Compounds and compositions for selective degradation of engineered proteins

Abstract

Provided herein are compounds and compositions thereof for degrading engineered peptides in cells. In some embodiments, the compounds and compositions provided are useful for the treatment of cancer.

Description

CROSS-REFERENCE TO RELATED APPLICATIONS

[0001] This application claims the benefit of priority to U.S. Provisional Application No. 63 / 444,208, filed February 8, 2023, the entirety of which is incorporated herein by reference for any purpose. Sequence Listing

[0002] This application contains a sequence listing, which has been submitted electronically in XML format. The XML copy was created on January 23, 2024, is named "01277-0037-00PCT.xml" and is 6,097 bytes in size. The entire text of the information in the electronic format of this sequence listing is incorporated herein by reference. Technical Field

[0003] The present disclosure relates to compounds and compositions thereof for degrading engineered proteins in cells. Background Art

[0004] Engineered cells containing engineered heterologous polypeptides, such as chimeric antigen receptor T (CAR-T) cells, have been developed for therapeutic use. Modulation of the expression level of these engineered heterologous polypeptides can improve the therapeutic benefits of engineered cells by, for example, reducing side effects and / or increasing the efficacy of the engineered cells.

[0005] Thus, in one aspect, provided herein are engineered polypeptides and degraders, wherein the engineered polypeptide comprises a degradation domain that, when bound to the degrader, mediates ubiquitination in a cell. Summary of the Invention

[0006] In certain embodiments, described herein are compounds and compositions thereof for modulating the levels of a heterologous polypeptide in a cell. In various embodiments, the compounds and compositions thereof can be used to reduce the levels of a heterologous polypeptide in a cell.

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

[0008] In some embodiments, provided herein are compounds of Formula (I): or a pharmaceutically acceptable salt thereof, wherein: R 1 is H or oxo; Each R 2 are independently H or halogen, wherein at least one R 2 For fluorine; 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 optionally substituted C5-C6 cycloalkyl, optionally substituted C5-C6 aryl, optionally substituted 6-10 membered heterocyclyl, or optionally substituted 5-9 membered heteroaryl, wherein the heterocyclyl or heteroaryl contains 1-3 heteroatoms selected from nitrogen, oxygen and sulfur. DETAILED DESCRIPTION definition

[0009] As used herein, the terms "comprising" and "including" can be used interchangeably. The terms "comprising" and "including" should be interpreted as specifying the presence of the features or components, but not excluding the presence or addition of one or more features, components, or groups thereof. Additionally, the terms "comprising" and "including" are intended to encompass examples encompassed by the term "consisting of." Therefore, the term "consisting of" can 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 features or components it consists of. In another embodiment, the term "consisting of excludes any other features or components from the scope of any subsequent recitation, unless those features or components 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 any 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." An exception to this definition occurs only when a combination of elements, functions, steps, or actions are inherently mutually exclusive in some way.

[0012] In this specification, unless otherwise indicated, any concentration range, percentage range, ratio range, or integer range should be understood to include any integer within the range, as well as fractions thereof (such as tenths and hundredths of integers) where appropriate. In addition, unless otherwise indicated, any numerical range described herein relating to any physical characteristic, such as polymer subunits, size, or thickness should be understood to include any integer within the range. As used herein, unless otherwise indicated, the terms "about" and "approximately" mean ±20%, ±10%, ±20%, ±10%, ±20%, ±10%, ±20%, ±10%, ±20%, ±20%, ±3 ... ±10%, ±5% or ±1%.

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

[0014] "Cyano" refers to a -CN group.

[0015] "Oxo" refers to a =0 group.

[0016] An "alkyl" group is a group having 1 to 10 carbon atoms (C1-C 10In some embodiments, the alkyl group is a saturated, partially saturated, or unsaturated straight or branched chain acyclic hydrocarbon 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; while saturated branched alkyl groups 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 defined 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 as described herein are referred to as "substituted," they may be substituted with any one or more of the substituents present in the exemplary compounds and embodiments disclosed herein, as well as halogen; hydroxy; alkoxy; cycloalkyloxy, aryloxy, heterocyclyloxy, heteroaryloxy, heterocyclylalkyloxy, cycloalkylalkyloxy, aralkyloxy, heterocyclylalkyloxy, heteroarylalkyloxy, heterocyclylalkyloxy; oxo (=O); amino, alkylamino, cycloalkylamino, arylamino, heterocyclylamino, heteroarylamino, heterocyclylalkylamino, cycloalkylalkylamino alkylamino, heterocyclylalkylamino, heteroarylalkylamino, heterocyclylalkylalkylamino; imino; imido; amidino; guanidino; enamino; acylamino; sulfonylamino; urea, nitrourea; oxime; hydroxyamino; alkoxyamino; aralkyloxyamino; hydrazine; hydrazide; hydrazono; azido; nitro; thio (-SH), alkylthio; =S; sulfinyl; sulfonyl; aminosulfonyl; phosphonate; phosphinyl; acyl; formyl; carboxyl; ester; carbamate; acylamino; cyano; isocyanato; isothiocyanato; cyanato; thiocyanato; or -B(OH)2.In certain embodiments, when alkyl groups described herein are referred to as "substituted," they may be substituted with any one or more of the substituents present in the exemplary compounds and embodiments disclosed herein, as well as halogen (chlorine, iodine, bromine, or fluorine); alkyl; hydroxy; alkoxy; alkoxyalkyl; amino; alkylamino; carboxyl; nitro; cyano; sulfhydryl; thioether; imino; imido; amidino; guanidino; enamino; aminocarbonyl; acylamino; phosphonate; phosphino; thiocarbonyl; sulfinyl; sulfone; sulfonamide; ketone; aldehyde; ester; urea; urethane; oxime; hydroxylamine; alkoxyamine; aralkyloxyamine; N-oxide; hydrazine; hydrazide; hydrazone; azide; isocyanate; isothiocyanate; cyanate; thiocyanate; B(OH)2 or O(alkyl)aminocarbonyl.

[0017] An "alkylene" group is a radical that is identical to an alkyl group but has a divalent radical. Specific alkylene groups are radicals having 1 to 10 carbon atoms (C1-C 10 Alkylene), typically those of 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 3 to 10 carbon atom (C3-C4) alkyl radical having a single ring or multiple fused or bridged rings which may be optionally substituted. 10In some embodiments, the cycloalkyl group is a saturated cycloalkyl. In some embodiments, the cycloalkyl group has 3 to 8 ring carbon atoms (C3-C8 cycloalkyl), but 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, the cycloalkyl group is a saturated cycloalkyl. These saturated cycloalkyls include, for example, monocyclic structures such as cyclopropyl, cyclobutyl, cyclopentyl, cyclohexyl, cycloheptyl, cyclooctyl, 1-methylcyclopropyl, 2-methylcyclopentyl, 2-methylcyclooctyl etc., or polycyclic structures 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 etc. In other embodiments, the cycloalkyl group is an unsaturated cycloalkyl group. Examples of unsaturated cycloalkyl groups include cyclohexenyl, cyclopentenyl, cyclohexadienyl, butadienyl, pentadienyl, hexadienyl, etc. The cycloalkyl group may be substituted or unsubstituted. These substituted cycloalkyl groups include, for example, cyclohexanol, etc.

[0019] "Heterocyclyl" is a non-aromatic cycloalkyl group in which 1 to 4 ring carbon atoms are independently replaced by heteroatoms selected from O, S, and N. In some embodiments, the heterocyclyl group includes 3 to 10 ring members, while other such groups have 3 to 5, 3 to 6, or 3 to 8 ring members. The heterocyclyl group may also be bonded to other groups on any ring atom (i.e., any carbon atom or heteroatom of the heterocycle). The heterocycloalkyl group may be substituted or unsubstituted. The heterocyclyl group encompasses saturated and partially saturated ring systems. In addition, the term heterocyclyl is intended to encompass any non-aromatic ring containing at least one heteroatom, which ring may be fused to an aryl or heteroaryl ring, whether or not it is attached to the rest of the molecule. The phrase also includes bridged polycyclic ring systems containing heteroatoms. Representative examples of heterocyclyl groups include, but are not limited to, aziridinyl, azetidinyl, azepanyl, pyrrolidinyl, imidazolidinyl (e.g., imidazolidin-4-one or imidazolidin-2,4-dione), pyrazolidinyl, thiazolidinyl, tetrahydrothienyl, tetrahydrofuranyl, piperidinyl, piperazinyl (e.g., piperazin-2-one), morpholinyl, thiomorpholinyl, tetrahydropyranyl (e.g., tetrahydro-2H-pyranyl), tetrahydrothiopyranyl, oxathianyl, dithianyl, 1,4-dioxaspiro[4.5]decyl, homopiperazinyl, quinuclidinyl, or tetrahydropyrimidin-2(1H)-one. Representative substituted heterocyclyl groups may be monosubstituted or substituted more than once, such as, but not limited to, pyridinyl or morpholinyl groups, which are 2-, 3-, 4-, 5-, or 6-substituted or disubstituted with various substituents as listed below.

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

[0021] An "aryl" group is a 6-14 carbon atom (C6-C14) group having a single ring (eg, phenyl) or multiple fused rings (eg, naphthyl or anthracenyl). 14 In some embodiments, the aryl group contains 6-14 carbon atoms (C6-C 14 aryl), and in other embodiments contains 6 to 12 (C6-C 12 aryl) or even 6 to 10 carbon atoms (C6-C 10 aryl). Specific aryl groups include phenyl, biphenyl, naphthyl, and the like. Aryl groups may 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, etc.).

[0022] A "heteroaryl" group is an aromatic ring system having from 1 to 4 heteroatoms as ring atoms in the heteroaryl ring system, with the remaining atoms being carbon atoms. In some embodiments, the heteroaryl group contains from 3 to 6 ring atoms, and in other embodiments contains from 6 to 9 or even 6 to 10 heteroatoms in the ring portion of the group. Suitable heteroatoms include oxygen, sulfur, and nitrogen. In certain embodiments, the heteroaryl ring system is monocyclic or bicyclic. Non-limiting examples include, but are not limited to, groups such as pyrrolyl, pyrazolyl, imidazolyl, triazolyl, tetrazolyl, oxazolyl, isoxazolyl, benzisoxazolyl (e.g., benzo[d]isoxazolyl), thiazolyl, pyrrolyl, pyridazinyl, pyrimidinyl, pyrazinyl, thienyl, benzothienyl, furanyl, benzofuranyl, indolyl (e.g., indolyl-2-one or isoindolin-1-one), azaindolyl (pyrrolopyridinyl or 1H-pyrrolo[2,3-b]pyridinyl), indazolyl, benzimidazolyl (e.g., 1H-benzo[d]imidazolyl), imidazopyridinyl (e.g., The heteroaryl groups include benzophenone, ...

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

[0024] When groups described herein are referred to as "substituted," they may be substituted with any suitable substituent or substituents. Illustrative examples of substituents are those present in the exemplary compounds and embodiments disclosed herein, as well as halogen (chlorine, iodine, bromine, or fluorine); alkyl; hydroxy; alkoxy; alkoxyalkyl; amino; alkylamino; carboxyl; nitro; cyano; mercapto; thioether; imino; imido; amidino; guanidino; enamino; aminocarbonyl; acylamino; phosphonate; phosphine; thiocarbonyl; sulfinyl; sulfone; sulfonamide; ketone; aldehyde; ester; urea; urethane; oxime; hydroxylamine; alkoxyamine; aralkyloxyamine; N-oxide; hydrazine; hydrazide; hydrazone; azide; isocyanate; isothiocyanate; cyanate; thiocyanate; oxo(=O); B(OH)2, O(alkyl)aminocarbonyl; cycloalkyl, which may be monocyclic or fused or non-fused polycyclic (e.g., cyclopropyl, cyclobutyl, cyclopentyl or cyclohexyl), or a heterocyclic group which may be monocyclic or fused or non-fused polycyclic (e.g., pyrrolidinyl, piperidinyl, piperazinyl, morpholinyl or thiazinyl); a monocyclic or fused or non-fused polycyclic aryl or heteroaryl group (e.g., phenyl, naphthyl, pyrrolyl, indolyl, furanyl, thienyl, imidazolyl, oxazolyl, isoxazolyl, thiazolyl, triazolyl, tetrazolyl, pyrazolyl, pyridyl, quinolyl, isoquinolyl, acridinyl, pyrazinyl, pyridazinyl, pyrimidinyl, benzimidazolyl, benzothienyl or benzofuranyl); an aryloxy group; an aralkyloxy group; a heterocyclyloxy group; and a heterocyclylalkoxy group.

[0025]

[00146] Embodiments of the present disclosure are intended to encompass pharmaceutically acceptable salts, tautomers, isotopomers, and stereoisomers of the compounds provided herein, such as compounds of Formula (I).

[0026] As used herein, the term "pharmaceutically acceptable salt" refers to a salt prepared from a pharmaceutically acceptable non-toxic acid or base, including inorganic acids and bases and organic acids and bases. Suitable pharmaceutically acceptable base addition salts of the compound of formula (I) include, but are not limited to, metal salts prepared from aluminum, calcium, lithium, magnesium, potassium, sodium and zinc, or organic salts prepared from lysine, N,N'-diphenylmethylethylenediamine, 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, aminobenzoic acid, benzenesulfonic acid, benzoic acid, camphorsulfonic acid, citric acid, ethylenesulfonic acid, formic acid, fumaric acid, furoic acid, galacturonic acid, gluconic acid, glucuronic acid, glutamic acid, glycolic acid, hydrobromic acid, hydrochloric acid, hydroxyethanesulfonic 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, sulfonic acid and methanesulfonic acid. Therefore, the example of specific salt includes hydrochloride, formate and mesylate. Others are known in the art, referring to, for example, Remington's Pharmaceutical Sciences, 18 th eds., Mack Publishing, Easton PA (1990) or Remington: The Science and Practice of Pharmacy, 19 th eds., Mack Publishing, Easton PA (1995).

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

[0028] The use of stereoisomerically pure forms of the compounds disclosed herein, as well as the use of mixtures of those forms, is encompassed within the embodiments disclosed herein. For example, mixtures comprising equal or unequal amounts of enantiomers of a particular compound can be used in the methods and compositions disclosed herein. These isomers can be asymmetric synthesized or resolved using standard techniques such as chiral columns or chiral resolving agents. See, 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. 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 also 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 certain embodiments, the compounds may be isolated as E or Z isomers. In other embodiments, the compounds are mixtures of E and Z isomers.

[0030] "Tautomers" refers to isomeric forms of a compound that are in equilibrium with each other. The concentrations of the isomeric forms will depend on the environment in which the compound is located and may differ, for example, depending on whether the compound is a solid or in an organic or aqueous solution. For example, in aqueous solution, pyrazole can exhibit the following isomeric forms, which are referred to as tautomers of each other:

[0031] As will be readily appreciated by those skilled in the art, various functional groups and other structures may exhibit tautomerism, and all tautomers of compounds of Formula (I) are within the scope of this 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 be substituted with radioactive isotopes such as, for example, tritium ( 3 H), iodine-125 ( 125 I), sulfur-35( 35 S) or carbon-14 ( 14 C)) may be radiolabeled, or may be isotopically enriched, such as with deuterium ( 2 H), carbon-13 ( 13 C) or nitrogen-15( 15 N) enrichment. As used herein, an "isotopologue" is an isotopically enriched compound. The term "isotopically enriched" refers to an atom having an isotopic composition other than the natural isotopic composition of the atom. "Isotopically enriched" may also refer to a compound containing at least one atom having an isotopic composition different from the natural isotopic composition of the atom. The term "isotopic composition" refers to the amount of each isotope present in a given atom. Radiolabeled and isotopically enriched compounds can be used as therapeutic agents, such as cancer therapeutics, research reagents, such as binding assay reagents, and diagnostic reagents, such as in vivo imaging agents. All isotopic variants of the compounds as described herein, whether radioactive or not, are intended to be encompassed within the scope of the embodiments provided herein. In some embodiments, isotopologues of the compounds disclosed herein are provided, for example, compounds enriched in deuterium, carbon-13, and / or nitrogen-15. As used herein, "deuterated" means a compound in which at least one hydrogen (H) has been replaced by deuterium (with D or 2 H represents) substituted compounds, ie, the compounds are enriched in deuterium at at least one position.

[0033] It will be understood that, independent of stereoisomer or isotopic composition, each compound disclosed herein can be provided in the form of any pharmaceutically acceptable salt discussed herein. Likewise, it will be understood that the isotopic composition can vary independently of the stereoisomer composition of each compound described herein. Furthermore, the isotopic composition, while limited to those elements present in the corresponding compound disclosed herein or a salt thereof, can also vary independently of the choice of a pharmaceutically acceptable salt of the corresponding compound.

[0034] It should be noted that if there is a discrepancy between a described structure and the name of the structure, the described structure takes precedence.

[0035] As used herein, "treating" means alleviating a disorder, disease, or condition, or one or more symptoms associated with a disorder, disease, or condition, in whole or in part, or slowing or stopping the further progression or worsening of these symptoms, or alleviating or eliminating one or more causes of the disorder, disease, or condition itself. In one embodiment, the disorder is a neurodegenerative disease or a symptom thereof as described herein.

[0036] As used herein, "prevention" means a method of delaying and / or preventing, in whole or in part, the onset, recurrence, or spread of a disorder, disease, or condition; preventing a subject from developing a disorder, disease, or condition; or reducing the risk of a subject developing a disorder, disease, or condition. In one embodiment, the disorder is a neurodegenerative disease or a symptom thereof as described herein.

[0037] The term "effective amount" in connection with a compound disclosed herein means an amount capable of treating or preventing a disorder, disease or condition disclosed herein, or a symptom thereof.

[0038] As used herein, the term "subject" or "patient" includes animals, including but not limited to animals such as cows, monkeys, horses, sheep, pigs, chickens, turkeys, quail, cats, dogs, mice, rats, rabbits, or guinea pigs, and in one embodiment, mammals, and in another embodiment, humans. In one embodiment, the subject is a human who has or is at risk of having an S1P5-mediated disease or a symptom thereof.

[0039] Although various features of the invention may be described in the context of a single embodiment, these features may also be provided separately or in any suitable combination. Conversely, although the invention may be described in the context of separate embodiments for clarity, the invention may also be implemented in a single embodiment. Compound

[0040] In one aspect, provided herein are compounds of formula (I): or a pharmaceutically acceptable salt thereof, wherein: R 1 is H or oxo; Each R 2 are independently H or halogen, wherein at least one R 2 For fluorine; 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 optionally substituted C5-C6 cycloalkyl, optionally substituted C5-C6 aryl, optionally substituted 6-10 membered heterocyclyl, or 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 In some embodiments, R 1 is H. In some embodiments, R 1 For oxygen.

[0042] In some embodiments, R 2 In some embodiments, R 2 is H. In some embodiments, R 2 In some embodiments, R 2 is F, Cl or Br. In some embodiments, R 2 For 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)NHCH2-.

[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, wherein the heterocyclyl or heteroaryl group 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 group, wherein the heterocyclyl or heteroaryl group contains 1-3 heteroatoms selected from nitrogen, oxygen, and sulfur. In some embodiments, Ring A is optionally substituted cyclohexyl, optionally substituted phenyl, 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), 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, 6-10 membered heterocyclyl 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 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, 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 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 a cyclohexyl optionally substituted with -CF3. In some embodiments, Ring A 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 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 group 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 C1-C3 alkyl, optionally substituted C4-C6 cycloalkyl, -C(O)(6-membered heteroaryl), or -C(O)(9-membered heterocyclyl), wherein the heterocyclyl or heteroaryl group 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 C1-C3 alkyl, C4-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 where R 3 is H or OH; R 4 is optionally substituted C1-C3 alkyl, optionally substituted C3-C6 cycloalkyl, -C(O)(6-membered heteroaryl) or -C(O)(9-membered heterocyclyl); R 5 is an optionally substituted C1-C3 alkyl group; and R 6 In some embodiments, Ring A is In some embodiments, Ring A Part of In some embodiments, Ring A Part of

[0054] In some embodiments, Ring A is an optionally substituted 5-9 membered heteroaryl group containing 1-3 heteroatoms selected from nitrogen, oxygen, and sulfur. In some embodiments, Ring A is a 5-9 membered heteroaryl group containing 1-3 heteroatoms selected from nitrogen, oxygen, and sulfur, which is optionally substituted with an optionally substituted C1-C3 alkyl group, an optionally substituted amine, or a C4-C6 cycloalkyl group. In some embodiments, Ring A is a 5-9 membered heteroaryl group containing 1-3 heteroatoms selected from nitrogen, oxygen, and sulfur, which is optionally substituted with a C1-C3 alkyl group, an amine, or a C4 cycloalkyl group. In some embodiments, Ring A is a 5-9 membered heteroaryl group containing 1-3 heteroatoms selected from nitrogen, oxygen, and sulfur, which is optionally substituted with a C1 alkyl group, an amine, or a C4 cycloalkyl group. In some embodiments, Ring A is a 5-9 membered heteroaryl group containing 1-3 heteroatoms selected from nitrogen, oxygen, and sulfur, which is optionally substituted with a -CH3, an amine, or a cyclobutyl group. In some embodiments, Ring A is where 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, Part of

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

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

[0057] In the description herein, it should be understood that each description, variation, embodiment or aspect of a section can be combined with each description, variation, embodiment or aspect of other sections, just as if each and every combination of descriptions were specifically and individually listed. For example, the descriptions provided herein for R of formula (I) 1Each description, variation, embodiment or aspect may be combined with R 2 , X, Ring A, R 3 、R 4 、R 5 、R 6 、R 7 and R 8 Each description, modification, embodiment or aspect combination of formula (I) is equally applicable to other formulae described in detail herein, and is also described as if each and each description, modification, embodiment or aspect of all formulae are separated and are also listed separately.For example, when applicable, all description, modification, embodiment or aspect of formula (I) are equally applicable to any other formulae described in detail herein, such as formula (IIa), (IIb), (IIc), (IId), (IIIa), (IIIb), (IIIc) and (IIId) and are also described as if each and each description, modification, embodiment or aspect of all formulae are separated and are also listed separately.

[0058] In some embodiments, a compound selected from Table 1 or a pharmaceutically acceptable salt thereof is provided. Although certain compounds described in the present disclosure, including those in Table 1, are represented as specific stereoisomers and / or are presented in non-stereochemical forms, it is understood that any and all stereochemical forms, including any enantiomeric or diastereomeric forms, as well as any tautomeric or other forms, of any compound of the present disclosure, including those in Table 1, are described herein. Table 1. *“&1” indicates the stereocenter *“abs” indicates chiral center or a pharmaceutically acceptable salt thereof.

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

[0060] In addition, all compounds of formula (I) that exist in free base or free acid form can be converted into their pharmaceutically acceptable salts by treating with appropriate inorganic bases or organic bases or inorganic or organic acids by methods known to those skilled in the art. The salts of compounds of formula (I) can be converted into their free base or free acid forms by standard techniques. Synthesis method

[0061] The compounds described herein can be prepared using conventional organic synthesis and commercially available starting materials or methods provided herein. By way of example and not limitation, compounds of formula (I) can be prepared according to the methods outlined in General Schemes 1-4 and according to the Examples listed herein. It should be noted that one skilled in the art will know how to modify the methods described in the illustrative schemes and examples to obtain the desired products. How to use

[0062] Embodiments of the present disclosure provide methods for degrading an engineered polypeptide in a cell, methods for reducing the level of an engineered polypeptide in a cell, and methods of treating a disease, such as cancer, in a subject in need thereof.

[0063] In some embodiments, there is provided a method for reducing the level of an engineered polypeptide comprising a degradation domain, comprising contacting the engineered polypeptide with a compound of formula (I). In some embodiments, the contact occurs in an intracellular space, and the compound of formula (I) is combined with a degradation domain and a ubiquitin ligase, resulting in ubiquitination and degradation of the engineered polypeptide. In some embodiments, the degradation of the engineered polypeptide results in a reduction in at least one activity of the cell and / or an increase in at least one activity of the cell and / or cell death. Non-limiting exemplary effects include reducing the threshold value of cell (such as T cell) activation, increasing the functional persistence of cells (such as T cells), promoting cell survival, and increasing cell proliferation. In some embodiments, the method comprises administering a compound of formula (I) to the subject, wherein the subject comprises a cell comprising the engineered polypeptide.

[0064] In some embodiments, the engineered polypeptide is degraded in the presence of a compound of formula (I). In some embodiments, the compound of formula (I) interacts with the degradation domain and with a ubiquitin ligase, such as cereblon. In some embodiments, the compound of formula (I) mediates a complex comprising the degradation domain, the compound of formula (I), and the ubiquitin ligase, resulting in ubiquitination of the engineered polypeptide.

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

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

[0067] In some embodiments, the compound of Formula (I) degrades the engineered 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 compounds of formula (I) degrade the engineered polypeptide in the cell 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%, 85-100%, 90-100%, 95-100%, 100-100%, 110-100%, 120-100%, 130-100%, 140-100%, 150-100%, 160-100%, 170-100%, 180-100%, 190-100%, 200-100%, 210-100%, 220-100%, 230-100%, 240-100%, 250-100%, 260-100%, 270-100%, 280-100%, 290-100%, 300-100%, 350-100%, 3 %, 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 an engineered polypeptide in a cell, comprising contacting the cell with an effective amount of a compound of formula (I). The reduction in the level of an engineered polypeptide in the cell can be evaluated and demonstrated by various methods known in the art. Kits and commercially available assays, including cell-based assays, can be used to determine whether the engineered polypeptide in the cell has been degraded or to what extent.

[0069] In some embodiments, the compound of Formula (I) reduces the level of the engineered polypeptide in the 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 compounds of formula (I) reduce the level of the engineered polypeptide in the cell 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%, 100-100%, 110-100%, 120-100%, 130-100%, 140-100%, 150-100%, 160-100%, 170-100%, 180-100%, 190-100%, 200-100%, 210-100%, 220-200%, 230-200%, 240-200%, 250-200%, 260-200%, 270-200%, 280-200%, 290-300%, 300-300%, 310-310%, 310-310%, 320-320%, 330-330%, 340-330%, 350-330%, 360-330%, 370-330%, 380-330%, 390-3 %, 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 compound of formula (I) has an EC 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 as measured in an engineered polypeptide degradation assay. 50 In some embodiments, the compound of formula (I) has an EC of about 0.05 μM to about 0.2 μM. 50 In some embodiments, the compound of formula (I) has an EC of about 0.2 μM to about 1 μM. 50 In some embodiments, the compound of formula (I) has an EC50 of less than about 1 μM. In some embodiments, the compound of formula (I) has an EC50 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, after administering a CAR cell comprising an engineered polypeptide (comprising a degradation domain), it may be necessary to reduce or eliminate the expression of CAR and therefore reduce or eliminate target cell killing. In some such embodiments, the method may further include administering a compound of formula (I) to the subject. Administration of a compound of formula (I) results in degradation of an engineered peptide (e.g., CAR), and reduces or eliminates the targeting of the modified cell to cells expressing an antigen by the antigen-binding domain of CAR. In this way, the therapeutic activity of CAR cells can be adjusted, and safety can be improved.

[0072] In some embodiments, the modified cell population is first administered to the subject, followed by administration of the compound of formula (I) a specified time period after administration of the modified cell population, e.g., 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 can 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 tumors, aesmoplastic small round cell tumors, endocrine tumors, Ewing sarcoma, peripheral primitive neuroectodermal tumors, solid germ cell tumors, hepatoblastoma, neuroblastoma, non-rhabdomyosarcoma soft tissue sarcomas, 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 macroglobulinemia, splenic marginal zone lymphoma, plasma cell myeloma, plasmacytoma, extranodal marginal zone B-cell lymphoma, mucosa-associated lymphoid tissue lymphoma (MALT lymphoma), nodal 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, and leukemia. lymphoma), T-lymphocyte prolymphocytic leukemia, T-lymphocyte large granular lymphocytic leukemia, aggressive NK-cell leukemia, adult T-lymphocytic leukemia / lymphoma, extranodal NK / T-lymphocyte lymphoma, nasal and enteropathy-type T-lymphocyte lymphoma, hepatosplenic T-lymphocyte lymphoma, blastic NK-cell lymphoma, mycosis fungoides, Sézary syndrome, primary cutaneous anaplastic large cell lymphoma, lymphomatoid papulosis, angioimmunoblastic T-lymphocytic lymphoma, peripheral T-lymphocyte lymphoma (not otherwise specified), anaplastic large cell lymphoma, Hodgkin lymphoma, and non-Hodgkin lymphoma.

[0074] Modified cells described herein, such as CAR cells, in the treatment of a disease or disorder, for example, in the treatment of an individual with cancer can be evaluated by one or more criteria known to those skilled in the art, indicating the progression of the disease or disorder, specific to a particular disease or disorder. Typically, when one or more of the indices are detectably, for example, significantly moving from a disease state value or range to a normal value or range, it is effective to administer CAR cells (e.g., CAR T lymphocytes) to an individual with a disease / disorder (e.g., cancer).

[0075] In some embodiments, the compound of formula (I) is used in the preparation of a medicament for reducing the level of an engineered polypeptide in a cell.

[0076] The methods and uses of the present disclosure may involve the use of a compound of formula (I) alone or in combination with one or more additional therapies (eg, non-drug therapies or therapeutic agents).

[0077] Formula (I) compound can be before one or more such additional therapies, after or with one or more such additional therapies simultaneously.When combined, the dosage of formula (I) compound and the dosage of one or more additional therapies (e.g., non-drug treatment or therapeutic agent) can provide a therapeutic effect (e.g., synergistic or additional therapeutic effect). Formula (I) compound and additional therapies, such as anticancer agents can be administered together, such as in a single pharmaceutical composition, or administered individually, and when administered alone, this can occur simultaneously or sequentially. This order can be used to approach or stay away in time.

[0078] In some embodiments, the additional therapy is the administration of a side effect limiting agent (e.g., an agent intended to reduce the occurrence or severity of a therapeutic side effect). For example, in some embodiments, the compound of formula (I) can be used in combination with a therapeutic agent for treating 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, one or more additional therapies include non-drug treatment (e.g., surgery or radiotherapy). In some embodiments, one or more additional therapies include therapeutic agents (e.g., compounds or biological agents, which are antiproliferative agents). In some embodiments, one or more additional therapies include non-drug treatment and therapeutic agents. In other embodiments, one or more additional therapies include two therapeutic agents. In other embodiments, one or more additional therapies include three therapeutic agents. In some embodiments, one or more additional therapies include four or more therapeutic agents. Pharmaceutical compositions and routes of administration

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

[0081] The compounds disclosed herein can be administered orally, topically or parenterally to a subject in the form of conventional formulations such as capsules, microcapsules, tablets, granules, powders, lozenges, pills, suppositories, injections, suspensions, syrups, patches, creams, lotions, ointments, gels, sprays, solutions and emulsions. Suitable formulations can be prepared by commonly used methods using 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., magnesium stearate, light anhydrous silicic acid, talc or sodium lauryl sulfate), flavorings (e.g., citric acid, menthol, glycine or orange powder), preservatives (e.g., sodium benzoate, sodium bisulfite, methylparaben or propylparaben), stabilizers (e.g., citric acid, sodium citrate or acetic acid), suspending agents (e.g., methylcellulose, polyvinylpyrrolidone or aluminum stearate), dispersants (e.g., hydroxypropyl methylcellulose), diluents (e.g., water) and base waxes (e.g., cocoa butter, white petrolatum or polyethylene glycol). The effective amount of the compound of formula (I) in the pharmaceutical composition can be at a level that will exert the desired effect; for example, a unit dose of about 0.005 mg / kg subject body weight to about 10 mg / kg subject body weight for both oral and parenteral administration.

[0082] The dosage of a compound of formula (I) administered to a subject varies widely and can be determined at the discretion of a healthcare practitioner. The amount of a compound of formula (I) administered in any given case will depend on factors such as the solubility of the active ingredient, the formulation used, and the route of administration.

[0083] In another embodiment, provided herein are unit dosage formulations comprising between about 0.1 mg and 500 mg, between about 1 mg and 250 mg, between about 1 mg and about 100 mg, between about 1 mg and about 50 mg, between about 1 mg and about 25 mg, or between about 1 mg and about 10 mg of a compound of Formula (I).

[0084] For reasons of convenience, the compound of formula (I) can be administered orally. In one embodiment, when administered orally, the compound of formula (I) is administered with a meal and water. In another embodiment, the compound of formula (I) is dispersed in water or fruit juice (e.g., apple juice or orange juice) or any other liquid and administered orally as a solution or suspension.

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

[0086] In one embodiment, provided herein are capsules containing a compound of Formula (I) without additional carriers, excipients, or vehicles.

[0087] In another embodiment, provided herein is a composition comprising an effective amount of a compound of formula (I) and a pharmaceutically acceptable carrier or vehicle, wherein the pharmaceutically acceptable carrier or vehicle may comprise an excipient, a diluent, or a mixture thereof. In one embodiment, the composition is a pharmaceutical composition.

[0088] The composition can be in the form of tablets, chewable tablets, capsules, solutions, parenteral solutions, lozenges, suppositories and suspensions. The composition can be formulated as a convenient portion containing a daily dose or a daily dose in a dosage unit, which can be a single tablet or capsule or a convenient volume of liquid. In one embodiment, the solution is prepared by a water-soluble salt such as hydrochloride. Generally, all compositions are prepared according to known methods in pharmaceutical chemistry. Capsules can be prepared by mixing the compound of formula (I) with a suitable carrier or diluent, and filling an appropriate amount of the mixture in a capsule. Common carriers and diluents include but are not limited to inert powdered substances such as many different types of starch, powdered cellulose (especially crystalline and microcrystalline cellulose), sugars (such as fructose, mannitol and sucrose), cereal flour and similar edible powders.

[0089] Tablets can be prepared by direct compression, wet granulation or dry granulation. Its preparation usually incorporates diluents, binders, lubricants and disintegrants as well as compounds. Typical diluents include, for example, various types of starch, lactose, mannitol, kaolin, calcium phosphate or calcium sulfate, inorganic salts (such as sodium chloride) and powdered sugar. Powdered cellulose derivatives can also be used. Typical tablet binders are substances such as starch, gelatin and sugars (such as lactose, fructose, glucose, etc.). Natural gums and synthetic gums are also suitable, including gum arabic, alginate, methylcellulose, polyvinyl pyrrolidone, etc. Polyethylene glycol, ethyl cellulose and wax can also be used as binders.

[0090] Lubricants may be necessary in tablet formulations to prevent tablets and punches from sticking to the die. Lubricants can be selected from smooth solids such as talc, magnesium stearate and calcium stearate, stearic acid and hydrogenated vegetable oils. Tablet disintegrants are materials that expand when wet and break the tablet and release the compound. They include starch, clay, cellulose, algin and gum. More specifically, for example, corn and potato starch, methylcellulose, agar, bentonite, lignocellulose, powdered natural sponges, cation exchange resins, alginic acid, guar gum, citrus pulp and carboxymethyl cellulose, and sodium lauryl sulfate can be used. Tablets can be used as sugar coatings for flavorings and sealants, or coated with film-forming protective agents to change the dissolution characteristics of tablets. Compositions can also be formulated as chewable tablets, for example, by using materials such as mannitol in the formulation.

[0091] When it is desired to administer the compound of formula (I) as a suppository, typical bases may be used. Cocoa butter is a traditional suppository base, modified by the addition of waxes to slightly increase its melting point. Water-miscible suppository bases (particularly 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 formulations. For example, slowly dissolving pellets of the compound of formula (I) can be prepared and incorporated into tablets or capsules, or as a sustained-release implantable device. The technology also includes preparing pellets with several different dissolution rates and filling capsules with a mixture of these pellets. Tablets or capsules can be coated with a film that resists dissolution over a predictable period of time. Parenteral formulations can even be made long-lasting by dissolving or suspending the compound of formula (I) in an oily or emulsifying vehicle that slowly disperses it in serum. Example

[0093] The following examples are intended to illustrate rather than limit the examples. The naming of the compounds was performed using the automatic name generation tool provided by ChemBiodraw Ultra (Cambridgesoft), which generates systematic names for chemical structures and supports the Cahn-Ingold-Prelog rule for stereochemistry. One skilled in the art can modify the procedures shown 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 (eg, TFA, formic acid, or HCl) in the mobile phase during chromatographic purification, or stirring the product with an acid solution (eg, aqueous HCl) after chromatographic purification.

[0095] The following abbreviations may be used in connection with 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]palladium(II) dichloride Pd(dppf)Cl2:[1,1′-bis(diphenylphosphino)ferrocene]palladium(II) dichloride Pd(dppf)Cl2.DCM:[1,1′-bis(diphenylphosphino)ferrocene]palladium(II) dichloride complex with dichloromethane Pd2(dba)3: Tris(dibenzylindeneacetone)dipalladium Pd / C: Palladium on carbon HATU: (1-[bis(dimethylamino)methylene]-1H-1,2,3-triazolo[4,5-b]pyridine 3-oxide hexafluorophosphate, azabenzotriazole tetramethyluronium hexafluorophosphate) Synthesis Example Analytical methods

[0096] Preparative HPLC method 1: XBridge C18, 200 mm x 19 mm, 5-μm particles; Mobile phase A: 5:95 acetonitrile:water containing 10 mM ammonium acetate; Mobile phase B: 95:5 acetonitrile:water containing 10 mM ammonium acetate; Gradient: hold at 15% B for 0 min, 15-50% B over 25 min, then hold at 100% B for 6 min; Flow rate: 20 mL / min; Column temperature: 25°C. Fraction collection was triggered by the MS signal.

[0097] Preparative HPLC method 2: 1-Phen Luna Axia C18 5u 30×100 mm; mobile phase A: 95% H₂O / 5% ACN / 0.05% TFA; mobile phase B: 5% H₂O / 95% ACN / 0.05% TFA; gradient: hold at 2% B for 0 min, 2-100% B over 12 min, then hold at 100% B for 5 min; flow rate: 25 mL / min; column temperature: 25°C. Fraction collection was triggered by UV (220) nm.

[0098] Analytical HPLC method 1: Waters XBridge C18, 2.1 mm x 50 mm, 1.7 μm particles; mobile phase A: 5:95 acetonitrile:water with 0.1% trifluoroacetic acid; mobile phase B: 95:5 acetonitrile:water with 0.1% trifluoroacetic acid; temperature: 50°C; gradient: 0% B to 100% B over 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) Solution 1

[0100] Step 1: at 0 DEG C under nitrogen atmosphere to 2,2,6,6-tetramethylpiperidine (17.95mL, 105mmol) in THF (150mL) stirred solution dropwise add n-BuLi (63.3mL, 101mmol), and 0 DEG C of gained mixture is stirred 30min.Then the reaction mixture is cooled to about-45 DEG C (using dry ice / MeCN bath) and dropwise add 4-bromo-3,5-difluorobenzoic acid (10.0g, 42.2mmol) dissolved in THF (25mL) and continue stirring at-45 DEG C.After 3h, DMF (4.88mL, 63.3mmol) is added dropwise and the reaction mixture is warmed to room temperature and stirred overnight.At 0 DEG C, the reaction mixture is quenched with 3M HCl aqueous solution (40mL) and extracted with DCM (x3).By the organic layer merged through NaSODry, filter and be concentrated to dryness. The crude product was purified via silica gel chromatography (eluting with 0-80% EtOAc / hexanes) to afford 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 (S)-tert-butyl 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 h. 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 h. The reaction was quenched by the addition of 10% aqueous LiCl solution, and the product was then extracted with EtOAc. The organic layer was washed with brine and dried over MgSO and then concentrated. The crude material was purified using an ISCO 120 g silica gel column eluting with 0-80% EtOAc / hexanes to afford (S)-tert-butyl 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, 1H) 6.11-6.26 (m, 1H) 5.39 (br s, 1H) 4.91 (dd, J=8.68, 6.15 Hz, 1H) 4.67-4.76 (m, 1H) 4.49-4.58 (m, 1H) 2.26-2.43 (m, 3H) 2.11-2.21 (m, 1H) 1.45 (s, 9H)

[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 (S)-tert-butyl 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 KCO (1.236 g, 8.94 mmol) dissolved in water (15 mL). PdCl(dppf).DCM (0.146 g, 0.179 mmol) was added and the air was replaced with nitrogen. The mixture was heated to 100° C. for 1 h. The mixture 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 eluting with 0-15% B / DCM [where B = 15% ethanol / EtOAc + 0.1% TEA] .

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

[0106] Step 4: Oxygen was bubbled into a solution of (S)-tert-butyl 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 (1.00 g, 1.867 mmol) in DCM (10 mL) and 2-propanol (40 mL) for 5 min. Then, tris(2,2,6,6-tetramethyl-3,5-heptanedione)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 under an oxygen balloon for 2 days. The reaction mixture was diluted with EtOAc and washed with sodium thiosulfate solution. The organic layer was dried and concentrated and purified by ISCO (using a 40 g silica gel column and 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 F2N3O7 calculated value 553.3].

[0108] Step 5: To a 20 mL microwave vial was added (S)-tert-butyl 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 hour. It was concentrated to dryness and the residue was washed with ether to remove excess pTsOH. The precipitate was air-dried to give 391 mg of the product as the mono-pTsOH salt.

[0109] LC / MS(ESI)m / z 380.3[(M+H) + ,C 18 H 19 Calcd. for F2N3O4 379.1]. The enantiomeric excess of this material and subsequent 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 2 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 the product.

[0111] LC / MS(ESI)m / z 538.1[(M+H) + ,C 26 H 24 F5N3O4 calculated value 537.2]; HPLC a T Ret =1.25min; 1H NMR (500MHz, DMSO-d6) δ11.01(s,1H),7.87-7.82(m,2H),7.76(br d,J=7.9Hz,2H),7.44(br d,J=10.4Hz,1H),5.11-5.05(m,1H),4.53(br d,J=17.1Hz,1H),4.41-4.35(m,1H),2.95-2.85(m,1H),2.62(br d,J=17.1Hz,1H),2.51(br s, 4H), 2.47-2.33 (m, 2H), 2.25 (brd, J = 13.7Hz, 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) Option 1a

[0113] Step 1: To a 200 mL round-bottom flask was added 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 stirred vigorously at room temperature under a hydrogen atmosphere for 16 h. 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 F2N3O6 calculated 537.3]

[0115] Step 2: To a 20 mL microwave vial was added 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 diethyl ether and the precipitate was collected by filtration and air dried to give the mono-PhS03H salt of the desired product. This intermediate was subjected to preparative HPLC method 2 to obtain its TFA salt form.

[0116] MS:C 18 H 19 F2N3O3[M+H] + 364,[M+H] + Measured value: 364. 1 H NMR (300MHz, DMSO-d6) δ11.02(s,1H),8.65(d,J=11.3Hz,1H),8.38(d,J=10.8Hz,1H),7.74-7 .55(m,3H),7.50(d,J=8.8Hz,1H),7.40-7.23(m,5H),5.12(dd,J=13.3,5.1Hz,1H),4.55(d,J= 17.3Hz,1H),4.38(d,J=17.3Hz,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=13.6Hz,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) dissolved in 1 mL of DMF were added two drops of AcOH and then stirred at room temperature. After 0.5 h, NaBH(OAc)3 (24.50 mg, 0.116 mmol) was added and the resulting solution was stirred at room temperature for 16 h. This was purified by preparative HPLC method 1 to give 7.2 mg of the title compound. LC / MS (ESI) m / z 522.1 [(M+H) + ,C 26 H 24 F5N3O3 calculated value 521.2]; HPLC a T Ret =1.30min; 1H NMR (500MHz, DMSO-d6) δ11.01(s,1H),7.65(br d,J=7.6Hz,2H),7.53(br d,J=7.9Hz,2H),7.40(br d,J=8.5Hz,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) Option 1c

[0119] Step 1: To a microwave oven vial containing (S)-5-amino-4-(5-bromo-4,6-difluoro-1-oxoisoindolin-2-yl)-5-oxopentanoic acid tert-butyl ester (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) was added DMF (20 mL). The air was replaced with argon and heated in a microwave oven at 130 ° C for 1 h. It was diluted with EtOAc, washed with 10% LiCl aqueous solution and brine, dried over MgSO4, and then concentrated. The crude product was purified by flash column chromatography (eluted with 5-80% EtOAc / hexane) to give 501 mg (57% yield) of the desired product.

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

[0121] Step 2: To a solution of (S)-tert-butyl 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) at 0°C, followed by NaBH4 (19.94 mg, 0.527 mmol) and the atmosphere was replaced with N2. 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 KH2PO4 aqueous solution. The organic layer was separated and washed with brine, dried over Na2SO4, filtered and concentrated. The crude product was purified by flash column chromatography (eluting with 0-50% B / DCM [where 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 for F2N3O4: 383.2

[0123] Step 3: To a 5 mL microwave vial was added (S)-tert-butyl 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 washed with ether to give the mono-PhS03H salt of the desired product (151 mg, 86% yield).

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

[0125] Step 4: By following the procedure outlined for the synthesis of Example S1, the title compound was obtained in 18% yield by reductive amination of 3-(5-(aminomethyl)-4,6-difluoro-1-oxoisoindolin-2-yl)piperidine-2,6-dione with 4-(difluoromethyl)benzaldehyde.

[0126] LC / MS(ESI)m / z 450.0[(M+H) + ,C 22H 19 F4N3O3 calculated value 449.1]; HPLC a T Ret =1.11min; 1 H NMR (500MHz, DMSO-d6) δ11.05-10.95(m,1H),7.55-7.46(m,5H),5.12(dd,J=13.2,5.2Hz,1H),4.58-4.50(m,1H),4.39(br d,J=17.6Hz,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] Table 2 lists example compounds prepared by following the procedures outlined in Schemes 1 and 1a-1c using the appropriate aldehyde.

[0128] Table 2

[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)

[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 h. 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 F2N4O4S calculated value 526.1]; HPLC a TRet =0.93min; 1 H NMR (500MHz, DMSO-d6) δ10.96(s,1H),9.34(s,1H),8.16-8.09(m,1H),8.04(br d,J=8.9Hz,1H),7.59-7.49(m,1H),7.36(br d,J=10.4Hz,1H),5.62-5.40(m,1H),5.05(br dd,J=13.3,5.0Hz,1H),4.47(br d,J=17.4Hz,1H),4.30(br d,J=17.4Hz,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(brs,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).

[0133] The title compound was obtained as an off-white solid in 34% yield by following the alkylation method outlined for the synthesis of Example S68 using 3-(4,6-difluoro-1-oxo-5-(piperidin-4-yl)isoindolin-2-yl)piperidine-2,6-dione as the amine coupling partner.

[0134] LC / MS(ESI)m / z 511.1[(M+H) + ,C 26 H 24 F2N4O3S calculated value 510.2]; HPLC a T Ret =0.93min; 1H NMR (500MHz, DMSO-d6) δ10.96(s,1H),9.47(s,1H),8.29(s,1H),8.16(d,J=8.2Hz,1H),7.66(br d,J=8.5Hz,1H),7.44(br d,J=8.9Hz,1H),5.06(br dd,J=13.6,5.0Hz,1H),4.53-4.41(m,2H),4.33(br d,J=17.1Hz,1H),2.92-2.80(m,1H),2.56(br d,J=17.1Hz,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).

[0136] The title compound was obtained as a white solid in 39% yield by following the amidation procedure outlined 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.

[0137] LC / MS(ESI)m / z 561.0[(M+H) + ,C 25 H 23 BrF2N4O4 calculated value 560.1]; HPLC a T Ret =1.52min; 1 H NMR (500MHz, DMSO-d6) δ10.96(s,1H),8.51(s,1H),8.02(s,1H),7.42(d,J=8.7Hz,1H),5.05(dd,J=13.3,4.9Hz,1H),4.63(br d,J=12.6Hz,1H),4.50(d,J=17.3Hz,1H),4.33(d,J=17.1Hz,1H),3.18-3.12(m,1H),2.93-2.80(m,2H),2.56(br d,J=17.5Hz,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=12.8Hz,1H)

[0138] 1.76 (m, 3H)

[0139] Table 3 lists examples of compounds prepared by following the alkylation procedures outlined for the synthesis of Example S68, Example S69, and Example S72 using the appropriate alkyl halide, and the amidation procedures outlined for the synthesis of Example S70 and Example S71 using the appropriate carboxylic acid.

[0140] Table 3

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

[0142] Step 1: To a solution of rac-(3S)-3-[4,6-difluoro-5-(4-hydroxy-4-piperidinyl)-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 h. 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), gradient 10% ACN to 70% ACN in 20 min; detector, UV 254 nm) to give N-[2-[4-[4,6-difluoro-1-oxo-2-[rac-(3S)-2,6-dioxo-3-piperidinyl]isoindolin-5-yl]-4-hydroxy-1-piperidinyl]ethyl]-N-methyl-carbamic acid tert-butyl ester (150 mg, 63.6% yield) as a light brown solid.

[0143] MS:m / z:C 26 H 34 F2N4O6[M+H] + Calculated value: 537; Measured 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-piperidinyl]isoindolin-5-yl]-4-hydroxy-1-piperidinyl]ethyl]-N-methyl-carbamate (150 mg, 0.28 mmol) in DCM (4 mL) was added 4M HCl (1 mL) in 1,4-dioxane. The resulting mixture was stirred at room temperature for 3 h. 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-piperidinyl]-1-oxo-isoindolin-2-yl]piperidine-2,6-dione (130 mg crude) as a brown solid.

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

[0146] Step 3: To a solution of rac-(3S)-3-[4,6-difluoro-5-[4-hydroxy-1-[2-(methylamino)ethyl]-4-piperidinyl]-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 h. The reaction was monitored by LCMS. The mixture was concentrated under reduced pressure. The product was purified by prep-HPLC to give N-[2-[4-[2-(2,6-dioxo-3-piperidinyl)-4,6-difluoro-1-oxo-isoindolin-5-yl]-4-hydroxy-1-piperidinyl]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. 1H NMR (300MHz, DMSO-d6) δ11.03(s,1H),9.47(s,1H),7.48(d,J=10.6Hz,1H),6.05(s ,1H),5.13(dd,J=13.2,5.1Hz,1H),4.55(d,J=17.4Hz,1H),4.37(d,J=17.4Hz,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 Ultimate AQ-C18, 50*250 mm*10 μm; mobile phase A: water (0.05% TFA), mobile phase B: ACN; flow rate: 100 mL / min; gradient: 20% B to 32% B, 32% B in 20 min; wavelength: 254 nm.

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

[0150] The title compound was prepared as a brown solid in 25.7% yield using benzenesulfonyl chloride in step 3 according to the preparation of Example S101.

[0151] MS:m / z:C 27 H 30 F2N4O6S[M+H] + Calculated value: 577; Measured value: 577. 1H NMR(300MHz,DMSO-d6)δ11.03(s,1H),9.55(s,1H),7.99-7.81(m,2H),7.80-7.59(m,3H ),7.49(d,J=10.6Hz,1H),6.05(s,1H),5.13(dd,J=13.2,5.0Hz,1H),4.55(d,J=17.4Hz ,1H),4.38(d,J=17.4Hz,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 Ultimate AQ-C18, 50*250 mm*10 μm; mobile phase A: water (0.05% TFA), mobile phase B: ACN; flow rate: 100 mL / min; gradient: 20% B to 48% B, 48% B in 20 min; wavelength: 254 nm.

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

[0154] Step 1: to a solution of (4S)-5-amino-4-(5-bromo-4,6-difluoro-1-oxo-isoindolin-2-yl)-5-oxo-pentanoic acid tert-butyl ester (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 three times with nitrogen 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, eluted with petroleum ether / EtOAc (2 / 1) to give (4S)-5-amino-4-[4,6-difluoro-5-(hydroxymethyl)-1-oxo-isoindolin-2-yl]-5-oxo-pentanoic acid tert-butyl ester (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 (4S)-5-amino-4-[4,6-difluoro-5-(hydroxymethyl)-1-oxo-isoindolin-2-yl]-5-oxo-pentanoic acid tert-butyl ester (180 mg, 0.47 mmol) in MeCN (2 mL) and CCl (2 mL) were added 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 x 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), gradient from 10% ACN to 30% ACN in 10 min; 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 x 50 mL), washed with water (3 x 50 mL) and dried over anhydrous Na2SO4. After filtration, the organic layer was concentrated under reduced pressure. The crude product was applied to a silica gel column and eluted with petroleum ether / EtOAc (1 / 3) to give (4S)-5-amino-4-[5-(7-cyano-3,4-dihydro-1H-isoquinoline-2-carbonyl)-4,6-difluoro-1-oxo-isoindolin-2-yl]-5-oxo-pentanoic acid tert-butyl ester (60 mg, 0.11 mmol, 88.7% yield) as a light 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 (4S)-5-amino-4-[5-(7-cyano-3,4-dihydro-1H-isoquinoline-2-carbonyl)-4,6-difluoro-1-oxo-isoindolin-2-yl]-5-oxo-pentanoic acid tert-butyl ester (55 mg, 0.09 mmol) in MeCN (3 mL) was added benzenesulfonic acid (44.1 mg, 0.28 mmol). The resulting mixture was degassed three times with nitrogen and then stirred at 60 ° C. under a nitrogen atmosphere overnight. The reaction was monitored by LCMS. The mixture was concentrated under reduced pressure. The pH of the solution was adjusted to 9 with sodium bicarbonate solution and extracted with EtOAc (3 x 50 mL). The combined organic layers were dried over Na2SO4 and concentrated under reduced pressure. The crude product was purified by preparative HPLC to give 2-[2-(2,6-dioxo-3-piperidinyl)-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 (300MHz, DMSO-d6) δ11.05(s,1H),7.90-7.56(m,3H),7.42(t,J=8.0Hz,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 preparative 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% B to 50% B, 50% B in 6 min; 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).

[0164] The title compound was prepared as a white solid in 39% overall yield using 5-chloro-1,2,3,4-tetrahydroisoquinoline in step 3 according to the preparation of Example S105. MS: m / z: C 23 H 18 ClF2N3O4,[M+H] + Calculated value: 474; measured value: 474. 1 H NMR(400MHz, DMSO-d6)δ11.06(d,J=4.8Hz,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. 9Hz, 1H), 2.61 (d, J = 17.8Hz, 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% B to 57% B, 57% B in 10 min; 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)

[0167] The title compound was prepared as a white solid in 22.2% overall yield using 6,7-difluoro-1,2,3,4-tetrahydroisoquinoline in step 3 according to the preparation of Example S105.

[0168] MS:m / z:C 23 H 17 F4N3O4,[M+H] + Calculated value: 476; measured value: 476. 1 H NMR(300MHz,DMSO-d6)δ11.05(d,J=3.1Hz,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),2.8 0-2.69(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% B to 53% B, 53% B in 10 min; wavelength: 254 nm.

[0170] 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) Option 6

[0171] 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 Boc2O (628.3 mg, 2.88 mmol) at 0 ° C. The solution was stirred at room temperature for 2 hours. The desired product could be detected by LCMS. After completion, the mixture was concentrated in vacuo and diluted with water (50 mL) and extracted with DCM (3 x 80 mL). The combined organic layers were dried over Na2SO4 and concentrated under reduced pressure. The residue was purified by silica gel chromatography (petroleum ether: ethyl acetate = 8: 1) to obtain 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.

[0172] MS:m / z:C 12 H 15 ClN2O2[M+H] + Calculated value: 255; measured value: 255.

[0173] Step 2: to 2-chloro-5,7-dihydropyrrolo-[3,4-b]pyridine-6-carboxylic acid tert-butyl ester (300mg, 1.18mmol) and 1-(4-methoxyphenyl)-N-methyl-methylamine (178.1mg, 1.18mmol) in 1,4-dioxane (20mL) add CsCO(1148.4mg, 3.53mmol), Ruphos (55mg, 0.12mmol) and RuphosPdG(109.8mg, 0.12mmol).Solution is degassed three times with nitrogen and stirred at 90 DEG C overnight.Expected product can be detected by LCMS.After completion, mixture is concentrated in vacuum and diluted with water (50mL) and extracted with DCM (3x 50mL).By the organic layer merged through NaSODry and under reduced pressure concentrate. 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.

[0174] MS:m / z:C 21 H 27 N3O3[M+H] + Calculated value: 370; measured value: 370.

[0175] Step 3: to 2- [(4- methoxyphenyl) methyl-methyl-amino] -5,7- dihydro pyrrolo [3,4-b] pyridine -6- acid tert-butyl ester (370mg, 1mmol) in DCM (20mL) is added 4MHCl in 1,4- dioxane. The solution is stirred at room temperature for 2 hours. The expected product can be detected by LCMS. After completion, N- [(4- methoxyphenyl) methyl] -N- methyl -6,7- dihydro -5H- pyrrolo [3,4-b] pyridine -2- amine (230mg thick material) as a yellow solid is obtained by concentration under reduced pressure.

[0176] MS:m / z:C 16 H 19 N3O[M+H] + Calculated value: 270; measured value: 270.

[0177] 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 for 3 hours under a nitrogen atmosphere. The desired product can be detected by LCMS. After completion, the mixture was concentrated in vacuo and diluted with water (50 mL) and extracted with DCM (3x80 mL). The combined organic layers were dried over Na SO and concentrated under reduced pressure. The residue was purified by silica gel chromatography (petroleum ether:ethyl acetate=8:1) to give (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-pentanoic acid tert-butyl ester (106 mg, 0.15 mmol, 61.3% yield) as a white solid.

[0178] MS:m / z:C 34 H 37 F2N5O6[M+H] + Calculated value: 650; measured value: 650.

[0179] Step 5: under nitrogen atmosphere at 60 DEG C by (4S) -5- amino -4- [4,6- difluoro -5- [2- [(4- methoxyphenyl) methyl -methyl -amino] -5,7- dihydro pyrrolo [3,4-b] pyridine -6- carbonyl] -1- oxo -isoindolin -2- base] -5- oxo - tert-butyl pentanoate (100mg, 0.15mmol) and benzenesulfonic acid (72.9mg, 0.46mmol) in MeCN (8mL) solution is stirred 24 hours. Expected product can be detected by LCMS. The mixture is concentrated in vacuo and diluted with water (50mL). The resulting mixture is basified to pH 8 with NaCO aqueous solution and extracted with DCM (3x 80mL). The combined organic layers were dried over Na2SO4 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) as a white solid.

[0180] MS:m / z:C 30 H 27 F2N5O5[M+H] + Calculated value 576; measured value 576.

[0181] Step 6: under nitrogen atmosphere at 60 DEG C by 3- [4,6- bis-fluoro -5- [2- [(4- methoxyphenyl) methyl -methyl -amino] -5,7- dihydro pyrrolo [3,4-b] pyridine -6- carbonyl] -1- oxo - isoindoline -2- base] piperidine -2,6- dione (75mg, 0.13mmol) in TFA (4mL) solution stirred for 2 hours. Expected product can be detected by LCMS. The resulting solution is concentrated under reduced pressure and purified by preparative HPLC to obtain 3- [4,6- bis-fluoro -5- [2- (methylamino) -5,7- dihydro pyrrolo [3,4-b] pyridine -6- carbonyl] -1- oxo - isoindoline -2- base] piperidine -2,6- dione (32.4mg, 54.4% yield) as a white solid.

[0182] MS:m / z:C 24 H 18 F2N4O4[M+H] + Calculated value: 456; measured value: 456. 1H NMR (400MHz, DMSO-d6) δ11.05(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=9.3Hz,2 H),4.70-4.41(m,4H),2.99-2.90(m,1H),2.85(s,2H),2.75(d,J=3.0Hz,1H),2.70-2.57(m,1H),2.50-2.44(m,1H),2.10-1.93(m,1H).

[0183] 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: 12% B to 15% B, 15% B in 6 min; wavelength: 254 / 210 nm.

[0184] Example S116. N-[[2-(2,6-dioxo-3-piperidinyl)-4,6-difluoro-1-oxo-isoindolin-5-yl]methyl]indolizine-2-carboxamide (Compound 69)

[0185] The title compound was prepared as a white solid in 21.6% yield in step 4 according to general procedure 2 using indolizine-2-carboxylic acid, TCFH and NMI.

[0186] MS:m / z:C 23 H 18 F2N4O4,[M+H] - Calculated value: 453; measured value: 453. 1 H NMR(400MHz,DMSO-d6)δ11.02(s,1H),8.66(t,J=5.3Hz,1H),8.24(dd,J=7.0,1.3Hz,1H), 7.95(d,J=1.6Hz,1H),7.48(d,J=7.8Hz,1H),7.41(d,J=9.1Hz,1H),6.78(s,1H),6.75-6. 66(m,1H),6.66-6.51(m,1H),5.13(dd,J=13.3,5.1Hz,1H),4.75-4.46(m,3H),4.39(d,J= 17.3Hz,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).

[0187] 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, 32% B in 6 min; wavelength: 254 nm.

[0188] Example S119. 3-(6-Fluoro-1-oxo-5-(1-(4-(trifluoromethyl)benzyl)piperidin-4-yl)isoindolin-2-yl)piperidine-2,6-dione (Compound 85) Option 9a

[0189] Step 1: To a 200 mL round-bottom flask was added 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 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.

[0190] LC / MS(ESI)m / z 520.5[(M+H) + ,C 27 H 38 FN3O6 calculated value 519.3].

[0191] Step 2: To a 100 mL round-bottom flask was added 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 washed with ether to remove excess PhSO 3 H and dried to give the mono-benzenesulfonate salt of the desired product (668 mg).

[0192] 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) was added 2 drops of AcOH and then stirred at room temperature. After 0.5 h, NaBH(OAc) (36.8 mg, 0.174 mmol) was added and the resulting solution was stirred at room temperature for 2 h. This was purified by preparative HPLC method 1 to give 6.0 mg of the title compound.

[0193] LC / MS(ESI)m / z 504.1[(M+H) + ,C 26 H 25 F4N3O3 calculated value 503.2]; HPLC a T Ret =1.29min; 1 H NMR(500MHz,DMSO-d6)δ10.99(s,1H),7.74-7.67(m,J=8.2Hz,2H),7.63(d,J=6.0Hz,1H), 7.61-7.55(m,J=8.2Hz,2H),7.47(d,J=9.2Hz,1H),5.10(dd,J=13.3,5.0Hz,1H),4.46-4.3 8(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),2.5 7-2.56(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)

[0194] Table 4 lists examples of compounds prepared by following the procedures outlined in Schemes 9 and 9a using the appropriate aldehydes.

[0195] Table 4.

[0196] 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)

[0197] To a 1-dram vial was added 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 h. It was cooled to room temperature and purified via preparative HPLC Method 1 to yield 6.0 mg of the title compound.

[0198] LC / MS(ESI)m / z 509.2[(M+H) + ,C 26 H 25 FN4O4S calculated value 508.2]; HPLC a T Ret =0.97min; 1 H NMR (500MHz, DMSO-d6) δ11.00(br s,1H),9.49(s,1H),8.36(br s,1H),8.20(br d,J=8.2Hz,1H),7.88(br d,J=6.1Hz,1H),7.74(br d,J=8.5Hz,1H),7.54-7.45(m,1H),5.08(br dd,J=12.4,4.4Hz,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=16.5Hz,1H),2.51(br s,6H),2.47-2.32(m,2H),2.06-1.99(m,1H),1.90-1.77(m,2H).

[0199] 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)

[0200] The title compound was obtained as an off-white solid in 51% yield by following the alkylation procedure outlined 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.

[0201] LC / MS(ESI)m / z 493.1[(M+H)+ ,C 26 H 25 FN4O3S calculated value 492.2]; HPLC a T Ret =1.07min; 1 H NMR (500MHz, DMSO-d6) δ10.95(s,1H),9.46(s,1H),8.30(br s,1H),8.16(d,J=8.4Hz,1H),7.67(br d,J=8.4Hz,1H),7.47(br d,J=9.3Hz,2H),5.06(br dd,J=13.0,4.8Hz,1H),4.51-4.42(m,2H),4.39(br d,J=17.2Hz,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).

[0202] Table 5 lists examples of compounds prepared by following the alkylation procedures outlined for the synthesis of Example S131 and Example S132 using the appropriate alkyl halide.

[0203] Table 5.

[0204] Example S140. 3-(5-(6-amino-4,5-dimethylpyridin-2-yl)-6-fluoro-1-oxoisoindolin-2-yl)piperidine-2,6-dione (Compound 34) Plan 11

[0205] Step 1: To a solution of 4,4,4',4',5,5,5',5'-octamethyl-2,2'-bis(1,3,2-dioxaborolane) (275 mg, 1.084 mmol), potassium acetate (213 mg, 2.167 mmol) and (S)-5-amino-4-(5-bromo-6-fluoro-1-oxoisoindolin-2-yl)-5-oxopentanoic acid tert-butyl ester (300 mg, 0.722 mmol) in dioxane (10 mL) was added PdCl2(dppf).DCM (59.0 mg, 0.072 mmol) and the air was replaced with N2. It was heated to 100 ° C. for 16 h. It was cooled to room temperature, diluted with EtOAc and quenched with brine and the organic layer was separated, dried over Na2SO4 and concentrated. The crude material was purified by flash column chromatography (eluting with 0-6% MeOH / DCM) to give 302 mg (90% yield) of the desired product.

[0206] LC / MS(ESI)m / z 325.1[(M-137) + ,C 23 H 32 BFN2O6 calculated value 462.2].

[0207] Step 2: A 5 mL microwave vial was charged with 6-chloro-3,4-dimethylpyridin-2-amine (16.5 mg, 0.105 mmol), (S)-tert-butyl 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). The vial was sealed and the air was replaced with nitrogen, and then heated in a microwave at 120° C. for 0.25 h. It was diluted with EtOAc, washed with brine, and the organic layer was separated, dried over MgSO 4 and concentrated to give 32 mg of desired product, which was used in the next step without further purification.

[0208] LC / MS(ESI)m / z 457.3[(M+H) + ,C 24 H 29 FN4O4 calculated value 456.2].

[0209] Step 3: To 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 1 mL of a solution of PhSO 3 H in MeCN (0.25 M) and microwaved at 120° C. for 15 min. This was purified by preparative HPLC method 1 to give 11.9 mg (28% yield) of the desired product.

[0210] LC / MS(ESI)m / z 383.1[(M+H) + ,C 20 H 19 FN4O3 calculated value 382.1]; HPLC a T Ret =1.00min; 1 HNMR(500MHz,DMSO-d6)δ11.00(s,1H),8.03(d,J=6.6Hz,1H),7.57(d,J=9.9Hz,1 H),6.93-6.88(m,1H),5.70(s,1H),5.10(dd,J=13.4,5.1Hz,1H),4.50(d,J=17.2 Hz,1H),4.37(d,J=17.0Hz,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)

[0211] Other compounds can be prepared by following the procedures outlined for Schemes 3 and 4 using the appropriate boronic acid / ester.

[0212] Example S146. 3-(5-(6-amino-4,5-dimethylpyridin-2-yl)-4,6-difluoro-1-oxoisoindolin-2-yl)piperidine-2,6-dione (Compound 93) Plan 12

[0213] Step 1: 6- chloro- 3,4- dimethylpyridine -2- amine (30.0mg, 0.192mmol), 1,1,1,2,2,2- hexamethyldistanane (69.0mg, 0.211mmol) and PdCl2 (dtbpf) (12.48mg, 0.019mmol) in toluene (5mL) solution and replace air with N2. It is heated to 110 DEG C and kept for 4h. After cooling to room temperature, it is washed with EtOAc and salt water, and the organic layer is separated, through Na2SO4 drying, filtering and concentrating. The crude material (38mg) is used for the next step without further purification.

[0214] LC / MS(ESI)m / z 287.1[(M+H) + ,C 10 H 18 N2Sn calculated value 286.0].

[0215] Step 2: To a 2-dram pressure vial was added 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 for 16 h under a nitrogen atmosphere. 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 material (28 mg) was used in the next step without further purification.

[0216] LC / MS(ESI)m / z 475.1[(M+H) + ,C 24 H 28 F2N4O4 calculated value 474.2].

[0217] 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) was added 1 mL of a solution of PhSO 3 H in MeCN (0.25 M) and microwaved at 120° C. for 15 min. This was purified by preparative HPLC method 1 to give 6.8 mg (28% yield) of the desired product.

[0218] LC / MS(ESI)m / z 401.0[(M+H) + ,C 20 H 18F2N4O3 calculated value 400.1]; HPLC a T Ret =1.04min; 1 H NMR(500MHz,DMSO-d6)δ11.03(s,1H),7.53(d,J=7.5Hz,1H),6.58(s,1H),5.80(s,2H),5.15(dd ,J=13.4,5.0Hz,1H),4.60(d,J=17.2Hz,1H),4.43(d,J=17.3Hz,1H),2.97-2.89(m,1H),2.62(br d,J=18.1Hz,1H),2.49-2.41(m,1H),2.22(s,3H),2.06(br s,1H),2.04(s,3H)

[0219] 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).

[0220] The title compound was synthesized from 7-chloro-1,2,3,4-tetrahydro-1,8-naphthyridine by following the route outlined for the synthesis of Example S146.

[0221] LC / MS(ESI)m / z 413.1[(M+H) + ,C 21 H 18 Calculated value of F2N4O3: 412.1]; HPLC a T Ret =1.05min; 1 H NMR (500MHz, DMSO-d6) δ11.02(s,1H),7.53(d,J=7.6Hz,1H),7.29(d,J=7.2Hz,1H),6.65(br s,1H),6.58(d,J=7.2Hz,1H),5.14(dd,J=13.2,4.9Hz,1H),4.60(d,J=17.2Hz,1H),4.43(d,J=17.2Hz,1H),2.97-2.89(m,1H),2.73(br t,J=6.1Hz,2H),2.62(br dd,J=15.6,2.2Hz,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)

[0222] Example S148 includes compounds prepared according to General Scheme 1. Table 6 Example S149 includes the following compounds, which were prepared according to the general procedure specified: Table 7. Biological Examples Example B1. Evaluation of Small Molecule-Induced Degradation of IKZF1 ZNF2_ZNF3 Q1F-Expressing Jurkat Cells

[0223] Compounds targeting the Q1F degrader were screened in Jurkat cells engineered to express IKZF1 ZNF2_ZNF3 Q1F. This cell line was generated using lentiviral vectors containing a CD19 CAR tagged with IKZF1 ZNF2_ZNF3 Q1F-Nluc, which were transduced into Jurkat cells. The CD19 CAR contains an anti-CD19 scFv, a CD28 transmembrane domain, a 4-1BB costimulatory domain, a CD3ζ signaling domain, and the ZNF2_ZNF3Q1F degrader (GERPF F CNQCGASFTQKGNL LRHIKLHSGE KPFKCHLCNY ACRRRDALTG HLRTHS; SEQ ID NO: 1; Q1F replaces the underline mark). The transduced cells were treated with titration of various small molecules or without drug treatment and then incubated at 37 ° C for 18 hours. The cells were washed and stained with appropriate staining reagents to measure CAR levels. The cells were incubated in the staining reagent at 4 ° C for 20 min and then washed 3 times, and then read on a flow cytometer. CAR levels were normalized to cells without drug treatment. The resulting titration curve was used to calculate EC50 and Ymin values. This identified small molecules that efficiently degrade Q1F degrader-tagged CAR (Table 1). Example B2. Evaluation of small molecule selectivity for Aiolos, Ikaros, CK1α, GSPT1, and Helios

[0224] Helios DF15 multiple myeloma cells stably expressing ePL-tagged Aiolos, Ikaros, or GSPT1, and MDS-L cells stably expressing ePL-tagged CK1a, were generated by infection with pLOC-ePL-Aiolos (or Ikaros, GSPT1, or CK1a) lentivirus. The sequences used for human Aiolos and Ikaros are shown below:

[0225] Aiolos: MEDIQTNAEL KSTQEQSVPA ESAAVLNDYS LTKSHEMENV DSGEGPANED EDIGDDSMKVKDEYSERDEN VLKSEPMGNA EEPEIPYSYS REYNEYENIK LERHVVSFDS SRPTSGKMNC DVCGLSCISFNVLMVHKRSH TGERP FQCNQ CGASFTQKGN LLRHIKLHTG EKPFKCHLCN YACQRRDALTGHLRTH SVEKPYKCEFCGRS YKQRSSLEEH KERCRTFLQS TDPGDTASAE ARHIKAEMGS ERALVLDRLA SNVAKRKSSMPQKFIGEKRH CFDVNYNSSY MYEKESELIQ TRMMDQAINN AISYLGAEAL RPLVQTPPAP TSEMVPVISSMYPIALTRAE MSNGAPQELE KKSIHLPEKS VPSERGLSPN NSGHDSTDTD SNHEERQNHI YQQNHMVLSRARNGMPLLKE VPRSYELLKP PPICPRDSVK VINKEGEVMD VYRCDHCRVL FLDYVMFTIH MGCHGFRDPFECNMCGYRSH DRYEFSSHIA RGEHRALLK (SEQ ID NO: 2; ZNF2 and ZNF3 are underlined).

[0226] Ikaros: MDADEGQDMS QVSGKESPPV SDTPDEGDEP MPIPEDLSTT SGGQQSSKSD RVVASNVKVETQSDEENGRA CEMNGEECAE DLRMLDASGE KMNGSHRDQG SSALSGVGGI RLPNGKLKCD ICGIICIGPNVLMVHKRSHT GERP FQCNQC GASFTQKGNLLRHIKLHSGEKPFKCHLCNY ACRRRDALTG HLRTHSVGKPHKCGYCGRSY KQRSSLEEHK ERCHNYLESM GLPGTLYPVI KEETNHSEMA EDLCKIGSER SLVLDRRLASNVAKRKSSMPQ KFLGDKGLSD TPYDSSASYE KENEMMKSHV MDQAINNAIN YLGAESLRPL VQTPPGGSEVVPVISPMYQL HKPLAEGTPR SNHSAQDSAV ENLLLLSKAK LVPSEREASP SNSCQDSTDT ESNNEEQRSGLIYLTNHIAP HARNGLSLKE EHRAYDLLRA ASENSQDALR VVSTSGEQMK VYKCEHCRVL FLDHVMYTIHMGCHGFRDPF ECNMCGYHSQ DRYEFSSHIT RGEHRFHMS (SEQ ID NO: 3; ZNF2 and ZNF3 are underlined).

[0227] DF15 multiple myeloma cells expressing Ikaros, Aiolos, and GSPT1 fused to an ePL tag (DiscoverX) and MDS-L cells expressing CK1a fused to an ePL tag were dispensed into pre-compounded 384-well plates (Corning No. 3570). Compounds were dispensed into the 384-well plates using an acoustic dispenser (ATS Acoustic Dispensing System from EDC Biosystems) using a 10-point dose-response curve with a 3-fold dilution starting at 10 μM and decreasing to 0.0005 μM. Next, 25 μL of culture 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. The assay plates were incubated at 37°C, 5% CO₂ for 4 hours, while the GSPT1 assay plates were incubated for 20 hours. Following incubation, 25 μL of InCELL Hunter Detection Reagent Working Solution (DiscoverX, catalog number 96-0002, Fremont, CA) was added to each well and incubated at room temperature in the dark for 60 minutes. After 60 minutes, fluorescence was read on an Envision or PHERAstar fluorescence plate reader.

[0228] For Helios, a stable Jurkat cell line was engineered using CRISPR / Cas9 to insert an in-frame HiBit tag into the carboxyl-terminal reading frame of the IKZF2 gene. The sequence of human Helios is shown below: METEAIDGYI TCDNELSPER EHSNMAIDLT SSTPNGQHAS PSHMTSTNSV KLEMQSDEECDRKPLSREDE IRGHDEGSSL EEPLIESSEV ADNRKVQELQ GEGGIRLPNG KLKCDVCGMV CIGPNVLMVHKRSHTGERP FHCNQCGASFTQKGNLLRHIK LHSGEKPFKC PFCSYACRRR DALTGHLRTH SVGKPHKCNYCGRSYKQRSS LEEHKERCHN YLQNVSMEAA GQVMSHHVPP MEDCKEQEPI MDNNISLVPF ERPAVIEKLTGNMGKRKSST PQKFVGEKLM RFSYPDIHFD MNLTYEKEAE LMQSHMMDQA INNAITYLGA EALHPLMQHPPSTIAEVAPV ISSAYSQVYH PNRIERPISR ETADSHENNM DGPISLIRPK SRPQEREASP SNSCLDSTDSESSHDDHQSY QGHPALNPKR KQSPAYMKED VKALDTTKAP KGSLKDIYKV FNGEGEQIRA FKCEHCRVLFLDHVMYTIHM GCHGYRDPLE CNICGYRSQD RYEFSSHIVR GEHTFH (SEQ ID NO: 4; ZNF2 and ZNF3 are underlined).

[0229] The test compound was transferred to a 1536-well plate using an acoustic dispenser and seeded at 10,000 cells / well of Jurkat / Helios / HiBit cells in DMEM / 10% FCS in a final volume of 5 μL. The cells were incubated at 37°C, 95% RH for 18 hours. Luciferase activity was measured by adding 2 μL / well of Nano-Glo reagent (Promega) and incubating for 30 min at RT. Fluorescence values ​​were read on a microtiter plate reader.

[0230] To determine the EC50 value (the concentration of compound that achieves half of the observed maximum degradation) of a compound for degradation of a given substrate, a four-parameter logistic model (sigmoidal dose-response model) (FIT = (A + {(B-A) / 1 + [(C / x)D]})) was used, where C was the turning point (EC50), D was the correlation coefficient, and A and B were the lower and upper limits of the fit, respectively. All substrate degradation curves were processed and evaluated using the data analysis software package ActivityBase (IDBS). Ymax is the % degraded protein (Ymin = 100 - Ymax and is the minimum percentage of remaining protein).

[0231] Some results are shown in Table 8. Table 8. *nd = not determined.

[0232] Although the present invention has been described in some detail by way of illustration and example for clarity of understanding, these descriptions and examples should not be construed as limiting the scope of the present invention. The disclosures of all patents and scientific literature cited herein are expressly incorporated herein by reference in their entirety.

Claims

1. Compound of formula (I): or a pharmaceutically acceptable salt thereof, wherein: R 1 is H or oxo; Each R 2 are independently H or halogen, wherein at least one R 2 For fluorine; 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 optionally substituted C5-C6 cycloalkyl, optionally substituted C5-C6 aryl, optionally substituted 6-10 membered heterocyclyl, or optionally substituted 5-9 membered heteroaryl, wherein the heterocyclyl or heteroaryl contains 1-3 heteroatoms selected from nitrogen, oxygen and sulfur.

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

3. The compound according to claim 1 or a pharmaceutically acceptable salt thereof, wherein: The compound of formula (I) is a compound of formula (Ib):

4. The compound according to claim 1 or a pharmaceutically acceptable salt thereof, wherein: R 1 For H.

5. The compound according to claim 1 or a pharmaceutically acceptable salt thereof, wherein: R 1 For oxygen.

6. The compound according to claim 1 or a pharmaceutically acceptable salt thereof, wherein: At least one R 2 For H.

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

8. The compound according to claim 7 or a pharmaceutically acceptable salt thereof, wherein: One or two R 2 For F.

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

10. The compound according to any one of claims 1 to 8, or a pharmaceutically acceptable salt thereof, wherein: X is -CH2-.

11. The compound according to any one of claims 1 to 8, or a pharmaceutically acceptable salt thereof, wherein: X is -C(O)NHCH2-.

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. The compound according to any one of claims 1 to 8, or a pharmaceutically acceptable salt thereof, wherein: X is -CH2NHCH2-.

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 cyclohexyl group.

16. The compound according to claim 15 or a pharmaceutically acceptable salt thereof, wherein: Ring A is 17. The compound according to any one of claims 1 to 14, or a pharmaceutically acceptable salt thereof, wherein: Ring A is an optionally substituted C6 aryl group.

18. The compound according to claim 17 or a pharmaceutically acceptable salt thereof, wherein: Ring A is 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 6-10 membered heterocyclic group containing 1-3 heteroatoms selected from nitrogen, oxygen and sulfur.

20. The compound according to claim 19 or a pharmaceutically acceptable salt thereof, wherein: Ring A is where R 3 is H or OH; R 4 is optionally substituted C1-C3 alkyl, optionally substituted C3-C6 cycloalkyl, -C(O)(6-membered heteroaryl) or -C(O)(9-membered heterocyclyl); R 5 is an optionally substituted C1-C3 alkyl group; and R 6 It is a cyano group or a halide group.

21. The compound according to any one of claims 1 to 14 and 20, or a pharmaceutically acceptable salt thereof, wherein: Ring A is 22. The compound according to claim 21, wherein: for 23. The compound according to claim 21, wherein: for 24. The compound according to any one of claims 1 to 14, or a pharmaceutically acceptable salt thereof, wherein: Ring A is an optionally substituted 5-9 membered heteroaryl group containing 1-3 heteroatoms selected from nitrogen, oxygen and sulfur.

25. The compound according to claim 24 or a pharmaceutically acceptable salt thereof, wherein: Ring A is where R 7 is an optionally substituted C1-C3 alkyl group or an optionally substituted amine, and n is 0, 1, 2, 3 or 4.

26. The compound according to claim 25 or a pharmaceutically acceptable salt thereof, wherein: for 27. The compound according to any one of claims 1-3 and 5-26, or a pharmaceutically acceptable salt thereof, wherein the compound is of formula (IIa):

28. The compound according to any one of claims 1-4 and 6-26, or a pharmaceutically acceptable salt thereof, wherein the compound is of formula (IIb):

29. The compound according to any one of claims 1-3 and 5-26, or a pharmaceutically acceptable salt thereof, wherein the compound is of formula (IIc):

30. The compound according to any one of claims 1-3 and 5-26, or a pharmaceutically acceptable salt thereof, wherein the compound is of formula (IId):

31. The compound according to any one of claims 1-3, 5-14 and 19-21, or a pharmaceutically acceptable salt thereof, wherein the compound is of formula (IIIa): where R 8 is an optionally substituted C1-C6 alkyl group or an optionally substituted C3-C6 cycloalkyl group.

32. The compound according to any one of claims 1-3, 5-14 and 19-21, or a pharmaceutically acceptable salt thereof, wherein the compound is of formula (IIIb): where R 8 is an optionally substituted C1-C6 alkyl group or an optionally substituted C3-C6 cycloalkyl group.

33. The compound according to any one of claims 1-3, 5-14 and 19-21, or a pharmaceutically acceptable salt thereof, wherein the compound is of formula (IIIc): where R 8 is an optionally substituted C1-C6 alkyl group or an optionally substituted C3-C6 cycloalkyl group.

34. The compound according to any one of claims 1-3, 5-14 and 19-21, or a pharmaceutically acceptable salt thereof, wherein the compound is of formula (IIId): where R 8 is an optionally substituted C1-C6 alkyl group or an optionally substituted C3-C6 cycloalkyl group.

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

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

38. The method of claim 37, wherein the cells are human effector cells.

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

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

41. The method of any one of claims 37-40, wherein the engineered peptide comprises a degradation domain.

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

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

44. The method of claim 43, wherein the cells are human effector cells.

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

46. ​​The method of any one of claims 43-45, wherein the engineered peptide comprises a degradation domain.

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

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

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

50. 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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