acid (2.0 mL) and dichloromethane (4.0 mL). The resulting mixture was stirred overnight at 40 °C under an air atmosphere. Then, the reaction mixture was concentrated under reduced pressure to give 2-[(6-[[5-chloro-2-(4-formylpiperidin-1-yl)pyrimidin-4-yl]amino]-1-isopropyl-2-oxoquinolin-3-yl)oxy]-N-methylacetamide (202 mg) as a yellow oil, which was used in the next step without further purification.

[0696] Step 3: 1-[(4E)-1-(tert-butoxycarbonyl)-3,3-difluoropiperidin-4-ylidene]-4-(4,4,5,5-tetramethyl-1,3,2-dioxaborolan-2-yl)-1,2,3,6-tetrahydro-1λ5-pyridin-1-ium

[0697]

[0698] To a stirred solution of 4-(4,4,5,5-tetramethyl-1,3,2-dioxaborolan-2-yl)-1,2,3,6-tetrahydropyridine (4.0 g, 19 mmol) in acetonitrile / toluene (20 mL / 40 mL) was added sodium acetate (5.6 g, 68 mmol) and acetic acid (4 mL). The resulting mixture was stirred for 15 minutes. Then 3,3-difluoro-4-oxopiperidine-1-carboxylic acid tert-butyl ester (5.76 g, 24 mmol) was added and the mixture was stirred at 100 °C for 4 hours. The mixture was filtered and the filter cake was washed with dichloromethane (20 mL × 3). The filtrate was concentrated under reduced pressure to give 1-[(4E)-1-(tert-butoxycarbonyl)-3,3-difluoropiperidin-4-ylidene]-4-(4,4,5,5-tetramethyl-1,3,2-dioxaborolan-2-yl)-1,2,3,6-tetrahydro-1λ5-pyridin-1-ium (1.1 g) as a white solid. MS (ESI): m / z 427.30 [M+H] + 。

[0699] Step 4: tert-butyl 3,3-difluoro-4-[4-(4,4,5,5-tetramethyl-1,3,2-dioxaborolan-2-yl)-3,6-dihydro-2H-pyridin-1-yl]piperidine-1-carboxylate

[0700]

[0701] A solution of 1-[(4E)-1-(tert-butoxycarbonyl)-3,3-difluoropiperidin-4-ylidene]-4-(4,4,5,5-tetramethyl-1,3,2-dioxaborolan-2-yl)-1,2,3,6-tetrahydro-1λ5-pyridin-1-ium (1.1 g, 2.5 mmol) and sodium triacetoxyborohydride (8.6 g, 40 mmol) in dichloroethane (10 mL) was stirred overnight at room temperature. It was diluted with water and extracted with dichloromethane (20 mL × 3). The extracts were combined and concentrated. The residue was purified by reverse-phase flash chromatography (column, C18 silica gel; mobile phase, aqueous acetonitrile (10 mmol / L NH4HCO3), 10% to 80% gradient in 30 minutes) to give tert-butyl 3,3-difluoro-4-[4-(4,4,5,5-tetramethyl-1,3,2-dioxaborolan-2-yl)-3,6-dihydro-2H-pyridin-1-yl]piperidine-1-carboxylate (1.1 g) as a white solid. MS (ESI): m / z 429.20 [M+H] + 。

[0702] Step 5: tert-Butyl 4-{4-[2-(2,6-dioxopiperidin-3-yl)-4-fluoro-1-oxo-3H-isoindol-5-yl]-3,6-dihydro-2H-pyridin-1-yl}-3,3-difluoropiperidine-1-carboxylate

[0703]

[0704] To a stirred solution of tert-butyl 3,3-difluoro-4-[4-(4,4,5,5-tetramethyl-1,3,2-dioxaborolan-2-yl)-3,6-dihydro-2H-pyridin-1-yl]piperidine-1-carboxylate (150 mg, 0.4 mmol) and 3-(5-bromo-4-fluoro-1-oxo-3H-isoindol-2-yl)piperidine-2,6-dione (119.5 mg, 0.4 mmol) in dioxane (2 mL) and water (0.2 mL) was added [1,1'-bis(di-tert-butylphosphino)ferrocene]dichloropalladium(II) (45 mg, 0.07 mmol) and cesium fluoride (159.59 mg, 1.0 mmol). The resulting mixture was stirred at 90 °C under a nitrogen atmosphere for 3 h, then diluted with water and extracted with ethyl acetate (20 mL × 3). The combined organic layers were washed with brine (20 mL × 3), dried over anhydrous sodium sulfate, filtered and concentrated under reduced pressure. The residue was purified by silica gel column chromatography (petroleum ether / ethyl acetate = 1:3) to afford tert-butyl 4-{4-[2-(2,6-dioxopiperidin-3-yl)-4-fluoro-1-oxo-3H-isoindol-5-yl]-3,6-dihydro-2H-pyridin-1-yl}-3,3-difluoropiperidine-1-carboxylate (118 mg, 59%) as a white solid. MS (ESI): m / z 563.40 [M+H] + 。

[0705] Step 6: tert-Butyl 4-[2-(2,6-dioxopiperidin-3-yl)-4-fluoro-1-oxo-3H-isoindol-5-yl]-3',3'-difluoro-[1,4'-bipiperidine]-1'-carboxylate

[0706]

[0707] A mixture of tert-butyl 4-{4-[2-(2,6-dioxopiperidin-3-yl)-4-fluoro-1-oxo-3H-isoindol-5-yl]-3,6-dihydro-2H-pyridin-1-yl}-3,3-difluoropiperidine-1-carboxylate (150 mg, 0.3 mmol) and Pd / C (70 mg) in tetrahydrofuran (1 mL) and isopropanol (5 mL) was degassed and purged with hydrogen, then stirred at 40 °C overnight under a hydrogen atmosphere. The resulting mixture was filtered through diatomaceous earth and washed with dichloromethane (10 mL × 3). The filtrate was concentrated under reduced pressure to afford tert-butyl 4-[2-(2,6-dioxopiperidin-3-yl)-4-fluoro-1-oxo-3H-isoindol-5-yl]-3',3'-difluoro-[1,4'-bipiperidine]-1'-carboxylate (118 mg, 78%) as a white solid. MS (ESI): m / z 565.40 [M+H] + 。

[0708] Step 7: 3-(5-{3',3'-Difluoro-[1,4'-dipiperidin]-4-yl}-4-fluoro-1-oxo-3H-isoindol-2-yl)piperidine-2,6-dione

[0709]

[0710] A solution of tert-butyl 4-[2-(2,6-dioxopiperidin-3-yl)-4-fluoro-1-oxo-3H-isoindol-5-yl]-3',3'-difluoro-[1,4'-dipiperidin]-1'-carboxylate (106 mg, 0.2 mmol) and trifluoroacetic acid (2 mL) in dichloromethane (3 mL) was stirred overnight at room temperature. The reaction mixture was concentrated under reduced pressure to afford 3-(5-{3',3'-difluoro-[1,4'-dipiperidin]-4-yl}-4-fluoro-1-oxo-3H-isoindol-2-yl)piperidine-2,6-dione trifluoroacetate (87 mg) as a white solid. MS(ESI): m / z 465.25 [M+H] + 。

[0711] Step 8: 2-{[6-({5-Chloro-2-[4-({4-[2-(2,6-dioxopiperidin-3-yl)-4-fluoro-1-oxo-3H-isoindol-5-yl]-3',3'-difluoro-[1,4'-dipiperidin]-1'-yl}methyl)piperidin-1-yl]pyrimidin-4-yl}amino)-1-isopropyl-2-oxoquinolin-3-yl]oxy}-N-methylacetamide

[0712]

[0713] To a stirred mixture of 3-(5-{3',3'-difluoro-[1,4'-bipiperidin]-4-yl}-4-fluoro-1-oxo-3H-isoindol-2-yl)piperidine-2,6-dione (87 mg, 0.2 mmol) in dimethyl sulfoxide (1 mL) and dichloroethane (10 mL) was added 2-[(6-{[5-chloro-2-(4-formylpiperidin-1-yl)pyrimidin-4-yl]amino}-1-isopropyl-2-oxoquinolin-3-yl)oxy]-N-methylacetamide (96 mg, 0.2 mmol) dissolved in dichloroethane (2 mL). The mixture was basified to pH 7 - 8 with N,N-diisopropylethylamine and stirred overnight at room temperature. Sodium triacetoxyborohydride (119 mg, 0.6 mmol) was added to the above mixture and stirred for 2 h at room temperature. The reaction mixture was filtered and the cake was washed with dichloromethane. The filtrate was concentrated under reduced pressure. The residue was purified by reverse-phase flash chromatography (column, C18 silica gel; mobile phase, acetonitrile / water (10 mmol / L NH4HCO3), 10% to 80% gradient in 30 min) to afford 2-{[6-({5-chloro-2-[4-({4-[2-(2,6-dioxopiperidin-3-yl)-4-fluoro-1-oxo-3H-isoindol-5-yl]-3',3'-difluoro-[1,4'-bipiperidin]-1'-yl}methyl)piperidin-1-yl]pyrimidin-4-yl}amino)-1-isopropyl-2-oxoquinolin-3-yl]oxy}-N-methylacetamide (50.9 mg) as a white solid. 1 H NMR (300 MHz, DMSO) δ 11.01 (s, 1H), 8.81 (s, 1H), 8.04 (s, 1H), 7.95 m, 2H), 7.69 (m, 2H), 7.54 (s, 2H), 7.15 (s, 1H), 5.11 (m, 1H), 4.61 - 4.36 (m, 6H), 3.01 - 2.89 (s, 8H), 2.83 (m, 5H), 2.71–2.54 (m, 2H), 2.21 - 2.02 (s, 6H), 1.73 (s, 9H), 1.57 (m, 7H), 1.02 (s, 2H); MS (ESI): m / z 961.50 [M + H] + 。

[0714] Example 11 - Synthesis of Compound 44

[0715] Step 1: tert-Butyl 3'-fluoro-3,6-dihydro-2H-[4,4'-bipyridine]-1-carboxylate

[0716]

[0717] To a stirred mixture of 4-bromo-3-fluoropyridine (5.0 g, 28 mmol) and tert-butyl 4-(4,4,5,5-tetramethyl-1,3,2-dioxaborolan-2-yl)-3,6-dihydro-2H-pyridine-1-carboxylate (8.78 g, 28 mmol) in dioxane (20 mL) and water (2 mL) was added sodium carbonate (9.03 g, 85 mmol) and [1,1'-bis(diphenylphosphino)ferrocene]dichloropalladium(II) (2.31 g, 2.8 mmol). The resulting mixture was stirred at 80 °C under a nitrogen atmosphere for 2 h, then diluted with water and extracted with ethyl acetate (3 × 10 mL). The combined organic layers were washed with brine (3 × 10 mL), dried over anhydrous sodium sulfate, filtered and concentrated under reduced pressure. The residue was purified by silica gel column chromatography (petroleum ether / ethyl acetate = 10:1) to afford tert-butyl 3'-fluoro-3,6-dihydro-2H-[4,4'-bipyridine]-1-carboxylate as a yellow oil (7.3 g, 92%). MS (ESI): m / z 279.2 [M+H] + 。

[0718] Step 2: 1-Benzyl-1'-(tert-butoxycarbonyl)-3-fluoro-3',6'-dihydro-2'H-[4,4'-bipyridine]-1-ium tert-butyl salt

[0719]

[0720] At 0 °C, benzyl bromide (5.38 g, 31 mmol) was added dropwise over 3 min to a solution of tert-butyl 3'-fluoro-3,6-dihydro-2H-[4,4'-bipyridine]-1-carboxylate (7.3 g, 26 mmol) and acetone (70 mL). The resulting mixture was stirred at 60 °C overnight and then concentrated in vacuo. The residue was triturated with petroleum ether (600 mL) to afford 1-benzyl-1'-(tert-butoxycarbonyl)-3-fluoro-3',6'-dihydro-2'H-[4,4'-bipyridine]-1-ium as a brown solid (9 g, 93%). MS (ESI): m / z 370.3 [M+H] + 。

[0721] Step 3: tert-butyl 1'-benzyl-3'-fluoro-2H,2'H,3H,3'H,6H,6'H-[4,4'-bipyridine]-1-carboxylate

[0722]

[0723] At room temperature, NaBH4 (4.61 g, 122 mmol) was added portionwise to a solution of 1-benzyl-1'-(tert-butoxycarbonyl)-3-fluoro-3',6'-dihydro-2'H-[4,4'-bipyridin]-1-ium (9 g) in methanol (200 mL). The resulting mixture was stirred at room temperature for 2 days, then diluted with water and extracted with ethyl acetate (3 × 10 mL). The combined organic layers were washed with brine (3 × 10 mL), dried over anhydrous sodium sulfate, filtered and concentrated under reduced pressure. The residue was purified by silica gel column chromatography (petroleum ether / ethyl acetate = 10:1) to give tert-butyl 1'-benzyl-3'-fluoro-2H,2'H,3H,3'H,6H,6'H-[4,4'-bipyridin]-1-carboxylate (3.5 g, 39%) as a yellow oil. MS (ESI): m / z 373.30 [M+H] + 。

[0724] Step 4: tert-butyl 3'-fluoro-[4,4'-dipiperidine]-1-carboxylate

[0725]

[0726] A mixture of tert-butyl 1'-benzyl-3'-fluoro-2H,2'H,3H,3'H,6H,6'H-[4,4'-bipyridin]-1-carboxylate (1.8 g, 4.8 mmol) and Pd(OH)2 / C (0.90 g, 6.4 mmol) in methanol (50 mL) was degassed and purged with hydrogen, then stirred overnight at room temperature under a hydrogen atmosphere. The resulting mixture was filtered through a pad of diatomaceous earth and washed with dichloromethane (3 × 10 mL). The filtrate was concentrated under reduced pressure to give tert-butyl 3'-fluoro-[4,4'-dipiperidine]-1-carboxylate (1.4 g) as a yellow oil. MS (ES + ): m / z 287.30 [M+H] + 。

[0727] Step 5: 1-benzyl 1'-tert-butyl 3-fluoro-[4,4'-dipiperidine]-1,1'-dicarboxylate

[0728]

[0729] At 0 °C, benzyl chloroformate (857.7 mg, 5 mmol) was added to a stirred solution of tert-butyl 3'-fluoro-[4,4'-bipiperidine]-1-carboxylate (1.2 g, 4.2 mmol) and triethylamine (848 mg, 8.4 mmol) in dichloromethane. The resulting mixture was stirred at room temperature for 2 hours and then concentrated in vacuo. The residue was purified by reverse-phase flash chromatography (column, C18 silica gel; mobile phase: aqueous acetonitrile (10 mmol / L NH4HCO3), 10% to 50% gradient in 10 minutes) to afford tert-butyl 3-fluoro-[4,4'-bipiperidine]-1,1'-dicarboxylate 1-benzyl ester 1'-tert-butyl ester (570 mg, 32%) as a white solid. MS (ESI): m / z 421.30 [M+H] + 。

[0730] Step 6: Benzyl 3-fluoro-[4,4'-bipiperidine]-1-carboxylate

[0731]

[0732] HCl (gas) was added to a solution of tert-butyl 3-fluoro-[4,4'-bipiperidine]-1,1'-dicarboxylate 1-benzyl ester 1'-tert-butyl ester (270 mg, 0.6 mmol) in 1,4-dioxane (5.0 ml) at room temperature. The resulting mixture was stirred at room temperature for 2 hours and then concentrated in vacuo to afford benzyl 3-fluoro-[4,4'-bipiperidine]-1-carboxylate (200 mg, 97%) as a white solid. MS (ESI): m / z 321.30 [M+H] + 。

[0733] Step 7: Benzyl 1'-[2-(2,6-dioxopiperidin-3-yl)-4-fluoro-1-oxo-3H-isoindol-5-yl]-3-fluoro-[4,4'-bipiperidine]-1-carboxylate

[0734]

[0735] To a stirred mixture of benzyl 3-fluoro-[4,4'-bipiperidine]-1-carboxylate (400 mg, 1.2 mmol) and 3-(5-bromo-4-fluoro-1-oxo-3H-isoindol-2-yl)piperidine-2,6-dione (426 mg, 1.2 mmol) in N,N-dimethylformamide was added dichloro[1,3-bis(2,6-di-3-pentylphenyl)imidazol-2-ylidene](3-chloropyridyl)palladium(II) (5.25 mg, 0.006 mmol) and cesium carbonate (814 mg, 2.5 mmol). The resulting mixture was stirred at 100 °C for 2 h under a nitrogen atmosphere. It was diluted with water and extracted with ethyl acetate (3 × 10 mL). The combined organic layers were washed with brine (3 × 10 mL), dried over anhydrous sodium sulfate, filtered and concentrated under reduced pressure. The residue was purified by reverse-phase flash chromatography (column, C18 silica gel; mobile phase, aqueous acetonitrile (10 mmol / L NH4HCO3), 10% to 50% gradient in 10 min) to give benzyl 1'-[2-(2,6-dioxopiperidin-3-yl)-4-fluoro-1-oxo-3H-isoindol-5-yl]-3-fluoro-[4,4'-bipiperidine]-1-carboxylate (210 mg, 29%). MS (ESI): m / z 581.30 [M+H] + 。

[0736] Step 8: 3-(4-Fluoro-5-{3'-fluoro-[4,4'-bipiperidin]-1-yl}-1-oxo-3H-isoindol-2-yl)piperidine-2,6-dione

[0737]

[0738] To 1'-[2-(2,6-dioxopiperidin-3-yl)-4-fluoro-1-oxo-3H-isoindol-5-yl]-3-fluoro-[4,4'-bipiperidine]-1-carboxylate (150 mg, 0.3 mmol) was added trifluoroacetic acid (3 mL, 0.03 mmol). The resulting mixture was stirred at 50 °C for 3 h and then concentrated in vacuo to give 3-(4-fluoro-5-{3'-fluoro-[4,4'-bipiperidin]-1-yl}-1-oxo-3H-isoindol-2-yl)piperidine-2,6-dione (100 mg, 87%) as a brown oil. MS (ESI): m / z 447.30 [M+H] + 。

[0739] Step 9: 2-{[6-({5-chloro-2-[4-({1'-[2-(2,6-dioxopiperidin-3-yl)-4-fluoro-1-oxo-3H-isoindol-5-yl]-3-fluoro-[4,4'-bipiperidin]-1-yl}methyl)piperidin-1-yl]pyrimidin-4-yl}amino)-1-isopropyl-2-oxoquinolin-3-yl]oxy}-N-methylacetamide

[0740]

[0741] To a stirred mixture of 3-(4-fluoro-5-{3'-fluoro-[4,4'-bipiperidin]-1-yl}-1-oxo-3H-isoindol-2-yl)piperidine-2,6-dione (70 mg, 0.2 mmol) and 2-[(6-{[5-chloro-2-(4-formylpiperidin-1-yl)pyrimidin-4-yl]amino}-1-isopropyl-2-oxoquinolin-3-yl)oxy]-N-methylacetamide (80.43 mg, 0.2 mmol) in dichloroethane (5 mL) was added dropwise N,N-diisopropylethylamine to adjust the pH to 8. The resulting mixture was stirred at 40 °C overnight. At room temperature, sodium triacetoxyborohydride (99.68 mg, 0.5 mmol) was added to the above mixture. The resulting mixture was stirred at room temperature for 2 h. The crude material was purified by reverse-phase flash chromatography (column, C18 silica gel; mobile phase, acetonitrile / water (10 mmol / L NH4HCO3), 10% to 55% gradient in 10 min) to afford 2-{[6-({5-chloro-2-[4-({1'-[2-(2,6-dioxopiperidin-3-yl)-4-fluoro-1-oxo-3H-isoindol-5-yl]-3-fluoro-[4,4'-bipiperidin]-1-yl}methyl)piperidin-1-yl]pyrimidin-4-yl}amino)-1-isopropyl-2-oxoquinolin-3-yl]oxy}-N-methylacetamide (41 mg, 25%) as an off-white solid. 11H NMR (300 MHz, DMSO) δ 10.97 (s, 1H), 7.62 (d, J = 7.9 Hz, 1H), 7.52–7.45 (m, 1H), 7.39 (dd, J = 7.9, 1.5 Hz, 1H), 7.32 (d, J = 4.4 Hz, 4H), 7.31–7.18 (m, 1H), 5.09 (dd, J = 13.2, 5.1 Hz, 1H), 4.41 (d, J = 17.2 Hz, 1H), 4.27 (d, J = 17.2 Hz, 1H), 3.49 (s, 2H), 2.90 (dq, J = 13.5, 6.6, 5.3 Hz, 3H), 2.70–2.51 (m, 2H), 2.37 (qd, J = 13.1, 4.3 Hz, 1H), 2.02 (dtd, J = 17.0, 11.4, 4.1 Hz, 3H), 1.74 (s, 3H), 1.69 (dd, J = 12.2, 3.5 Hz, 1H); MS (ESI): m / z 943.39 [M+H] + 。

[0742] Example 12: Synthesis of Compound 1

[0743] Step 1: tert-Butyl 4-[(1r,3r)-3-{4-[1-(2,6-dioxopiperidin-3-yl)-3-methyl-2-oxo-1,3-benzodiazol-4-yl]-3,6-dihydro-2H-pyridin-1-yl}cyclobutoxy]piperidine-1-carboxylate

[0744]

[0745] At 90 °C under a nitrogen atmosphere, CsF (336.90 mg, 2.217 mmol, 3 eq) and Pd(dtbpf)Cl2 (48.18 mg, 0.074 mmol, 0.1 eq) were added dropwise / batchwise to a stirred solution / mixture of tert-butyl 4-[(1r,3r)-3-[4-(4,4,5,5-tetramethyl-1,3,2-dioxaborolan-2-yl)-3,6-dihydro-2H-pyridin-1-yl]cyclobutoxy]piperidine-1-carboxylate (Example 5, Step 6; 512.82 mg, 1.109 mmol, 1.5 eq) and 3-(4-bromo-3-methyl-2-oxo-1,3-benzoxazol-1-yl)piperidine-2,6-dione (WO2019060693, 250 mg, 0.739 mmol, 1.00 eq) in 1,4-dioxane and water. The resulting mixture was stirred at 90 °C under a nitrogen atmosphere for 3 h. The resulting mixture was extracted with EtOAc (10 × mL). The combined organic layers were washed with brine (3 × 10 mL), dried over anhydrous Na2SO4. After filtration, the filtrate was concentrated under reduced pressure. The residue was purified by reverse-phase flash chromatography under the following conditions: column, C18 silica gel; mobile phase, MeCN / water (10 mmol / L NH4HCO3), 10% to 50% gradient, within 10 min; detector, UV 254 nm. This gave tert-butyl 4-[(1r,3r)-3-{4-[1-(2,6-dioxopiperidin-3-yl)-3-methyl-2-oxo-1,3-benzoxazol-4-yl]-3,6-dihydro-2H-pyridin-1-yl}cyclobutoxy]piperidine-1-carboxylate (230 mg, 52.40%) as a white solid. LC-MS (ES + ): m / z 594.37 [MH + , t R = 1.119 min (2.0 min run).

[0746] Step 2: 2-({6-[(5-chloro-2-{4-[(1r,3r)-3-{4-[4-fluoro-1-oxo-2-(2-oxopiperidin-3-yl)-3H-isoindol-5-yl]piperidin-1-yl}cyclobutoxy]piperidin-1-yl}pyrimidin-4-yl)amino]-1-isopropyl-2-oxoquinolin-3-yl}oxy)-N-methylacetamide

[0747]

[0748] Compound 1 was prepared using the procedure of Example 5 / Compound 37, Steps 8 - 10 as a 45 mg off-white solid (100%). 11H NMR (300 MHz, DMSO-d6, ppm): δ 11.08 (s, 1H), 8.84 (s, 1H), 8.05 (s, 1H), 7.97 (d, J = 10.4 Hz, 2H), 7.70 (d, J = 1.4 Hz, 2H), 7.06–6.89 (m, 4H), 5.37 (dd, J = 12.8, 5.3 Hz, 1H), 4.55 (s, 2H), 4.13 (d, J = 14.8 Hz, 3H), 3.57 (s, 3H), 3.53 (s, 1H), 3.22 (d, J = 10.0 Hz, 3H), 3.00 (d, J = 9.9 Hz, 4H), 2.87 (d, J = 14.8 Hz, 2H), 2.69 (d, J = 4.6 Hz, 3H), 2.22–2.12 (m, 2H), 2.00 (d, J = 5.6 Hz, 3H), 1.82 (d, J = 11.3 Hz, 8H), 1.72 (d, J = 11.4 Hz, 1H), 1.58 (d, J = 6.8 Hz, 5H), 1.38 (d, J = 8.7 Hz, 3H).

[0749] LC-MS (ES + ): m / z 895.39 [MH + , t R = 1.235 min (run time 3.0 min).

[0750] Example 13: Synthesis of Compound 5

[0751] Step 1: 3-[(4-Bromophenyl)amino]piperidine-2,6-dione

[0752]

[0753] At room temperature, DIEA (4.5 g, 34.9 mmol, 3.0 equiv) was added to a stirred solution of 4-bromoaniline (2.0 g, 11.6 mmol, 1.0 equiv) and 3-bromopiperidine-2,6-dione (2.2 g, 11.6 mmol, 1.0 equiv) in DMF (50 mL). The resulting mixture was stirred at 100 °C overnight. The residue was purified by reverse-phase flash chromatography under the following conditions: column, C18 silica gel; mobile phase, CH3CN / H2O (10 mmol / L FA), 5% to 50% gradient over 45 min; detector, UV 254 nm. This gave 3-[(4-bromophenyl)amino]piperidine-2,6-dione (260 mg) as a grayish-brown solid. LC-MS (ES + ): m / z 279.00 [MH + , t R = 0.908 min (run time 2.0 min).

[0754] Step 2: tert-Butyl 4-[(1r,3r)-3-(4-{4-[(2,6-dioxopiperidin-3-yl)amino]phenyl}piperazin-1-yl)cyclobutoxy]piperidine-1-carboxylate

[0755]

[0756] A mixture of 3-[(4-bromophenyl)amino]piperidine-2,6-dione (260 mg, 0.8 mmol, 1.0 equiv), tert-butyl 4-[(1r,3r)-3-(piperazin-1-yl)cyclobutoxy]piperidine-1-carboxylate (WO2022221673, 299.8 mg, 0.8 mmol, 1.0 equiv) and Cs2CO3 (863.10 mg, 2.649 mmol, 3.0 equiv), Pd-PEPPSI-IPentCl (Khadra A, Mayer S, Organ M G. Pd-PEPPSI-IPent Cl : A Useful Catalyst for the Coupling of 2-Aminopyridine Derivatives. Chemistry. Mar 2, 2017; 23(13): 3206-3212.), 2-methylpyridine (o-methylpyridine) (67.28 mg, 0.08 mmol, 0.1 equiv) in DMF (5.0 mL) was degassed with nitrogen three times. The mixture was stirred at 110 °C overnight. The mixture was cooled to room temperature. The resulting mixture was diluted with ethyl acetate (300 mL). The resulting mixture was extracted with EtOAc (200 mL). The combined organic layers were washed with brine (100 mL) and dried over anhydrous Na2SO4. After filtration, the filtrate was concentrated under reduced pressure. The residue was purified by reverse-phase flash chromatography under the following conditions: column, C18 silica gel; mobile phase, MeCN / water (10 mmol / L NH4HCO3), 5% to 50% gradient, within 30 minutes; detector, UV 254 nm. This gave tert-butyl 4-[(1r,3r)-3-(4-{4-[(2,6-dioxopiperidin-3-yl)amino]phenyl}piperazin-1-yl)cyclobutoxy]piperidine-1-carboxylate as a black solid (136 mg, 28%). LC-MS (ES + ): m / z 542.40 [M+H + , t R = 0.704 min (1.2 min run).

[0757] Step 3:

[0758]

[0759] Compound 5 (33.8 mg, white solid) was prepared using the procedure of Example 5 / Compound 37, Steps 9 - 10. 1 H NMR (300 MHz, DMSO - d6, ppm): δ 8.83 (s, 1H), 8.04 (s, 1H), 7.97 (s, 2H), 7.69 (d, J = 10.2 Hz, 3H), 7.28 (d, J = 8.5 Hz, 2H), 7.16 (s, 1H), 7.02 (s, 1H), 6.89 (d, J = 8.8 Hz, 2H), 4.54 (d, J = 7.3 Hz, 2H), 4.18 - 4.10 (m, 3H), 3.52 (s, 1H), 3.20 (d, J = 10.8 Hz, 2H), 3.08 (s, 3H), 2.78 (s, 1H), 2.66 (d, J = 4.5 Hz, 3H), 2.39 (s, 4H), 2.32 (s, 1H), 2.17 (s, 2H), 2.02 - 1.96 (m, 3H), 1.80 (s, 2H), 1.56 (d, J = 6.8 Hz, 6H), 1.37 (s, 2H), 1.22 (s, 4H), 0.82 (s, 2H). LC - MS (ES + ): m / z 841.30 [M + H + , t R = 7.599 min (run time 13.0 min).

[0760] Example 14: Synthesis of Compound 17

[0761] Step 1: 3-(5 - Bromoindol - 1 - yl)piperidine - 2,6 - dione

[0762]

[0763] At room temperature, 5-bromoindole (1.02 g, 5.20 mmol, 2.0 eq) dissolved in THF (10 mL) and NaH (156 mg, 3.90 mmol, 1.5 eq, 60%) were added to a 30 mL sealed tube. The resulting mixture was stirred at room temperature for 1 h under a nitrogen atmosphere. At 0 °C, 3-bromopiperidine-2,6-dione (0.50 g, 2.60 mmol, 1.0 eq) dissolved in THF (5 mL) was added dropwise to the above mixture. The resulting mixture was stirred at 60 °C overnight. The resulting mixture was extracted with 4:1 CH2Cl2 / IPA (3 × 20 mL). The combined organic layers were dried over anhydrous Na2SO4. After filtration, the filtrate was concentrated under reduced pressure. The residue was purified by silica gel column chromatography, eluting with petroleum ether / ethyl acetate (5:1) to give 3-(5-bromoindol-1-yl)piperidine-2,6-dione (420 mg, 52.56%) as a white solid. LC-MS (ES + ): m / z 306 [MH + , t R = 0.91 min (2.00 min run).

[0764] Step 2: 2-({6-[(5-chloro-2-{4-[(1r,3r)-3-{4-[1-(2,6-dioxopiperidin-3-yl)indol-5-yl]piperazin-1-yl}cyclobutoxy]piperidin-1-yl}pyrimidin-4-yl)amino]-1-isopropyl-2-oxoquinolin-3-yl}oxy)-N-methylacetamide

[0765]

[0766] Compound 17 was prepared analogously to Example 13 / Compound 5 to give 65.9 mg (46.80%) of a white solid. 11H NMR (300 MHz, DMSO-d6) δ 11.04 (s, 1H), 8.83 (s, 1H), 8.05 (s, 1H), 8.00–7.92 (m, 2H), 7.70 (m, 2H), 7.35–7.24 (m, 2H), 7.06–6.99 (m, 2H), 6.91 (m, 1H), 6.36 (m, 1H), 5.53 (m, 1H), 5.33 (s, 1H), 4.55 (s, 2H), 4.25–4.02 (m, 3H), 3.54 (s, 1H), 3.24 (m, 2H), 3.06 (s, 4H), 2.95–2.80 (m, 2H), 2.69 (m, 4H), 2.60 (s, 1H), 2.45 (s, 4H), 2.40 (s, 3H), 2.15 (m, 2H), 1.83 (m, 2H), 1.58 (m, 6H), 1.39 (m, 2H). LC-MS (ES + ): m / z 864.38 [MH + , t R = 1.13 min (run time 2.00 min).

[0767] Example 15: Synthesis of Compound 21

[0768] Step 1: 3-(5-Bromo-1H-indazol-1-yl)piperidine-2,6-dione and 3-(5-bromo-1H-indazol-2-yl)piperidine-2,6-dione

[0769]

[0770] At room temperature, 5-bromo-1H-indazole (1970.23 mg, 9.999 mmol, 2.00 equiv) and THF (40 mL) were added to a 100 mL three-necked round-bottom flask. NaH (239.97 mg, 10.000 mmol, 2.00 equiv) was added portionwise to the above mixture at 0 °C. The resulting mixture was stirred for an additional 30 minutes at 60 °C. At 60 °C, 3-bromopiperidine-2,6-dione (960 mg, 5.000 mmol, 1.00 equiv) dissolved in THF was added dropwise to the above mixture. The resulting mixture was stirred overnight at 60 °C. The resulting mixture was diluted with ethyl acetate (200 mL). The resulting mixture was added to 200 mL of 20% HCl (aqueous solution). The aqueous layer was extracted with EtOAc (2 × 80 mL). The residue was purified by silica gel column chromatography, eluting with petroleum ether / ethyl acetate (2:3) to isolate two regioisomers.

[0771] 3-(5-Bromo-1H-indazol-1-yl)piperidine-2,6-dione (1 g, 64.91%), a grayish-white solid. LC-MS (ES +): m / z 309.95 [MH + , t R = 0.721 min (run time 2.0 min).

[0772] 3-(5-Bromoindazol-2-yl)piperidine-2,6-dione (600 mg, 38.95%), a grayish-white solid.

[0773] LC-MS (ES + ): m / z 307.85 [MH + , t R = 0.687 min (run time 2.0 min).

[0774] Step 2: Synthesis of 2-({6-[(5-chloro-2-{4-[(1r,3r)-3-{4-[1-(2,6-dioxopiperidin-3-yl)indazol-5-yl]piperazin-1-yl}cyclobutoxy]piperidin-1-yl}pyrimidin-4-yl)amino]-1-isopropyl-2-oxoquinolin-3-yl}oxy)-N-methylacetamide

[0775]

[0776] Compound 21 was prepared similar to Example 13 / Compound 5 by substituting 3-(5-bromoindazol-1-yl)piperidine-2,6-dione for 3-[(4-bromophenyl)amino]piperidine-2,6-dione in Step 1, giving 55 mg (35.47%) of a light brown solid. 1 HNMR (300 MHz, DMSO-d6, ppm) δ 11.08 (s, 1H), 8.85 (s, 1H), 8.04 - 7.91 (m, 4H), 7.69 (m, 2H), 7.49 (m, 1H), 7.26 (m, 1H), 7.10 (m, 1H), 7.02 (s, 1H), 5.76 (m, 1H), 5.32 (s, 1H), 4.55 (s, 2H), 4.25 - 4.07 (m, 3H), 3.23 (m, 1H), 3.20 (s, 2H), 3.09 (s, 4H), 2.82 (s, 3H), 2.77 - 2.64 (m, 1H), 2.46 (s, 4H), 2.31 - 2.19 (m, 3H), 2.19 (s, 3H), 2.00 (m, 2H), 1.83 (m, 2H), 1.57 (m, 6H), 1.38 (m, 2H). LC-MS (ES + ): m / z 866.45 [MH + , t R = 6.315 min (run time 13.0 min).

[0777] Example 16: Synthesis of Compound 22

[0778]

[0779] Compound 22 was prepared in a similar manner to Example 13 / Compound 5 by substituting 3-[(4-bromophenyl)amino]piperidine-2,6-dione with 3-(5-bromo-1H-indazol-2-yl)piperidine-2,6-dione in Step 1, yielding 27 mg (16%) of a light gray solid. 1 H NMR (300 MHz, DMSO-d6, ppm) δ 8.84 (s, 1H), 8.20 (s, 1H), 8.05 (s, 1H), 7.97 (m, 2H), 7.70 (s, 2H), 7.47 (m, 1H), 7.17 (m, 1H), 7.03 (s, 1H), 6.90 (s, 1H), 5.64 (m, 1H), 4.55 (s, 2H), 4.11 (m, 3H), 3.54 (s, 1H), 3.26 - 3.13 (m, 2H), 3.07 (s, 2H), 3.07 (s, 4H), 2.77 - 2.65 (m, 2H), 2.50 (s, 1H), 2.45 (s, 3H), 2.31 (s, 1H), 2.19 (s, 4H), 1.82 (s, 5H), 1.57 (m, 6H), 1.39 (s, 3H), 1.24 (s, 1H). LC-MS (ES + ): m / z 866.45 [MH + , t R = 5.598 min (10.0 min run).

[0780] Example 17: Synthesis of Compound 23

[0781]

[0782] Compound 23 was prepared in a similar manner to Compound 21 by substituting 3-(5-bromo-1H-indazol-1-yl)piperidine-2,6-dione with 3-(5-bromo-1,3-benzodiazol-1-yl)piperidine-2,6-dione in the first step, yielding the title compound as 42 mg (23%) of an off-white solid. 11H NMR (300 MHz, DMSO-d6) δ 11.15 (s, 1H), 8.86 (s, 1H), 8.14 (s, 1H), 8.05 (s, 1H), 7.97 (m, 2H), 7.70 (m, 2H), 7.38 (m, 1H), 7.13 (m, 1H), 7.03 (s, 1H), 5.62 (m, 1H), 5.32 (s, 1H), 4.55 (s, 2H), 4.24–4.07 (m, 2H), 3.54 (s, 1H), 3.23 (m, 2H), 3.10 (s, 3H), 2.75 (m, 6H), 2.45 (s, 3H), 2.25–2.15 (m, 3H), 1.99 (m, 0H), 1.84 (m, 2H), 1.57 (m, 6H), 1.38 (m, 1H). LC-MS (ES + ): m / z 866.55 [MH + , t R = 2.694 min (9 min run).

[0783] Example 18: Synthesis of Compound 24

[0784]

[0785] Compound 24 was prepared similar to Compound 21 by substituting 3-(6-bromo-1,3-benzodiazol-1-yl)piperidine-2,6-dione for 3-(5-bromoindazol-1-yl)piperidine-2,6-dione in the first step to give the title compound as a 52.6 mg (44%) white solid. 1 1H NMR (300 MHz, DMSO-d6) δ 11.15 (s, 1H), 8.86 (s, 1H), 8.14 (s, 1H), 8.05 (s, 1H), 7.97 (m, 2H), 7.70 (m, 2H), 7.38 (m, 1H), 7.13 (m, 1H), 7.03 (s, 1H), 5.62 (m, 1H), 5.32 (s, 1H), 4.55 (s, 2H), 4.24–4.07 (m, 2H), 3.54 (s, 1H), 3.23 (m, 2H), 3.10 (s, 3H), 2.75 (m, 6H), 2.45 (s, 3H), 2.25–2.15 (m, 3H), 1.99 (m, 0H), 1.84 (m, 2H), 1.57 (m, 6H), 1.38 (m, 1H). LC-MS (ES + ): m / z 866.55 [MH + , t R = 2.694 min (9 min run).

[0786] Compound Characterization (Mass Spectrometry)

[0787] The mass spectrometry data of the compounds of the present disclosure are provided in Table 4 below.

[0788] Table 4. Mass spectrometry data of the compounds of the present disclosure

[0789]

[0790]

[0791]

[0792]

[0793]

[0794]

[0795]

[0796] Biological activity data of the compounds of the present disclosure

[0797] Protein synthesis. The BCL6 protein was expressed by transforming Invitrogen One Shot cells with the GS63525 pET24a-His-SUMO-TEV-BCLm-Avitag plasmid according to the manufacturer's instructions. In addition, biotin at a final concentration of 50 μM and IPTG at a final concentration of 1 mM were added to the culture and shaken overnight at room temperature.

[0798] Immunofluorescence protocol for BCL6 high-content imaging

[0799] T47D cells were seeded in 100 μl volumes of RPMI 1640 - 10% FBS in 96-well black / clear bottom plates for adherent lines (Corning #3904).

[0800] Day 1. T47D breast cancer epithelial cells were seeded at a density such that the confluence at the end point was approximately 70% - 90%. Cells were seeded at 7K / 0.1 mL / well one morning one day before adding the exemplary bifunctional degrading compound.

[0801] Compound treatment

[0802] Day 2. Prepare an 11-point 3-fold serial dilution of the exemplary bifunctional compound in DMSO and aliquot appropriate volumes into the cell growth medium to yield 2-fold final concentrations of the exemplary bifunctional compound. Add an equal volume (0.1 ml) of the 2-fold exemplary bifunctional compound / media mixture to the previously plated cells such that the final highest concentration in the aqueous cell growth medium is 0.1 μM or 1 μM. Incubate at 37 °C, 5% CO2 for 3 days.

[0803] Day 5 Immunofluorescence. Discard the cell medium. Rinse each well with 200 μl of phosphate-buffered saline (PBS) at room temperature. Prepare 4% paraformaldehyde (PFA) from 16% PFA (Electron Microscopy Sciences #15710) using 1X PBS. Add fifty μL of 4% PF to each well and incubate at room temperature for 15 minutes to fix the cells. Aspirate the PFA and wash the cells twice with PBS (200 μL).

[0804] Prepare PBS containing 0.1% Triton X-100 using a 10% triton X-100 stock solution. Permeabilize the cells by adding 100 μL of PBS containing 0.1% Triton X-100 to each well and incubating at room temperature for 15 minutes. Wash the cells twice with PBS.

[0805] Prepare 3% BSA / PBS (from TBS containing Thermofisher #37515 blocking agent BSA, 10%) and add 100 μL to each well. Incubate the cells at room temperature for at least 1 hour.

[0806] Prepare 1% BSA / PBS using the blocking agent BSA / PBS and remove the 3% BSA / PBS from the wells.

[0807] For the no primary antibody control, add 50 μL of 1% BSA / PBS.

[0808] Dilute the primary antibody (BCL6Rb Ab, CST-14895, Cell Signaling) 1:300 in 1% BSA / PBS using the blocking agent BSA / PBS.

[0809] Add fifty μl of the primary antibody to all remaining wells (i.e., all wells except the primary antibody control) and incubate the cells overnight at 4 °C with slow orbital shaking.

[0810] Day 6. Remove the contents of the wells and wash the cells four times with 200 μL PBS. Prepare 1% BSA / PBS using PBS containing the blocking agent BSA.

[0811] Dilute the secondary antibody goat anti-Rb IgG Alexa-488 1:1000 and 1% BSA / PBS containing CellMask-Alexa-647 1:3000 in the same mixture. Add 50 μL to each well and incubate for 1 hour at room temperature in the dark.

[0812] Wash the cells three times with 200 μL PBS and then incubate with 100 μL Hoechst dye (20 mM stock solution) at 1 μg / mL for 10 minutes to stain the cell nuclei. Then wash the wells with 200 μl PBS, add 100 μL PBS to each well, and cover the plate with an opaque plastic lid. Store the plate at 4 °C, covered in aluminum foil, until imaging.

[0813] Before reading, equilibrate the plate to room temperature. Immediately before imaging, wipe the bottom of the plate with 70% isopropanol.

[0814] Imaging:

[0815] 10x, 4 fields / well, including Top hat smoothing in the analysis protocol.

[0816] Supplies / Reagents:

[0817] 16% Paraformaldehyde: Electron Microscopy Sciences #15710

[0818] Hoechst: Thermo Fisher Scientific #62249

[0819] PBS containing blocking agent BSA, 10%: Thermo Fisher Scientific #37515

[0820] TBS containing blocking agent BSA, 10%: Thermo Fisher Scientific #37520

[0821] Goat anti-rabbit or mouse AlexaFluor-488: Thermo Fisher Scientific #A11008

[0822] CellMask Deep Red AlexaFluor-647: Thermo Fisher Scientific #C10046 Wash buffer, PBS: 20x PBS, Thermo Fisher Scientific

[0823] Table 5. Bioactivity data of the compounds of the present disclosure

[0824]

[0825]

[0826] nd = undetermined; *DC 50 (nM) Range: A < 10; 10 ≤ B < 50; 50 ≤ C < 100; D ≥ 100; **D MAX (%) Range: A ≥ 70; 50 ≤ B < 70; C < 50

[0827] Enumerated embodiments

[0828] Aspects of the present disclosure are further illustrated with reference to the following numbered embodiments:

[0829] 1A. A bifunctional compound of formula (I), formula (II) or formula (III):

[0830]

[0831]

[0832] Or a pharmaceutically acceptable salt thereof, wherein:

[0833] R 1 is H or C1-C6 alkyl;

[0834] Q is

[0835] X is N or CH;

[0836] Y1, Y2 and Y3 are each independently N or CR 3 ;

[0837] Z1 and Z2 are each independently N or CH;

[0838] R 2 is H or C1-C6 alkyl;

[0839] Each R 3 is independently H, halogen, C1-C6 alkyl, C1-C6 haloalkyl, -O-(C1-C6 alkyl), -O-(C1-C6 haloalkyl);

[0840] Of Q Indicates the point of attachment to X or succinimide;

[0841] R 1a is H or halogen;

[0842] R 2a is H or C1-C3 alkyl;

[0843] X 3a is CHR 3aor C(O);

[0844] R 3a is H or C1-C3 alkyl;

[0845] X 4a and X 6a are each independently CH or N; and

[0846] R 5a is H, C1-C3 alkyl or halogen;

[0847] L is

[0848] X 6b is CHR 6b or C(O);

[0849] R 6b is H or C1-C3 alkyl;

[0850] R 1b , R 2b , R 3b and R 4b are each independently H or halogen, where at least one of R 1b , R 2b , R 3b , R 4b is halogen;

[0851] R 5b is H or halogen;

[0852] R 6b is H or C1-C3 alkyl;

[0853] X 1b and X 2b are each independently CH or N, where at least one of X 1b and X 2b is N; and

[0854] where each of L indicates a point of attachment.

[0855] 1B. A bifunctional compound of formula (I):

[0856]

[0857] or a pharmaceutically acceptable salt thereof, wherein:

[0858] R 1 is H or C1-C6 alkyl;

[0859] Q is

[0860] X is N or CH;

[0861] Y1, Y2 and Y3 are each independently N or CR 3 ;

[0862] Z1 and Z2 are each independently N or CH;

[0863] R 2 is H or C1-C6 alkyl;

[0864] Each R 3 is independently H, halogen, C1-C6 alkyl, C1-C6 haloalkyl, -O-(C1-C6 alkyl), -O-(C1-C6 haloalkyl); and

[0865] The indication of Q is the point of attachment to X or succinimide.

[0866] 1C. A bifunctional compound of formula (II):

[0867]

[0868] or a pharmaceutically acceptable salt thereof, wherein:

[0869] R 1a is H or halogen;

[0870] R 2a is H or C1-C3 alkyl;

[0871] X 3a is CHR 3a or C(O);

[0872] R 3a is H or C1-C3 alkyl;

[0873] X 4a and X 6a are each independently CH or N; and

[0874] R 5a is H, C1-C3 alkyl or halogen.

[0875] 1D. A bifunctional compound of formula (III):

[0876]

[0877] or a pharmaceutically acceptable salt thereof, wherein:

[0878] L is

[0879] X 6b is CHR 6bor C(O);

[0880] R 6b is H or C1-C3 alkyl;

[0881] R 1b 、R 2b 、R 3b and R 4b are each independently H or halogen, provided that at least one of R 1b 、R 2b 、R 3b 、R 4b is halogen;

[0882] R 5b is H or halogen;

[0883] R 6b is H or C1-C3 alkyl;

[0884] X 1b and X 2b are each independently CH or N, provided that at least one of X 1b and X 2b is N; and

[0885] where each of the L's denotes a point of attachment.

[0886] 1E. The bifunctional compound or a pharmaceutically acceptable salt thereof according to Example 1A or 1C, wherein the compound is a compound of formula (II-a):

[0887]

[0888] or a pharmaceutically acceptable salt thereof.

[0889] 1F. The bifunctional compound or a pharmaceutically acceptable salt thereof according to Example 1A or 1C, wherein the compound is a compound of formula (II-b), formula (II-c), formula (II-d), formula (II-e), formula (II-f), formula (II-g), formula (II-h), formula (II-i) or formula (II-j):

[0890]

[0891]

[0892]

[0893] or a pharmaceutically acceptable salt thereof.

[0894] 1G. A bifunctional compound or a pharmaceutically acceptable salt thereof according to Embodiment 1A or 1D, wherein the compound is a compound of Formula (III-a), Formula (III-c), Formula (III-d), Formula (III-f), Formula (III-g), Formula (III-h) or Formula (III-i):

[0895]

[0896]

[0897] or a pharmaceutically acceptable salt thereof.

[0898] 1H. A bifunctional compound of Formula (IV-b) or Formula (IV-e):

[0899]

[0900]

[0901] or a pharmaceutically acceptable salt thereof.

[0902] 2A. A bifunctional compound of Formula (I), Formula (II) or Formula (III):

[0903]

[0904]

[0905] Wherein:

[0906] R 1 is H or C1-C6 alkyl;

[0907] Q is

[0908] X is N or CH;

[0909] Y1, Y2 and Y3 are each independently N or CR 3 ;

[0910] Z1 and Z2 are each independently N or CH;

[0911] R 2 is H or C1-C6 alkyl;

[0912] Each R 3 is independently H, halogen, C1-C6 alkyl, C1-C6 haloalkyl, -O-(C1-C6 alkyl), -O-(C1-C6 haloalkyl);

[0913] The designation of Q indicates the point of attachment to X or the succinimide

[0914] R 1a is H or a halogen;

[0915] R 2a is H or a C1-C3 alkyl;

[0916] X 3a is CHR 3a or C(O);

[0917] R 3a is H or a C1-C3 alkyl;

[0918] X 4a and X 6a each independently is CH or N; and

[0919] R 5a is H, a C1-C3 alkyl or a halogen;

[0920] L is

[0921] X 6b is CHR 6b or C(O);

[0922] R 6b is H or a C1-C3 alkyl;

[0923] R 1b 、R 2b 、R 3b and R 4b each independently is H or a halogen, wherein R 1b 、R 2b 、R 3b 、R 4b in at least one of which is a halogen;

[0924] R 5b is H or a halogen;

[0925] R 6b is H or a C1-C3 alkyl;

[0926] X 1b and X 2b each independently is CH or N, wherein X 1b and X 2b in at least one of which is N; and

[0927] wherein each of L denotes a point of attachment.

[0928] 2B. A bifunctional compound of formula (I):

[0929]

[0930] Wherein:

[0931] R 1 is H or a C1-C6 alkyl group;

[0932] Q is

[0933] X is N or CH;

[0934] Y1, Y2, and Y3 are each independently N or CR 3 ;

[0935] Z1 and Z2 are each independently N or CH;

[0936] R 2 is H or a C1-C6 alkyl group;

[0937] Each R 3 is independently H, halogen, a C1-C6 alkyl group, a C1-C6 haloalkyl group, -O-(C1-C6 alkyl), -O-(C1-C6 haloalkyl); and

[0938] The designation of Q indicates the point of attachment to X or the succinimide.

[0939] 2C. A bifunctional compound of formula (II):

[0940]

[0941] Wherein:

[0942] R 1a is H or halogen;

[0943] R 2a is H or a C1-C3 alkyl group;

[0944] X 3a is CHR 3a or C(O);

[0945] R 3a is H or a C1-C3 alkyl group;

[0946] X 4a and X 6a are each independently CH or N; and

[0947] R 5a is H, a C1-C3 alkyl group, or halogen.

[0948] 2D. A bifunctional compound of formula (III):

[0949]

[0950] Wherein:

[0951] L is

[0952] X 6b is CHR 6b or C(O);

[0953] R 6b is H or C1-C3 alkyl;

[0954] R 1b , R 2b R 3b and R 4b are each independently H or halogen, wherein at least one of R 1b R 2b R 3b R 4b is halogen;

[0955] R 5b is H or halogen;

[0956] R 6b is H or C1-C3 alkyl;

[0957] X 1b and X 2b are each independently CH or N, wherein at least one of X 1b and X 2b is N; and

[0958] wherein each of the L's denotes a point of attachment.

[0959] 2E. The bifunctional compound according to embodiment 2A or 2C, wherein the compound is a compound of formula (II-a):

[0960]

[0961] 2F. The bifunctional compound according to embodiment 2A or 2C, wherein the compound is a compound of formula (II-b), formula (II-c), formula (II-d), formula (II-e), formula (II-f), formula (II-g), formula (II-h), formula (II-i) or formula (II-j):

[0962]

[0963]

[0964]

[0965]

[0966] 2G. A bifunctional compound according to Example 2A or 2D, wherein the compound is a compound of formula (III-a), formula (III-c), formula (III-d), formula (III-f), formula (III-g), formula (III-h) or formula (III-i):

[0967]

[0968]

[0969]

[0970] 2H. A bifunctional compound of formula (IV-b) or formula (IV-e):

[0971]

[0972] 3. The bifunctional compound or a pharmaceutically acceptable salt thereof according to any one of Examples 1A, 1B, 2A or 2B, wherein Q is

[0973] 4. The bifunctional compound or a pharmaceutically acceptable salt thereof according to any one of Examples 1A, 1B, 2A or 2B, wherein each of Y1, Y2 and Y3 is CH.

[0974] 5. The bifunctional compound or a pharmaceutically acceptable salt thereof according to any one of Examples 11A, 1B, 2A or 2B, wherein one of Y1, Y2 and Y3 is N, and the other two of Y1, Y2 or Y3 are CH.

[0975] 6. The bifunctional compound or a pharmaceutically acceptable salt thereof according to any one of Examples 1A, 1B, 2A or 2B, wherein two of Y1, Y2 and Y3 are N, and the other one of Y1, Y2 or Y3 is CH.

[0976] 7. The bifunctional compound or a pharmaceutically acceptable salt thereof according to any one of Examples 1A, 1B, 2A or 2B, wherein R 1 is H or CH3.

[0977] 8. The bifunctional compound or a pharmaceutically acceptable salt thereof according to any one of Examples 1A, 1B, 2A or 2B, wherein X is N.

[0978] 9. The bifunctional compound or a pharmaceutically acceptable salt thereof according to any one of Examples 1A, 1B, 2A or 2B, wherein X is CH.

[0979] 10. The bifunctional compound according to any one of Embodiment 1A, 1B, 12A or 2B, or a pharmaceutically acceptable salt thereof, wherein R 2 is methyl, ethyl or isopropyl.

[0980] 11. The bifunctional compound according to any one of Embodiment 1A, 1B, 2A or 2B, or a pharmaceutically acceptable salt thereof, wherein each R 3 is independently H, methyl, fluorine or methoxy.

[0981] 12. The bifunctional compound according to any one of Embodiment 1A, 1C, 1E, 2A, 2C or 2E, or a pharmaceutically acceptable salt thereof, wherein R 1a is F.

[0982] 13. The bifunctional compound according to any one of Embodiment 1A, 1C, 1E, 2A, 2C or 2E, or a pharmaceutically acceptable salt thereof, wherein R 1a is Cl.

[0983] 14. The bifunctional compound according to any one of Embodiment 1A, 1C, 1E, 2A, 2C or 2E, or a pharmaceutically acceptable salt thereof, wherein R 1a is H.

[0984] 15. The bifunctional compound according to any one of Embodiment 1A, 1C, 1E, 1F, 2A, 2C, 2E or 2F, or a pharmaceutically acceptable salt thereof, wherein X 4a and X 6a are each N.

[0985] 16. The bifunctional compound according to any one of Embodiment 1A, 1C, 1E, 1F, 2A, 2C, 2E or 2F, or a pharmaceutically acceptable salt thereof, wherein one of X 4a and X 6a is N and the other is CH.

[0986] 17. The bifunctional compound according to any one of Embodiment 1A, 1C, 1E, 2A, 2C or 2E, or a pharmaceutically acceptable salt thereof, wherein R 3a is CH3.

[0987] 18. The bifunctional compound according to any one of Embodiment 1A, 1C, 1E, 2A, 2C or 2E, or a pharmaceutically acceptable salt thereof, wherein R 3a is H.

[0988] 19. The bifunctional compound according to any one of Embodiment 1A, 1C, 1E, 1F, 2A, 2C, 2E or 2F, or a pharmaceutically acceptable salt thereof, wherein R 5a is CH3.

[0989] The bifunctional compound or a pharmaceutically acceptable salt thereof according to any one of embodiments 1A, 1C, 1E, 1F, 2A, 2C, 2E or 2F, wherein R 5a is H.

[0990] The bifunctional compound or a pharmaceutically acceptable salt thereof according to any one of embodiments 1A, 1C, 1E, 1F, 2A, 2C, 2E or 2F, wherein R 5a is F.

[0991] The bifunctional compound or a pharmaceutically acceptable salt thereof according to any one of embodiments 1A, 1C, 1E, 2A, 2C or 2E, wherein R 2a is CH3.

[0992] The bifunctional compound or a pharmaceutically acceptable salt thereof according to any one of embodiments 1A, 1C, 1E, 2A, 2C or 2E, R 2a is H.

[0993] The bifunctional compound or a pharmaceutically acceptable salt thereof according to any one of embodiments 1A, 1D, 1G, 1E, 2A, 2D, 2G or 2E, wherein L is

[0994] The bifunctional compound or a pharmaceutically acceptable salt thereof according to any one of embodiments 1A, 1D, 1G, 1E, 2A, 2D, 2G or 2E, wherein L is

[0995] The bifunctional compound or a pharmaceutically acceptable salt thereof according to any one of embodiments 1A, 1D, 1G, 1E, 2A, 2D, 2G or 2E, wherein L is

[0996] The bifunctional compound or a pharmaceutically acceptable salt thereof according to any one of embodiments 1A, 1D, 1G, 1E, 2A, 2D, 2G or 2E, wherein L is

[0997] The bifunctional compound or a pharmaceutically acceptable salt thereof according to any one of embodiments 1A, 1D, 1G, 1E, 2A, 2D, 2G or 2E or 27, wherein L is

[0998] The bifunctional compound or a pharmaceutically acceptable salt thereof according to any one of embodiments 1A, 1D, 1G, 1E, 2A, 2D, 2G or 2E or 27, wherein L is

[0999] 30. A bifunctional compound selected from any one of the compounds in Table 1 or a pharmaceutically acceptable salt thereof.

[1000] 31. A bifunctional compound selected from any one of the compounds in Table 1.

[1001] 32. A pharmaceutical composition comprising the bifunctional compound or a pharmaceutically acceptable salt thereof according to any one of Examples 1A to 1D or 3 to 31, and one or more pharmaceutically acceptable excipients.

[1002] 33. A pharmaceutical composition comprising the bifunctional compound according to any one of Examples 2A to 2D or 3 to 31, and one or more pharmaceutically acceptable excipients.

[1003] 34A. A pharmaceutical composition comprising the bifunctional compound or a pharmaceutically acceptable salt thereof according to Example 1A or 2A, and one or more pharmaceutically acceptable excipients.

[1004] 34B. A pharmaceutical composition comprising the bifunctional compound or a pharmaceutically acceptable salt thereof according to Example 1B or 2B, and one or more pharmaceutically acceptable excipients.

[1005] 34C. A pharmaceutical composition comprising the bifunctional compound or a pharmaceutically acceptable salt thereof according to Example 1C or 2C, and one or more pharmaceutically acceptable excipients.

[1006] 34D. A pharmaceutical composition comprising the bifunctional compound or a pharmaceutically acceptable salt thereof according to Example 1D or 2D, and one or more pharmaceutically acceptable excipients.

[1007] 34E. A pharmaceutical composition comprising the bifunctional compound according to Example 2A, and one or more pharmaceutically acceptable excipients.

[1008] 34F. A pharmaceutical composition comprising the bifunctional compound according to Example 2B, and one or more pharmaceutically acceptable excipients.

[1009] 34G. A pharmaceutical composition comprising the bifunctional compound according to Example 2C, and one or more pharmaceutically acceptable excipients.

[1010] 34H. A pharmaceutical composition comprising the bifunctional compound according to Example 2D, and one or more pharmaceutically acceptable excipients.

[1011] 36. The pharmaceutical composition according to any one of embodiments 31 to 34H, wherein the composition further comprises an effective amount of at least one additional anti-cancer agent.

[1012] 37A. A method of treating a disease or disorder in a subject, the method comprising administering to a subject in need thereof a therapeutically effective amount of a bifunctional compound according to any one of embodiments 1 to 31, or a therapeutically effective amount of a pharmaceutical composition according to any one of embodiments 32 to 36.

[1013] 37B. A method of treating cancer in a subject, the method comprising administering to a subject in need thereof a therapeutically effective amount of a bifunctional compound according to any one of embodiments 1 to 31, or a therapeutically effective amount of a pharmaceutical composition according to any one of embodiments 32 to 36.

[1014] 38. The method according to embodiment 37A or 37B, wherein the therapeutically effective amount of the bifunctional compound or the pharmaceutical composition is orally administered to the subject.

[1015] 39. The method according to embodiment 37A, 37B or 38, wherein the therapeutically effective amount of the bifunctional compound or the pharmaceutical composition is administered to the subject once, twice, three times or four times a day.

[1016] 40. The method according to embodiment 37A, 37B or any one of 38 to 39, wherein the therapeutically effective amount of the bifunctional compound or the pharmaceutical composition is administered to the subject once a day.

[1017] 41. The method according to embodiment 37A, 37B or any one of 38 to 39, wherein the therapeutically effective amount of the bifunctional compound or the pharmaceutical composition is administered to the subject all at once, or in two, three or four divided doses.

[1018] 42. The method according to embodiment 37A, 37B or any one of 38 to 41, wherein the therapeutically effective amount of the bifunctional compound is about 1 mg to about 1000 mg.

[1019] 43. The method according to embodiment 37A, 37B or any one of 38 to 41, wherein the therapeutically effective amount of the bifunctional compound is about 5 mg to about 750 mg.

[1020] 44. The method according to embodiment 37A, 37B or any one of 38 to 41, wherein the therapeutically effective amount of the bifunctional compound is about 10 mg to about 500 mg.

[1021] 45. The method according to any one of embodiments 37A, 37B, or 38 to 41, wherein the therapeutically effective amount of the bifunctional compound is from about 20 mg to about 250 mg.

[1022] 46. The method according to any one of embodiments 37A, 37B, or 38 to 45, wherein the subject is in a fed state at the time of administration.

[1023] 47. The method according to any one of embodiments 37A, 37B, or 38 to 45, wherein the subject is in a fasted state at the time of administration.

[1024] 48. The method according to any one of embodiments 37A, 37B, or 38 to 47, further comprising administering to the subject in need thereof an effective amount of at least one additional anti - cancer agent.

[1025] 49. The method according to any one of embodiments 37A, 37B, or 38 to 48, wherein the disease or disorder is associated with abnormal BCL6 expression and / or activity.

[1026] 50. The method according to any one of embodiments 37A, 37B, or 38 to 48, wherein the disease or disorder is a cancer associated with abnormal BCL6 expression and / or activity.

[1027] 51. The method according to any one of embodiments 37A, 37B, or 38 to 50, wherein the disease or disorder is breast cancer, ovarian cancer, leukemia, lymphoma, benign lymphoma, malignant lymphoma, Burkitt's lymphoma, non - Hodgkin's lymphoma, B - cell non - Hodgkin's lymphoma, sarcoma, Ewing's sarcoma, angiosarcoma, Kaposi's sarcoma, liposarcoma, myosarcoma, synovial sarcoma, meningeal sarcoma, carcinosarcoma, acute myeloid leukemia (AML), acute lymphoblastic leukemia (ALL), T - lineage acute lymphoblastic leukemia (T - ALL), T - lineage lymphoblastic lymphoma (T - LL), peripheral T - cell lymphoma, adult T - cell leukemia, precursor B acute lymphoblastic leukemia, precursor B lymphoma, B - cell lymphoma, large B - cell lymphoma, diffuse large B - cell lymphoma, B - cell acute lymphoblastic leukemia (ALL), Philadelphia chromosome - positive acute lymphoblastic leukemia (ALL), Philadelphia chromosome - positive chronic myeloid leukemia (CML), follicular lymphoma, intravascular large B - cell lymphoma, angioimmunoblastic T - cell lymphoma (AITL), T - cell lymphoma, B - cell leukemia, chronic myeloid leukemia, non - small cell lung cancer, systemic lupus erythematosus (SLE), brain tumor or central nervous system cancer.

[1028] 52. The method according to any one of embodiments 37A, 37B or 38 to 50, wherein the disease or disorder is diffuse large B-cell lymphoma, Burkitt lymphoma, follicular lymphoma or angioimmunoblastic T-cell lymphoma (AITL).

Claims

1. A bifunctional compound of formula (I), formula (II) or formula (III): or a pharmaceutically acceptable salt thereof, wherein: R 1 is H or a C1-C6 alkyl group; Q is X is N or CH; Y1, Y2, and Y3 are each independently N or CR 3 ; Z1 and Z2 are each independently N or CH; R 2 is H or a C1-C6 alkyl group; Each R 3 independently is H, halogen, C1-C6 alkyl, C1-C6 haloalkyl, -O-(C1-C6 alkyl), -O-(C1-C6 haloalkyl); Of Q Indicates the point of attachment to X or succinimide; R 1a is H or a halogen; R 2a is H or a C1-C3 alkyl group; X 3a is CHR 3a or C(O); R 3a is H or a C1-C3 alkyl group; X 4a and X 6a each independently is CH or N; and R 5a is H, C1-C3 alkyl or halogen; L is X 6b is CHR 6b or C(O); R 6b is H or a C1-C3 alkyl group; R 1b 、R 2b 、R 3b and R 4b each independently is H or a halogen, where at least one of R 1b 、R 2b 、R 3b 、R 4b is a halogen; R 5b is H or a halogen; R 6b is H or a C1-C3 alkyl group; X 1b and X 2b are each independently CH or N, where at least one of 1b and X 2b is N; and Each of which L indicates a connection point.

2. The bifunctional compound according to claim 1, wherein the compound is a compound of formula (I) or a pharmaceutically acceptable salt thereof.

3. The bifunctional compound according to claim 1, wherein the compound is a compound of formula (II) or a pharmaceutically acceptable salt thereof.

4. The bifunctional compound according to claim 1, wherein the compound is a compound of formula (III) or a pharmaceutically acceptable salt thereof.

5. A bifunctional compound of formula (I), formula (II) or formula (III): wherein: R 1 is H or a C1-C6 alkyl group; Q is X is N or CH; Y1, Y2, and Y3 are each independently N or CR 3 ; Z1 and Z2 are each independently N or CH; R 2 is H or a C1-C6 alkyl group; Each R 3 is independently H, halogen, C1-C6 alkyl, C1-C6 haloalkyl, -O-(C1-C6 alkyl) or -O-(C1-C6 haloalkyl); Of Q Indicates the point of attachment to X or glutarimide; R 1a is H or a halogen; R 2a is H or a C1-C3 alkyl group; X 3a is CHR 3a or C(O); R 3a is H or a C1-C3 alkyl group; X 4a and X 6a each independently is CH or N; and R 5a is H, C1-C3 alkyl or halogen; L is X 6b is CHR 6b or C(O); R 6b is H or a C1-C3 alkyl group; R 1b , R 2b , R 3b and R 4b are each independently H or halogen, wherein R 1b , R 2b , R 3b , R 4b at least one of which is a halogen; R 5b is H or a halogen; R 6b is H or a C1-C3 alkyl group; X 1b and X 2b are each independently CH or N, where at least one of 1b and X 2b is N; and Each of which L indicates a connection point.

6. The bifunctional compound or a pharmaceutically acceptable salt thereof according to any one of claims 1, 3 or 5, wherein the compound is a compound of formula (II-a):

7. The bifunctional compound or a pharmaceutically acceptable salt thereof according to any one of claims 1, 2 or 5, wherein Q is 8. The bifunctional compound or a pharmaceutically acceptable salt thereof according to any one of claims 1, 2, 5, 6 or 7, wherein each of Y1, Y2 and Y3 is CH.

9. The bifunctional compound or a pharmaceutically acceptable salt thereof according to any one of claims 1, 2, 5 or 6 to 8, wherein R 1 is H or CH3.

10. The bifunctional compound or a pharmaceutically acceptable salt thereof according to any one of claims 1, 2, 5 or 6 to 9, wherein X is N.

11. The bifunctional compound or a pharmaceutically acceptable salt thereof according to any one of claims 1, 2, 5 or 6 to 10, wherein R 2 is methyl, ethyl or isopropyl.

12. A bifunctional compound or a pharmaceutically acceptable salt thereof according to any one of claims 1, 2, 5 or 6 to 11, wherein each R 3 is independently H, methyl, fluoro or methoxy.

13. The bifunctional compound or a pharmaceutically acceptable salt thereof according to any one of claims 1, 3 or 5, wherein R 1a is F.

14. The bifunctional compound or a pharmaceutically acceptable salt thereof according to any one of claims 1, 3 or 5, wherein X 4a and X 6a are each N.

15. The bifunctional compound or a pharmaceutically acceptable salt thereof according to any one of claims 1, 3, 5 or 6, wherein R 5a is CH3.

16. The bifunctional compound or a pharmaceutically acceptable salt thereof according to any one of claims 1, 3, 5 or 6, wherein R 2a is H.

17. The bifunctional compound or a pharmaceutically acceptable salt thereof according to any one of claims 1, 4 or 5, wherein L is 18. The bifunctional compound or a pharmaceutically acceptable salt thereof according to any one of claims 1, 4 or 5, wherein L is 19. The bifunctional compound or a pharmaceutically acceptable salt thereof according to any one of claims 1, 4 or 5, wherein L is 20. A bifunctional compound or a pharmaceutically acceptable salt thereof, which is any one of the compounds selected from the compounds in Table 1.

21. A bifunctional compound, which is any one of the compounds selected from the compounds in Table 1.

22. A pharmaceutical composition comprising the bifunctional compound or a pharmaceutically acceptable salt thereof according to any one of claims 1 to 21, and one or more pharmaceutically acceptable excipients.

23. The pharmaceutical composition according to claim 22, wherein the composition further comprises an effective amount of at least one additional anti-cancer agent.

24. A method for treating a disease or disorder in a subject, the method comprising administering to a subject in need thereof a therapeutically effective amount of the bifunctional compound according to any one of claims 1 to 21, or a therapeutically effective amount of the pharmaceutical composition according to claim 22 or 23.

25. The method according to claim 24, which further comprises administering to the subject in need thereof an effective amount of at least one additional anti-cancer agent.

26. The method according to claim 24 or 25, wherein the disease or disorder is related to abnormal BCL6 expression and / or activity.

27. The method according to claim 26, wherein the disease or disorder is a cancer related to abnormal BCL6 expression and / or activity.

28. The method according to any one of claims 24 to 27, wherein the disease or disorder is breast cancer, ovarian cancer, leukemia, lymphoma, benign lymphoma, malignant lymphoma, Burkitt's lymphoma, non-Hodgkin's lymphoma, B-cell non-Hodgkin's lymphoma, sarcoma, Ewing's sarcoma, angiosarcoma, Kaposi's sarcoma, liposarcoma, rhabdomyosarcoma, synovial sarcoma, meningeal sarcoma, carcinosarcoma, acute myeloid leukemia (AML), acute lymphoblastic leukemia (ALL), T-lineage acute lymphoblastic leukemia (T-ALL), T-lineage lymphoblastic lymphoma (T-LL), peripheral T-cell lymphoma, adult T-cell leukemia, precursor B acute lymphoblastic leukemia, precursor B lymphoma, B-cell lymphoma, large B-cell lymphoma, diffuse large B-cell lymphoma, B-cell acute lymphoblastic leukemia (ALL), Philadelphia chromosome-positive acute lymphoblastic leukemia (ALL), Philadelphia chromosome-positive chronic myeloid leukemia (CML), follicular lymphoma, intravascular large B-cell lymphoma, angioimmunoblastic T-cell lymphoma (AITL), T-cell lymphoma, B-cell leukemia, chronic myeloid leukemia, non-small cell lung cancer, systemic lupus erythematosus (SLE), brain tumor or central nervous system cancer.

29. The method according to any one of claims 24 to 27, wherein the disease or disorder is large B-cell lymphoma, diffuse large B-cell lymphoma, Burkitt's lymphoma, follicular lymphoma or angioimmunoblastic T-cell lymphoma (AITL).

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