Compounds and compositions for the selective degradation of modified proteins

Abstract

The present application provides compounds and compositions for degrading modified polypeptides in cells. In some embodiments, the compounds and compositions are provided for treating cancer.

Description

[Technical Field]

[0001] CROSS-REFERENCE TO RELATED APPLICATIONS This application claims priority to U.S. Provisional Application No. 63 / 444,208, filed February 8, 2023, the entire contents of which are incorporated herein by reference for all purposes.

[0002] Sequence Listing

[0001] This application contains a Sequence Listing that has been submitted electronically in XML format. The XML copy was created on January 23, 2024, is entitled "01277-0037-00PCT.xml," and is 6,097 bytes in size. The information in the electronic format of the Sequence Listing is incorporated herein by reference in its entirety.

[0003] The present disclosure relates to compounds and compositions thereof for degrading modified proteins within cells. [Background technology]

[0004] Engineered cells containing modified heterologous polypeptides, such as chimeric antigen receptor T (CAR-T) cells, have been developed for therapeutic use. Modulating the expression levels of such modified heterologous polypeptides may improve the therapeutic benefits of the modified cells, for example, by reducing side effects and / or enhancing the efficacy of the modified cells.

[0005] Thus, in one aspect, provided herein are modified polypeptides and degradation agents, wherein the modified polypeptide comprises a degradation domain that catalyzes ubiquitination in a cell when the degradation domain is bound to the degradation agent. Summary of the Invention

[0006] Described herein, in some embodiments, are compounds and compositions thereof for modulating heterologous polypeptide levels in cells. In various embodiments, the compounds and compositions thereof may be used to reduce the level of heterologous polypeptides in cells.

[0007] Embodiments of the present invention may be more fully understood by reference to the detailed description and examples, which are intended to be illustrative of non-limiting embodiments.

[0008] In some embodiments, provided herein are compounds of formula (I): [ka] or a pharmaceutically acceptable salt thereof is provided. [In formula: R 1 is H or oxo; R 2 are each independently H or halo, where at least one R 2 is fluoro; X is a bond, C1-C3 alkylene, -C(O)NHCH2-, -NHC(O)-, -C(O)-, or -(C1-C3 alkylene)NH(C1-C3 alkylene)-; Ring A is an optionally substituted C5-C6 cycloalkyl, an optionally substituted C5-C6 aryl, an optionally substituted 6-10 membered heterocyclyl, or an optionally substituted 5-9 membered heteroaryl, wherein the heterocyclyl or heteroaryl contains 1-3 heteroatoms selected from nitrogen, oxygen, and sulfur. DETAILED DESCRIPTION OF THE INVENTION

[0009] Detailed Description definition As used herein, the terms "comprising" and "including" may be used interchangeably. The terms "comprising" and "including" should be interpreted as specifying the presence of a described feature or referenced component, but do not exclude the presence or addition of one or more features or components, or groups thereof. In addition, the terms "comprising" and "including" are intended to include examples encompassed by the term "consisting of." As a result, the term "consisting of" may be used in place of the terms "comprising" and "including" to provide more specific embodiments of the present invention.

[0010] The term "consisting of" means that the subject matter has at least 90%, 95%, 97%, 98%, or 99% of the recited features or components thereof. In other embodiments, the term "consisting of" excludes from the scope of any succeeding recitation any other features or components, excluding those that are not essential to the technical effect to be achieved.

[0011] As used herein, the term "or" should be interpreted as an inclusive "or" meaning either one or any combination. Thus, "A, B, or C" means any of the following: "A; B; C; A and B; A and C; B and C; A, B, and C." Exceptions to this definition will occur only where combinations of elements, features, steps, or acts are, in some way, mutually exclusive by nature.

[0012] Any concentration range, percentage range, proportion range, or integer range herein should be understood to include any integer value in the stated range and, where appropriate, fractions thereof (e.g., tenths and hundredths of an integer), unless otherwise indicated. Also, any numerical range for any physical characteristic (e.g., polymer subunits, size, or thickness) described herein should be understood to include any integer within the stated range, unless otherwise indicated. As used herein, the terms "about" and "approximately" mean ±20%, ±10%, ±5%, or ±1% of the indicated range, value, or structure, unless otherwise indicated.

[0013] "Amino" refers to the group -NH2.

[0014] "Cyano" refers to the radical -CN.

[0015] "Oxo" refers to the group =O.

[0016] An "alkyl" group is a saturated, partially saturated, or unsaturated, straight or branched chain, acyclic hydrocarbon having from 1 to 10 carbon atoms (C1 to C 10alkyl), typically having 1 to 8 carbon atoms (C1-C8 alkyl), or in some embodiments, 1 to 6 (C1-C6 alkyl), 1 to 4 (C1-C4 alkyl), 1 to 3 (C1-C3 alkyl), or 2 to 6 (C2-C6 alkyl) carbon atoms. In some embodiments, the alkyl group is a saturated alkyl group. Representative saturated alkyl groups include -methyl, -ethyl, -n-propyl, -n-butyl, -n-pentyl, and -n-hexyl; saturated branched alkyls include -isopropyl, -sec-butyl, -isobutyl, tert-butyl, -isopentyl, -neopentyl, tert-pentyl, -2-methylpentyl, -3-methylpentyl, -4-methylpentyl, -2,3-dimethylbutyl, and the like. In some embodiments, the alkyl group is an unsaturated alkyl group, also referred to as an alkenyl or alkynyl group. An "alkenyl" group is an alkyl group containing one or more carbon-carbon double bonds. An "alkynyl" group is an alkyl group containing one or more carbon-carbon triple bonds. Examples of unsaturated alkyl groups include, but are not limited to, vinyl, allyl, CH=CH(CH), -CH=C(CH), -C(CH)=CH, -C(CH)=CH(CH), -C(CHCH)=CH, -C≡CH, -C≡C(CH), -C≡C(CHCH), -CHC≡CH, CHC≡C(CH), and CHC≡C(CHCH). Alkyl groups can be substituted or unsubstituted. When alkyl groups described herein are referred to as "substituted," they can be substituted with any of the substituents found in the exemplary compounds and embodiments disclosed herein, as well as those found below: Halogen; Hydroxy; Alkoxy; Cycloalkyloxy, Aryloxy, Heterocyclyloxy, Heteroaryloxy, Heterocycloalkyloxy, Cycloalkylalkyloxy, Aralkyloxy, Heterocyclylalkyloxy, Heteroarylalkyloxy, Heterocycloalkylalkyloxy; Oxo(=O); Amino, Alkylamino, Cycloalkylamino, Arylamino, Heterocyclylamino, Heteroarylamino, Heterocycloalkylamino, Cycloalkylalkylamino, Aralkylamino, Heterocyclylalkylamino, Heteroaralkyl alkylamino, heterocycloalkylalkylamino; imino; imido; amidino; guanidino; enamino; acylamino; sulfonylamino; urea, nitrourea; oxime; hydroxylamino; alkoxyamino; aralkoxyamino; hydrazino; hydrazide; hydrazono; azide; nitro; thio (-SH), alkylthio; =S; sulfinyl; sulfonyl; aminosulfonyl; phosphonate; phosphinyl; acyl; formyl; carboxy; ester; carbamate; amido; cyano; isocyanate; isothiocyanate; cyanate; thiocyanate; or -B(OH)2. In some embodiments, when alkyl groups described herein are referred to as "substituted," they can be substituted with any of the substituents found in the exemplary compounds and embodiments disclosed herein, as well as those found below: Halogen (chloro, iodo, bromo, or fluoro); alkyl; hydroxyl; alkoxy; alkoxyalkyl; amino; alkylamino; carboxy; nitro; cyano; thiol; thioether; imine; imide; amidine; guanidine; enamine; aminocarbonyl; acylamino; phosphonate; phosphine; thiocarbonyl; sulfinyl; sulfone; sulfonamide; ketone; aldehyde; ester; urea; urethane; oxime; hydroxylamine; alkoxyamine; aralkoxyamine; N-oxide; hydrazine; hydrazide; hydrazone; azide; isocyanate; isothiocyanate; cyanate; thiocyanate; B(OH)2, or O(alkyl)aminocarbonyl.

[0017] An "alkylene" group refers to a group having the same remaining moieties as alkyl, but which is divalent. Particular alkylene groups have 1 to 10 carbon atoms (C1-C 10 alkylene), typically having 1 to 8 carbon atoms (C1-C8 alkylene), or in some embodiments 1 to 6 (C1-C6 alkylene) or 1 to 3 (C1-C3 alkylene) carbon atoms. Examples of alkylene include, but are not limited to, groups such as methylene (-CH2-), ethylene (-CH2CH2-), propylene (-CH2CH2CH2-), isopropylene (-CH2CH(CH3)-), butylene (-CH2(CH2)2CH2-), isobutylene (-CH2CH(CH3)CH2-), pentylene (-CH2(CH2)3CH2-), hexylene (-CH2(CH2)4CH2-), heptylene (-CH2(CH2)5CH2-), octylene (-CH2(CH2)6CH2-), and the like.

[0018] A "cycloalkyl" group is a saturated or partially saturated cyclic alkyl group having 3 to 10 carbon atoms (C 10Cycloalkyl groups (cycloalkyls) have a single ring or multiple fused or bridged rings, which may be optionally substituted. In some embodiments, cycloalkyl groups have 3 to 8 ring carbon atoms (C3-C8 cycloalkyl), while in other embodiments, the number of ring carbon atoms ranges from 3 to 5 (C3-C5 cycloalkyl), 3 to 6 (C3-C6 cycloalkyl), or 3 to 7 (C3-C7 cycloalkyl). In some embodiments, cycloalkyl groups are saturated cycloalkyl groups. Such saturated cycloalkyl groups include, by way of example, single ring structures such as cyclopropyl, cyclobutyl, cyclopentyl, cyclohexyl, cycloheptyl, cyclooctyl, 1-methylcyclopropyl, 2-methylcyclopentyl, 2-methylcyclooctyl, and the like, or multiple ring or bridged ring structures such as 1-bicyclo[1.1.1]pentyl, bicyclo[2.1.1]hexyl, bicyclo[2.2.1]heptyl, bicyclo[2.2.2]octyl, adamantyl, and the like. In another embodiment, the cycloalkyl group is an unsaturated cycloalkyl group. Examples of unsaturated cycloalkyl groups include cyclohexenyl, cyclopentenyl, cyclohexadienyl, butadienyl, pentadienyl, hexadienyl, and the like. The cycloalkyl group can be substituted or unsubstituted. Examples of such substituted cycloalkyl groups include cyclohexanol, and the like.

[0019] A "heterocyclyl" is a non-aromatic cycloalkyl in which one to four ring carbon atoms are independently replaced with a heteroatom selected from O, S, and N. In some embodiments, a heterocyclyl group contains 3 to 10 ring atoms, while other such groups contain 3 to 5, 3 to 6, or 3 to 8 ring atoms. A heterocyclyl can be attached to another group at any ring atom (i.e., at any carbon atom or heteroatom of the heterocycle). A heterocyclyl group can be substituted or unsubstituted. Heterocyclyl groups include saturated and partially saturated ring systems. Furthermore, the term "heterocyclyl" is intended to include any non-aromatic ring containing at least one heteroatom, which ring may be fused to an aryl or heteroaryl ring, regardless of its attachment to the rest of the molecule. The term also includes bridged polycyclic ring systems containing heteroatoms. Representative examples of heterocyclyl groups include, but are not limited to, aziridinyl, azetidinyl, azepanyl, pyrrolidyl, imidazolidinyl (e.g., imidazolidin-4-onyl or imidazolidin-2,4-dionyl), pyrazolidinyl, thiazolidinyl, tetrahydrothiophenyl, tetrahydrofuranyl, piperidyl, piperazinyl (e.g., piperazin-2-onyl), morpholinyl, thiomorpholinyl, tetrahydropyranyl (e.g., tetrahydro-2H-pyranyl), tetrahydrothiopyranyl, oxathianyl, dithianyl, 1,4-dioxaspiro[4.5]decanyl, homopiperazinyl, quinuclidinyl, or tetrahydropyrimidin-2(1H)-one. Representative substituted heterocyclyl groups may be mono-substituted or di- or higher-substituted, for example, but not limited to, pyridyl or morpholinyl groups that are di-, tri-, tetra-, pentapi-, or hexa-substituted, or di-substituted with various substituents as listed below.

[0020] A "heterocyclylene" group refers to a divalent "heterocyclyl" group.

[0021] An "aryl" group is an aromatic carbocyclic group having 6 to 14 carbon atoms (C6-C 14 aryl) having a single ring (e.g., phenyl) or multiple fused rings (e.g., naphthyl or anthryl). In some embodiments, aryl groups contain 6 to 14 carbons (C6-C8) in the ring portion of the group. 14 aryl), and in other cases, 6 to 12 (C6 to C 12 aryl) or 6 to 10 carbon atoms (C6 to C 10 Particular aryl groups include phenyl, biphenyl, naphthyl, and the like. Aryl groups can be substituted or unsubstituted. The phrase "aryl group" also includes groups containing fused rings, such as fused aromatic-aliphatic ring systems (e.g., indanyl, tetrahydronaphthyl, and the like).

[0022] A "heteroaryl" group is an aromatic ring system having 1 to 4 heteroatoms as ring atoms, with the remaining atoms being carbon atoms. In some embodiments, heteroaryl groups contain 3 to 6 ring atoms, and in other cases 6 to 9 or 6 to 10 atoms, in the ring portion of the group. Suitable heteroatoms include oxygen, sulfur, and nitrogen. In some embodiments, the heteroaryl ring system is monocyclic or bicyclic. Non-limiting examples include, but are not limited to, pyrrolyl, pyrazolyl, imidazolyl, triazolyl, tetrazolyl, oxazolyl, isoxazolyl, benzisoxazolyl (e.g., benzo[d]isoxazolyl), thiazolyl, pyrrolyl, pyridazinyl, pyrimidyl, pyrazinyl, thiophenyl, benzothiophenyl, furanyl, benzofuranyl, indolyl (e.g., indolyl-2-onyl or isoindolin-1-onyl), azaindolyl (pyrrolopyridyl or 1H-pyrrolo[2,3-b]pyridyl), indazolyl, benzimidazolyl (e.g., 1H-benzo[d]imidazolyl), imidazolyl, Examples of heteroaryl groups include pyridyl (e.g., azabenzimidazolyl or 1H-imidazo[4,5-b]pyridyl), pyrazolopyridyl, triazolopyridyl, benzotriazolyl (e.g., 1H-benzo[d][1,2,3]triazolyl), benzoxazolyl (e.g., benzo[d]oxazolyl), benzothiazolyl, benzothiadiazolyl, isoxazolopyridyl, thianaphthalenyl, purinyl, xanthinyl, adeninyl, guaninyl, quinolinyl, isoquinolinyl (e.g., 3,4-dihydroisoquinolin-1(2H)-onyl), tetrahydroquinolinyl, quinoxalinyl, and quinazolinyl groups. Heteroaryl groups can be substituted or unsubstituted.

[0023] "Halogen" or "halo" means fluorine, chlorine, bromine or iodine.

[0024] When groups described herein are referred to as "substituted," they may be substituted with any suitable substituent(s). Specific examples of substituents are found in the exemplary compounds and embodiments disclosed herein, as well as below: Halogen (chloro, iodo, bromo, or fluoro); alkyl; hydroxyl; alkoxy; alkoxyalkyl; amino; alkylamino; carboxy; nitro; cyano; thiol; thioether; imine; imide; amidine; guanidine; enamine; aminocarbonyl; acylamino; phosphonate; phosphine; thiocarbonyl; sulfinyl; sulfone; sulfonamide; ketone; aldehyde; ester; urea; urethane; oxime; hydroxylamine; alkoxyamine; aralkoxyamine; N-oxide; hydrazine; hydrazide; hydrazone; azide; isocyanate; isothiocyanate; cyanate; thiocyanate; oxygen (=O); B(OH)2, O(alkyl)aminocarbonyl; cycloalkyl, which may be monocyclic or fused or non-fused polycyclic (e.g., heterocyclyl, which may be monocyclic or fused or non-fused polycyclic (e.g., pyrrolidyl, piperidyl, piperazinyl, morpholinyl, or thiazinyl); aryl or heteroaryl, which may be monocyclic or fused or non-fused polycyclic (e.g., phenyl, naphthyl, pyrrolyl, indolyl, furanyl, thiophenyl, imidazolyl, oxazolyl, isoxazolyl, thiazolyl, triazolyl, tetrazolyl, pyrazolyl, pyridyl, quinolinyl, isoquinolinyl, acridinyl, pyrazinyl, pyridazinyl, pyrimidyl, benzimidazolyl, benzothiophenyl, or benzofuranyl), aryloxy; aralkyloxy; heterocyclyloxy; and heterocyclylalkoxy.

[0025] Embodiments of the present disclosure are intended to include pharmaceutically acceptable salts, tautomers, isotopic substitutions, and stereoisomers of the compounds provided herein, e.g., compounds of Formula (I).

[0026] As used herein, the term "pharmaceutically acceptable salt(s)" refers to salts prepared from pharmaceutically acceptable non-toxic acids or bases, including inorganic acids and bases, and organic acids and bases. Suitable pharmaceutically acceptable base addition salts of compounds of formula (I) include, but are not limited to, metallic salts made from aluminum, calcium, lithium, magnesium, potassium, sodium, and zinc, or organic salts made from lysine, N,N'-dibenzylethylenediamine, chloroprocaine, choline, diethanolamine, ethylenediamine, meglumine (N-methyl-glucamine), and procaine. Suitable non-toxic acids include, but are not limited to, inorganic and organic acids such as acetic acid, alginic acid, anthranilic acid, benzenesulfonic acid, benzoic acid, camphorsulfonic acid, citric acid, ethenesulfonic acid, formic acid, fumaric acid, furoic acid, galacturonic acid, gluconic acid, glucuronic acid, glutamic acid, glycolic acid, hydrobromic acid, hydrochloric acid, isethionic acid, lactic acid, maleic acid, malic acid, mandelic acid, methanesulfonic acid, mucic acid, nitric acid, pamoic acid, pantothenic acid, phenylacetic acid, phosphoric acid, propionic acid, salicylic acid, stearic acid, succinic acid, sulfanilic acid, sulfuric acid, tartaric acid, and p-toluenesulfonic acid. Specific non-toxic acids include hydrochloric acid, hydrobromic acid, maleic acid, phosphoric acid, sulfuric acid, and methanesulfonic acid. Accordingly, specific examples of salts include hydrochloride, formate, and mesylate salts. Other salts are known in the art; see, for example, Remington's Pharmaceutical Sciences, 18th ed., Mack Publishing, Easton PA (1990) or Remington: The Science and Practice of Pharmacy, 19th ed., Mack Publishing, Easton PA (1995).

[0027] As used herein, and unless otherwise specified, the term "stereoisomer" or "stereoisomerically pure" means one stereoisomer of a particular compound that is substantially free of other stereoisomers of that compound. For example, a stereomerically pure compound having one chiral center will be substantially free of the other enantiomer of that compound. A stereomerically pure compound having two chiral centers will be substantially free of other diastereomers of that compound. A typical stereoisomerically pure compound contains more than about 80% by weight of one stereoisomer of the compound and less than about 20% by weight of the other stereoisomer of the compound; more than about 90% by weight of one stereoisomer of the compound and less than about 10% by weight of the other stereoisomer of the compound; more than about 95% by weight of one stereoisomer of the compound and less than about 5% by weight of the other stereoisomer of the compound; or more than about 97% by weight of one stereoisomer of the compound and less than about 3% by weight of the other stereoisomer of the compound. The compounds disclosed herein may have chiral centers and may occur as racemates, individual enantiomers or diastereomers, and mixtures thereof. All of these isomeric forms, including mixtures thereof, are included in the embodiments disclosed herein.

[0028] The use of stereomerically pure forms of the compounds disclosed herein, as well as mixtures of these forms, are encompassed by the embodiments disclosed herein. For example, mixtures containing equal or unequal amounts of the enantiomers of a particular compound may be used in the methods and compositions disclosed herein. These isomers may be asymmetrically synthesized or resolved using standard techniques, such as chiral columns or chiral resolving agents.For example, Jacques, J., et al., Enantiomers, Racemates and Resolutions (Wiley Interscience, New York, 1981); Wilen, SH, et al., Tetrahedron 33:2725 (1977); Eliel, EL, Stereochemistry of Carbon Compounds (McGraw Hill, NY, 1962); Wilen, SH, Tables of Resolving Agents and Optical Resolutions p. 268 (EL Eliel, Ed., Univ. of Notre Dame Press, Notre Dame, IN, 1972); Todd, M., Separation Of Enantiomers : Synthetic Methods (Wiley-VCH Verlag GmbH & Co. KGaA, Weinheim, Germany, 2014); Toda, F., Enantiomer Separation: Fundamentals and Practical Methods (Springer Science & Business Media, 2007); Subramanian, G. See Chiral Separation Techniques: A Practical Approach (John Wiley & Sons, 2008); Ahuja, S., Chiral Separation Methods for Pharmaceutical and Biotechnological Products (John Wiley & Sons, 2011).

[0029] It should be noted that the compounds disclosed herein may include E and Z isomers, or mixtures thereof, as well as cis and trans isomers, or mixtures thereof. In some embodiments, the compounds are isolated as either the E or Z isomer. In other embodiments, the compounds are mixtures of E and Z isomers.

[0030] "Tautomer" refers to isomers of a compound that are in equilibrium with each other. The concentration of isomers depends on the conditions in which the compound is found, and may vary depending on, for example, whether the compound is a solid or in an organic or aqueous solution. For example, in aqueous solution, pyrazole may exhibit the following isomers, which are referred to as tautomers of each other: [ka]

[0031] As will be readily understood by one of ordinary skill in the art, various functional groups and other structures may exhibit tautomerism, and all tautomers of the compounds of formula (I) are within the scope of the present disclosure.

[0032] It should also be noted that the compounds disclosed herein may contain unnatural proportions of atomic isotopes at one or more atoms. For example, the compounds may contain radioactive isotopes, such as tritium ( 3 H), iodine-125( 125 I), sulfur-35( 35 S), or carbon-14 ( 14 C), or may be radiolabeled with deuterium ( 2 H), carbon-13( 13 C), or nitrogen-15( 15The compound may be isotopically enriched with, for example, N. As used herein, an "isotopologue" is an isotopically enriched compound. The term "isotopically enriched" refers to an atom having an isotopic composition different from the natural isotopic composition of that atom. "Isotopically enriched" may refer to a compound containing at least one atom having an isotopic composition different from the natural isotopic composition of that atom. The term "isotopic composition" refers to the amount of each isotope present in a given atom. Radiolabeled, isotopically enriched compounds are useful as therapeutic agents, e.g., cancer therapeutic agents, research reagents, e.g., binding assay reagents, and diagnostic agents, e.g., in vivo imaging agents. All isotopic variations of the compounds described herein, whether radioactive or non-radioactive, are intended to be encompassed within the scope of the embodiments provided herein. In some embodiments, isotopic substitutions of the compounds disclosed herein are provided, e.g., isotopic substitutions are compounds enriched in deuterium, carbon-13, and / or nitrogen-15. As used herein, "deuterated" refers to a compound in which at least one hydrogen (H) is replaced with a deuterium (D or 2 H), i.e., the compound is enriched in deuterium at at least one position.

[0033] It is understood that each compound disclosed herein can be provided in the form of any of the pharmaceutically acceptable salts discussed herein, regardless of stereoisomerism or isotopic composition.Similarly, it is understood that the isotopic composition can vary independently of the stereoisomeric composition of each compound mentioned herein.Furthermore, although the isotopic composition is limited to the elements present in each compound or its salt discussed herein, it can also vary independently of the choice of pharmaceutically acceptable salt of each compound.

[0034] It should be noted that if there is a discrepancy between the depicted structure and the name of that structure, the depicted structure should be given more weight.

[0035] "Treating," as used herein, means to completely or partially alleviate a disorder, disease, or condition, or one or more symptoms associated with a disorder, disease, or condition, or to slow or halt the progression or worsening of those symptoms, or to reduce or eradicate the cause(s) of the disorder, disease, or condition. In certain embodiments, the disorder is a neurodegenerative disease described herein, or a symptom thereof.

[0036] "Preventing," as used herein, means delaying and / or eliminating, completely or partially, the onset, recurrence, or spread of a disorder, disease, or condition; preventing a subject from acquiring a disorder, disease, or condition; or reducing a subject's risk of acquiring a disorder, disease, or condition. In certain embodiments, the disorder is a neurodegenerative disease described herein, or a symptom thereof.

[0037] The term "effective amount" means an amount that can be used in combination with a compound disclosed herein to treat or prevent a disorder, disease or condition, or a symptom thereof, as disclosed herein.

[0038] The terms "subject" or "patient," as used herein, include animals, including, but not limited to, cows, monkeys, horses, sheep, pigs, chickens, turkeys, quail, cats, dogs, mice, rats, rabbits, or guinea pigs, and in certain embodiments, mammals, and in other embodiments, humans. In certain embodiments, the subject is a human having or at risk for a disease or symptom thereof mediated by IRAK3.

[0039] While various features of the invention may be described in the context of a single embodiment, the features may also be provided separately or in any suitable subcombination. Conversely, although the invention may, for clarity, be described herein in the context of separate embodiments, the invention may also be practiced in a single embodiment.

[0040] compound In one embodiment, the compound of formula (I): [ka] or a pharmaceutically acceptable salt thereof, as provided herein [In formula: R 1 is H or oxo; R 2 are each independently H or halo, where at least one R 2 is fluoro; X is a bond, C1-C3 alkylene, -C(O)NHCH2-, -NHC(O)-, -C(O)-, or -(C1-C3 alkylene)NH(C1-C3 alkylene)-; Ring A is an optionally substituted C5-C6 cycloalkyl, an optionally substituted C5-C6 aryl, an optionally substituted 6-10 membered heterocyclyl, or an optionally substituted 5-9 membered heteroaryl, wherein the heterocyclyl or heteroaryl contains 1-3 heteroatoms selected from nitrogen, oxygen, and sulfur.

[0041] In some embodiments, R 1 is H or oxo. In some embodiments, R 1 is H. In some embodiments, R 1 is oxo.

[0042] In some embodiments, R 2 is H or halo. In some embodiments, R 2 is H. In some embodiments, R 2 is halo. In some embodiments, R 2 is F, Cl, or Br. In some embodiments, R 2 is F.

[0043] In some embodiments, X is a bond, C1-C3 alkylene, -C(O)NHCH2-, -NHC(O)-, -C(O)-, or -(C1-C3 alkylene)NH(C1-C3 alkylene)-. In some embodiments, X is a bond, C1 alkylene, -C(O)NHCH2-, -NHC(O)-, -C(O)-, or -(C1 alkylene)NH(C1 alkylene)-. In some embodiments, X is a bond, -CH2-, -C(O)NHCH2-, -NHC(O)-, -C(O)-, or -CH2NHCH2-.

[0044] In some embodiments, X is a bond.

[0045] In some embodiments, X is C1-C3 alkylene. In some embodiments, X is C1 alkylene. In some embodiments, X is -CH2-.

[0046] In some embodiments, X is —C(O)NHCH 2 —.

[0047] In some embodiments, X is —NHC(O)—.

[0048] In some embodiments, X is —C(O)—.

[0049] In some embodiments, X is -(C1-C3 alkylene)NH(C1-C3 alkylene)-. In some embodiments, X is -(C1 alkylene)NH(C1 alkylene)-. In some embodiments, X is -CH2NHCH2-.

[0050] In some embodiments, ring A is an optionally substituted C5-C6 cycloalkyl, an optionally substituted C5-C6 aryl, an optionally substituted 6-10-membered heterocyclyl, or an optionally substituted 5-9-membered heteroaryl, where the heterocyclyl or heteroaryl contains 1-3 heteroatoms selected from nitrogen, oxygen, and sulfur. In some embodiments, ring A is an optionally substituted C6 cycloalkyl, an optionally substituted C6 aryl, an optionally substituted 6-10-membered heterocyclyl, or a 5-9-membered heteroaryl optionally substituted with an optionally substituted C1-C3 alkyl, an optionally substituted amine, or a C4-C6 cycloalkyl, where the heterocyclyl or heteroaryl contains 1-3 heteroatoms selected from nitrogen, oxygen, and sulfur. In some embodiments, Ring A is optionally substituted cyclohexyl, optionally substituted phenyl, or 6-10 membered heterocyclyl optionally substituted with H, OH, cyano, halo, optionally substituted C1-C3 alkyl, optionally substituted C3-C6 cycloalkyl, -C(O)(6-membered heteroaryl), or -C(O)(9-membered heterocyclyl), or 5-9 membered heteroaryl optionally substituted with C1-C3 alkyl, amine, or C4 cycloalkyl, wherein the heterocyclyl or heteroaryl contains 1-3 heteroatoms selected from nitrogen, oxygen, and sulfur. In some embodiments, Ring A is cyclohexyl substituted with -CF3, phenyl substituted with -CHF2, a 6-10 membered heterocyclyl optionally substituted with H, OH, cyano, F, Cl, Br, optionally substituted C1-C3 alkyl, optionally substituted C4-C6 cycloalkyl, -C(O)(6-membered heteroaryl), or -C(O)(9-membered heterocyclyl), or a 5-9 membered heteroaryl optionally substituted with C1 alkyl, amine, or C4 cycloalkyl, wherein the heterocyclyl or heteroaryl contains 1-3 heteroatoms selected from nitrogen, oxygen, and sulfur.In some embodiments, Ring A is cyclohexyl substituted with -CF3, phenyl substituted with -CHF2, a 6-10 membered heterocyclyl substituted with H, OH, cyano, F, Cl, optionally substituted C1-C3 alkyl, C4-C6 cycloalkyl, -C(O)(6-membered heteroaryl), or -C(O)(9-membered heterocyclyl), or a 5-9 membered heteroaryl optionally substituted with -CH3, amine, or cyclobutyl, wherein the heterocyclyl or heteroaryl contains 1-3 heteroatoms selected from nitrogen, oxygen, and sulfur.

[0051] In some embodiments, ring A is an optionally substituted C5-C6 cycloalkyl. In some embodiments, ring A is an optionally substituted C6 cycloalkyl. In some embodiments, ring A is an optionally substituted cyclohexyl. In some embodiments, ring A is cyclohexyl substituted with -CF3. In some embodiments, ring A has the structure: [ka] is.

[0052] In some embodiments, ring A is an optionally substituted C5-C6 aryl. In some embodiments, ring A is an optionally substituted C6 aryl. In some embodiments, ring A is an optionally substituted phenyl. In some embodiments, ring A is a phenyl substituted with -CHF2. In some embodiments, ring A has the structure: [ka] is.

[0053] In some embodiments, ring A is an optionally substituted 6-10 membered heterocyclyl containing 1-3 heteroatoms selected from nitrogen, oxygen, and sulfur. In some embodiments, ring A is a 6-10 membered heterocyclyl substituted with H, OH, cyano, halo, optionally substituted C1-C3 alkyl, optionally substituted C3-C6 cycloalkyl, -C(O)(6-membered heteroaryl), or -C(O)(9-membered heterocyclyl), wherein the heterocyclyl or heteroaryl contains 1-3 heteroatoms selected from nitrogen, oxygen, and sulfur. In some embodiments, Ring A is a 6-10 membered heterocyclyl substituted with H, OH, cyano, F, Cl, Br, optionally substituted C-C alkyl, optionally substituted C-C cycloalkyl, -C(O)(6-membered heteroaryl), or -C(O)(9-membered heterocyclyl), where the heterocyclyl or heteroaryl contains 1-3 heteroatoms selected from nitrogen, oxygen, and sulfur. In some embodiments, Ring A is a 6-10 membered heterocyclyl substituted with H, OH, cyano, F, Cl, optionally substituted C-C alkyl, C-C cycloalkyl, -C(O)(6-membered heteroaryl), or -C(O)(9-membered heterocyclyl), where the heterocyclyl or heteroaryl contains 1-3 heteroatoms selected from nitrogen, oxygen, and sulfur. In some embodiments, ring A has the structure: [ka] is [In the formula, R 3 is H or OH; R 4 is an optionally substituted C1-C3 alkyl, an optionally substituted C3-C6 cycloalkyl, —C(O)(6-membered heteroaryl), or —C(O)(9-membered heterocyclyl); R 5 is optionally substituted C1-C3 alkyl; and R 6is cyano or halo. In some embodiments, ring A has the structure: [ka] [ka] [ka] [ka] In some embodiments, the structure of Ring A is: [ka] The parts are structured as follows: [ka] In some embodiments, the structure of Ring A is: [ka] The parts are structured as follows: [ka] is.

[0054] In some embodiments, ring A is an optionally substituted 5-9 membered heteroaryl containing 1-3 heteroatoms selected from nitrogen, oxygen, and sulfur. In some embodiments, ring A is a 5-9 membered heteroaryl containing 1-3 heteroatoms selected from nitrogen, oxygen, and sulfur, optionally substituted with an optionally substituted C1-C3 alkyl, an optionally substituted amine, or a C4-C6 cycloalkyl. In some embodiments, ring A is a 5-9 membered heteroaryl containing 1-3 heteroatoms selected from nitrogen, oxygen, and sulfur, optionally substituted with a C1-C3 alkyl, an amine, or a C4 cycloalkyl. In some embodiments, ring A is a 5-9 membered heteroaryl containing 1-3 heteroatoms selected from nitrogen, oxygen, and sulfur, optionally substituted with a C1-C3 alkyl, an amine, or a C4 cycloalkyl. In some embodiments, Ring A is a 5-9 membered heteroaryl containing 1-3 heteroatoms selected from nitrogen, oxygen, and sulfur, and optionally substituted with -CH, amine, or cyclobutyl. In some embodiments, Ring A has the structure: [ka] and [In the formula, R 7 is an optionally substituted C1-C3 alkyl or an optionally substituted amine, and n is 0, 1, 2, 3, or 4. In some embodiments, the structure of ring A: [ka] The parts are structured as follows: [ka] is.

[0055] In some embodiments, the compound of Formula (I) is a compound of Formula (IIa), (IIb), (IIc), or (IId): [ka] wherein X and ring A are as described for formula (I).

[0056] In some embodiments, the compound of Formula (I) is a compound of Formula (IIIa), (IIIb), (IIIc), or (IIId): [ka] [In the formula, R 8 is optionally substituted C1-C6 alkyl or optionally substituted C3-C6 cycloalkyl.

[0057] In the description herein, it is understood that any description, variation, embodiment, or aspect of one moiety may be combined with any description, variation, embodiment, or aspect of any other moiety, just as if each combination of descriptions were specifically and individually listed. For example, R 1 Any description, variation, embodiment, or aspect provided herein with respect to R is intended to be used interchangeably with any other description, variation, embodiment, or aspect provided herein with respect to R as if each and every combination were specifically and individually listed. 2 ,X,ring A,R 3 , R 4 , R 5 , R 6 , R 7 , and R 8It is also understood that all descriptions, variations, embodiments, or aspects of formula (I) apply equally to, and are equally described, other formulas detailed herein, where applicable, as if every description, variation, embodiment, or aspect were listed separately and individually for every formula. For example, all descriptions, variations, embodiments, or aspects of formula (I) apply equally to, and are equally described, every formula detailed herein, where applicable, as if every description, variation, embodiment, or aspect were listed separately and individually for every formula, e.g., formulas (IIa), (IIb), (IIc), (IId), (IIIa), (IIIb), (IIIc), and (IIId).

[0058] In some embodiments, provided is a compound selected from the compounds set forth in Table 1, or a pharmaceutically acceptable salt thereof. Although some compounds described in this disclosure, including in Table 1, are presented as particular stereoisomeric and / or non-stereochemical forms, it is understood that all stereochemical forms (including all enantiomeric or diastereomeric forms, and all tautomeric forms) or other forms of any of the disclosed compounds are described herein, including in Table 1. [Table 1] [Table 2] [Table 3] [Table 4] [Table 5] [Table 6] [Table 7] [Table 8] [Table 9] [Table 10] [Table 11] [Table 12] [Table 13] [Table 14] [Table 15] [Table 16] [Table 17] [Table 18] [Table 19] [Table 20] [Table 21] [Table 22] [Table 23] * "&1" represents a stereocenter. *"abs" represents a chiral center. or a pharmaceutically acceptable salt thereof.

[0059] It is understood that combinations of substituents and / or variables of the depicted formulae herein are permissible only if such combinations result in stable compounds.

[0060] Furthermore, all compounds of formula (I) that exist in free base or free acid form can be converted into their pharmaceutically acceptable salts by treatment with an appropriate inorganic or organic base or acid in a manner well known to those skilled in the art. Salts of compounds of formula (I) can be converted into their free base or free acid form by standard techniques.

[0061] Synthesis method The compounds described herein can be made using conventional organic synthesis and commercially available starting materials or using the methods provided herein. By way of example and not limitation, compounds of formula (I) can be prepared as shown in general schemes 1-4 and in the examples provided herein. It should be noted that those skilled in the art will know how to modify the procedures presented in the illustrative schemes and examples to obtain the desired products. [ka] [ka] [ka] [ka]

[0062] How to use Embodiments of the present disclosure provide methods for degrading modified polypeptides in cells, methods for reducing modified polypeptide levels in cells, and methods for treating diseases, such as cancer, in a subject in need of treatment.

[0063] In some embodiments, a method for reducing the level of a modified polypeptide comprising a degradation domain is provided, comprising contacting the modified polypeptide with a compound of Formula (I). In some embodiments, the contacting occurs intracellularly, and the compound of Formula (I) binds to the degradation domain and a ubiquitin ligase, causing ubiquitination and degradation of the modified polypeptide. In some embodiments, degradation of the modified polypeptide results in a decrease in at least one cellular activity, and / or an increase in at least one cellular activity, and / or cell death. Non-limiting exemplary effects include lowering the threshold for cellular (e.g., T cell) activation, improving the functional persistence of cells (e.g., T cells), promoting cell survival, and increasing cell proliferation. In some embodiments, the method comprises administering a compound of Formula (I) to a subject, wherein the subject comprises cells comprising the modified polypeptide.

[0064] In some embodiments, the modified polypeptide is degraded in the presence of a compound of Formula (I). In some embodiments, the compound of Formula (I) interacts with a degradation domain and a ubiquitin ligase (e.g., cereblon). In some embodiments, the compound of Formula (I) causes ubiquitination of the modified polypeptide via a complex comprising the degradation domain, the compound of Formula (I), and the ubiquitin ligase.

[0065] The engineered cells provided herein (e.g., CAR cells), such as T lymphocytes (i.e., T cells) engineered to contain / express an engineered polypeptide, can be used to treat an individual who would benefit from the engineered cells, for example, because the individual has a cancer that expresses the target of the CAR. In some embodiments, the cells are effector T cells. In some embodiments, the cells are CD4+ T cells or CD8+ T cells. In some embodiments, either the T cells, effector T cells, CD4+ T cells, or CD8+ T cells comprise the engineered polypeptide.

[0066] In one aspect, provided herein is a method for degrading a modified polypeptide comprising a degradation domain in a cell in a subject in need of treatment, the method comprising contacting the cell with an effective amount of a compound of Formula (I). Degradation of the modified polypeptide in a cell can be assessed and performed by various methods known in the art. Kits and commercially available assays (including cell-based assays) can be used to determine whether and to what extent the modified polypeptide in the cell has been degraded. In some embodiments, the compound of Formula (I) partially degrades the modified polypeptide in the cell. In some embodiments, the compound of Formula (I) completely degrades the modified polypeptide in the cell.

[0067] In some embodiments, the compound of Formula (I) degrades about 1%, 5%, 10%, 15%, 20%, 25%, 30%, 35%, 40%, 45%, 50%, 55%, 60%, 65%, 70%, 75%, 80%, 85%, 90%, 95%, or 100% of the modified polypeptide in the cell. In some embodiments, the compound of formula (I) inhibits the expression of modified polypeptide in cells by about 1-100%, 5-100%, 10-100%, 15-100%, 20-100%, 25-100%, 30-100%, 35-100%, 40-100%, 45-100%, 50-100%, 55-100%, 60-100%, 65-100%, 70-100%, 75-100%, 80-100%, 90-100%, 95-100%, 100-100%, 110-100%, 120-100%, 130-100%, 140-100%, 150-100%, 160-100%, 170-100%, 180-100%, 190-100%, 210-100%, 220-100%, 230-100%, 240-100%, 250-100%, 260-100%, 270-100%, 280-100%, 290-100%, 300-100%, 310-100%, 320-100%, 330-100%, 340-100%, 350-100%, 360-100%, 370-100%, 380-100%, 390-100%, 400-100%, 410-100%, 420-100%, 430-100%, 440-100%, 450-100%, 460-100%, 470-100%, 480-1 Decomposes 00%, 85-100%, 90-100%, 95-100%, 5-95%, 5-90%, 5-85%, 5-80%, 5-75%, 5-70%, 5-65%, 5-60%, 5-55%, 5-50%, 5-45%, 5-40%, 5-35%, 5-30%, 5-25%, 5-20%, 5-15%, 5-10%, 10-90%, 20-80%, 30-70%, or 40-60%.

[0068] In some embodiments, provided herein are methods for reducing the level of a modified polypeptide in a cell, the methods comprising contacting a cell with an effective amount of a compound of Formula (I). Reduction of the level of a modified polypeptide in a cell can be assessed and performed by various methods known in the art. Kits and commercially available assays (cell-based assays) can be used to determine whether and to what extent the level of a kinase protein has been reduced.

[0069] In some embodiments, the compound of Formula (I) reduces the level of modified polypeptide in a cell by about 1%, 5%, 10%, 15%, 20%, 25%, 30%, 35%, 40%, 45%, 50%, 55%, 60%, 65%, 70%, 75%, 80%, 85%, 90%, 95%, or 100%. In some embodiments, the compound of formula (I) increases the level of modified polypeptide in cells by about 1-100%, 5-100%, 10-100%, 15-100%, 20-100%, 25-100%, 30-100%, 35-100%, 40-100%, 45-100%, 50-100%, 55-100%, 60-100%, 65-100%, 70-100%, 75-100%, 80-100%, 90-100%, 95-100%, 100-100%, 110-100%, 120-100%, 130-100%, 140-100%, 150-100%, 160-100%, 170-100%, 180-100%, 190-100%, 210-100%, 220-100%, 230-100%, 240-100%, 250-100%, 260-100%, 270-100%, 280-100%, 290-100%, 300-100%, 310-100%, 320-100%, 330-100%, 340-100%, 350-100%, 360-100%, 370-100%, 380-100%, 390-100%, 410-100%, 420-100%, 430-100%, 440-100%, 450-100%, 460-100%, 470-100%, 480-100%, 490-10 Reduced by 100%, 85-100%, 90-100%, 95-100%, 5-95%, 5-90%, 5-85%, 5-80%, 5-75%, 5-70%, 5-65%, 5-60%, 5-55%, 5-50%, 5-45%, 5-40%, 5-35%, 5-30%, 5-25%, 5-20%, 5-15%, 5-10%, 10-90%, 20-80%, 30-70%, or 40-60%.

[0070] In some embodiments, the compounds of Formula (I) have an EC 20 or EC 30 or EC 40 or EC 50 or EC 60 or EC 70 or EC 80 or EC 90 or EC 100 or EC 110 or EC 120 or EC 130 or EC 140 or EC 150 or EC 160 or EC 170 or EC 180 or EC 50 In some embodiments, the compounds of Formula (I) have an EC value of about 0.0003 μM to about 1 μM, or about 0.0003 μM to about 0.2 μM, or about 0.0003 μM to about 0.05 μM. In some embodiments, the compounds of Formula (I) have an EC value of about 0.05 μM to about 0.2 μM. 50 In some embodiments, the compounds of Formula (I) have an EC 50 In some embodiments, compounds of Formula (I) have an EC 50 In some embodiments, the compound of formula (I) has EC 50 has a value of less than 0.2 μM, less than 0.05 μM, less than 0.001 μM, or less than about 0.0003 μM.

[0071] In some embodiments, administration of CAR cells containing a modified polypeptide comprising a degradation domain may desirably reduce or eliminate CAR expression, thereby reducing or eliminating target cell killing. In some such embodiments, the method may further comprise administering a compound of Formula (I) to a subject. Administration of a compound of Formula (I) causes degradation of the modified polypeptide (e.g., CAR), reducing or eliminating targeting of the modified cells to cells expressing an antigen bound to the antigen-binding domain of the CAR. In this way, the therapeutic activity of the CAR cells may be modulated and safety may be improved.

[0072] In some embodiments, the modified cell population is first administered to a subject, followed by administration of a compound of Formula (I) a specific period of time after administration of the modified cell population, for example, 30 minutes, 1 hour, 6 hours, 12 hours, 1 day, 2 days, 3 days, 4 days, 5 days, 6 days, or 1 week after administration of the cell population.

[0073] A non-limiting list of cancers that may be treated according to the methods of treatment described herein includes lymphoma, leukemia, lung cancer, breast cancer, prostate cancer, adrenocortical carcinoma, thyroid cancer, nasopharyngeal carcinoma, melanoma, skin cancer, colorectal cancer, desmoid tumor, dysplastic small round cell tumor, endocrine tumor, Ewing's sarcoma, peripheral primitive neuroectodermal tumor, solid germinoma, hepatoblastoma, neuroblastoma, non-rhabdomyosarcoma soft tissue sarcoma, osteosarcoma, retinoblastoma, rhabdomyosarcoma, Wilms' tumor, glioma, glioblastoma, myxoma, fibroma, and lipoma. Exemplary lymphomas and leukemias include, but are not limited to, chronic lymphocytic leukemia (small lymphocytic lymphoma), B-cell prolymphocytic leukemia, lymphoplasmacytic lymphoma, Waldenstrom's macroglobulinemia, marginal zone lymphoma, plasma cell myeloma, plasmacytoma, extramuscular marginal zone B-cell lymphoma, MALT lymphoma, muscular marginal zone B-cell lymphoma, follicular lymphoma, mantle cell lymphoma, diffuse large B-cell lymphoma, mediastinal (thymic) large B-cell lymphoma, intravascular large B-cell lymphoma, primary effusion lymphoma, Burkitt's lymphoma, T-cell lymphoma, These include prolymphocytic leukemia, T-cell lymphocytic large granular lymphocytic leukemia, aggressive NK-cell leukemia, adult T-cell lymphocytic leukemia / lymphoma, extramuscular NK / T-cell lymphocytic lymphoma, nasal type, enteropathy type T-cell lymphocytic lymphoma, hepatosplenic T-cell lymphocytic lymphoma, blastic NK-cell lymphoma, mycosis fungoides, Sézary syndrome, primary cutaneous anaplastic large cell lymphoma, lymphomatoid papulosis, angioimmunoblastic T-cell lymphocytic lymphoma, peripheral T-cell lymphocytic lymphoma (not otherwise specified), anaplastic large cell lymphoma, Hodgkin's lymphoma, or non-Hodgkin's lymphoma.

[0074] The efficacy of the modified cells described herein, such as CAR cells, in treating a disease or disorder (e.g., treating an individual with cancer) can be assessed by one or more criteria specific to the particular disease or disorder, known to those skilled in the art, and which indicate the progression of the disease or disorder. Generally, administration of CAR cells (e.g., CAR T cell lymphocytes) to an individual with a disease / disorder (e.g., cancer) is efficacious if one or more of said criteria detectably (e.g., significantly) shifts from a disease-state value or range to or toward a normal value or range.

[0075] In some embodiments, the compounds of Formula (I) are useful in the manufacture of a medicament for reducing the level of an altered polypeptide in a cell.

[0076] The methods and uses of the disclosure may involve compounds of formula (I) used alone or in combination with one or more additional therapies (e.g., non-drug treatments or therapeutic agents).

[0077] The compound of formula (I) may be administered before, after, or simultaneously with one or more such additional therapies. When used in combination, the dose of the compound of formula (I) and the dose of one or more additional therapies (e.g., non-drug treatments or therapeutic agents) may provide a therapeutic effect (e.g., a synergistic or additive therapeutic effect). The compound of formula (I) and the additional therapies (e.g., anticancer agents) may be administered together, for example, in a single pharmaceutical composition, or separately, and when administered separately, they may be administered simultaneously or sequentially. Such sequential administration may be administered at a short or long interval.

[0078] In some embodiments, the additional therapy is the administration of an agent that limits side effects (e.g., an agent intended to reduce the occurrence or severity of side effects of treatment). For example, in some embodiments, a compound of Formula (I) can be used in combination with a therapeutic agent that treats nausea. Examples of agents that can be used to treat nausea include, but are not limited to, dronabinol, granisetron, metoclopramide, ondansetron, prochlorperazine, and pharmaceutically acceptable salts thereof.

[0079] In some embodiments, the one or more additional therapies include a non-drug treatment (e.g., surgery or radiation therapy). In some embodiments, the one or more additional therapies include a therapeutic agent (e.g., a chemotherapeutic or biotherapeutic agent that is an anti-proliferative agent). In some embodiments, the one or more additional therapies include a non-drug treatment and a therapeutic agent. In other embodiments, the one or more additional therapies include two therapeutic agents. In yet other embodiments, the one or more additional therapies include three therapeutic agents. In some embodiments, the one or more additional therapies include four or more therapeutic agents.

[0080] Pharmaceutical Compositions and Routes of Administration The compounds provided herein can be administered to a subject orally, topically, or parenterally in conventional preparations such as capsules, microcapsules, tablets, granules, powders, troches, pills, suppositories, injections, suspensions, syrups, patches, creams, lotions, ointments, gels, sprays, solutions, and emulsions.

[0081] The compounds disclosed herein can be administered to a subject orally, topically, or parenterally in conventional preparation forms, such as capsules, microcapsules, tablets, granules, powders, lozenges, pills, suppositories, injections, suspensions, syrups, patches, creams, lotions, ointments, gels, sprays, solutions, and emulsions. Suitable formulations may contain conventional organic or inorganic additives, such as excipients (e.g., sucrose, starch, mannitol, sorbitol, lactose, glucose, cellulose, talc, calcium phosphate, or calcium carbonate), binders (e.g., cellulose, methylcellulose, hydroxymethylcellulose, polypropylpyrrolidone, polyvinylpyrrolidone, gelatin, gum arabic, polyethylene glycol, sucrose, or starch), disintegrants (e.g., starch, carboxymethylcellulose, hydroxypropyl starch, low-substituted hydroxypropyl cellulose, sodium bicarbonate, calcium phosphate, or calcium citrate), lubricants (e.g., stearic acid, The pharmaceutical compositions can be prepared by a commonly used method using additives such as cereals such as cereals, dairy products, and pharmaceutical preparations, such as cereals containing glutamic acid, cereals containing glutamic acid, and / or cereals containing glutamic acid. The pharmaceutical compositions can be prepared by a commonly used method using additives ...

[0082] The dose of a compound of formula (I) administered to a subject may be relatively variable and subject to the judgment of a health care practitioner. In any given case, the amount of a compound of formula (I) administered will depend on factors such as the solubility of the active ingredient, the formulation used, and the route of administration.

[0083] In other embodiments, provided herein are unit dose formulations comprising about 0.1 mg to 500 mg, about 1 mg to 250 mg, about 1 mg to about 100 mg, about 1 mg to about 50 mg, about 1 mg to about 25 mg, or about 1 mg to about 10 mg of a compound of Formula (I).

[0084] The compound of formula (I) can be administered orally for convenience.In some embodiments, when administered orally, the compound of formula (I) is administered with food and water.In other embodiments, the compound of formula (I) is dispersed in water or juice (e.g., apple juice or orange juice) or any other liquid, and is orally administered as a solution or suspension.

[0085] The compounds disclosed herein may also be administered intradermally, intramuscularly, intraperitoneally, transdermally, intravenously, subcutaneously, intranasally, epidurally, sublingually, intracerebrally, intravaginally, transdermally, rectally, mucosally, by inhalation, or topically to the ophthalm, nose, eye, or skin. The mode of administration is left to the discretion of the health care practitioner and may depend, in part, on the site of the condition.

[0086] In one embodiment, a capsule is provided that contains a compound of formula (I) and no additional carriers, excipients, or vehicles.

[0087] In another embodiment, a composition is provided comprising an effective amount of a compound of Formula (I) and a pharmaceutically acceptable carrier or vehicle, wherein the pharmaceutically acceptable carrier or vehicle comprises an excipient, a diluent, or a mixture thereof. In some embodiments, the composition is a pharmaceutical composition.

[0088] The compositions may be in the form of tablets, chewable tablets, capsules, solutions, parenteral solutions, lozenges, suppositories, and suspensions. The compositions may be formulated into dosage units containing a daily dose, or a convenient fraction of the daily dose, such as a single tablet or capsule, or a convenient amount of liquid. In some embodiments, solutions are prepared from water-soluble salts, such as hydrochlorides. Generally, all compositions are prepared according to methods well known in medicinal chemistry. Capsules may be prepared by mixing a compound of Formula (I) with a suitable carrier or diluent and filling the appropriate amount of the mixture into capsules. Typical carriers and diluents include, but are not limited to, inert powdered substances such as various starches, powdered cellulose, especially crystalline and microcrystalline cellulose, sugars such as fructose, mannitol, and sucrose, wheat flour, and similar edible powders.

[0089] Tablets can be prepared by direct compression, wet granulation, or dry granulation. These formulations usually contain diluents, binders, lubricants, and disintegrants and compounds. Typical diluents include, for example, various types of starch, lactose, mannitol, kaolin, calcium phosphate or sulfate, inorganic salts such as sodium chloride, and powdered sugar. Powdered cellulose derivatives are also useful. Typical tablet binders include starch, gelatin, and sugars such as lactose, fructose, and glucose. Natural and synthetic gums are also useful, including acacia, alginate, methylcellulose, polyvinylpyrrolidine, and the like. Polyethylene glycol, ethylcellulose, and waxes can also serve as binders.

[0090] Lubricants may be necessary in tablet formulations to prevent colorants from adhering to the tablet and punch. Lubricants can be selected from slippery solids such as talc, magnesium and calcium stearate, stearic acid, and hydrogenated vegetable oils. Tablet disintegrants are substances that swell upon absorption of water, breaking the tablet and releasing the compound. These include starch, clay, cellulose, algin, and gums. Examples of suitable disintegrants include corn starch and potato starch, methylcellulose, agar, bentonite, wood cellulose, natural sponge powder, cation exchange resins, alginic acid, guar gum, citrus fruit extract, and carboxymethylcellulose, as well as sodium lauryl sulfate. Tablets can be coated with sugars for flavoring and sealing, or with film-forming protective agents to modify tablet dissolution. The composition can also be formulated as a chewable tablet, for example, using substances such as mannitol during the manufacturing process.

[0091] When it is desired to administer the compound of formula (I) as a suppository, typical bases can be used.Cocoa butter is a traditional suppository base, and the melting point can be slightly increased by adding wax.In particular, water-miscible suppository bases, including polyethylene glycols of various molecular weights, are widely used.

[0092] The effect of the compound of formula (I) can be delayed or prolonged by appropriate formulation. For example, slowly dissolving pellets of the compound of formula (I) can be prepared into tablets or capsules or incorporated into sustained-release implantable devices. Techniques include creating pellets with several different dissolution rates and filling capsules with a mixture of the pellets. The tablets or capsules can be coated with a film that prevents dissolution for a predictable period of time. The duration of effect can be extended even in parenteral preparations by dissolving or suspending the compound of formula (I) in an oily or emulsion vehicle that allows it to be slowly dispersed in the blood. [Example]

[0093] The following examples are presented by way of non-limiting example. Compounds are named using the automated naming tool provided in ChemBiodraw Ultra (Cambridgesoft), which generates systematic names for chemical structures according to the Cahn-Ingold-Prelog rules of stereochemistry. Those skilled in the art can modify the procedures presented in the illustrative examples to obtain the desired products.

[0094] Salts of the compounds described herein can be prepared by standard methods, such as including an acid (e.g., TFA, formic acid, or HCl) in the mobile phase during chromatographic purification, or by stirring the product with an acidic solution (e.g., HCl solution) after chromatographic purification.

[0095] The following abbreviations may be relevant to this application: Abbreviation EtOAc: ethyl acetate DCM: dichloromethane DMF: dimethylformamide DMSO: dimethyl sulfoxide MeCN: acetonitrile THF: tetrahydrofuran MeOH: Methanol EtOH: Ethanol AcOH: acetic acid TFA: trifluoroacetic acid NaBH4: sodium borohydride NaBH(OAc)3: Sodium triacetoxyborohydride DIPEA: N,N-diisopropylethylamine TEA: Triethylamine TMP: 2,2,6,6-tetramethylpiperidine n-BuLi: n-butyllithium N2: Nitrogen Na2SO4: Sodium sulfate MgSO4: Magnesium sulfate PdCl2(dtbpf): [1,1'-bis(di-tert-butylphosphino)ferrocene]dichloropalladium(II) Pd(dppf)Cl2: [1,1'-bis(diphenylphosphino)ferrocene]dichloropalladium(II) Pd(dppf)Cl2.DCM: Dichloromethane complex of [1,1'-bis(diphenylphosphino)ferrocene]dichloropalladium(II) Pd2(dba)3: Tris(dibenzylideneacetone)dipalladium Pd / C: Palladium on carbon HATU: (1-[bis(dimethylamino)methylene]-1H-1,2,3-triazolo[4,5-b]pyridinium 3-oxide hexafluorophosphate, hexafluorophosphate azabenzotriazole tetramethyl uronium)

[0096] Synthesis Examples Analysis method Preparative HPLC Method 1: XBridge C18, 200 mm x 19 mm, 5 μm particle size; Mobile Phase A: 5:95 acetonitrile:water (10 mM ammonium acetate); Mobile Phase B: 95:5 acetonitrile:water (10 mM ammonium acetate); Gradient: 15% B at 0 min, 15–50% B over 25 min, then 100% B for 6 min; Flow Rate: 20 mL / min; Column Temperature: 25°C. Fraction collection was based on MS signal.

[0097] Preparative HPLC Method 2: 1-Phen Luna Axia C18 5u 30 x 100mm; Mobile Phase A: 95% HO / 5% ACN / 0.05% TFA; Mobile Phase B: 5% HO / 95% ACN / 0.05% TFA; Gradient: 2% B at 0 min, 2-100% B over 12 min, then 100% B for 5 min; Flow Rate: 25 mL / min; Column Temperature: 25 °C. Fraction collection was based on UV (220) nm.

[0098] Analytical HPLC Method 1: Waters XBridge C18, 2.1 mm x 50 mm, 1.7 μm particle size; Mobile Phase A: 5:95 acetonitrile:water (0.1% trifluoroacetic acid); Mobile Phase B: 95:5 acetonitrile:water (0.1% trifluoroacetic acid); Temperature: 50 °C; Gradient: 0% to 100% B in 3 min, then hold at 100% B for 0.50 min; Flow Rate: 1 mL / min; Detection: MS and UV (220 nm).

[0099] Example S1. 3-(4,6-Difluoro-5-(4-hydroxy-1-(4-(trifluoromethyl)benzyl)piperidin-4-yl)-1-oxoisoindolin-2-yl)piperidine-2,6-dione (Compound No. 11) [ka] [ka]

[0100] Step 1: To a stirred solution of 2,2,6,6-tetramethylpiperidine (17.95 mL, 105 mmol) in THF (150 mL) at 0°C under a nitrogen atmosphere, n-BuLi (63.3 mL, 101 mmol) was added dropwise, and the resulting mixture was stirred at 0°C for 30 minutes. The reaction mixture was then cooled to approximately −45°C (using a dry ice / MeCN bath), and 4-bromo-3,5-difluorobenzoic acid (10.0 g, 42.2 mmol) dissolved in THF (25 mL) was added dropwise, and stirring was continued at −45°C. After 3 hours, DMF (4.88 mL, 63.3 mmol) was added dropwise, and the reaction mixture was allowed to warm to room temperature and stir overnight. The reaction mixture was quenched with 3 M aqueous HCl (40 mL) at 0°C and extracted with DCM (×3). The combined organic layers were dried over NaSO, filtered, and concentrated to dryness. The crude product was purified by silica gel chromatography (eluting with 0-80% EtOAc / hexanes) to give 5-bromo-4,6-difluoro-3-hydroxyisobenzofuran-1(3H)-one (4.612 g, 33.0% yield).

[0101] 1 H NMR (400 MHz, chloroform-d) δ 10.27 (s, 1H), 7.53 (dd, J = 8.2, 1.7 Hz, 1H).

[0102] Step 2: To a solution of 5-bromo-4,6-difluoro-3-hydroxyisobenzofuran-1(3H)-one (4.0 g, 15.09 mmol) in DMF (100 mL) was added tert-butyl (S)-4,5-diamino-5-oxopentanoate, HCl (3.60 g, 15.09 mmol), followed by NaBH(OAc) (4.80 g, 22.64 mmol). This was stirred at room temperature for 16 hours. To this mixture was added HATU (7.17 g, 18.87 mmol) and triethylamine (8.42 mL, 60.4 mmol) and stirred at room temperature for 2 hours. The reaction was quenched by the addition of 10% aqueous LiCl, and the product was then extracted with EtOAc. The organic layer was washed with brine, dried over MgSO, and then concentrated. The crude product was purified using a 120 g silica gel column (ISCO) (eluting with 0–80% EtOAc / hexanes) to give (S)-5-amino-4-(5-bromo-4,6-difluoro-1-oxoisoindolin-2-yl)-5-oxopentanoate (2.01 g, 56.1% yield).

[0103] 1 H NMR (400 MHz, chloroform-d) δ ppm 7.40 - 7.51 (m, 1 H) 6.11 - 6.26 (m, 1 H) 5.39 (br s, 1 H) 4.91 (dd, J = 8.68, 6.15 Hz, 1 H) 4.67 - 4.76 (m, 1 H) 4.49 - 4.58 (m, 1 H) 2.26 - 2.43 (m, 3 H) 2.11 - 2.21 (m, 1 H) 1.45 (s, 9 H)

[0104] Step 3: To a solution of tert-butyl 4-(4,4,5,5-tetramethyl-1,3,2-dioxaborolan-2-yl)-3,6-dihydropyridine-1(2H)-carboxylate (1.383 g, 4.47 mmol) and tert-butyl (S)-5-amino-4-(5-bromo-4,6-difluoro-1-oxoisoindolin-2-yl)-5-oxopentanoate (1.55 g, 3.58 mmol) in dioxane (30 mL) was added K2CO3 (1.236 g, 8.94 mmol) dissolved in water (15 mL). PdCl2(dppf).DCM (0.146 g, 0.179 mmol) was added, and the atmosphere was replaced with nitrogen. The mixture was heated to 100 °C for 1 hour. It was cooled to room temperature, diluted with EtOAc, quenched with brine, and the organic layer was separated. The organic layer was washed with brine, dried over Na2SO4, filtered, and concentrated. The crude product was purified by flash column chromatography (eluted with 0-15% B / DCM [B = 15% ethanol / EtOAc + 0.1% TEA]).

[0105] LC / MS (ESI) m / z 536.5 [(M+H) + , C 27 H 35 Calculated value for F2N3O6: 535.2].

[0106] Step 4: A solution of tert-butyl (S)-4-(2-(1-amino-5-(tert-butoxy)-1,5-dioxopent-2-yl)-4,6-difluoro-1-oxoisoindolin-5-yl)-3,6-dihydropyridine-1(2H)-carboxylate (1.00 g, 1.867 mmol) in DCM (10 mL) and 2-propanol (40 mL) was bubbled with oxygen for 5 minutes. Then, tris(2,2,6,6-tetramethyl-3,5-heptanedionato)manganese(III) (0.113 g, 0.187 mmol) and phenylsilane (0.404 g, 3.73 mmol) were added. The reaction mixture was stirred at room temperature with an oxygen balloon attached for 2 days. The reaction mixture was diluted with EtOAc and washed with sodium thiosulfate solution. The organic layer was dried, concentrated and purified by ISCO using a 40 g silica gel column (eluting with 0-5% MeOH / DCM) to give 417 mg of the desired product as a white solid.

[0107] LC / MS (ESI) m / z 498.4 [(M-55) + , C 27 H 37 Calculated value for F2N3O7: 553.3].

[0108] Step 5: A 20 mL microwave vial was charged with (tert-butyl (S)-4-(2-(1-amino-5-(tert-butoxy)-1,5-dioxopentan-2-yl)-4,6-difluoro-1-oxoisoindolin-5-yl)-4-hydroxypiperidine-1-carboxylate (410 mg, 0.741 mmol), 4-methylbenzenesulfonic acid (255 mg, 1.481 mmol) and acetonitrile (10 mL). It was heated to 120° C. in a microwave for 1 h. It was concentrated to dryness and the residue was rinsed with ether to remove excess pTsOH. The precipitate was air-dried to give 391 mg of the product as the pTsOH monohydrate salt.

[0109] LC / MS (ESI) m / z 380.3 [(M+H) + , C 18 H 19Calculated value for F2N3O4 379.1]. The enantiomeric excess of this material and the following compounds was not determined.

[0110] Step 6: To a solution of 4-(trifluoromethyl)benzaldehyde (12.85 mg, 0.074 mmol) and (S)-3-(4,6-difluoro-5-(4-hydroxypiperidin-4-yl)-1-oxoisoindolin-2-yl)piperidine-2,6-dione (14 mg, 0.037 mmol) in DMF (1 mL) was added two drops of AcOH and then stirred at room temperature. After 0.5 h, NaBH(OAc) (23.46 mg, 0.111 mmol) was added, and the resulting solution was stirred at room temperature for 16 h. The reaction mixture was purified by preparative HPLC Method 1 to give 7.7 mg of product.

[0111] LC / MS (ESI) m / z 538.1 [(M+H)+,C 26 H 24 Calculated for F5N3O4 537.2]; HPLC a retention time = 1.25 minutes; 1 H NMR (500 MHz,DMSO-d6) δ 11.01 (s,1H),7.87 - 7.82 (m,2H),7.76 (br d,J=7.9 Hz,2H),7.44 (br d,J=10.4 Hz,1H),5.11 - 5.05 (m,1H),4.53 (br d,J=17.1 Hz,1H),4.41 - 4.35 (m,1H),2.95 - 2.85 (m,1H),2.62 (br d,J=17.1 Hz,1H),2.51 (br s,4H),2.47 - 2.33 (m,2H),2.25 (br d,J=13.7 Hz,2H),2.06 - 1.99 (m,1H).

[0112] Example S2. 3-(4,6-Difluoro-1-oxo-5-(1-(4-(trifluoromethyl)benzyl)piperidin-4-yl)isoindolin-2-yl)piperidine-2,6-dione (Compound No. 52) [ka]

[0113] Step 1: A 200 mL round-bottom flask was charged with tert-butyl (S)-4-(2-(1-amino-5-(tert-butoxy)-1,5-dioxopentan-2-yl)-4,6-difluoro-1-oxoisoindolin-5-yl)-3,6-dihydropyridine-1(2H)-carboxylate (1.20 g, 2.241 mmol), MeOH (50 mL), and Pd / C (0.119 g, 0.112 mmol). The air was replaced with hydrogen and the mixture was stirred vigorously at room temperature under a hydrogen atmosphere for 16 hours. It was filtered through a pad of Celite and concentrated to give the desired product.

[0114] LC / MS (ESI) m / z 538.5 [(M+H) + , C 27 H 37 Calculated value of F2N3O6: 537.3

[0115] Step 2: A 20 mL microwave vial was charged with tert-butyl (S)-4-(2-(1-amino-5-(tert-butoxy)-1,5-dioxopentan-2-yl)-4,6-difluoro-1-oxoisoindolin-5-yl)piperidine-1-carboxylate (770 mg, 1.432 mmol), benzenesulfonic acid (453 mg, 2.86 mmol), and MeCN (15 mL). It was heated to 130° C. in a microwave for 0.5 h. It was diluted with ether, and the precipitate was collected by filtration and then air-dried to give the PhSO3H monohydrate salt of the desired product. Preparative HPLC Method 2 gave the TFA salt form of this intermediate.

[0116] MS: C 18 H 19 F2N3O3[M+H] + 364, Measured value [M+H] + 364. 1H NMR (300 MHz,DMSO-d6) δ 11.02 (s,1H),8.65 (d,J = 11.3 Hz,1H),8.38 (d,J = 10.8 Hz,1H),7.74 - 7.55 (m,3H),7.50 (d,J = 8.8 Hz,1H),7.40 - 7.23 (m,5H),5.12 (dd,J = 13.3,5.1 Hz,1H),4.55 (d,J = 17.3 Hz,1H),4.38 (d,J = 17.3 Hz,1H),3.39 - 3.32(m,2H),3.09 (m,2H),2.98 - 2.84 (m,1H),2.63 - 2.50 (m,1H),2.41 (m,2H),2.29 - 2.10 (m,2H),2.07 - 1.94 (m,1H),1.94 - 1.82 (d,J = 13.6 Hz,2H).

[0117] Step 3: To a solution of 4-(trifluoromethyl)benzaldehyde (13.42 mg, 0.077 mmol) and (S)-3-(4,6-difluoro-1-oxo-5-(piperidin-4-yl)isoindolin-2-yl)piperidine-2,6-dione, benzenesulfonate (20 mg, 0.038 mmol) in DMF (1 mL) was added two drops of AcOH and then stirred at room temperature. After 0.5 h, NaBH(OAc) (24.50 mg, 0.116 mmol) was added, and the resulting solution was stirred at room temperature for 16 h. Purification by preparative HPLC Method 1 afforded 7.2 mg of the title compound. LC / MS (ESI) m / z 522.1 [(M+H)+, C 26 H 24 Calculated value for F5N3O3: 521.2; HPLC a retention time = 1.30 minutes; 1H NMR (500 MHz,DMSO-d6) δ 11.01 (s,1H),7.65 (br d,J=7.6 Hz,2H),7.53 (br d,J=7.9 Hz,2H),7.40 (br d,J=8.5 Hz,1H),5.10 - 5.00 (m,1H),4.54 - 4.45 (m,1H),4.37 - 4.28 (m,1H),3.63 - 3.48 (m,1H),3.01 - 2.80 (m,4H),2.63 - 2.54 (m,1H),2.46 (br s,4H),2.41 - 2.29 (m,1H),2.09 - 1.89 (m,5H),1.70 - 1.60 (m,2H).

[0118] Example S4. 3-(5-(((4-(difluoromethyl)benzyl)amino)methyl)-4,6-difluoro-1-oxoisoindolin-2-yl)piperidine-2,6-dione (Compound 79) [ka] [ka]

[0119] Step 1: A microwave vial was charged with tert-butyl (S)-5-amino-4-(5-bromo-4,6-difluoro-1-oxoisoindolin-2-yl)-5-oxopentanoate (1.00 g, 2.308 mmol), dicyanozinc (0.271 g, 2.308 mmol), Xantphos (0.033 g, 0.057 mmol), and Pd2(dba)3 (0.106 g, 0.115 mmol) in DMF (20 mL). The atmosphere was replaced with argon and heated in a microwave at 130 °C for 1 h. It was diluted with EtOAc, washed with 10% aqueous LiCl and brine, dried over MgSO4, and then concentrated. The crude product was purified by flash column chromatography (eluting with 5-80% EtOAc / hexanes) to give 501 mg of the desired product (57% yield).

[0120] LC / MS (ESI) m / z 324.2 [(M-55)+, C 18 H 19 Calculated value of F2N3O4: 379.1

[0121] Step 2: To a solution of tert-butyl (S)-5-amino-4-(5-cyano-4,6-difluoro-1-oxoisoindolin-2-yl)-5-oxopentanoate (200 mg, 0.527 mmol) in methanol (20 mL) was added cobalt(II) chloride (137 mg, 1.054 mmol) followed by NaBH (19.94 mg, 0.527 mmol) at 0 °C, and the atmosphere was replaced with N. After 10 min, the cooling bath was removed and the reaction was allowed to warm to room temperature. After 2 h, it was concentrated to dryness, and the residue was suspended in EtOAc and washed with 1.5 M aqueous KHPO. The organic layer was separated, washed with brine, dried over NaSO, filtered, and concentrated. The crude product was purified by flash column chromatography (eluting with 0–50% B / DCM [B = 15% EtOH / EtOAc + 0.1% TEA]) to give tert-butyl 5-amino-4-(5-(aminomethyl)-4,6-difluoro-1-oxoisoindolin-2-yl)-5-oxopentanoate (150 mg, 74.2% yield).

[0122] LC / MS (ESI) m / z 384.2 [(M+H) + , C 18 H 23 Calculated value of F2N3O4: 383.2

[0123] Step 3: A 5 mL microwave vial was charged with tert-butyl (S)-5-amino-4-(5-(aminomethyl)-4,6-difluoro-1-oxoisoindolin-2-yl)-5-oxopentanoate (149 mg, 0.389 mmol), benzenesulfonic acid (123 mg, 0.777 mmol), and MeCN (3 mL). It was heated to 130 °C for 0.5 h. It was concentrated to dryness, and the residue was rinsed with ether to give the PhSO3H monohydrate salt of the desired product (151 mg, 86% yield).

[0124] LC / MS (ESI) m / z 310.2 [(M+H) + , C 14 H 13 Calculated value of F2N3O3: 309.3

[0125] Step 4: 3-(5-(aminomethyl)-4,6-difluoro-1-oxoisoindolin-2-yl)piperidine-2,6-dione was reacted with 4-(difluoromethyl)benzaldehyde by reductive amination to give the title compound in 18% yield according to the procedure shown in Example S1.

[0126] LC / MS (ESI) m / z 450.0 [(M+H) + ,C 22 H 19 Calculated F4N3O3 449.1]; HPLC a retention time = 1.11 min; 1 H NMR (500 MHz,DMSO-d6) δ 11.05 - 10.95 (m,1H),7.55 - 7.46 (m,5H),5.12 (dd,J=13.2,5.2 Hz,1H),4.58 - 4.50 (m,1H),4.39 (br d,J=17.6 Hz,1H),3.91 - 3.79 (m,4H),2.97 - 2.87 (m,1H),2.66 - 2.57 (m,1H),2.49 - 2.39 (m,1H),2.08 - 1.98 (m,1H)

[0127] Examples of compounds prepared according to the procedures shown in Scheme 1 and Schemes 1a-1c using the appropriate aldehyde are listed in Table 2.

[0128] [Table 24] [Table 25] [Table 26] [Table 27] [Table 28] [Table 29]

[0129] Example S68. 3-(5-(1-(benzo[d]thiazol-5-ylmethyl)-4-hydroxypiperidin-4-yl)-4,6-difluoro-1-oxoisoindolin-2-yl)piperidine-2,6-dione (Compound 12) [ka]

[0130] To 6-(chloromethyl)benzo[d]thiazole (7.5 mg, 0.041 mmol) was added 3-(4,6-difluoro-5-(4-hydroxypiperidin-4-yl)-1-oxoisoindolin-2-yl)piperidine-2,6-dione (14 mg, 0.037 mmol) dissolved in 1 mL of DMF, followed by Hunig's base (0.045 mL, 0.258 mmol). The resulting mixture was heated at 85° C. for 2 hours. It was cooled to room temperature and purified by preparative HPLC Method 1 to give 6.0 mg of the title compound.

[0131] LC / MS (ESI) m / z 527.0 [(M+H) + , C 26 H 24 Calculated for F2N4O4S 526.1]; HPLCa retention time = 0.93 min; 1H NMR (500 MHz,DMSO-d6) δ 10.96 (s,1H),9.34 (s,1H),8.16 - 8.09 (m,1H),8.04 (br d,J=8.9 Hz,1H),7.59 - 7.49 (m,1H),7.36 (br d,J=10.4 Hz,1H),5.62 - 5.40 (m,1H),5.05 (br dd,J=13.3,5.0 Hz,1H),4.47 (br d,J=17.4 Hz,1H),4.30 (br d,J=17.4 Hz,1H),3.52 - 3.30 (m,1H),2.90 - 2.81 (m,1H),2.59 - 2.53 (m,1H),2.46 (br s,5H),2.43 - 2.32 (m,1H),2.22 (br s,1H),1.99 - 1.93 (m,2H).

[0132] Example S69. 3-(5-(1-(benzo[d]thiazol-6-ylmethyl)piperidin-4-yl)-4,6-difluoro-1-oxoisoindolin-2-yl)piperidine-2,6-dione (Compound 18) [ka]

[0133] Following the alkylation procedure shown for the synthesis of Example S68, 3-(4,6-difluoro-1-oxo-5-(piperidin-4-yl)isoindolin-2-yl)piperidine-2,6-dione was used as the amine coupling partner to afford the title compound as an off-white solid in 34% yield.

[0134] LC / MS (ESI) m / z 511.1 [(M+H) + , C 26 H 24 Calculated value for F2N4O3S: 510.2; HPLC a retention time = 0.93 min; 1H NMR (500 MHz,DMSO-d6) δ 10.96 (s,1H),9.47 (s,1H),8.29 (s,1H),8.16 (d,J=8.2 Hz,1H),7.66 (br d,J=8.5 Hz,1H),7.44 (br d,J=8.9 Hz,1H),5.06 (br dd,J=13.6,5.0 Hz,1H),4.53 - 4.41 (m,2H),4.33 (br d,J=17.1 Hz,1H),2.92 - 2.80 (m,1H),2.56 (br d,J=17.1 Hz,1H),2.46 (br s,5H),2.43 - 2.31 (m,1H),2.29 - 2.16 (m,1H), 1.99 - 1.89 (m,2H).

[0135] Example S71. 3-(5-(1-(5-bromo-3-methylpicolinoyl)piperidin-4-yl)-4,6-difluoro-1-oxoisoindolin-2-yl)piperidine-2,6-dione (Compound 66) [ka]

[0136] Following the amidation procedure shown in General Scheme 1, using 3-(4,6-difluoro-1-oxo-5-(piperidin-4-yl)isoindolin-2-yl)piperidine-2,6-dione as the amine coupling partner, the title compound was obtained in 39% yield as a white solid.

[0137] LC / MS (ESI) m / z 561.0 [(M+H) + ,C 25 H 23 Calculated for BrF2N4O4 560.1]; HPLCa retention time = 1.52 min; 1H NMR (500 MHz,DMSO-d6) δ 10.96 (s,1H),8.51 (s,1H),8.02 (s,1H),7.42 (d,J=8.7 Hz,1H),5.05 (dd,J=13.3,4.9 Hz,1H),4.63 (br d,J=12.6 Hz,1H),4.50 (d,J=17.3 Hz,1H),4.33 (d,J=17.1 Hz,1H),3.18 - 3.12 (m,1H),2.93 - 2.80 (m,2H),2.56 (br d,J=17.5 Hz,1H),2.46 (s,4H),2.43 - 2.33 (m,1H),2.23 (s,3H),2.01 - 1.79 (m,3H),1.63 (br d,J=12.8 Hz,1H)

[0138] 1.76 (m,3H)

[0139] Examples of compounds prepared using the appropriate alkyl halide according to the alkylation procedures shown for the synthesis of Examples S68, S69, and S72, and also using the appropriate carboxylic acid according to the amidation procedures shown for the synthesis of Examples S70 and S71, are listed in Table 3.

[0140] [Table 30] [Table 31] [Table 32]

[0141] Example S101. N-[2-[4-[2-(2,6-dioxo-3-piperidyl)-4,6-difluoro-1-oxo-isoindolin-5-yl]-4-hydroxy-1-piperidyl]ethyl]-N,2,4-trimethyl-thiazole-5-sulfonamide (Compound 39) [ka] [ka]

[0142] Step 1: To a solution of rac-(3S)-3-[4,6-difluoro-5-(4-hydroxy-4-piperidyl)-1-oxo-isoindolin-2-yl]piperidine-2,6-dione (150 mg, 0.4 mmol) in DCM (5 mL) was added tert-butyl N-methyl-N-(2-oxoethyl)carbamate (205.4 mg, 1.19 mmol) and NaBH(OAc) (251.4 mg, 1.19 mmol). The resulting solution was stirred at room temperature for 2 hours. The reaction was monitored by LCMS. The mixture was concentrated under reduced pressure. The product was purified by reverse-phase flash chromatography (column, C18 silica gel; mobile phase, ACN and water (0.05% TFA), 10% ACN to 70% ACN (20 min gradient); detector, UV 254 nm) to give tert-butyl N-[2-[4-[4,6-difluoro-1-oxo-2-[rac-(3S)-2,6-dioxo-3-piperidyl]isoindolin-5-yl]-4-hydroxy-1-piperidyl]ethyl]-N-methyl-carbamate (150 mg, 63.6% yield) as a light brown solid.

[0143] MS: m / z: C 26 H 34 Calculated value of F2N4O6 [M+H] + 537; Actual value 537.

[0144] Step 2: To a solution of tert-butyl N-[2-[4-[4,6-difluoro-1-oxo-2-[rac-(3S)-2,6-dioxo-3-piperidyl]isoindolin-5-yl]-4-hydroxy-1-piperidyl]ethyl]-N-methyl-carbamate (150 mg, 0.28 mmol) in DCM (4 mL) was added 4 M HCl (1 mL) in 1,4-dioxane. The resulting mixture was stirred at room temperature for 3 hours. The reaction was monitored by LCMS. The mixture was concentrated under reduced pressure to give rac-(3S)-3-[4,6-difluoro-5-[4-hydroxy-1-[2-(methylamino)ethyl]-4-piperidyl]-1-oxo-isoindolin-2-yl]piperidine-2,6-dione (130 mg crude product) as a brown solid.

[0145] MS: m / z: C 21 H 26 Calculated value of F2N4O4 [M+H] + 437; Actual value 437.

[0146] Step 3: To a solution of rac-(3S)-3-[4,6-difluoro-5-[4-hydroxy-1-[2-(methylamino)ethyl]-4-piperidyl]-1-oxo-isoindolin-2-yl]piperidine-2,6-dione (65 mg, 0.15 mmol) in DCM (2 mL) was added 2,4-dimethylthiazole-5-sulfonyl chloride (63.8 mg, 0.3 mmol) and TEA (58.4 mg, 0.45 mmol). The resulting solution was stirred at room temperature for 2 hours. The reaction was monitored by LCMS. The mixture was concentrated under reduced pressure. The product was purified by preparative HPLC to give N-[2-[4-[2-(2,6-dioxo-3-piperidyl)-4,6-difluoro-1-oxo-isoindolin-5-yl]-4-hydroxy-1-piperidyl]ethyl]-N,2,4-trimethyl-thiazole-5-sulfonamide (34.7 mg, 37.4% yield) as a light brown solid.

[0147] MS: m / z: C 26 H 31 F2N5O6S2[M+H] +Calculated value 610; measured value 610. 1 H NMR (300 MHz,DMSO-d6) δ 11.03 (s,1H),9.47 (s,1H),7.48 (d,J = 10.6 Hz,1H),6.05 (s,1H),5.13 (dd,J = 13.2,5.1 Hz,1H),4.55 (d,J = 17.4 Hz,1H),4.37 (d,J = 17.4 Hz,1H),3.62-3.32 (m,8H),3.04-2.78 (m,4H),2.69 (s,3H),2.68-2.63 (m,1H),2.62-2.53 (m,4H),2.47-2.45(m,1H),2.43-2.39 (m,1H),2.37-2.27 (m,2H),2.08-1.96 (m,1H).

[0148] Preparative HPLC conditions: Column: Welch Utimate AQ-C18, 50 * 250mm * 10 μm; Mobile phase A: water (0.05% TFA), Mobile phase B: ACN; Flow rate: 100 mL / min; Gradient: 20% to 32% B in 20 min, 32% B; Wavelength: 254 nm.

[0149] Example S102. N-[2-[4-[2-(2,6-dioxo-3-piperidyl)-4,6-difluoro-1-oxo-isoindolin-5-yl]-4-hydroxy-1-piperidyl]ethyl]-N-methyl-benzenesulfonamide (Compound 40) [ka]

[0150] Prepare the title compound as a brown solid (25.7% yield) according to the preparation of Example S101, using benzenesulfonyl chloride in step 3.

[0151] MS: m / z: C 27 H 30 F2N4O6S [M+H] + Calculated value 577; measured value 577. 1H NMR (300 MHz,DMSO-d6) δ 11.03 (s,1H),9.55 (s,1H),7.99-7.81 (m,2H),7.80-7.59 (m,3H),7.49 (d,J = 10.6 Hz,1H),6.05 (s,1H),5.13 (dd,J = 13.2,5.0 Hz,1H),4.55 (d,J = 17.4 Hz,1H),4.38 (d,J = 17.4 Hz,1H),3.62-3.49 (m,2H),3.4-3.15 (m,6H),3.10-2.84 (m,1H),2.74 (s,3H),2.67-2.53 (m,2H),2.47-2.39 (m,2H),2.39-2.26 (m,2H),2.15-1.86 (m,1H).

[0152] Preparative HPLC conditions: Column: Welch Utimate AQ-C18, 50 * 250mm * 10 μm; mobile phase A: water (0.05% TFA), mobile phase B: ACN; flow rate: 100 mL / min; gradient: 20% to 48% B in 20 min, 48% B; wavelength: 254 nm.

[0153] Example S105. 2-[2-(2,6-dioxo-3-piperidyl)-4,6-difluoro-1-oxo-isoindoline-5-carbonyl]-3,4-dihydro-1H-isoquinoline-7-carbonitrile (Compound 70) [ka] [ka]

[0154] Step 1: To a stirred solution of tert-butyl (4S)-5-amino-4-(5-bromo-4,6-difluoro-1-oxo-isoindolin-2-yl)-5-oxo-pentanoate (500 mg, 1.15 mmol) in 1,4-dioxane (5 mL) was added tributylstannylmethanol (741.1 mg, 2.31 mmol), XPhos Pd G3 (105.68 mg, 0.12 mmol), XPhos (55.1 mg, 0.12 mmol), and TEA (298.3 mg, 2.31 mmol). The resulting solution was degassed with nitrogen three times and stirred at 60 °C overnight. The reaction was monitored by LCMS. The mixture was concentrated under reduced pressure and applied to a silica gel column (petroleum ether / EtOAc (2 / 1)) to give tert-butyl (4S)-5-amino-4-[4,6-difluoro-5-(hydroxymethyl)-1-oxo-isoindolin-2-yl]-5-oxo-pentanoate (205 mg, 0.53 mmol, 46.2% yield) as a white solid.

[0155] MS: m / z: C 18 H 22 F2N2O5[M+H] + Calculated value 385; measured value 385.

[0156] Step 2: To a stirred solution of tert-butyl (4S)-5-amino-4-[4,6-difluoro-5-(hydroxymethyl)-1-oxo-isoindolin-2-yl]-5-oxo-pentanoate (180 mg, 0.47 mmol) in MeCN (2 mL) and CCl (2 mL) was added a solution of NaIO (251 mg, 1.4 mmol) and RuCl (23.9 mg, 0.09 mmol) in water (1 mL). The resulting solution was stirred at room temperature overnight. The reaction was monitored by LCMS. The mixture was concentrated under reduced pressure. The residue was diluted with DCM (50 mL), poured into ice-water (50 mL), and extracted with DCM (3 × 50 mL). The combined organic layers were dried over NaSO and concentrated under reduced pressure. The crude product was purified by reverse-phase flash chromatography under the following conditions: column, C18 silica gel; mobile phase, ACN and water (0.05% TFA), 10 min gradient: ACN 10% to 30%; detector, UV 254 nm) to give 2-[(1S)-4-tert-butoxy-1-carbamoyl-4-oxo-butyl]-4,6-difluoro-1-oxo-isoindoline-5-carboxylic acid (110 mg, 0.28 mmol, 58.9% yield) as a white solid.

[0157] MS: m / z: C 18 H 20 F2N2O6[M+H] + Calculated value 399; measured value 399.

[0158] Step 3: To a solution of 2-[(1S)-4-tert-butoxy-1-carbamoyl-4-oxo-butyl]-4,6-difluoro-1-oxo-isoindoline-5-carboxylic acid (50 mg, 0.13 mmol) in DMF (2 mL) was added DIEA (75 mg, 0.63 mmol) and HATU (71.6 mg, 0.19 mmol). To the above mixture was added 1,2,3,4-tetrahydroisoquinoline-7-carbonitrile (39.7 mg, 0.25 mmol). The resulting mixture was stirred at 25 °C overnight. The reaction was monitored by LCMS. After completion of the reaction, the final reaction solution was extracted with ethyl acetate (3 × 50 mL), washed with water (3 × 50 mL), and dried over anhydrous NaSO. After filtration, the organic layer was concentrated under reduced pressure. The crude product was applied to a silica gel column (petroleum ether / EtOAc (1 / 3)) to give tert-butyl (4S)-5-amino-4-[5-(7-cyano-3,4-dihydro-1H-isoquinoline-2-carbonyl)-4,6-difluoro-1-oxo-isoindolin-2-yl]-5-oxo-pentanoate (60 mg, 0.11 mmol, 88.7% yield) as a pale yellow solid.

[0159] MS: m / z: C 28 H 28 F2N4O5[M+H] + Calculated value 539; measured value 539.

[0160] Step 4: To a solution of tert-butyl (4S)-5-amino-4-[5-(7-cyano-3,4-dihydro-1H-isoquinoline-2-carbonyl)-4,6-difluoro-1-oxo-isoindolin-2-yl]-5-oxo-pentanoate (55 mg, 0.09 mmol) in MeCN (3 mL) was added benzenesulfonic acid (44.1 mg, 0.28 mmol). The resulting mixture was degassed with nitrogen three times and then stirred overnight at 60 °C under a nitrogen atmosphere. The reaction was monitored by LCMS. The mixture was concentrated under reduced pressure. The pH of the solution was adjusted to 9 using saturated sodium bicarbonate solution and extracted with EtOAc (3 × 50 mL). The combined organic layers were dried over Na SO and concentrated under reduced pressure. The crude product was purified by preparative HPLC to give 2-[2-(2,6-dioxo-3-piperidyl)-4,6-difluoro-1-oxo-isoindoline-5-carbonyl]-3,4-dihydro-1H-isoquinoline-7-carbonitrile (12.3 mg, 0.026 mmol, 28.4% yield) as a white solid.

[0161] MS: m / z: C 24 H 18 F2N4O4[M+H] + Calculated value 465; measured value 465. 1 H NMR (300 MHz,DMSO-d6) δ 11.05 (s,1H),7.90 - 7.56 (m,3H),7.42 (t,J = 8.0 Hz,1H),5.28 - 5.04 (m,1H),4.92 (s,1H),4.68 - 4.56 (m,2H),4.54 - 4.36 (m,1H),4.06 - 3.84 (m,1H),3.64 - 3.56 (m,1H),3.07 - 2.95 (m,1H),2.94 - 2.85 (m,2H),2.68 - 2.56 (m,1H),2.49 - 2.33 (m,1H),2.10 - 1.93 (m,1H).

[0162] Preparative HPLC conditions: Column: SunFire Prep C18 OBD Column, 19 *150 mm, 5 μm; Mobile phase A: water (0.05% TFA), Mobile phase B: ACN; Flow rate: 20 mL / min; Gradient: 30% to 50% B in 6 min, 50% B; Wavelength: 254 nm.

[0163] Example S106. 3-[5-(5-chloro-3,4-dihydro-1H-isoquinoline-2-carbonyl)-4,6-difluoro-1-oxo-isoindolin-2-yl]piperidine-2,6-dione (Compound 71) [ka]

[0164] Following the preparation of Example S105, in step 3, 5-chloro-1,2,3,4-tetrahydroisoquinoline was used to give the title compound as a white solid (39% final yield). MS: m / z: C 23 H 18 ClF2N3O4, [M+H] + Calculated value 474; measured value 474. 1 H NMR (400 MHz,DMSO-d6) δ 11.06 (d,J = 4.8 Hz,1H),7.71 - 7.61 (m,1H),7.45 - 7.04 (m,3H),5.23 - 5.09 (m,1H),5.00 - 4.83 (m,1H),4.68 - 4.53 (m,3H),4.09 - 3.90 (m,1H),3.65 - 3.59 (m,1H),2.91 - 2.85 (m,2H),2.78 (d,J = 4.9 Hz,1H),2.61 (d,J = 17.8 Hz,1H),2.46 - 2.43 (m,1H),2.06 - 1.97 (m,1H).

[0165] Preparative HPLC purification conditions: Column: Xselect CSH C18 OBD Column 30 * 150 mm 5 μm, n; Mobile phase A: water (0.05% TFA), Mobile phase B: ACN; Flow rate: 60 mL / min; Gradient: 27% to 57% B in 10 min, 57% B; Wavelength: 254 nm.

[0166] Example S107. 3-[5-(6,7-difluoro-3,4-dihydro-1H-isoquinoline-2-carbonyl)-4,6-difluoro-1-oxo-isoindolin-2-yl]piperidine-2,6-dione (Compound 72) [ka]

[0167] Following the preparation of Example S105, 6,7-difluoro-1,2,3,4-tetrahydroisoquinoline was used in step 3 to afford the title compound as a white solid (22.2% final yield).

[0168] MS: m / z: C 23 H 17 F4N3O4,[M+H] + Calculated value 476; measured value 476. 1 H NMR (300 MHz,DMSO-d6) δ 11.05 (d,J = 3.1 Hz,1H),7.66 - 7.63 (m,1H),7.52 - 7.14 (m,2H),5.24 - 5.07 (m,1H),4.84 (s,1H),4.68 - 4.55 (m,2H),4.49 - 4.43 (m,1H),4.04 - 3.79 (m,1H),3.66 - 3.49 (m,1H),3.05 - 2.82 (m,2H),2.80 - 2.69 (m,1H),2.69 - 2.50 (m,1H),2.50 - 2.35 (m,1H),2.09 - 1.90 (m,1H).

[0169] Preparative HPLC purification conditions: Column: Xselect CSH C18 OBD Column 30 * 150 mm 5 μm, n; mobile phase A: water (0.05% TFA), mobile phase B: ACN; flow rate: 60 mL / min; gradient: 23% to 53% B in 10 min, 53% B; wavelength: 254 nm.

[0170]

[0171] Example S108. 3-[4,6-Difluoro-5-[2-(methylamino)-5,7-dihydropyrrolo[3,4-b]pyridine-6-carbonyl]-1-oxo-isoindolin-2-yl]piperidine-2,6-dione (Compound 73) [ka] [ka]

[0172] Step 1: To a solution of 2-chloro-6,7-dihydro-5H-pyrrolo[3,4-b]pyridine hydrochloride (500 mg, 2.62 mmol) and TEA (1.37 mL, 7.85 mmol) in THF (25 mL) was added BocO (628.3 mg, 2.88 mmol) at 0 °C. The solution was stirred at room temperature for 2 h. The desired product was detected by LCMS. Upon completion, the mixture was concentrated in vacuo, diluted with water (50 mL), and extracted with DCM (3 × 80 mL). The combined organic layers were dried over NaSO and concentrated under reduced pressure. The residue was purified by silica gel chromatography (petroleum ether:ethyl acetate = 8:1) to give tert-butyl 2-chloro-5,7-dihydropyrrolo[3,4-b]pyridine-6-carboxylate (469 mg, 1.84 mmol, 70.36% yield) as a brown solid.

[0173] MS: m / z: C 12 H 15 ClN2O2[M+H] + Calculated value 255; measured value 255.

[0174] Step 2: To a solution of tert-butyl 2-chloro-5,7-dihydropyrrolo[3,4-b]pyridine-6-carboxylate (300 mg, 1.18 mmol) and 1-(4-methoxyphenyl)-N-methyl-methanamine (178.1 mg, 1.18 mmol) in 1,4-dioxane (20 mL) was added CsCO (1148.4 mg, 3.53 mmol), Ruphos (55 mg, 0.12 mmol), and Ruphos Pd G (109.8 mg, 0.12 mmol). The solution was degassed with nitrogen three times and stirred at 90 °C overnight. The desired product could be detected by LCMS. Upon completion, the mixture was concentrated in vacuo, diluted with water (50 mL), and extracted with DCM (3 × 50 mL). The combined organic layers were dried over NaSO and concentrated under reduced pressure. The residue was purified by silica gel chromatography (petroleum ether:ethyl acetate=10:1) to give tert-butyl 2-[(4-methoxyphenyl)methyl-methyl-amino]-5,7-dihydropyrrolo[3,4-b]pyridine-6-carboxylate (370 mg, 1 mmol, 85% yield) as a yellow solid.

[0175] MS: m / z: C 21 H 27 N3O3[M+H] + Calculated value 370; measured value 370.

[0176] Step 3: To a solution of tert-butyl 2-[(4-methoxyphenyl)methyl-methyl-amino]-5,7-dihydropyrrolo[3,4-b]pyridine-6-carboxylate (370 mg, 1 mmol) in DCM (20 mL) was added 4 M HCl / 1,4-dioxane. The solution was stirred at room temperature for 2 hours. The desired product was detected by LCMS. After completion, concentration under reduced pressure gave N-[(4-methoxyphenyl)methyl]-N-methyl-6,7-dihydro-5H-pyrrolo[3,4-b]pyridin-2-amine (230 mg crude product) as a yellow solid.

[0177] MS: m / z: C 16 H 19 NO [M+H] +Calculated value 270; measured value 270.

[0178] Step 4: To a solution of N-[(4-methoxyphenyl)methyl]-N-methyl-6,7-dihydro-5H-pyrrolo[3,4-b]pyridin-2-amine (81.1 mg, 0.3 mmol) and 2-[(1S)-4-tert-butoxy-1-carbamoyl-4-oxo-butyl]-4,6-difluoro-1-oxo-isoindoline-5-carboxylic acid (100 mg, 0.25 mmol) in DMF (5 mL) was added HATU (143.2 mg, 0.38 mmol) and DIEA (0.06 mL, 0.75 mmol). The solution was stirred at 30 °C under a nitrogen atmosphere for 3 hours. The desired product could be detected by LCMS. Upon completion, the mixture was concentrated in vacuo, diluted with water (50 mL), and extracted with DCM (3 × 80 mL). The combined organic layers were dried over NaSO and concentrated under reduced pressure. The residue was purified by silica gel chromatography (petroleum ether:ethyl acetate=8:1) to give tert-butyl (4S)-5-amino-4-[4,6-difluoro-5-[2-[(4-methoxyphenyl)methyl-methyl-amino]-5,7-dihydropyrrolo[3,4-b]pyridine-6-carbonyl]-1-oxo-isoindolin-2-yl]-5-oxo-pentanoate (106 mg, 0.15 mmol, 61.3% yield) as a white solid.

[0179] MS: m / z: C 34 H 37 F2N5O6[M+H] + Calculated value 650; measured value 650.

[0180] Step 5: A solution of tert-butyl (4S)-5-amino-4-[4,6-difluoro-5-[2-[(4-methoxyphenyl)methyl-methyl-amino]-5,7-dihydropyrrolo[3,4-b]pyridine-6-carbonyl]-1-oxo-isoindolin-2-yl]-5-oxo-pentanoate (100 mg, 0.15 mmol) and benzenesulfonic acid (72.9 mg, 0.46 mmol) in MeCN (8 mL) was stirred at 60 °C under a nitrogen atmosphere for 24 hours. The desired product was detected by LCMS. The mixture was concentrated in vacuo and diluted with water (50 mL). The resulting mixture was basified to pH 8 with aqueous NaCO solution and extracted with DCM (3 × 80 mL). The combined organic layers were dried over NaSO and concentrated under reduced pressure to give 3-[4,6-difluoro-5-[2-[(4-methoxyphenyl)methyl-methyl-amino]-5,7-dihydropyrrolo[3,4-b]pyridine-6-carbonyl]-1-oxo-isoindolin-2-yl]piperidine-2,6-dione (80 mg crude product) as a white solid.

[0181] MS: m / z: C 30 H 27 F2N5O5[M+H] + Calculated value of 576; Measured value 576.

[0182] Step 6: A solution of 3-[4,6-difluoro-5-[2-[(4-methoxyphenyl)methyl-methyl-amino]-5,7-dihydropyrrolo[3,4-b]pyridine-6-carbonyl]-1-oxo-isoindolin-2-yl]piperidine-2,6-dione (75 mg, 0.13 mmol) in TFA (4 mL) was stirred at 60 °C under a nitrogen atmosphere for 2 hours. The desired product was detected by LCMS. The resulting solution was concentrated under reduced pressure and purified by preparative HPLC to give 3-[4,6-difluoro-5-[2-(methylamino)-5,7-dihydropyrrolo[3,4-b]pyridine-6-carbonyl]-1-oxo-isoindolin-2-yl]piperidine-2,6-dione (32.4 mg, 54.4% yield) as a white solid.

[0183] MS: m / z: C 24 H 18 F2N4O4[M+H] + Calculated value 456; measured value 456. 1 H NMR (400 MHz,DMSO-d6) δ 11.05 (s,1H),7.73 - 7.65 (m,1H),7.58 - 7.37 (m,1H),6.65 - 6.50 (m,1H),5.24 - 5.09 (m,1H),4.76 (d,J = 9.3 Hz,2H),4.70 - 4.41 (m,4H),2.99 - 2.90 (m,1H),2.85 (s,2H),2.75 (d,J = 3.0 Hz,1H),2.70 - 2.57 (m,1H),2.50 - 2.44 (m,1H),2.10 - 1.93 (m,1H).

[0184] Preparative HPLC conditions: Column: SunFire Prep C18 OBD Column, 19 * 150 mm, 5 μm; Mobile phase A: water (0.05% TFA), Mobile phase B: ACN; Flow rate: 20 mL / min; Gradient: Bga 12% to 15% in 6 min, B 15%; Wavelength: 254 / 210 nm.

[0185] Example S116. N-[[2-(2,6-dioxo-3-piperidyl)-4,6-difluoro-1-oxo-isoindolin-5-yl]methyl]indolizine-2-carboxamide (Compound 69) [ka]

[0186] According to general scheme 2, in step 4, indolizine-2-carboxylic acid, TCFH and NMI were used to give the title compound as a white solid (yield 21.6%).

[0187] MS: m / z: C 23 H 18 F2N4O4,[M+H] - Calculated value 453; measured value 453. 1H NMR (400 MHz,DMSO-d6) δ 11.02 (s,1H),8.66 (t,J = 5.3 Hz,1H),8.24 (dd,J = 7.0,1.3 Hz,1H),7.95 (d,J = 1.6 Hz,1H),7.48 (d,J = 7.8 Hz,1H),7.41 (d,J = 9.1 Hz,1H),6.78 (s,1H),6.75 - 6.66 (m,1H),6.66 - 6.51 (m,1H),5.13 (dd,J = 13.3,5.1 Hz,1H),4.75 - 4.46 (m,3H),4.39 (d,J = 17.3 Hz,1H),3.01 - 2.82 (m,1H),2.67 - 2.57 (m,1H),2.49 - 2.36 (m,1H),2.09 - 1.95 (m,1H).

[0188] Preparative HPLC purification conditions: Column: SunFire Prep C18 OBD Column, 19 * 150 mm, 5 μm; Mobile phase A: water (0.05% TFA), Mobile phase B: ACN; Flow rate: 20 mL / min; Gradient: 32% B to 32% B in 6 min; Wavelength: 254 nm.

[0189] Example S119. 3-(6-Fluoro-1-oxo-5-(1-(4-(trifluoromethyl)benzyl)piperidin-4-yl)isoindolin-2-yl)piperidine-2,6-dione (Compound 85) [ka]

[0190] Step 1: A 200 mL round-bottom flask was charged with tert-butyl (S)-4-(2-(1-amino-5-(tert-butoxy)-1,5-dioxopentan-2-yl)-6-fluoro-1-oxoisoindolin-5-yl)-3,6-dihydropyridine-1(2H)-carboxylate (1.000 g, 1.932 mmol), MeOH (50 mL), and Pd / C (0.822 g, 0.386 mmol). The air was replaced with hydrogen and the mixture was stirred vigorously under a hydrogen atmosphere overnight. It was filtered through a pad of Celite and concentrated to give 1.0 g (100% yield) of the desired product.

[0191] LC / MS (ESI) m / z 520.5 [(M+H) + , C 27 H 38 Calculated value for FN3O6 519.3].

[0192] Step 2: A 100 mL round-bottom flask was charged with tert-butyl (S)-4-(2-(1-amino-5-(tert-butoxy)-1,5-dioxopentan-2-yl)-6-fluoro-1-oxoisoindolin-5-yl)piperidine-1-carboxylate (1.00 g, 1.925 mmol), benzenesulfonic acid (0.609 g, 3.85 mmol), and MeCN (35 mL). It was heated to 80° C. for 4 h. It was concentrated to dryness, and the residue was rinsed with ether to remove excess PhSO3H and dried to give the mono-benzenesulfonate salt of the desired product (668 mg).

[0193] Step 3: To a solution of 4-(trifluoromethyl)benzaldehyde (20.17 mg, 0.116 mmol) and (S)-3-(6-fluoro-1-oxo-5-(piperidin-4-yl)isoindolin-2-yl)piperidine-2,6-dione (20 mg, 0.058 mmol) in DMF (1 mL), two drops of AcOH were added and then stirred at room temperature. After 0.5 hours, NaBH(OAc) (36.8 mg, 0.174 mmol) was added, and the resulting solution was stirred at room temperature for 2 hours. It was purified by preparative HPLC method 1 to give 6.0 mg of the title compound.

[0194] LC / MS (ESI) m / z 504.1[(M+H)+, C 26 H 25 Calculated F4N3O3 503.2]; HPLC a retention time = 1.29 minutes; 1 H NMR (500 MHz,DMSO-d6) δ 10.99 (s,1H),7.74 - 7.67 (m,J=8.2 Hz,2H),7.63 (d,J=6.0 Hz,1H),7.61 - 7.55 (m,J=8.2 Hz,2H),7.47 (d,J=9.2 Hz,1H),5.10 (dd,J=13.3,5.0 Hz,1H),4.46 - 4.38 (m,1H),4.34 - 4.26 (m,1H),3.66 - 3.58 (m,1H),2.97 - 2.87 (m,3H),2.64 - 2.57 (m,1H),2.57 - 2.56 (m,1H),2.52 - 2.50 (m,7H),2.46 - 2.33 (m,1H),2.22 - 2.09 (m,2H),2.04 - 1.96 (m,1H)

[0195] Examples of compounds prepared according to the procedures shown for Scheme 9 and Scheme 9a using the appropriate aldehyde are listed in Table 4.

[0196] [Table 33]

[0197] Example S131. 3-(5-(1-(benzo[d]thiazol-6-ylmethyl)-4-hydroxypiperidin-4-yl)-6-fluoro-1-oxoisoindolin-2-yl)piperidine-2,6-dione (compound 35) [ka]

[0198] A 1-dram vial was charged with 3-(6-fluoro-5-(4-hydroxypiperidin-4-yl)-1-oxoisoindolin-2-yl)piperidine-2,6-dione, benzenesulfonate (20 mg, 0.038 mmol) dissolved in 1 mL of DMF. To this was added 6-(chloromethyl)benzo[d]thiazole (10.60 mg, 0.058 mmol), followed by Hunig's base (0.034 mL, 0.192 mmol). The resulting mixture was heated at 80° C. for 1 hour. It was cooled to room temperature and purified by preparative HPLC, Method 1, to give 6.0 mg of the title compound.

[0199] LC / MS (ESI) m / z 509.2 [(M+H) + , C 26 H 25 Calculated value for FN4O4S: 508.2]; HPLC a retention time = 0.97 min; 1 H NMR (500 MHz,DMSO-d6) δ 11.00 (br s,1H),9.49 (s,1H),8.36 (br s,1H),8.20 (br d,J=8.2 Hz,1H),7.88 (br d,J=6.1 Hz,1H),7.74 (br d,J=8.5 Hz,1H),7.54 - 7.45 (m,1H),5.08 (br dd,J=12.4,4.4 Hz,1H),4.63 - 4.39 (m,3H),4.37 - 4.27 (m,1H),3.66 - 3.49 (m,2H),2.95 - 2.83 (m,1H),2.62 (br d,J=16.5 Hz,1H),2.51 (br s,6H),2.47 - 2.32 (m,2H),2.06 - 1.99 (m,1H),1.90 - 1.77 (m,2H).

[0200] Example S132. 3-(5-(1-(benzo[d]thiazol-6-ylmethyl)piperidin-4-yl)-6-fluoro-1-oxoisoindolin-2-yl)piperidine-2,6-dione (Compound 81) [ka]

[0201] Following the alkylation procedure given for the synthesis of Example S131, using 3-(6-fluoro-1-oxo-5-(piperidin-4-yl)isoindolin-2-yl)piperidine-2,6-dione as the coupling partner, the title compound was obtained as an off-white solid (51% yield).

[0202] LC / MS (ESI) m / z 493.1 [(M+H) + , C 26 H 25 Calculated value for FN4O3S 492.2]; HPLC a retention time = 1.07 min; 1 H NMR (500 MHz,DMSO-d6) δ 10.95 (s,1H),9.46 (s,1H),8.30 (br s,1H),8.16 (d,J=8.4 Hz,1H),7.67 (br d,J=8.4 Hz,1H),7.47 (br d,J=9.3 Hz,2H),5.06 (br dd,J=13.0,4.8 Hz,1H),4.51 - 4.42 (m,2H),4.39 (br d,J=17.2 Hz,1H),4.31 - 4.23 (m,1H),3.69 - 3.45 (m,2H),3.21 - 3.04 (m,2H),2.91 - 2.81 (m,1H),2.59 - 2.52 (m,1H),2.46 (br s,4H),2.39 - 2.28 (m,2H),2.09 - 1.99 (m,1H),1.90 - 1.77 (m,2H).

[0203] Listed in Table 5 are example compounds that were prepared using the appropriate alkyl halides following the alkylation procedures shown for the synthesis of Example S131 and Example S132.

[0204] [Table 34]

[0205] Example S140. 3-(5-(6-amino-4,5-dimethylpyridin-2-yl)-6-fluoro-1-oxoisoindolin-2-yl)piperidine-2,6-dione (compound 34) [ka] [ka]

[0206] Step 1: To a solution of 4,4,4',4',5,5,5',5'-octamethyl-2,2'-bi(1,3,2-dioxaborolane) (275 mg, 1.084 mmol), potassium acetate (213 mg, 2.167 mmol), and tert-butyl (S)-5-amino-4-(5-bromo-6-fluoro-1-oxoisoindolin-2-yl)-5-oxopentanoate (300 mg, 0.722 mmol) in dioxane (10 mL), PdCl(dppf).DCM (59.0 mg, 0.072 mmol) was added and the atmosphere was replaced with N. It was heated to 100 °C for 16 h. It was cooled to room temperature, diluted with EtOAc, quenched with brine, and the organic layer was separated, dried over NaSO, and concentrated. The crude product was purified by flash column chromatography (eluting with 0-6% MeOH / DCM) to give 302 mg (90% yield) of the desired product.

[0207] LC / MS (ESI) m / z 325.1 [(M-137) + , C 23 H 32 Calculated value for BFN2O6: 462.2].

[0208] Step 2: A 5 mL microwave vial was charged with 6-chloro-3,4-dimethylpyridin-2-amine (16.5 mg, 0.105 mmol), tert-butyl (S)-5-amino-4-(6-fluoro-1-oxo-5-(4,4,5,5-tetramethyl-1,3,2-dioxaborolan-2-yl)isoindolin-2-yl)-5-oxopentanoate (60.9 mg, 0.132 mmol), PdCl(dtbpf) (3.43 mg, 5.27 μmol), and aqueous KPO (0.176 mL, 0.527 mmol) dissolved in dioxane (3 mL). It was sealed, the air was replaced with nitrogen, and then heated in a microwave at 120 °C for 0.25 h. It was diluted with EtOAc and washed with brine, the organic layer was separated, dried over MgSO4 and concentrated to give 32 mg of the desired product which was used in the next step without further purification.

[0209] LC / MS (ESI) m / z 457.3 [(M+H)+, C 24 H 29 Calculated value for FN4O4 456.2].

[0210] Step 3: tert-Butyl (S)-5-amino-4-(5-(6-amino-4,5-dimethylpyridin-2-yl)-6-fluoro-1-oxoisoindolin-2-yl)-5-oxopentanoate (27 mg, 0.059 mmol) was added to a solution of PhSOH in MeCN (0.25 M, 1 mL) and heated in the microwave at 120 °C for 15 min. It was purified by preparative HPLC method 1 to give 11.9 mg (28% yield) of the desired product.

[0211] LC / MS (ESI) m / z 383.1 [(M+H)+, C 20 H 19 Calculated value for FN4O3: 382.1; HPLC a Hold time = 1.00 min; 1H NMR (500 MHz,DMSO-d6) δ 11.00 (s,1H),8.03 (d,J=6.6 Hz,1H),7.57 (d,J=9.9 Hz,1H),6.93 - 6.88 (m,1H),5.70 (s,1H),5.10 (dd,J=13.4,5.1 Hz,1H),4.50 (d,J=17.2 Hz,1H),4.37 (d,J=17.0 Hz,1H),3.62 - 3.53 (m,1H),2.95 - 2.84 (m,1H),2.68 - 2.58 (m,1H),2.47 - 2.32 (m,1H),2.22 (s,3H),2.08 - 2.04 (m,1H),2.03 (s,3H),1.22 (br s,1H)

[0212] Additional compounds may be prepared following the procedures shown for Scheme 3 and Scheme 4 using the appropriate boronic acid / ester.

[0213] Example S146. 3-(5-(6-amino-4,5-dimethylpyridin-2-yl)-4,6-difluoro-1-oxoisoindolin-2-yl)piperidine-2,6-dione (Compound 93) [ka]

[0214] Step 1: A solution of 6-chloro-3,4-dimethylpyridin-2-amine (30.0 mg, 0.192 mmol), 1,1,1,2,2,2-hexamethyldistannane (69.0 mg, 0.211 mmol), and PdCl(dtbpf) (12.48 mg, 0.019 mmol) in toluene (5 mL) was prepared and the air was replaced with N. It was heated to 110 °C for 4 h. After cooling to room temperature, it was diluted with EtOAc and brine, and the organic layer was separated, dried over NaSO, filtered, and concentrated. The crude product (38 mg) was used in the next step without further purification.

[0215] LC / MS (ESI) m / z 287.1 [(M+H) + , C10 H 18 Calculated value of N2Sn 286.0].

[0216] Step 2: A 2-dram pressure vial was charged with tert-butyl (S)-5-amino-4-(5-bromo-4,6-difluoro-1-oxoisoindolin-2-yl)-5-oxopentanoate (38.0 mg, 0.088 mmol), 3,4-dimethyl-6-(trimethylstannyl)pyridin-2-amine (25 mg, 0.088 mmol), Pd(PPh3)4 (10.14 mg, 8.77 μmol), and toluene (6 mL). The mixture was heated at 100 °C under a nitrogen atmosphere for 16 h. After cooling to room temperature, it was diluted with EtOAc and brine, and the organic layer was separated, dried over MgSO4, filtered, and concentrated. The crude product (28 mg) was used in the next step without further purification.

[0217] LC / MS (ESI) m / z 475.1 [(M+H)+,C 24 H 28 Calculated value of F2N4O4 474.2].

[0218] Step 3: To tert-butyl (S)-5-amino-4-(5-(6-amino-4,5-dimethylpyridin-2-yl)-4,6-difluoro-1-oxoisoindolin-2-yl)-5-oxopentanoate (28 mg, 0.059 mmol), a solution of PhSOH in MeCN (1 mL, 0.25 M) was added and heated in the microwave at 120 °C for 15 min. It was purified by preparative HPLC method 1 to give 6.8 mg (28% yield) of the desired product.

[0219] LC / MS (ESI) m / z 401.0 [(M+H) + ,C 20 H 18 Calculated F2N4O3 400.1]; HPLC a retention time = 1.04 min; 1H NMR (500 MHz,DMSO-d6) δ 11.03 (s,1H),7.53 (d,J=7.5 Hz,1H),6.58 (s,1H),5.80 (s,2H),5.15 (dd,J=13.4,5.0 Hz,1H),4.60 (d,J=17.2 Hz,1H),4.43 (d,J=17.3 Hz,1H),2.97 - 2.89 (m,1H),2.62 (br d,J=18.1 Hz,1H),2.49 - 2.41 (m,1H),2.22 (s,3H),2.06 (br s,1H),2.04 (s,3H)

[0220] Example S147. 3-(4,6-Difluoro-1-oxo-5-(5,6,7,8-tetrahydro-1,8-naphthyridin-2-yl)isoindolin-2-yl)piperidine-2,6-dione (Compound 94) [ka]

[0221] The title compound was synthesized from 7-chloro-1,2,3,4-tetrahydro-1,8-naphthyridine according to the route shown for the synthesis of Example S146.

[0222] LC / MS (ESI) m / z 413.1 [(M+H) + ,C 21 H 18 Calculated F2N4O3 412.1]; HPLC a retention time = 1.05 min; 1H NMR (500 MHz,DMSO-d6) δ 11.02 (s,1H),7.53 (d,J=7.6 Hz,1H),7.29 (d,J=7.2 Hz,1H),6.65 (br s,1H),6.58 (d,J=7.2 Hz,1H),5.14 (dd,J=13.2,4.9 Hz,1H),4.60 (d,J=17.2 Hz,1H),4.43 (d,J=17.2 Hz,1H),2.97 - 2.89 (m,1H),2.73 (br t,J=6.1 Hz,2H),2.62 (br dd,J=15.6,2.2 Hz,1H),2.56 - 2.53 (m,2H),2.49 - 2.41 (m,1H),2.08 - 2.01 (m,1H),1.86 - 1.79 (m,2H)

[0223] Example S148 includes compounds made according to general scheme 1. [Table 35] [Table 36] Example S149 includes the following compounds that were made according to certain general procedures: [Table 37] [Table 38] [Table 39] [Table 40] [Table 41] [Table 42] [Table 43] [Table 44]

[0224] Biological Examples Example B1. Evaluation of small molecule-induced degradation in Jurkat cells expressing IKZF1 ZNF2_ZNF3 Q1F Compounds targeting the Q1F degron were screened in Jurkat cells engineered to express IKZF1 ZNF2_ZNF3 Q1F. The cell line was generated using a lentiviral vector containing a CD19 CAR tagged with IKZF1 ZNF2_ZNF3 Q1F Nluc and transduced into Jurkat cells. The CD19 CAR contained an anti-CD19 scFv, a CD28 transmembrane domain, a 4-1BB costimulatory domain, a CD3ζ signaling domain, and the ZNF2_ZNF3 Q1F degron (GERPF). F CNQC GASFTQKGNL LRHIKLHSGE KPFKCHLCNY ACRRRDALTG HLRTHS; SEQ ID NO: 1; Q1F substitutions are underlined). Transfected cells were titrated with each small molecule or no drug and then incubated at 37°C for 18 hours. Cells were washed and stained with the appropriate stain to measure CAR levels. Cells were incubated in the stain at 4°C for 20 minutes, then washed three times and read on a flow cytometer. CAR levels were normalized to cells without drug treatment. The resulting titration curves were used to calculate EC 50 and Y min In this way, small molecules that potentially degrade Q1F degron-tagged CAR were identified (Table 1).

[0225] Example B2. Evaluation of selectivity of small molecules on Aiolos, Ikaros, CK1α, GSPT1, and Helios DF15 multiple myeloma cells stably expressing ePL-tagged Aiolos, Ikaros, or GSPT1, and MDS-L cells stably expressing ePL-tagged CK1a were generated via lentiviral infection with pLOC-ePL-Aiolos (or Ikaros, GSPT1, or CK1a). The sequences of human Aiolos and Ikaros are shown below:

[0226] Human Aiolos: [ka] (SEQ ID NO: 2; ZNF2 and ZNF3 are underlined).

[0227] Human Ikaros: [ka] (SEQ ID NO: 3; ZNF2 and ZNF3 are underlined).

[0228] DF15 multiple myeloma cells (DiscoverX) expressing Ikaros, Aiolos, and ePL-tagged GSPT1 and MDS-L cells expressing ePL-tagged CK1a were dispensed into a 384-well plate (Corning no. 3570) pre-spotted with compounds. Compounds were dispensed into the 384-well plate using an acoustic dispenser (EDC Biosystems ATS acoustic transfer system) in a 10-point dose-response curve, with 3-fold dilutions starting at 10 μM and extending to 0.0005 μM. Then, 25 μL of medium (RPMI-1640 + 10% heat-inactivated FBS + 25 mM Hepes + 1 mM sodium pyruvate + 1× NEAA + 1× penicillin, streptomycin, glutamine) containing 5,000 DF15 or MSD-L cells was dispensed into each well. Assay plates were incubated at 37°C in 5% CO for 4 hours, and GSPT1 for 20 hours. After incubation, 25 μL of InCell Hunter detection reagent working solution (DiscoverX, Catalog No. 96-0002, Fremont, CA) was added to each well and incubated at room temperature for 60 minutes, protected from light. Luminescence was read after 60 minutes on an Envision or PHERAstar luminescence reader.

[0229] For Helios, a stable Jurkat cell line was modified using CRISPR / Cas9 to insert a HiBit tag into the carboxy-terminal reading frame of the IKZF2 gene. The sequence of human Helios is shown below: [ka] (SEQ ID NO: 4; ZNF2 and ZNF3 are underlined).

[0230] Test compounds were transferred to 1536-well plates using an acoustic dispenser, and Jurkat / Helios / HiBit cells in DMEM / 10% FCS were plated at 10,000 cells per well in a final volume of 5 μL. Cells were incubated at 37°C and 95% RH for 18 hours. 2 μL of Nano-Glo reagent (Promega) was added per well, incubated at room temperature for 30 minutes, and luciferase activity was measured by reading luminescence on a microtiter plate reader.

[0231] To determine the EC50 value (the compound concentration that achieves half of the observed maximal degradation) of a compound to degrade a given substrate, a four-parameter logistic model (sigmoidal dose-response model) (FIT = (A + {(B A) / 1 + [(C / x)D]})) was used, where C is the inflection point (EC 50 ), where D is the correlation coefficient, and A and B are the lower and upper limits of the fit. All substrate degradation curves were processed and evaluated using the data analysis software package ActivityBase (IDBS). Ymax is the % protein degraded (Ymin = 100 - Ymax, where Ymin is the minimum percentage of protein remaining).

[0232] Some results are shown in Table 8. [Table 45] [Table 46] [Table 47] [Table 48] [Table 49] [Table 50] * nd = not determined.

[0233] Although the present invention has been described in some detail by way of illustration and example, for purposes of clarity of understanding, the descriptions and examples should not be construed as limiting the scope of the invention. The disclosures of all patent and scientific literature cited herein are expressly incorporated herein by reference in their entireties.

Claims

1. Compounds of formula (I): 【Chemistry 1】 or a pharmaceutically acceptable salt thereof [In the formula, R １ is H or oxo; R ２ are each independently H or halo, where at least one R ２ is fluoro; X is a bond, C １ ~C ３ Alkylene, —C(O)NHCH ２ -, -NHC(O)-, -C(O)-, or -(C １ ~C ３ alkylene)NH(C １ ~C ３ alkylene)-; Ring A is an optionally substituted C ５ ~C ６ cycloalkyl, optionally substituted C ５ ~C ６ aryl, optionally substituted 6- to 10-membered heterocyclyl, or optionally substituted 5- to 9-membered heteroaryl, wherein the heterocyclyl or heteroaryl contains 1 to 3 heteroatoms selected from nitrogen, oxygen, and sulfur.

2. The compound of formula (I) may be a compound of formula (Ia): 【Chemistry 2】 2. The compound of claim 1, wherein:

3. The compound of formula (I) may be a compound of formula (Ib): 【Transformation 3】 2. The compound of claim 1, wherein:

4. R １ 2. The compound of claim 1, wherein is H, or a pharmaceutically acceptable salt thereof.

5. R １ 2. The compound of claim 1, or a pharmaceutically acceptable salt thereof, wherein is oxo.

6. At least one R ２ 2. The compound of claim 1, wherein is H, or a pharmaceutically acceptable salt thereof.

7. One or two R ２ 2. The compound of claim 1, or a pharmaceutically acceptable salt thereof, wherein is F, Cl, or Br.

8. One or two R ２ 8. The compound of claim 7, wherein is F, or a pharmaceutically acceptable salt thereof.

9. 9. The compound of any one of claims 1 to 8, or a pharmaceutically acceptable salt thereof, wherein X is a bond.

10. X is -CH ２ The compound according to any one of claims 1 to 8, or a pharmaceutically acceptable salt thereof, wherein

11. X is -C(O)NHCH ２ The compound according to any one of claims 1 to 8, or a pharmaceutically acceptable salt thereof, wherein

12. The compound according to any one of claims 1 to 8, or a pharmaceutically acceptable salt thereof, wherein X is -NHC(O)-.

13. The compound according to any one of claims 1 to 8, or a pharmaceutically acceptable salt thereof, wherein X is -C(O)-.

14. X is -CH ２ NHCH ２ The compound according to any one of claims 1 to 8, or a pharmaceutically acceptable salt thereof, wherein

15. The compound according to any one of claims 1 to 14, or a pharmaceutically acceptable salt thereof, wherein ring A is optionally substituted cyclohexyl.

16. Ring A has the structure: 【Chemistry 4】 16. The compound of claim 15, wherein:

17. Ring A is optionally substituted C ６ 15. The compound of any one of claims 1 to 14, or a pharmaceutically acceptable salt thereof, which is aryl.

18. Ring A has the structure: 【Transformation 5】 18. The compound of claim 17, wherein:

19. The compound according to any one of claims 1 to 14, or a pharmaceutically acceptable salt thereof, wherein ring A is an optionally substituted 6- to 10-membered heterocyclyl containing 1 to 3 heteroatoms selected from nitrogen, oxygen, and sulfur.

20. Ring A has the structure: 【Transformation 6】 20. The compound of claim 19, wherein: [In the formula, R ３ is H or OH; R ４ is an optionally substituted C １ ~C ３ alkyl, optionally substituted C ３ ~C ６ cycloalkyl, —C(O)(6-membered heteroaryl), or —C(O)(9-membered heterocyclyl); R ５ is an optionally substituted C １ ~C ３ alkyl; R ６ is cyano or halo].

21. The compound according to any one of claims 1 to 14 and 20, wherein ring A has the following structure, or a pharmaceutically acceptable salt thereof: 【Transformation 7】 【Transformation 8】 【Chemistry 9】 【Chemistry 10】

22. structure: 【Chemistry 11】 But the structure: 【Chemistry 12】 22. The compound of claim 21, wherein:

23. structure: 【Chemistry 13】 But the structure: 【Chemistry 14】 22. The compound of claim 21, wherein:

24. 15. The compound according to any one of claims 1 to 14, or a pharmaceutically acceptable salt thereof, wherein Ring A is an optionally substituted 5- to 9-membered heteroaryl containing 1 to 3 heteroatoms selected from nitrogen, oxygen, and sulfur.

25. Ring A has the structure: 【Chemistry 15】 where R ７ optionally substituted C １ ~C ３ 25. The compound of claim 24, wherein n is alkyl or an optionally substituted amine and n is 0, 1, 2, 3, or 4, or a pharmaceutically acceptable salt thereof.

26. structure: 【Chemistry 16】 The structure: 【Chemistry 17】 26. The compound of claim 25, wherein:

27. The compound has the formula (IIa): [Chemistry 18] 27. The compound according to any one of claims 1 to 3 and 5 to 26, wherein:

28. The compound has the formula (IIb): 【Chemistry 19】 27. The compound according to any one of claims 1 to 4 and 6 to 26, or a pharmaceutically acceptable salt thereof, wherein:

29. The compound has the formula (IIc): 【Chemistry 20】 27. The compound according to any one of claims 1 to 3 and 5 to 26, wherein:

30. The compound has the formula (IId): 【Chemistry 21】 27. The compound according to any one of claims 1 to 3 and 5 to 26, wherein:

31. The compound has the formula (IIIa): 【Chemistry 22】 22. The compound according to any one of claims 1 to 3, 5 to 14, and 19 to 21, or a pharmaceutically acceptable salt thereof, [In the formula, R ８ is an optionally substituted C １ ~C ６ Alkyl or optionally substituted C ３ ~C ６ is cycloalkyl.

32. The compound has the formula (IIIb): 【Chemistry 23】 22. The compound according to any one of claims 1 to 3, 5 to 14, and 19 to 21, or a pharmaceutically acceptable salt thereof, [In the formula, R ８ is an optionally substituted C １ ~C ６ Alkyl or optionally substituted C ３ ~C ６ is cycloalkyl.

33. The compound has the formula (IIIc): 【Chemistry 24】 22. The compound according to any one of claims 1 to 3, 5 to 14, and 19 to 21, or a pharmaceutically acceptable salt thereof, [In the formula, R ８ is an optionally substituted C １ ~C ６ Alkyl or optionally substituted C ３ ~C ６ is cycloalkyl.

34. The compound has the formula (IIId): 【Chemistry 25】 22. The compound according to any one of claims 1 to 3, 5 to 14, and 19 to 21, or a pharmaceutically acceptable salt thereof, [In the formula, R ８ is an optionally substituted C １ ~C ６ Alkyl or optionally substituted C ３ ~C ６ is cycloalkyl.

35. A compound selected from the compounds set forth in Table 1, Table 2, Table 3, Table 4, Table 5, Table 6, and Table 7, or a pharmaceutically acceptable salt thereof.

36. A pharmaceutical composition comprising a compound according to any one of claims 1 to 35, or a pharmaceutically acceptable salt thereof, and a pharmaceutically acceptable excipient.

37. 37. A method of reducing the level of an altered polypeptide in a cell, comprising contacting the cell with an effective amount of a compound of any one of claims 1 to 35, or a pharmaceutically acceptable salt thereof, or an effective amount of a pharmaceutical composition of claim 36.

38. 38. The method of claim 37, wherein the cell is a human effector cell.

39. 39. The method of claim 37 or 38, wherein the cell is a T cell or an NK cell.

40. 40. The method of claim 39, wherein the cell is a CAR-T cell.

41. The method of any one of claims 37 to 40, wherein the modified polypeptide comprises a degradation domain.

42. The method of any one of claims 37 to 41, further comprising administering the cells to a subject.

43. 37. A method of reducing the level of an intracellular modified polypeptide in a subject, comprising administering to the subject an effective amount of a compound according to any one of claims 1 to 35, or a pharmaceutically acceptable salt thereof, or an effective amount of a pharmaceutical composition according to claim 36.

44. 44. The method of claim 43, wherein the cell is a human effector cell.

45. 45. The method of claim 43 or claim 44, wherein the cell is a T cell or an NK cell.

46. 46. ​​The method of any one of claims 43 to 45, wherein the modified polypeptide comprises a degradation domain.

47. The method of any one of claims 42 to 46, wherein the subject has cancer.

48. 48. The method of claim 47, wherein the cancer is selected from a hematological cancer or a solid cancer.

49. 49. The method of claim 48, wherein the hematological cancer is acute myeloid leukemia (AML), acute lymphoblastic leukemia (ALL), chronic myelogenous leukemia (CML), lymphoma, non-Hodgkin's lymphoma (NHL), Hodgkin's disease, multiple myeloma, or myelodysplastic syndrome.

50. 49. The method of claim 48, wherein the solid cancer is brain cancer, prostate cancer, breast cancer, lung cancer, colon cancer, uterine cancer, skin cancer, liver cancer, bone cancer, pancreatic cancer, ovarian cancer, testicular cancer, bladder cancer, kidney cancer, head and neck cancer, stomach cancer, cervical cancer, rectal cancer, laryngeal cancer, and esophageal cancer.

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