Substituted imidazole-based compounds as ligand directed degradation agents for IRAK3

Abstract

Provided herein are compounds and compositions thereof for modulating IRAK3. 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 priority to U.S. Provisional Application No. 63 / 434,197, filed on December 21, 2022, which is incorporated herein by reference in its entirety for all purposes. Technical Field

[0002] The present disclosure generally relates to compounds, compositions, and methods of making and using the compounds and compositions for treating cancer. Background Art

[0003] The recruitment of immune cells to sites of injury involves the coordinated interplay of numerous soluble mediators. Several cytokines, including interleukin-1 (IL-1), appear to play a key role in these processes. IL-1 generates a proinflammatory response and contributes to the tissue degeneration processes observed in chronic inflammatory states. IL-1 has also been implicated in bone resorption and the regulation of adipose tissue. Thus, IL-1 plays a key role in a variety of pathological conditions, including rheumatoid arthritis, inflammatory bowel disease, multiple sclerosis, diabetes, obesity, cancer, and sepsis.

[0004] IL-1 treatment of cells results in the formation of a complex consisting of two IL-1 receptor chains (IL-1R1 and IL-1RAcP), and the resulting dimer recruits an adaptor molecule called MyD88, which binds to the IL-1 receptor-associated kinase (IRAK) (Wesche et al., J. Biol. Chem. 1999, 274, 19403-19410; O'Neill et al., J. Leukoc. Biol. 1998, 63, 650-657; Auron, Cytokine Growth Factor Rev. 1998, 9: 221-237; and O'Neill, Biochem. Soc. Trans. 2000, 28, 557-563). Four members of the IRAK family have been identified: IRAK1, IRAK2, IRAK3, and IRAK4. These proteins are characterized by a typical N-terminal death domain that mediates interaction with the MyD88 family adaptor proteins and a centrally located kinase domain. Of the four members of the mammalian IRAK family, IRAK2 and IRAK3 are considered to be catalytically inactive pseudokinases (Wesche et al., J.Biol.Chem.1999,274,19403-19410), but the specific roles of these two kinases are still largely unknown (Lagne et al., Structure 2021,29,238-251). Nevertheless, there are reports that IRAK3 is associated with the negative regulation of TLR (toll-like receptor) signaling pathways involved in microbial detection and protection of multicellular organisms from infection (Kobayashi et al., Cell 2002,110,191-202). More recent studies have revealed associations between mutations or high expression levels of IRAK3 and various diseases such as asthma and cancer (Balaci et al., Am. J. Hum. Genet. 2007, 80(6), 1103-1114; Kesselring, R. Cancer Cell 2016, 29(5), 685-696), indicating the potential of IRAK3 as a drug target and the need for small molecules that bind to IRAK3.

[0005] Protein degradation is a highly regulated process crucial for maintaining cellular homeostasis. Selective recognition and removal of damaged, misfolded, or excess proteins is achieved through the ubiquitin-proteasome pathway (UPP). The UPP is key to the regulation of virtually all cellular processes. Protein ubiquitination is accomplished by E3 ubiquitin ligases, which bind to proteins and add ubiquitin molecules to them, thereby marking them for proteasomal degradation.

[0006] The use of UPP for therapeutic applications has received significant attention (Zhou et al., Mol. Cell 2000, 6, 751-756). A promising treatment uses the proteolytic action of targeted chimeras (commonly referred to as PROTACs) to effectively remove unwanted proteins by protein degradation (Scheepstra et al., Comp. Struct. Biotech. J. 2019, 17, 160-176). PROTACS are ligand-directed degraders that bring together E3 ligases and target proteins to be degraded. These bivalent molecules are generally composed of E3 ligase ligands connected to small molecules that bind to target proteins through a connecting portion. PROTAC places E3 ligases at an appropriate distance and direction from the target protein, thereby ubiquitinating the target protein. The ubiquitinated target protein is then recognized and degraded by the proteasome.

[0007] Thus, in one aspect, provided herein are compounds that target IRAK3 for degradation. Summary of the Invention

[0008] In certain embodiments, described herein are compounds and compositions thereof for use in degrading IRAK3. In various embodiments, the compounds and compositions thereof can be used to treat cancer.

[0009] A more complete understanding of the embodiments of the present invention may be obtained by reference to the detailed description and examples which are intended to illustrate and not limit the embodiments.

[0010] Embodiment A1 is a compound of formula (I): or a pharmaceutically acceptable salt thereof, wherein: A is a C1-C6 alkyl group, a phenyl group, a C3-C6 cycloalkyl group, a 5- to 6-membered heteroaryl group, or a 6- to 10-membered heterocyclic group, wherein the phenyl group, the cycloalkyl group, the heteroaryl group, and the heterocyclic group are substituted by x R 1 Group substituted, and wherein the heteroaryl and heterocyclic groups contain 1-3 heteroatoms selected from N and O; Each R 1 independently halo, C1-C6 alkyl, C3-C6 cycloalkyl, C1-C6 alkoxy, C1-C6 haloalkyl or -SO2(C1-C6 alkyl); or two R on adjacent carbon atoms 1 The groups together form a fused C3-C6 cycloalkyl or a fused group; R a and R b Each is H or together form an oxo group; R c is H or C1-C6 alkyl; x is 0-5; R 2 is H or C1-C6 alkyl; R 3 is H or C1-C6 alkyl; R 4 is H or C1-C6 alkyl; X 1 is CH or N; X 2 N or CH2; Ring B is a C3-C6 cycloalkylene group or a 5- to 7-membered heterocyclylene group containing 1 or 2 nitrogen atoms; Each R 5 are independently halo, C1-C6 alkyl or C1-C6 haloalkyl; w is 0-5; L 1 -C(O)(CH2) n -、-(CH2) n -or-(CH2) n C(O)-; n is 1-6; Ring C is a 5- to 10-membered heterocyclylene group containing 1 or 2 nitrogen atoms; Each R 6 are independently halo, C1-C6 haloalkyl or C1-C6 alkyl; y is 0-5; Ring D is R 7a and R 7b Each is H or together form an oxo group; Each R 8 are independently halo, C1-C6 alkyl, C1-C6 haloalkyl or C1-C6 alkoxy; z is 0-4; X 3 N or CR 9 ; R 9 is H or C1-C6 alkyl; R 10 is H or C1-C6 alkyl; Each R 11 are independently halo, C1-C6 alkyl or C1-C6 haloalkyl; v is 0-4; and Each are independently a single bond or a double bond.

[0011] Embodiment A2 is a compound according to Embodiment A1 or a pharmaceutically acceptable salt thereof, wherein: A is C1-C3 alkyl, phenyl, C3-C5 cycloalkyl, 5- to 6-membered heteroaryl, or 8- to 10-membered heterocyclyl, wherein the phenyl, cycloalkyl, heteroaryl, and heterocyclyl groups are substituted by x R 1 group substitution; x is 0-3; and Each R 1 independently halo, C1-C3 alkyl, C3-C5 cycloalkyl, C1-C3 alkoxy, C1-C3 haloalkyl or -SO2(C1-C3 alkyl); or two R on adjacent carbon atoms 1 The groups together form a fused C3-C5 cycloalkyl or a fused group; R a and R b are each H or together form an oxo group; and R c is H or C1-C3 alkyl.

[0012] Embodiment A3 is a compound according to Embodiment A1 or A2, or a pharmaceutically acceptable salt thereof, wherein: A is

[0013] Embodiment A4 is a compound according to any one of Embodiments A1-A3, or a pharmaceutically acceptable salt thereof, wherein: R 2 is H or C1-C3 alkyl; R 3 is H or C1-C3 alkyl; and R 4 is H or C1-C3 alkyl.

[0014] Embodiment A5 is a compound according to any one of Embodiments A1-A4, or a pharmaceutically acceptable salt thereof, wherein: X 1 is N.

[0015] Embodiment A6 is a compound according to any one of Embodiments A1-A5, or a pharmaceutically acceptable salt thereof, wherein: for

[0016] Embodiment A7 is a compound according to any one of Embodiments A1-A6, or a pharmaceutically acceptable salt thereof, wherein: Ring B is a C4-C6 cycloalkylene group or a 6- to 7-membered heterocyclylene group containing one nitrogen atom; w is 0-2; and Each R 5 are independently halo, C1-C3 alkyl or C1-C3 haloalkyl.

[0017] Embodiment A8 is a compound according to any one of Embodiments A1-A7, or a pharmaceutically acceptable salt thereof, wherein: for

[0018] Embodiment A9 is a compound according to any one of Embodiments A1-A8, or a pharmaceutically acceptable salt thereof, wherein: L 1 -C(O)CH2-, -(CH2) n - or -CH2C(O)-; and n is 1-5.

[0019] Embodiment A10 is a compound according to any one of Embodiments A1-A9, or a pharmaceutically acceptable salt thereof, wherein: Ring C is a 6- to 8-membered heterocyclylene group containing 1 or 2 nitrogen atoms; y is 0-3; and Each R 6 are independently halo, C1-C3 haloalkyl or C1-C3 alkyl.

[0020] Embodiment A11 is a compound according to any one of Embodiments A1-A10, or a pharmaceutically acceptable salt thereof, wherein: for

[0021] Embodiment A12 is a compound according to any one of Embodiments A1-A11, or a pharmaceutically acceptable salt thereof, wherein: Ring D is

[0022] Embodiment A13 is a compound according to any one of Embodiments A1-A12, or a pharmaceutically acceptable salt thereof, wherein: X 3 CR 9 ; R 9 is H or C1-C3 alkyl; R 10 is H or C1-C3 alkyl; v is 0-2; and Each R 11 are independently halo, C1-C3 alkyl or C1-C3 haloalkyl.

[0023] Embodiment A14 is a compound according to any one of Embodiments A1-A13, or a pharmaceutically acceptable salt thereof, wherein: for

[0024] Embodiment A15 is a compound according to any one of Embodiments A1-A14, or a pharmaceutically acceptable salt thereof, wherein the compound is of Formula (IIIa), (IIIb) or (IIIc):

[0025] Embodiment A16 is a compound according to any one of Embodiments A1-A15, or a pharmaceutically acceptable salt thereof, wherein the compound is of Formula (IVa) or (IVb):

[0026] Embodiment A17 is a compound selected from the compounds of Table 1 or a pharmaceutically acceptable salt thereof.

[0027] Embodiment A18 is a pharmaceutical composition comprising a compound according to any one of Embodiments A1 to A17 or a pharmaceutically acceptable salt thereof and a pharmaceutically acceptable excipient.

[0028] Embodiment A19 is a method of modulating interleukin-1 receptor-associated kinase 3 (IRAK3), comprising contacting IRAK3 with an effective amount of a compound according to any one of embodiments A1-A17 or a pharmaceutically acceptable salt thereof, or a pharmaceutical composition according to embodiment A18.

[0029] Embodiment A20 is (i) a method of treating cancer in a subject in need thereof, comprising administering to the subject an effective amount of a compound according to any one of Embodiments A1-A17, or a pharmaceutically acceptable salt thereof, or a pharmaceutical composition according to Embodiment A18, optionally wherein the cancer is selected from bladder cancer, breast cancer, esophageal cancer, colon cancer, head and neck cancer, kidney cancer, lung cancer, pancreatic cancer, prostate cancer, melanoma, and gastric cancer; or (ii) a method of enhancing immunity in a vaccinated subject, comprising administering to the subject an effective amount of a compound according to any one of Embodiments A1-A17, or a pharmaceutically acceptable salt thereof, or a pharmaceutical composition according to Embodiment A18. DETAILED DESCRIPTION definition

[0030] As used herein, the terms "include" and "comprising" are used interchangeably. The terms "include" and "comprising" 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 "include" and "comprising" are intended to encompass examples encompassed by the term "consisting of." Therefore, the term "consisting of" can be used in place of the terms "include" and "comprising" to provide more specific embodiments of the present invention.

[0031] The term "consisting of means that the subject matter has at least 90%, 95%, 97%, 98%, or 99% of the features or components of which it is composed. In another embodiment, the term "consisting of excludes any other features or components from the scope of any subsequent recitation, except those that are not essential for the technical effect to be achieved.

[0032] 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.

[0033] 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, "about" and "approximately" mean ±20%, ±10%, ±5%, or ±1% of the referenced range, value, or structure.

[0034] 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 non-cyclic hydrocarbon having 1 to 8 carbon atoms (C1-C8 alkyl), or in some embodiments, 1 to 6 (C1-C6 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, etc. 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 an alkyl group as described herein is referred to as "substituted," it 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 (=0); amino, alkylamino, cycloalkylamino, arylamino, heterocyclylamino, heteroarylamino, heterocyclylalkylamino, cycloalkylalkylamino , arylalkylamino, heterocyclylalkylamino, heteroarylalkylamino, heterocyclylalkylalkylamino; imino; imido; amidino; guanidino; enamino; acylamino; sulfonylamino; urea, nitrourea; oxime; hydroxyamino; alkoxyamino; arylalkyloxyamino; 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 an alkyl group described herein is referred to as "substituted," it 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.

[0035] 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. 10 In 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, and hexadienyl. The cycloalkyl group may be substituted or unsubstituted. These substituted cycloalkyl groups include, for example, cyclohexanol, etc.

[0036] A "cycloalkylene" group refers to a divalent "cycloalkyl" group.

[0037] "Heterocyclyl" is a non-aromatic cycloalkyl group in which 1 to 4 of the 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 rings 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 connected 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.

[0038] A "heterocyclylene" group refers to a divalent "heterocyclyl" group.

[0039] 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, etc. The aryl group 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.).

[0040] "Heteroaryl" groups are aromatic ring systems having 1 to 4 heteroatoms as ring atoms in the heteroaromatic ring system, with the remaining atoms being carbon atoms. In some embodiments, the heteroaryl group contains 3 to 6 ring atoms, and in other embodiments contains 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, ...

[0041] "Halogen" or "halo" is fluorine, chlorine, bromine or iodine.

[0042] An "alkoxy" group is an -O-(alkyl) group wherein alkyl is as defined above.

[0043] "Haloalkyl" refers to an alkyl group as defined above, which is substituted with one or more halo substituents as defined above, for example, trifluoromethyl, difluoromethyl, trichloromethyl, 2,2,2-trifluoroethyl, 1,2-difluoroethyl, 3-bromo-2-fluoropropyl, 1,2-dibromoethyl, etc. In some embodiments, the haloalkyl group has 1 to 6 carbon atoms and is substituted with one or more halo groups (C1-C6 haloalkyl), or the haloalkyl group has 1 to 3 carbon atoms and is substituted with one or more halo groups (C1-C3 haloalkyl). The halo groups may be all the same or the halo groups may be different. Unless otherwise specifically stated, the haloalkyl group may be optionally substituted.

[0044] When groups described herein (other than alkyl groups) 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, It 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); monocyclic or fused or non-fused polycyclic or heteroaryl (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); aryloxy; aralkyloxy; heterocyclyloxy; and heterocyclylalkoxy.

[0045]

[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).

[0046] 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 inorganic bases and organic acids and organic 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 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).

[0047] As used herein and unless otherwise indicated, the term "stereoisomer" or "stereomerically pure" means 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 are included in the embodiments disclosed herein, including mixtures thereof.

[0048] 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 OpticalResolutions 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).

[0049] 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.

[0050] "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:

[0051] 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.

[0052] 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 radiolabeled with a radioactive isotope, such as, for example, tritium ( 3 H), iodine-125 ( 125 I), sulfur-35( 35 S) or carbon-14 ( 14 C) 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.

[0053] 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.

[0054] 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.

[0055] 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.

[0056] 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.

[0057] 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.

[0058] 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, in one embodiment mammals, in another embodiment humans. In one embodiment, the subject is a human having or at risk of having an IRAK3-mediated disease or a symptom thereof.

[0059] 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

[0060] In one aspect, provided herein are compounds of formula (I): or a pharmaceutically acceptable salt thereof, wherein: A is a C1-C6 alkyl group, a phenyl group, a C3-C6 cycloalkyl group, a 5- to 6-membered heteroaryl group, or a 6- to 10-membered heterocyclic group, wherein the phenyl group, the cycloalkyl group, the heteroaryl group, and the heterocyclic group are substituted by x R 1 Group substituted, and wherein the heteroaryl and heterocyclic groups contain 1-3 heteroatoms selected from N and O; Each R 1 independently halo, C1-C6 alkyl, C3-C6 cycloalkyl, C1-C6 alkoxy, C1-C6 haloalkyl or -SO2(C1-C6 alkyl); or two R on adjacent carbon atoms 1 The groups together form a fused C3-C6 cycloalkyl or a fused group; R a and R b Each is H or together form an oxo group; R c is H or C1-C6 alkyl; x is 0-5; R 2 is H or C1-C6 alkyl; R 3 is H or C1-C6 alkyl; R 4 is H or C1-C6 alkyl; X 1 is CH or N; X 2 N or CH2; Ring B is a C3-C6 cycloalkylene group or a 5- to 7-membered heterocyclylene group containing 1 or 2 nitrogen atoms; Each R 5 are independently halo, C1-C6 alkyl or C1-C6 haloalkyl; w is 0-5; L 1 -C(O)(CH2) n -、-(CH2) n -or-(CH2) n C(O)-; n is 1-6; Ring C is a 5- to 10-membered heterocyclylene group containing 1 or 2 nitrogen atoms; Each R 6 are independently halo, C1-C6 haloalkyl or C1-C6 alkyl; y is 0-5; Ring D is R 7a and R 7b Each is H or together form an oxo group; Each R 8 are independently halo, C1-C6 alkyl, C1-C6 haloalkyl or C1-C6 alkoxy; z is 0-4; X 3 N or CR 9 ; R 9 is H or C1-C6 alkyl; R 10 is H or C1-C6 alkyl; Each R 11 are independently halo, C1-C6 alkyl or C1-C6 haloalkyl; v is 0-4; and Each are independently a single bond or a double bond.

[0061] In some embodiments, A is C1-C6 alkyl, phenyl, C3-C6 cycloalkyl, 5- to 6-membered heteroaryl, or 6- to 10-membered heterocyclyl, wherein the phenyl, cycloalkyl, heteroaryl, and heterocyclyl are substituted by x R 1 In some embodiments, A is C1-C3 alkyl, phenyl, C3-C5 cycloalkyl, 5- to 6-membered heteroaryl, or 9- to 10-membered heterocyclyl, wherein the phenyl, cycloalkyl, heteroaryl, and heterocyclyl are substituted with x R 1 Group substitution.

[0062] In some embodiments, A is C1-C6 alkyl. In some embodiments, A is C1-C3 alkyl. In some embodiments, A is methyl, ethyl, n-propyl, or isopropyl. In some embodiments, A is methyl. In some embodiments, A is ethyl. In some embodiments, A is n-propyl. In some embodiments, A is isopropyl. In some embodiments, A is methyl or ethyl.

[0063] In some embodiments, A is x R 1 substituted phenyl groups.

[0064] In some embodiments, A is x R 1 In some embodiments, A is a C3-C6 cycloalkyl group substituted with a C3-C5 cycloalkyl group. In some embodiments, A is a cyclopropyl group, a cyclobutyl group, a cyclopentyl group, or a cyclohexyl group. In some embodiments, R 0In some embodiments, A is a saturated C3-C6 cycloalkyl. In some embodiments, A is a partially unsaturated C3-C6 cycloalkyl. In some embodiments, A is a cyclopropylene, cyclobutylene, cyclopentylene, or cyclohexylene. In some embodiments, A is a cyclopentylene or cyclohexylene. In some embodiments, A is a cyclopentylene. In some embodiments, A is a fused bicyclic C4-C6 cycloalkyl. In some embodiments, A is a fused bicyclic C4 cycloalkyl. In some embodiments, A is a fused bicyclic C5 cycloalkyl. In some embodiments, A is a fused bicyclic C6 cycloalkyl. In some embodiments, A is a cyclopropyl fused to a cyclobutyl or cyclopentyl ring. In some embodiments, A is a cyclopropyl fused to a cyclopentyl ring. In any of these variations, the cycloalkyl group is fused to a cyclobutyl or cyclopentyl ring. 1 Group substitution.

[0065] In some embodiments, A is a 5- to 6-membered heteroaryl group containing 1-3 heteroatoms selected from N and O and substituted by x R 1 In some embodiments, A is a 5-membered heteroaryl group containing 1-3 heteroatoms selected from N and O. In some embodiments, A is a 6-membered heteroaryl group containing 1-3 heteroatoms selected from N and O. In some embodiments, the heteroaryl group contains 1-2 heteroatoms selected from N and O. In some embodiments, the heteroaryl group contains 1 heteroatom selected from N and O. In some embodiments, the heteroaryl group contains one oxygen atom. In some embodiments, the heteroaryl group contains one nitrogen atom. In some embodiments, the heteroaryl group contains two heteroatoms selected from N and O. In some embodiments, the heteroaryl group contains one nitrogen atom and one oxygen atom. In some embodiments, the heteroaryl group contains two nitrogen atoms. In some embodiments, A is pyridyl, pyrazinyl, pyrimidinyl, pyridazinyl, triazinyl, pyrrolyl, imidazolyl, pyrazolyl, triazolyl, oxazolyl, or isoxazolyl. In some embodiments, A is pyrazolyl. In any of these variations, the heteroaryl group is substituted with x R 1 Group substitution.

[0066] In some embodiments, A is a 6- to 10-membered heterocyclyl containing 1-3 heteroatoms selected from N and O and substituted by x R 1 In some embodiments, A is an 8- to 10-membered heterocyclic group containing 1-3 heteroatoms selected from N and O and substituted by x R 1 In some embodiments, A is an 8- to 9-membered heterocyclic group containing 1-3 heteroatoms selected from N and O and substituted by x R 1 In some embodiments, A is a 9- to 10-membered heterocyclic group containing 1-3 heteroatoms selected from N and O and substituted by x R1 In some embodiments, A is an 8-membered heterocyclic group containing 1-3 heteroatoms selected from N and O and substituted by x R 1 In some embodiments, A is a 9-membered heterocyclic group containing 1-3 heteroatoms selected from N and O and substituted by x R 1 In some embodiments, A is a 10-membered heterocyclic group containing 1-3 heteroatoms selected from N and O and substituted by x R 1 In some embodiments, the heterocyclyl is a monocyclic heterocyclyl. In some embodiments, the heterocyclyl is a fused bicyclic heterocyclyl. In some embodiments, the fused bicyclic heterocyclyl contains a heterocyclyl fused to an aryl group. In some embodiments, the fused bicyclic heterocyclyl contains a heterocyclyl fused to a heteroaryl group. In any of these variations, the heterocyclyl is substituted with x R 1 Group substitution.

[0067] In some embodiments, x is 0-5. In some embodiments, x is 0-3. In some embodiments, x is 0. In some embodiments, x is 1. In some embodiments, x is 2. In some embodiments, x is 3. In some embodiments, x is 4. In some embodiments, x is 5.

[0068] In some embodiments, each R 1 is independently halo, C1-C6 alkyl, C3-C6 cycloalkyl, C1-C6 alkoxy, C1-C6 haloalkyl or -SO2(C1-C6 alkyl). 1 is independently halo, C1-C3 alkyl, C3-C5 cycloalkyl, C1-C3 alkoxy, C1-C3 haloalkyl, or -SO2(C1-C3 alkyl). 1 independently F, -CH3, cyclopropyl, -OCH3 or -SO2(CH3).

[0069] In some embodiments, R 1 In some embodiments, R 1 is Cl, F or Br. In some embodiments, R 1 In some embodiments, R 1 is F. In some embodiments, R 1 For Br.

[0070] In some embodiments, R 1 In some embodiments, R 1 In some embodiments, R 1is methyl, ethyl, n-propyl or isopropyl. In some embodiments, R 1 In some embodiments, R 1 In some embodiments, R 1 In some embodiments, R 1 It is isopropyl.

[0071] In some embodiments, R 1 is a C3-C6 cycloalkyl group. 1 In some embodiments, R 1 is cyclopropyl, cyclobutyl, cyclopentyl or cyclohexyl. In some embodiments, R 1 It is cyclopropyl.

[0072] In some embodiments, R 1 In some embodiments, R 1 In some embodiments, R 1 is -OCH3, -OCH2CH3, -OCH2CH2CH3 or -OCH(CH3)2. In some embodiments, R 1 In some embodiments, R 1 It is -OCH2CH3.

[0073] In some embodiments, R 1 is a C1-C6 haloalkyl group. 1 is a C1-C6 haloalkyl group containing 1 to 13 halogen atoms. 1 is a C1-C3 haloalkyl group. 1 is a C1-C3 haloalkyl group containing 1-7 halogen atoms. 1 In some embodiments, R 1 In some embodiments, R 1 is -CHF2.

[0074] In some embodiments, R 1 In some embodiments, R 1 is -SO2(C1-C3 alkyl). In some embodiments, R 1is -SO2(CH3), -SO2(CH2CH3) or -SO2(CH2CH2CH3). In some embodiments, R 1 It is -SO2(CH3).

[0075] In some embodiments, two R 1 The groups together form a fused C3-C6 cycloalkyl or a fused Group, where R a and R b Each is H or together form an oxo group, and R c is H or C1-C6 alkyl. In some embodiments, two R on adjacent carbon atoms 1 The groups together form a fused C3-C5 cycloalkyl or a fused Group, where R a and R b Each is H or together form an oxo group, and R c Is H or C1-C3 alkyl. In some embodiments, two R on adjacent carbon atoms 1 The groups together form fused cyclopropyl, cyclobutyl,

[0076] In some embodiments, two R 1 The groups together form a fused C3-C6 cycloalkyl group. In some embodiments, two R 1 The groups together form a fused C3-C5 cycloalkyl group. In some embodiments, two R 1 The groups together form a fused cyclopropyl, cyclobutyl or cyclopentyl. In some embodiments, two R 1 The groups together form a fused cyclopropyl or cyclobutyl group. In some embodiments, two R 1 In some embodiments, two R on adjacent carbon atoms 1 The groups are taken together to form a fused cyclobutyl group.

[0077] In some embodiments, two R 1 The groups are fused together Group, where R a and R b are each H or together form an oxo group, and R c is H or C1-C6 alkyl. In some embodiments, two R on adjacent carbon atoms 1 The groups are fused together Group, where R a and R bare each H or together form an oxo group, and R c is H or C1-C3 alkyl. In some embodiments, R a and R b Each is H. In some embodiments, R a and R b Together they form an oxo group. In some embodiments, R c is H. In some embodiments, R c is a C1-C6 alkyl group, such as a C1-C3 alkyl group. c is methyl, ethyl, n-propyl or isopropyl. In some embodiments, R c In some embodiments, two R on adjacent carbon atoms are 1 The groups are fused together

[0078] In some embodiments, A is: -CH3, or -CH2CH3.

[0079] In some embodiments, R 2 is H or C1-C6 alkyl. In some embodiments, R 2 is H or C1-C3 alkyl. In some embodiments, R 2 It is H or -CH3.

[0080] In some embodiments, R 2 For H.

[0081] In some embodiments, R 2 In some embodiments, R 2 In some embodiments, R 2 is methyl, ethyl, n-propyl or isopropyl. In some embodiments, R 2 In some embodiments, R 2 In some embodiments, R 2 In some embodiments, R 2 In some embodiments, R 2 It is methyl or ethyl.

[0082] In some embodiments, R 3 is H or C1-C6 alkyl. In some embodiments, R 3 is H or C1-C3 alkyl. In some embodiments, R 3 It is H or -CH3.

[0083] In some embodiments, R 3 For H.

[0084] In some embodiments, R 3 In some embodiments, R 3 In some embodiments, R 3 is methyl, ethyl, n-propyl or isopropyl. In some embodiments, R 3 In some embodiments, R 3 In some embodiments, R 3 In some embodiments, R 3 In some embodiments, R 3 It is methyl or ethyl.

[0085] In some embodiments, R 4 is H or C1-C6 alkyl. In some embodiments, R 4 is H or C1-C3 alkyl. In some embodiments, R 4 It is H or -CH3.

[0086] In some embodiments, R 4 For H.

[0087] In some embodiments, R 4 In some embodiments, R 4 In some embodiments, R 4 is methyl, ethyl, n-propyl or isopropyl. In some embodiments, R 4 In some embodiments, R 4 In some embodiments, R 4 In some embodiments, R 4 In some embodiments, R 4 It is methyl or ethyl.

[0088] In some embodiments, X 1 is CH or N. In some embodiments, X 1 In some embodiments, X 1 is N.

[0089] In some embodiments, X 2 is CH2 or N. In some embodiments, X 2 In some embodiments, X 2 is N.

[0090] In some embodiments, each is independently a single bond or a double bond. In some embodiments, Is a single bond. In some embodiments, For double bonds.

[0091] In some embodiments,

[0092] In some embodiments, ring B is C3-C6 cycloalkylene or a 5- to 7-membered heterocyclylene containing 1 or 2 nitrogen atoms. In some embodiments, ring B is C4-C6 cycloalkylene or a 6- to 7-membered heterocyclylene containing one nitrogen atom.

[0093] In some embodiments, Ring B is C3-C6 cycloalkylene. In some embodiments, Ring B is C4 cycloalkylene. In some embodiments, Ring B is C5 cycloalkylene. In some embodiments, Ring B is C6 cycloalkylene. In some embodiments, Ring B is cyclobutylene, cyclopentylene, or cyclohexylene. In some embodiments, Ring B is

[0094] In some embodiments, Ring B is a 5- to 7-membered heterocyclylene containing 1 or 2 nitrogen atoms. In some embodiments, Ring B is a 5-membered heterocyclylene containing 1 or 2 nitrogen atoms. In some embodiments, Ring B is a 6-membered heterocyclylene containing 1 or 2 nitrogen atoms. In some embodiments, Ring B is a 7-membered heterocyclylene containing 1 or 2 nitrogen atoms. In some embodiments, Ring B is a 6- to 7-membered heterocyclylene containing 1 nitrogen atom. In some embodiments, Ring B is a 6- to 7-membered heterocyclylene containing two nitrogen atoms. In some embodiments, Ring B is

[0095] In some embodiments, Ring B is:

[0096] In some embodiments, w is 0-5. In some embodiments, w is 0-2. In some embodiments, w is 0. In some embodiments, w is 1. In some embodiments, w is 2. In some embodiments, w is 3. In some embodiments, w is 4. In some embodiments, w is 5.

[0097] In some embodiments, each R 5 is independently halo, C1-C6 alkyl or C1-C6 haloalkyl. 5 is independently halo, C1-C3 alkyl or C1-C3 haloalkyl. 5 independently F, -CF3 or -CH3.

[0098] In some embodiments, R 5 In some embodiments, R 5 is Cl, F or Br. In some embodiments, R 5 In some embodiments, R 5 is F. In some embodiments, R 5 For Br.

[0099] In some embodiments, R 5 In some embodiments, R 5 In some embodiments, R 5 is methyl, ethyl, n-propyl or isopropyl. In some embodiments, R 5 In some embodiments, R 5 In some embodiments, R 5 In some embodiments, R 5 It is isopropyl.

[0100] In some embodiments, R 5 is a C1-C6 haloalkyl group. 5 is a C1-C6 haloalkyl group containing 1 to 13 halogen atoms. 5 is a C1-C3 haloalkyl group. 5 is a C1-C3 haloalkyl group containing 1-7 halogen atoms. 5 In some embodiments, R 5 In some embodiments, R 5 is -CHF2.

[0101] In some embodiments, for:

[0102] In some embodiments, L 1 -C(O)(CH2) n -、-(CH2) n -or-(CH2) n C(O)-. In some embodiments, L 1 -C(O)CH2-, -(CH2) n - or -CH2C(O)-, wherein n is 1-5. In some embodiments, L1 It is -C(O)CH2-, -CH2-, -CH2CH2-, -CH2CH2CH2-, -CH2CH2CH2CH2- or -CH2C(O)-.

[0103] In some embodiments, L 1 -C(O)(CH2) n -, wherein n is 1-6. In some embodiments, L 1 is -C(O)(CH2)-. In some embodiments, L 1 is -C(O)(CH2)2-. In some embodiments, L 1 is -C(O)(CH2)3-. In some embodiments, L 1 is -C(O)(CH2)4-. In some embodiments, L 1 is -C(O)(CH2)5-. In some embodiments, L 1 It is -C(O)(CH2)6-.

[0104] In some embodiments, L 1 -(CH2) n -, wherein n is 1-6. In some embodiments, L 1 In some embodiments, L 1 In some embodiments, L 1 is -(CH2)3-. In some embodiments, L 1 is -(CH2)4-. In some embodiments, L 1 is -(CH2)5-. In some embodiments, L 1 It is -(CH2)6-.

[0105] In some embodiments, L 1 -(CH2) n C(O)-, wherein n is 1-6. In some embodiments, L 1 is -(CH2)C(O)-. In some embodiments, L 1 is -(CH2)2C(O)-. In some embodiments, L 1 is -(CH2)3C(O)-. In some embodiments, L 1 is -(CH2)4C(O)-. In some embodiments, L 1 is -(CH2)5C(O)-. In some embodiments, L 1 It is -(CH2)6C(O)-.

[0106] In some embodiments, Ring C is a 5- to 10-membered heterocyclylene containing 1 or 2 nitrogen atoms. In some embodiments, Ring C is a 6- to 8-membered heterocyclylene containing 1 or 2 nitrogen atoms. In some embodiments, the heterocyclylene contains one nitrogen atom. In some embodiments, the heterocyclylene contains two nitrogen atoms. In some embodiments, the heterocyclylene is monocyclic. In some embodiments, the heterocyclylene is piperazinylene, piperidinylene, or pyrrolidinylene. In some embodiments, the heterocyclylene is polycyclic. In some embodiments, the heterocyclylene is spirocyclic.

[0107] In some embodiments, Ring C is:

[0108] In some embodiments, y is 0-5. In some embodiments, y is 0-3. In some embodiments, y is 0. In some embodiments, y is 1. In some embodiments, y is 2. In some embodiments, y is 3. In some embodiments, y is 4. In some embodiments, y is 5.

[0109] In some embodiments, each R 6 is independently halo, C1-C6 haloalkyl or C1-C6 alkyl. 6 is independently halo, C1-C3 haloalkyl or C1-C3 alkyl. 6 independently Cl, -CF3 or -CH3.

[0110] In some embodiments, R 6 In some embodiments, R 6 is Cl, F or Br. In some embodiments, R 6 In some embodiments, R 6 is F. In some embodiments, R 6 For Br.

[0111] In some embodiments, R 6 is a C1-C6 haloalkyl group. 6 is a C1-C6 haloalkyl group containing 1 to 13 halogen atoms. 6 is a C1-C3 haloalkyl group. 6 is a C1-C3 haloalkyl group containing 1-7 halogen atoms. 6In some embodiments, R 6 In some embodiments, R 6 is -CHF2.

[0112] In some embodiments, R 6 In some embodiments, R 6 In some embodiments, R 6 is methyl, ethyl, n-propyl or isopropyl. In some embodiments, R 6 In some embodiments, R 6 In some embodiments, R 6 In some embodiments, R 6 It is isopropyl.

[0113] In some embodiments, for:

[0114] In some embodiments, Ring D is: In some embodiments, Ring D is In some embodiments, Ring D is In some embodiments, Ring D is

[0115] In some embodiments, Ring D is where R 7a and R 7b Each is H or together form an oxo group. In some embodiments, ring D is where R 7a and R 7b Each is H. In some embodiments, Ring D is where R 7a and R 7b Together they form an oxo group. In some embodiments, ring D is In some embodiments, Ring D is

[0116] In some embodiments, Ring D is In some embodiments, Ring D is

[0117] In some embodiments, Ring D is

[0118] In some embodiments, z is 0-4. In some embodiments, z is 0-2. In some embodiments, z is 0. In some embodiments, z is 1. In some embodiments, z is 2. In some embodiments, z is 3. In some embodiments, z is 4.

[0119] In some embodiments, each R 8 is independently halo, C1-C6 alkyl, C1-C6 haloalkyl, or C1-C6 alkoxy. 8 R is independently halo, C1-C3 alkyl, C1-C3 haloalkyl or C1-C3 alkoxy. 8 independently F, Cl, -CH3, -OCH3 or -CF3.

[0120] In some embodiments, R 8 In some embodiments, R 8 is Cl, F or Br. In some embodiments, R 8 In some embodiments, R 8 is F. In some embodiments, R 8 For Br.

[0121] In some embodiments, R 8 In some embodiments, R 8 In some embodiments, R 8 is methyl, ethyl, n-propyl or isopropyl. In some embodiments, R 8 In some embodiments, R 8 In some embodiments, R 8 In some embodiments, R 8 It is isopropyl.

[0122] In some embodiments, R 8 is a C1-C6 haloalkyl group. 8 is a C1-C6 haloalkyl group containing 1 to 13 halogen atoms. 8 is a C1-C3 haloalkyl group. 8 is a C1-C3 haloalkyl group containing 1-7 halogen atoms. 8 In some embodiments, R 8In some embodiments, R 8 is -CHF2.

[0123] In some embodiments, R 8 In some embodiments, R 8 In some embodiments, R 8 is -OCH3, -OCH2CH3, -OCH2CH2CH3 or -OCH(CH3)2. In some embodiments, R 8 In some embodiments, R 8 It is -OCH2CH3.

[0124] In some embodiments, Ring D is:

[0125] In some embodiments, X 3 N or CR 9 In some embodiments, X 3 is N. In some embodiments, X 3 CR 9 .

[0126] In some embodiments, R 9 is H or C1-C6 alkyl. In some embodiments, R 9 is H or C1-C3 alkyl. In some embodiments, R 9 It is H or -CH3.

[0127] In some embodiments, R 9 For H.

[0128] In some embodiments, R 9 In some embodiments, R 9 In some embodiments, R 9 is methyl, ethyl, n-propyl or isopropyl. In some embodiments, R 9 In some embodiments, R 9 In some embodiments, R 9 In some embodiments, R 9 It is isopropyl.

[0129] In some embodiments, R 10 is H or C1-C6 alkyl. In some embodiments, R 10 is H or C1-C3 alkyl. In some embodiments, R 10 It is H or -CH3.

[0130] In some embodiments, R 10 For H.

[0131] In some embodiments, R 10 In some embodiments, R 10 In some embodiments, R 10 is methyl, ethyl, n-propyl or isopropyl. In some embodiments, R 10 In some embodiments, R 10 In some embodiments, R 10 In some embodiments, R 10 It is isopropyl.

[0132] In some embodiments, v is 0-4. In some embodiments, v is 0-2. In some embodiments, v is 0. In some embodiments, v is 1. In some embodiments, v is 2. In some embodiments, v is 3. In some embodiments, v is 4.

[0133] In some embodiments, each R 11 is independently halo, C1-C6 alkyl or C1-C6 haloalkyl. 11 is independently halo, C1-C3 alkyl or C1-C3 haloalkyl. 11 independently F, -CH3 or -CF3.

[0134] In some embodiments, R 11 In some embodiments, R 11 is Cl, F or Br. In some embodiments, R 11 In some embodiments, R 11 is F. In some embodiments, R 11 For Br.

[0135] In some embodiments, R 11 In some embodiments, R 11 In some embodiments, R 11 is methyl, ethyl, n-propyl or isopropyl. In some embodiments, R 11 In some embodiments, R 11 In some embodiments, R 11 In some embodiments, R 11 It is isopropyl.

[0136] In some embodiments, R 11 is a C1-C6 haloalkyl group. 11 is a C1-C6 haloalkyl group containing 1 to 13 halogen atoms. 11 is a C1-C3 haloalkyl group. 11 is a C1-C3 haloalkyl group containing 1-7 halogen atoms. 11 In some embodiments, R 11 In some embodiments, R 11 is -CHF2.

[0137] In some embodiments, for:

[0138] In some embodiments, the compound of formula (I) is a compound of formula (II): Among them, A, R 5 、R 6 ,w,y,L 1 , Ring B, Ring C and Ring D are as described in Formula (I).

[0139] In some embodiments, the compound of formula (I) is of formula (IIIa), (IIIb), or (IIIc): Among them, A, R 5 、R 6 、R 8 ,w,y,z,L 1 As described in formula (I).

[0140] In some embodiments, the compound of formula (I) is a compound of formula (IVa) or (IVb): where R 1 、R 5 、R 6 、R 8 , x, w, y, z, and L 1 As described in formula (I).

[0141] 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) 1 Each description, variation, embodiment or aspect may be combined with R 1 、R a 、R b 、R c 、R 2 、R 3 、R 4 、R 5 、R 6 、R 7a 、R 7b 、R 8 、R 9 、R 10 、R 11 、X 1 、X 2 、X 3 、L 1 , ring B, ring C, ring D, n, v, w, x, y and z each description, variation, embodiment or aspect combination, as each and each combination is specifically and individually listed.It should also be understood that, where applicable, all descriptions, variations, embodiments or aspects of formula (I) are equally applicable to other formulae described in detail herein, and are also described, as each and each description, variation, embodiment or aspect of all formulae are separated and individually listed.For example, where applicable, all descriptions, variations, embodiments or aspects of formula (I) are equally applicable to other formulae described in detail herein, such as formula (Ia), (II), (IIIa), (IIIb), (IIIc), (IVa) and (IVb) are also described, as each and each description, variation, embodiment or aspect of all formulae are separated and individually listed.

[0142] 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. or a pharmaceutically acceptable salt thereof.

[0143] 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.

[0144] In addition, all compounds of formula (I) that exist in free base or 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

[0145] 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 in the manner outlined in Schemes 1-14 and in the Examples listed herein. It should be noted that one skilled in the art will know how to modify the methods listed in the illustrative schemes and Examples to obtain the desired products.

[0146] Compounds of formula (I) can be prepared from two main building blocks: a target binding moiety (TBM) and a Cereblon-binding moiety (CBM). General synthetic routes for preparing TBM and CBM are outlined in Schemes 1-8. General synthetic routes for reacting TBM and CBM to form heterobifunctional molecules (i.e., ligand-directed degraders, LDDs), which are compounds of formula (I), are provided in Schemes 9-14. Intermediate compounds Solution 1. wherein Ring D is phenyl or pyrazole; G is N or CH; X is a halide such as Br; and Bn is benzyl.

[0147] Scheme 1 illustrates a method for synthesizing intermediate compound CBM-A. Intermediates Aa and Ab can be coupled in the presence of a palladium catalyst to produce intermediate Ac. Conversion of intermediate Ac to Ad, followed by deprotection, yields intermediate compound CBM-A. Option 2. wherein X is a halogen group such as Br; and R 6 、R 8 , y and z are as described in formula (I).

[0148] Scheme 2 illustrates a method for synthesizing intermediate compound CBM-B. Intermediate Ba and tert-butyl acrylate can be coupled in the presence of a base such as CsCO or KCO to produce intermediate Bb, which can be further coupled with intermediate Bc in the presence of a palladium catalyst to produce intermediate Bd. Bd forms an internal ring under acidic conditions, yielding intermediate compound CBM-B. Option 3. wherein ring D is phenyl or indazole, and G is N or C(CH3).

[0149] Scheme 3 illustrates the synthesis of intermediate compound CBM-C. Intermediate Ca and tert-butyl bis(2-oxoethyl)carbamate can be coupled in the presence of a reducing agent to produce intermediate Cb. Deprotection of intermediate Cb under acidic conditions yields intermediate compound CBM-C. Solution 4. Wherein Z is CH3, Cl or Br; X is a halogen such as Cl, Br or I; R is a chiral A, R 2 、R 3 、R 5 、X 1 , Ring B and w are as described in Formula (I).

[0150] Scheme 4 illustrates a method for synthesizing intermediate compound TBM-D. Intermediates Da and Db can be coupled under acidic conditions to produce intermediate Dc. Palladium-catalyzed coupling of intermediates Dc and Dd subsequently produces intermediate De, which is subsequently deprotected under acidic conditions to yield intermediate compound TBM-D. Option 5. wherein X is a halide such as Br; and A is as described in formula (I).

[0151] Scheme 5 illustrates a method for synthesizing intermediate compound TBM-E. Intermediates Ea and Eb can be coupled in the presence of a palladium catalyst to give intermediate compound TBM-E. Option 6. wherein each X is independently a halide such as Br or Cl; and A and ring B are as described for formula (I).

[0152] Scheme 6 illustrates a method for synthesizing the intermediate compound TBM-F. Intermediates Fa and Fb can be coupled under acidic conditions to produce intermediate Fc. Palladium-catalyzed coupling of intermediate Fc and Fd produces intermediate Fe, which is then deprotected under acidic conditions to yield intermediate compound TBM-F. Plan 7. wherein each X is independently a halide such as Br or Cl; R is an alkyl group; m is 0 or 1; Ring B is a cycloalkylene group; and A is as described in Formula (I).

[0153] Scheme 7 illustrates a method for synthesizing the intermediate compound TBM-G. Intermediate Ga can be reduced in the presence of a palladium catalyst to produce intermediate Gb. Intermediates Gb and Gc couple under acidic conditions to produce intermediate Gd, which then undergoes a palladium-catalyzed coupling reaction with intermediate Ge to produce intermediate Gf. The acid group of intermediate Gf is converted to an alcohol group to yield intermediate compound TBM-G. Plan 8. wherein X is a halide such as Br; and A and ring B are as described in formula (I).

[0154] Scheme 8 illustrates a method for synthesizing the intermediate compound TBM-H. Intermediates Ha and Hb can be coupled with a palladium catalyst under basic conditions to give the intermediate compound TBM-H. Final compound Plan 9. wherein A and ring D are as described in formula (I).

[0155] Scheme 9 illustrates a method for synthesizing compound LDD-a. Coupling of TBM-D' (a derivative of intermediate TBM-D of Scheme 4) and 2-bromoacetic acid under basic conditions can produce TBM-D", which can be further coupled with CBM-A' (a derivative of intermediate CBM-A of Scheme 1) under basic conditions to provide compound LDD-a, which is a compound of formula (I). Plan 10. wherein A and ring D are as described in formula (I).

[0156] Scheme 10 illustrates a method for synthesizing compound LDD-b. TBM-D' (a derivative of intermediate TBM-D in Scheme 4) and 2,2-dimethoxyacetaldehyde can be coupled under alkaline conditions with a reducing agent to produce TBM-D1. Treatment of TBM-D1 with a strong acid produces TBM-D2, which can be coupled with CBM-A' (a derivative of intermediate CBM-A in Scheme 1) under alkaline conditions with a reducing agent to provide compound LDD-b, which is a compound of formula (I). Plan 11. wherein A and ring D are as described in formula (I).

[0157] Scheme 11 illustrates an alternative synthesis of compound LDD-b. TBM-D1 can be reacted with a strong acid and subsequently coupled with CBM-A' under basic conditions and with a reducing agent to give compound LDD-b, which is a compound of formula (I). Plan 12. wherein A, ring B and ring D are as described in formula (I).

[0158] Scheme 12 illustrates a method for synthesizing compound LDD-c. The alcohol group of TBM-F (see Scheme 6) can be converted into a leaving group and then coupled with CBM-A' (a derivative of intermediate CBM-A in Scheme 1) under basic conditions and with a reducing agent to give compound LDD-c, which is a compound of formula (I). Plan 13. wherein A and ring D are as described in formula (I).

[0159] Scheme 13 illustrates a method for synthesizing compound LDD-d. CBM-A' (a derivative of intermediate CBM-A of Scheme 1) and 2-bromoacetic acid can be coupled under basic conditions to produce CBM-A". CBM-A" is then coupled with TBM-D' (a derivative of intermediate TBM-D of Scheme 4) under basic conditions to provide compound LDD-d, which is a compound of Formula (I). Plan 14. Among them, A, R 3 and Ring D are as described in Formula (I).

[0160] Scheme 14 illustrates a method for synthesizing compound LDD-e. CBM-A' (a derivative of intermediate CBM-A in Scheme 1) and 2,2-dimethoxyacetaldehyde can be coupled in the presence of a reducing agent, followed by reaction with a strong acid to produce CBM-A1. CBM-A1 is then coupled with TBM-D3 (a derivative of intermediate TBM-D in Scheme 4) in the presence of a reducing agent to provide compound LDD-e, which is a compound of Formula (I). How to use

[0161] Embodiments of the present disclosure provide methods for regulating IRAK3 in a subject in need thereof, comprising administering to the subject an effective amount of a compound of formula (I). Regulation (e.g., inhibition or activation) of IRAK3 can be evaluated and demonstrated by various methods known in the art. Kits and commercially available assays can be used to determine whether IRAK3 is regulated and to what extent (e.g., inhibition or activation).

[0162] In one aspect, provided herein are methods of modulating IRAK3, comprising contacting IRAK3 with an effective amount of a compound of formula (I), or any embodiment or variant thereof. In some embodiments, the compound of formula (I) inhibits IRAK3. In other embodiments, the compound of formula (I) activates IRAK3. In some embodiments, the compound of formula (I) causes degradation of IRAK3.

[0163] In some embodiments, provided herein are methods for targeting IRAK3 for degradation, comprising contacting IRAK3 with an effective amount of a compound of Formula (I), or any embodiment, or variant thereof.

[0164] In some embodiments, the compound of Formula (I) modulates the activity of IRAK3 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) modulate the activity of IRAK3 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%, 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%.

[0165] In certain embodiments of the present disclosure, a method for degrading IRAK3 in a subject in need thereof is also provided, comprising administering to the subject an effective amount of a compound of formula (I). IRAK3 degradation 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 IRAK3 is degraded and to what extent.

[0166] In one aspect, provided herein are methods of degrading IRAK3, comprising contacting IRAK3 with an effective amount of a compound of formula (I), or any embodiment or variant thereof. In some embodiments, the compound of formula (I) partially degrades IRAK3. In some embodiments, the compound of formula (I) completely degrades IRAK3.

[0167] In some embodiments, the compound of formula (I) degrades IRAK3 by about 1%, 5%, 10%, 15%, 20%, 25%, 30%, 35%, 40%, 45%, 50%, 55%, 60%, 65%, 70%, 75%, 80%, 85%, 90%, 95%, or 100%. In some embodiments, the compound of formula (I) degrades IRAK3 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%, 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%.

[0168] In another aspect, provided herein are methods for treating cancer in a subject in need thereof, comprising administering to the subject an effective amount of a compound of formula (I). In some embodiments, provided herein are methods for preventing cancer in a subject in need thereof, comprising administering to the subject an effective amount of a compound of formula (I). Non-limiting examples of cancer include bladder cancer, breast cancer, esophageal cancer, head and neck cancer, kidney cancer, lung cancer, pancreatic cancer, melanoma, and gastric cancer.

[0169] In some embodiments, administering a compound of formula (I) to a subject susceptible to cancer prevents the subject from developing any symptoms of the cancer (such as tumor growth or metastasis). In some embodiments, administering a compound of formula (I) to a subject who has not yet shown symptoms of cancer prevents the subject from developing any symptoms of the cancer. In some embodiments, administering a compound of formula (I) to a subject in need reduces the extent of the cancer in the subject. In some embodiments, administering a compound of formula (I) to a subject in need stabilizes the cancer (prevents or delays the worsening of the cancer). In some embodiments, administering a compound of formula (I) to a subject in need delays the appearance or recurrence of the cancer. In some embodiments, administering a compound of formula (I) to a subject in need slows down the progression of the cancer. In some embodiments, administering a compound of formula (I) to a subject in need provides partial relief of the cancer. In some embodiments, administering a compound of formula (I) to a subject in need provides complete relief of the cancer. In some embodiments, administering a compound of formula (I) to a subject in need reduces the dosage of one or more other drugs needed to treat the cancer. In some embodiments, administering a compound of formula (I) to a subject in need enhances the effect of another drug for treating the cancer. In some embodiments, administering a compound of formula (I) to a subject in need thereof delays the progression of the cancer. In some embodiments, administering a compound of formula (I) to a subject in need thereof improves the quality of life of a subject with cancer. In some embodiments, administering a compound of formula (I) to a subject in need thereof prolongs the survival of a subject with cancer.

[0170] In one aspect, provided herein are methods of preventing a subject susceptible to cancer from developing cancer, the methods comprising administering to the subject a compound of Formula (I).

[0171] In some aspects, provided herein are methods for reducing the extent of cancer in a subject, comprising administering a compound of formula (I) to the subject. In some embodiments, provided herein are methods for stabilizing cancer in a subject, comprising administering a compound of formula (I) to the subject. In some embodiments, the methods prevent worsening of the cancer.

[0172] In another aspect, provided herein are methods of delaying the onset or recurrence of cancer in a subject, comprising administering to the subject a compound of Formula (I).

[0173] In some embodiments, provided herein are methods of slowing the progression of a subject's cancer, comprising administering to the subject a compound of Formula (I). In some embodiments, the method provides partial remission of the cancer. In some embodiments, the method provides complete remission of the cancer.

[0174] In other aspects, provided herein are methods of reducing the dose of one or more other drugs used to treat cancer in a subject, the methods comprising administering to the subject a compound of formula (I). In some embodiments, provided herein are methods of enhancing the effect of another drug used to treat cancer in a subject, the methods comprising administering to the subject a compound of formula (I).

[0175] Also provided herein are methods for delaying the progression of a subject's cancer, comprising administering a compound of formula (I) to the subject. In some embodiments, the methods improve the quality of life of a subject suffering from cancer. In some embodiments, the methods prolong the survival of a subject suffering from cancer.

[0176] In some embodiments, compounds of Formula (I) are used in a method of treating a cancer selected from bladder cancer, breast cancer, esophageal cancer, colon cancer, head and neck cancer, kidney cancer, lung cancer, pancreatic cancer, prostate cancer, melanoma, and gastric cancer.

[0177] In some embodiments, provided herein are methods for enhancing the immunity of a subject receiving a vaccine, the methods comprising administering an effective amount of a compound of formula (I) to the subject. In some embodiments, the compound of formula (I) is administered to the subject before the vaccine is administered. In some embodiments, the compound of formula (I) is administered to the subject while the vaccine is administered. In some embodiments, the compound of formula (I) is administered to the subject after the vaccine is administered. In some embodiments, the compound of formula (I) is formulated as a component of the vaccine. In some embodiments, the compound of formula (I) is formulated separately from the vaccine. Pharmaceutical compositions and routes of administration

[0178] 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.

[0179] 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), Lubricant (e.g., magnesium stearate, light anhydrous silicic acid, talc or sodium lauryl sulfate), flavoring (e.g., citric acid, menthol, glycine or orange powder), preservative (e.g., sodium benzoate, sodium bisulfite, methylparaben or propylparaben), stabilizer (e.g., citric acid, sodium citrate or acetic acid), suspending agent (e.g., methylcellulose, polyvinylpyrrolidone or aluminum stearate), dispersant (e.g., hydroxypropyl methylcellulose), diluent (e.g., water) and base wax (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 produce 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.

[0180] The dosage of the compound of formula (I) applied to the subject varies greatly and can be judged by health care practitioners. Generally speaking, the compound disclosed herein can be applied 1 to 4 times a day with a dosage of about 0.001mg / kg subject weight to about 10mg / kg subject weight, but the above dosage can be appropriately changed according to the age, weight and medical condition of the subject and the type of application. In one embodiment, the dosage is about 0.001mg / kg subject weight to about 5mg / kg subject weight, about 0.01mg / kg subject weight to about 5mg / kg subject weight, about 0.05mg / kg subject weight to about 1mg / kg subject weight, about 0.1mg / kg subject weight to about 0.75mg / kg subject weight or about 0.25mg / kg subject weight to about 0.5mg / kg subject weight. In one embodiment, a dosage is given every day. In any given case, the amount of the compound of formula (I) applied will depend on factors such as the solubility of the active ingredient, the preparation used and the route of administration.

[0181] In some embodiments, the compound of Formula (I) is administered to a subject at a dose of about 0.01 mg / day to about 750 mg / day, about 0.1 mg / day to about 375 mg / day, about 0.1 mg / day to about 150 mg / day, about 0.1 mg / day to about 75 mg / day, about 0.1 mg / day to about 50 mg / day, about 0.1 mg / day to about 25 mg / day, or about 0.1 mg / day to about 10 mg / day.

[0182] In another embodiment, provided herein are unit dose formulations comprising about 0.1 mg to 500 mg, about 1 mg to 250 mg, about 1 mg to about 100 mg, about 1 mg to about 50 mg, about 1 mg to about 25 mg, or about 1 mg to about 10 mg of a compound of Formula (I).

[0183] In a specific embodiment, provided herein are unit dosage formulations comprising about 0.1 mg or 100 mg of a compound of Formula (I).

[0184] In another embodiment, provided herein are unit dose formulations comprising 0.5 mg, 1 mg, 5 mg, 10 mg, 15 mg, 20 mg, 30 mg, 35 mg, 50 mg, 70 mg, 100 mg, 125 mg, 140 mg, 175 mg, 200 mg, 250 mg, 280 mg, 350 mg, 500 mg, 560 mg, 700 mg, 750 mg, 1000 mg or 1400 mg of a compound of Formula (I).

[0185] The compound of formula (I) can be administered once, twice, three times, four times or more daily. In a specific embodiment, a dose of 100 mg or less is administered as a once daily dose, and a dose exceeding 100 mg is administered twice daily in an amount equal to half the total daily dose.

[0186] 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.

[0187] 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.

[0188] In one embodiment, provided herein are capsules containing a compound of Formula (I) without additional carriers, excipients, or vehicles.

[0189] In another embodiment, the compositions provided herein include an effective amount of a compound of formula (I) and a pharmaceutically acceptable carrier or vehicle, wherein the pharmaceutically acceptable carrier or vehicle may include an excipient, a diluent, or a mixture thereof. In one embodiment, the composition is a pharmaceutical composition.

[0190] 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 an amount of the mixture is filled 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.

[0191] 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.

[0192] 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 a type of material that expands when wet and breaks up tablets and releases compounds. 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 coated with sugar as a flavoring and sealant, or coated with a film-forming protective agent to change the dissolution characteristics of the tablet. Compositions can also be formulated as chewable tablets, for example, by using a material such as mannitol in the formulation.

[0193] 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.

[0194] The effect of the compound of formula (I) can be delayed or prolonged by appropriate formulations. For example, slowly dissolving microspheres 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 film-coated to resist 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 medium that slowly disperses in serum. Exemplary embodiments

[0195] The present disclosure is further described by the following embodiments.

[0196] Embodiment 1. Compound of formula (I): or a pharmaceutically acceptable salt thereof, wherein: A is a C1-C6 alkyl group, a phenyl group, a C3-C6 cycloalkyl group, a 5- to 6-membered heteroaryl group, or a 6- to 10-membered heterocyclic group, wherein the phenyl group, the cycloalkyl group, the heteroaryl group, and the heterocyclic group are substituted by x R 1 Group substituted, and wherein the heteroaryl and heterocyclic groups contain 1-3 heteroatoms selected from N and O; Each R 1 independently halo, C1-C6 alkyl, C3-C6 cycloalkyl, C1-C6 alkoxy, C1-C6 haloalkyl, or -SO2(C1-C6 alkyl); or two R on adjacent carbon atoms 1 The groups together form a fused C3-C6 cycloalkyl or a fused group; R a and R b Each is H or together form an oxo group; R c is H or C1-C6 alkyl; x is 0-5; R 2 is H or C1-C6 alkyl; R 3 is H or C1-C6 alkyl; R 4is H or C1-C6 alkyl; X 1 is CH or N; X 2 N or CH2; Ring B is a C3-C6 cycloalkylene group or a 5- to 7-membered heterocyclylene group containing 1 or 2 nitrogen atoms; Each R 5 are independently halo, C1-C6 alkyl or C1-C6 haloalkyl; w is 0-5; L 1 -C(O)(CH2) n -、-(CH2) n -or-(CH2) n C(O)-; n is 1-6; Ring C is a 5- to 10-membered heterocyclylene group containing 1 or 2 nitrogen atoms; Each R 6 are independently halo, C1-C6 haloalkyl or C1-C6 alkyl; y is 0-5; Ring D is R 7a and R 7b Each is H or together form an oxo group; Each R 8 are independently halo, C1-C6 alkyl, C1-C6 haloalkyl or C1-C6 alkoxy; z is 0-4; X 3 N or CR 9 ; R 9 is H or C1-C6 alkyl; R 10 is H or C1-C6 alkyl; Each R 11 are independently halo, C1-C6 alkyl or C1-C6 haloalkyl; v is 0-4; and Each are independently a single bond or a double bond.

[0197] Embodiment 2. The compound according to embodiment 1 or a pharmaceutically acceptable salt thereof, wherein: A is C1-C3 alkyl, phenyl, C3-C5 cycloalkyl, 5- to 6-membered heteroaryl, or 8- to 10-membered heterocyclyl, wherein the phenyl, cycloalkyl, heteroaryl, and heterocyclyl groups are substituted by x R 1 Group substitution.

[0198] Embodiment 3. The compound according to Embodiment 1 or 2, or a pharmaceutically acceptable salt thereof, wherein: x is 0-3.

[0199] Embodiment 4. The compound according to any one of Embodiments 1-3, or a pharmaceutically acceptable salt thereof, wherein: Each R 1 independently halo, C1-C3 alkyl, C3-C5 cycloalkyl, C1-C3 alkoxy, C1-C3 haloalkyl or -SO2(C1-C3 alkyl); or two R on adjacent carbon atoms 1 The groups together form a fused C3-C5 cycloalkyl or a fused group; R a and R b are each H or together form an oxo group; and R c is H or C1-C3 alkyl.

[0200] Embodiment 5. The compound according to embodiment 4 or a pharmaceutically acceptable salt thereof, wherein: Each R 1 are independently F, -CH3, cyclopropyl, -OCH3 or -SO2(CH3); or two R on adjacent carbon atoms 1 The groups together form fused cyclopropyl, cyclobutyl,

[0201] Embodiment 6. The compound according to any one of Embodiments 1-5, or a pharmaceutically acceptable salt thereof, wherein: A is -CH3 or -CH2CH3.

[0202] Embodiment 7. The compound according to any one of Embodiments 1-6, or a pharmaceutically acceptable salt thereof, wherein: R 2 is H or C1-C3 alkyl.

[0203] Embodiment 8. The compound according to embodiment 7 or a pharmaceutically acceptable salt thereof, wherein: R 2 It is H or -CH3.

[0204] Embodiment 9. The compound according to any one of Embodiments 1-8, or a pharmaceutically acceptable salt thereof, wherein: R 3 is H or C1-C3 alkyl.

[0205] Embodiment 10. The compound according to embodiment 9 or a pharmaceutically acceptable salt thereof, wherein: R 3 It is H or -CH3.

[0206] Embodiment 11. The compound according to any one of Embodiments 1-10, or a pharmaceutically acceptable salt thereof, wherein: R 4 is H or C1-C3 alkyl.

[0207] Embodiment 12. The compound according to embodiment 11 or a pharmaceutically acceptable salt thereof, wherein: R 4 It is H or -CH3.

[0208] Embodiment 13. The compound according to any one of Embodiments 1-12, or a pharmaceutically acceptable salt thereof, wherein: X 1 For CH.

[0209] Embodiment 14. The compound according to any one of Embodiments 1-12, or a pharmaceutically acceptable salt thereof, wherein: X 1 is N.

[0210] Embodiment 15. The compound according to any one of Embodiments 1-14, or a pharmaceutically acceptable salt thereof, wherein: X 2 is N.

[0211] Embodiment 16. The compound according to any one of Embodiments 1-14, or a pharmaceutically acceptable salt thereof, wherein: X 2 For CH2.

[0212] Embodiment 17. The compound according to any one of Embodiments 1-16, or a pharmaceutically acceptable salt thereof, wherein: for

[0213] Embodiment 18. The compound according to any one of Embodiments 1-17, or a pharmaceutically acceptable salt thereof, wherein: Ring B is a C4-C6 cycloalkylene group or a 6- to 7-membered heterocyclylene group containing one nitrogen atom.

[0214] Embodiment 19. The compound according to embodiment 18 or a pharmaceutically acceptable salt thereof, wherein: Ring B is

[0215] Embodiment 20. The compound according to any one of Embodiments 1-19, or a pharmaceutically acceptable salt thereof, wherein: w is 0-2.

[0216] Embodiment 21. The compound according to any one of Embodiments 1-20, or a pharmaceutically acceptable salt thereof, wherein: Each R 5 are independently halo, C1-C3 alkyl or C1-C3 haloalkyl.

[0217] Embodiment 22. The compound according to embodiment 21 or a pharmaceutically acceptable salt thereof, wherein: Each R 5 are independently F, -CF3, or -CH3.

[0218] Embodiment 23. The compound according to any one of Embodiments 1-22, or a pharmaceutically acceptable salt thereof, wherein: for

[0219] Embodiment 24. The compound according to any one of Embodiments 1-23, or a pharmaceutically acceptable salt thereof, wherein: L 1 -C(O)CH2-, -(CH2) n - or -CH2C(O)-; and n is 1-5.

[0220] Embodiment 25. The compound of embodiment 24 or a pharmaceutically acceptable salt thereof, wherein: L 1 It is -C(O)CH2-, -CH2-, -CH2CH2-, -CH2CH2CH2-, -CH2CH2CH2CH2- or -CH2C(O)-.

[0221] Embodiment 26. The compound according to any one of Embodiments 1-25, or a pharmaceutically acceptable salt thereof, wherein: Ring C is a 6- to 8-membered heterocyclylene group containing 1 or 2 nitrogen atoms.

[0222] Embodiment 27. The compound according to embodiment 26, or a pharmaceutically acceptable salt thereof, wherein: Ring C is

[0223] Embodiment 28. The compound according to any one of Embodiments 1-27, or a pharmaceutically acceptable salt thereof, wherein: y is 0-3.

[0224] Embodiment 29. The compound according to any one of Embodiments 1-28, or a pharmaceutically acceptable salt thereof, wherein: Each R 6 are independently halo, C1-C3 haloalkyl or C1-C3 alkyl.

[0225] Embodiment 30. The compound of embodiment 29 or a pharmaceutically acceptable salt thereof, wherein: Each R 6 independently Cl, -CF3 or -CH3.

[0226] Embodiment 31. A compound according to any one of Embodiments 1-30, or a pharmaceutically acceptable salt thereof, wherein: for

[0227] Embodiment 32. The compound according to any one of Embodiments 1-31, or a pharmaceutically acceptable salt thereof, wherein: Ring D is

[0228] Embodiment 33. The compound according to any one of Embodiments 1-31, or a pharmaceutically acceptable salt thereof, wherein: Ring D is

[0229] Embodiment 34. The compound of any one of Embodiments 1-31, or a pharmaceutically acceptable salt thereof, wherein: Ring D is

[0230] Embodiment 35. The compound of embodiment 34 or a pharmaceutically acceptable salt thereof, wherein: z is 0-2.

[0231] Embodiment 36. The compound of embodiment 34 or 35, or a pharmaceutically acceptable salt thereof, wherein: Each R 8 are independently halo, C1-C3 alkyl, C1-C3 haloalkyl or C1-C3 alkoxy.

[0232] Embodiment 37. The compound according to embodiment 36, or a pharmaceutically acceptable salt thereof, wherein: Each R 8independently F, Cl, -CH3, -OCH3 or -CF3.

[0233] Embodiment 38. The compound of any one of Embodiments 1-37, or a pharmaceutically acceptable salt thereof, wherein: Ring D is

[0234] Embodiment 39. The compound of any one of Embodiments 1-38, or a pharmaceutically acceptable salt thereof, wherein: X 3 is N.

[0235] Embodiment 40. The compound of any one of Embodiments 1-38, or a pharmaceutically acceptable salt thereof, wherein: X 3 CR 9 ;as well as R 9 is H or C1-C3 alkyl.

[0236] Embodiment 41. The compound of embodiment 40, or a pharmaceutically acceptable salt thereof, wherein: X 3 CR 9 ;as well as R 9 It is H or -CH3.

[0237] Embodiment 42. The compound of any one of Embodiments 1-41, or a pharmaceutically acceptable salt thereof, wherein: R 10 is H or C1-C3 alkyl.

[0238] Embodiment 43. The compound of embodiment 42 or a pharmaceutically acceptable salt thereof, wherein: R 10 It is H or -CH3.

[0239] Embodiment 44. The compound of any one of Embodiments 1-43, or a pharmaceutically acceptable salt thereof, wherein: v is 0-2.

[0240] Embodiment 45. The compound of any one of Embodiments 1-44, or a pharmaceutically acceptable salt thereof, wherein: Each R 11 are independently halo, C1-C3 alkyl or C1-C3 haloalkyl.

[0241] Embodiment 46. The compound of embodiment 45 or a pharmaceutically acceptable salt thereof, wherein: Each R 11 independently F, -CH3 or -CF3.

[0242] Embodiment 47. The compound of any one of Embodiments 1-46, or a pharmaceutically acceptable salt thereof, wherein: for

[0243] Embodiment 48. A compound according to any one of Embodiments 1-12, 14, 15, 17-38 and 40-47, or a pharmaceutically acceptable salt thereof, wherein the compound is of formula (II):

[0244] Embodiment 49. A compound according to any one of Embodiments 1-12, 14, 15, 17-32, 34-38 and 40-47, or a pharmaceutically acceptable salt thereof, wherein the compound is of Formula (IIIa), (IIIb) or (IIIc):

[0245] Embodiment 50. A compound according to any one of Embodiments 1-12, 14, 15, 17-31, 34-38 and 40-47, or a pharmaceutically acceptable salt thereof, wherein the compound is of Formula (IVa) or (IVb):

[0246] Embodiment 51. A compound selected from the compounds of Table 1, or a pharmaceutically acceptable salt thereof.

[0247] Embodiment 52. A pharmaceutical composition comprising a compound according to any one of Embodiments 1-51 or a pharmaceutically acceptable salt thereof and a pharmaceutically acceptable excipient.

[0248] Embodiment 53. A method of modulating interleukin-1 receptor associated kinase 3 (IRAK3), the method comprising contacting IRAK3 with an effective amount of a compound according to any one of embodiments 1-51 or a pharmaceutically acceptable salt thereof, or a pharmaceutical composition according to embodiment 52.

[0249] Embodiment 54. A method of treating cancer in a subject in need thereof, comprising administering to the subject an effective amount of a compound according to any one of Embodiments 1-51 or a pharmaceutically acceptable salt thereof, or a pharmaceutical composition according to Embodiment 52.

[0250] Embodiment 55. The method of embodiment 54, wherein the cancer is selected from bladder cancer, breast cancer, esophageal cancer, colon cancer, head and neck cancer, kidney cancer, lung cancer, pancreatic cancer, prostate cancer, melanoma and gastric cancer.

[0251] Embodiment 56. A method of enhancing immunity in a vaccinated subject, the method comprising administering to the subject an effective amount of a compound according to any one of embodiments 1-51 or a pharmaceutically acceptable salt thereof, or a pharmaceutical composition according to embodiment 52.

[0252] Embodiment 57. The method of embodiment 56, wherein the vaccine is administered to the subject before, simultaneously with, or after administration of the compound of any one of embodiments 1-51 or a pharmaceutically acceptable salt thereof, or the pharmaceutical composition of embodiment 52. Example

[0253] The following examples are intended to illustrate rather than limit the present invention. 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.

[0254] 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.

[0255] The following abbreviations may be used in connection with this application. abbreviation Synthesis Example Program Overview

[0256] The synthesized compounds are prepared from two main building blocks: the target binding moiety (TBM) and the Cereblon-binding moiety (CBM). The methods for these building blocks can be found in their respective sections. The TBM and CBM are then formed into heterobifunctional molecules (i.e., ligand-directed degraders, LDDs) through a 2- or 3-step method. Table 2 lists the building blocks used to synthesize each example and the sections in which these methods are located. Table 2. Methods used to synthesize LDD from CBM and TBM Methods for Cereblon-Binding Moieties (CBMs)

[0257] Table 3 lists the general procedures used to synthesize each CBM molecule. Table 3. General procedures used for CBM Example S1. General Procedure for CBM-A

[0258] Synthesis of CBM-A: Step 1 To a sealed tube, tert-butyl 4-(4-bromo-2-methoxy-phenyl)piperazine-1-carboxylate intermediate A-1b (250 mg, 0.67 mmol, 1 eq), K 3 PO 4 (324.85 mg, 1.53 mmol, 2.3 eq), 2,6-dibenzyloxy-3-(4,4,5,5-tetramethyl-1,3,2-dioxaborolan-2-yl)pyridine intermediate A-2 (337.2 mg, 0.81 mmol, 1.2 eq), and Pd(PPh 3 ) 4 (77.81 mg, 0.07 mmol, 0.1 eq) were added. The tube was then flushed with nitrogen for 5 minutes. To the 4- (4- (2,6-bis (benzyloxy) pyridin-3-yl) -2- methoxyphenyl) piperazine-1-carboxylic acid tert-butyl ester intermediate A-3b (252mg, 64% yield) was added 1,4-dioxane (2.5857mL, 0.2M) and water (0.6464mL, 0.2M) (by degassing with nitrogen bubbling for 10 minutes), and the resulting mixture was bubbling with nitrogen for another 5 minutes. The bottle was sealed and heated at 90 ° C overnight for 16h. LCMS shows that it is completely converted into intermediate A-3b. The crude mixture is passed through diatomaceous earth, washed with DCM and concentrated. The crude residue is purified by silica gel chromatography (0% to 25% EtOAc / heptane). Fractions are merged and concentrated to obtain tert-butyl 4- (4- (2,6-bis (benzyloxy) pyridin-3-yl) -2- methoxyphenyl) piperazine-1-carboxylate intermediate A-3b (252mg, 64% yield) as a yellow solid.

[0259] LCMS: [M+H] + =582.3.

[0260] 1H NMR (400MHz, chloroform-d) δppm 1.50(s,9H),3.03(t,J=4.8Hz,4H),3.62(t,J=4.8Hz,4H),3.73(s,3H),5.41(d,J=7.1Hz,4H),6.48(d,J=8.1Hz,1H), 6.92(d,J=8.3Hz,1H),7.08(dd,J=8.1,1.7Hz,1H),7.18(d,J=1.7Hz,1H),7.29-7.48(m,10H),7.64(d,J=8.1Hz,1H). Table 4. CBM-A: List of intermediates used in step 1 Table 5. List of intermediate A-3 compounds generated by CBM-A general procedure: Step 1

[0261] Synthesis of 2,6-bis(benzyloxy)-3-(4,4,5,5-tetramethyl-1,3,2-dioxaborolan-2-yl)pyridine (Intermediate A-2) In a sealed tube, 2,6-dibenzyloxy-3-bromo-pyridine (1.0 g, 2.701 mmol, 1.0 equiv), bis(pinacolato)diboron (1.0 g, 4.051 mmol, 1.5 equiv), KOAc (795.2 mg, 8.103 mmol, 3.0 equiv), and Pd(dppf)Cl2·DCM (220.57 mg, 0.270 mmol, 0.1 equiv) were dissolved in 1,4-dioxane (4.0 mL, 0.6 M) and sparged with nitrogen for 10 minutes. The tube was sealed and heated at 90°C overnight. The reaction mixture was cooled to room temperature. The reaction was filtered through celite, washed with 2-MeTHF, and the filtrate was evaporated. The residue was purified by silica gel chromatography (0% to 10% EtOAc / heptane) to give Intermediate A-2, which was contaminated with bis(pinacolato)diboron. A second purification was performed by silica gel chromatography (0% to 4.5% EtOAc) to give 2,6-dibenzyloxy-3-(4,4,5,5-tetramethyl-1,3,2-dioxaborolan-2-yl)pyridine intermediate A-2 as a white solid (210 mg, 19% yield).

[0262] LCMS: [M-pin+H] + =336.2,[M+H] + =418.2.

[0263] 1 H NMR(400MHz,DMSO-d6)δppm 1.28(s,12H),5.38(d,J=5.9Hz,4H),6.42(d,J=7.8Hz,1H),7.24-7.40(m,6H),7.40-7.44(m,2H),7.53(d,J=7.1Hz,2H),7.84(d,J=7.8Hz,1H).

[0264] Synthesis of CBM-A: Step 2 A solution of 4-[4-(2,6-dibenzyloxy-3-pyridyl)-2-methoxy-phenyl]piperazine-1-carboxylic acid tert-butyl ester intermediate A-3b (155 mg, 0.27 mmol, 1 equivalent) in THF (1.5 mL, 0.09 M) and ethanol (1.5 mL, 0.09 M) was degassed for 15 minutes, followed by addition of Pd(OH)2 (37.42 mg, 0.05 mmol, 0.2 equivalent) and bubbling was resumed for 5 minutes. Hydrogen was then bubbled through the reaction mixture for 5 minutes and the mixture was stirred in a hot water bath at 50 ° C under a hydrogen atmosphere. After 2 h, LCMS showed complete conversion to compound intermediate A-4b. The crude mixture was passed through diatomaceous earth, washed with DCM and concentrated. The crude product was purified by reverse phase column chromatography (5% to 50% MeCN / water, containing 0.1% formic acid). The fractions were combined and concentrated to give tert-butyl 4-(4-(2,6-dioxopiperidin-3-yl)-2-methoxyphenyl)piperazine-1-carboxylate intermediate A-4b (84 mg, 78% yield) as a yellow solid.

[0265] LCMS: [M+H] + =404.4.

[0266] 1 H NMR(400MHz,DMSO-d6)δppm 1.50(s,9H),3.03(t,J=4.8Hz,4H),3.62(t,J=4.8Hz,4H),3.73(s,3H),5.41(d,J=7.1Hz,4H),6.48(d,J=8.1Hz,1H), 6.92(d,J=8.3Hz,1H),7.08(dd,J=8.1,1.7Hz,1H),7.18(d,J=1.7Hz,1H),7.29-7.48(m,10H),7.64(d,J=8.1Hz,1H). Table 6. List of intermediate A-4 compounds generated by CBM-A general procedure: Step 2

[0267] Synthesis of CBM-A General Procedure: Step 3 To a solution of tert-butyl 4-[4-(2,6-dioxo-3-piperidinyl)-2-methoxy-phenyl]piperazine-1-carboxylate intermediate A-4b (130 mg, 0.32 mmol, 1 eq) in DCM (1.4 mL, 0.23 M) was added 4M HCl in 1,4-dioxane (1.2 mL, 4.82 mmol, 15 eq). The reaction mixture was stirred at room temperature overnight. LCMS showed complete conversion to compound CBM-12. The reaction mixture was concentrated under reduced pressure and the residue was co-evaporated with MeOH (2x) and MTBE (2x) to give 3-(3-methoxy-4-(piperazin-1-yl)phenyl)piperidine-2,6-dione CBM-12 (103 mg, 85% yield) as a double HCl salt as a brown solid.

[0268] LCMS: [M+H] + =304.2.

[0269] 1 H NMR (400MHz, DMSO-d6) δppm 1.94-2.06(m,1H),2.14-2.29(m,1H),2.43-2.49(m,1H),2.58-2.71(m,1H),3.17(br s,4H),3.18-3.25(m,4H),3.75-3.82(m,4H),6.71-6.77(m,1H),6.84-6.91(m,2H),9.08(br s,2H),10.80(s,1H). Table 7. List of CBM-A compounds generated by the general procedure CBM-A: Step 3 Example S2. General Procedure for CBM-B

[0270] Synthesis of CBM-B: Step 1 To a solution of 2-(4-bromophenyl)acetonitrile intermediate B-1a (25.0 g, 127.5 mmol, 1.0 equiv) in toluene (255 mL) was added tert-butyl prop-2-enoate intermediate B-2 (18.7 mL, 127.5 mmol, 1.0 equiv), N-benzyl-N,N-diethylethaneammonium chloride (BTEAC) (2.9 g, 12.7 mmol, 0.1 equiv) and potassium carbonate (17.6 g, 127.5 mmol, 1.0 equiv). The mixture was stirred at 65 ° C. under nitrogen for 3 hours, then cooled to room temperature and filtered through a glass sintered funnel. The mixture was concentrated to give a crude mixture which was purified by reverse phase flash chromatography (5% to 100% MeCN / water with 0.1% formic acid) to yield 12.8 g (27% yield) of tert-butyl 4-(4-bromophenyl)-4-cyano-butyrate intermediate B-3a as a light yellow oil.

[0271] LCMS: [M+H]+ = 324.2.

[0272] 1 H NMR(400MHz,DMSO-d6)δppm 1.37(s,3H),1.97-2.15(m,2H),2.28(t,J=7.5Hz,2H),4.25(t,J=7.4Hz,1H),7.35(d,J=8.3Hz,2H),7.62(d,J=8.3Hz,2H). Table 8. List of intermediate B-1 compounds used in CBM-B: Step 1 Table 9. List of intermediate B-3 compounds generated by general procedure CBM-B: Step 1

[0273] Synthesis of CBM-B: Step 2 To a solution of (2S)-2-methylpiperazine-1-carboxylic acid tert-butyl ester intermediate B-4a (340 mg, 1.7 mmol, 1.1 equivalents) in 1,4-dioxane (3.00 mL, 0.5 M) was added CsCO (1.01 g, 3.08 mmol, 2.0 equivalents), 4-(4-bromophenyl)-4-cyano-butyric acid tert-butyl ester intermediate B-3a (500. mg, 1.54 mmol, 1 equivalent), Xphos (147 mg, 0.31 mmol, 0.2 equivalents), Pd(dba) (141 mg, 0.15 mmol, 0.1 equivalents). The reaction mixture was degassed with N for 15 min and stirred at 90 ° C. After 18 h, LCMS showed complete conversion to intermediate B-5a. The reaction mixture was filtered through a celite pad and washed with EtOAc. The filtrate was concentrated under vacuum and the residue was purified by silica gel chromatography (0% to 80% EtOAc / heptane). The fractions were combined and concentrated to give tert-butyl (2S)-4-(4-(4-(tert-butoxy)-1-cyano-4-oxobutyl)phenyl)-2-methylpiperazine-1-carboxylate intermediate B-5a (452 ​​mg, 61% yield) as a brown oil.

[0274] LCMS: 93.4% purity at 215 nm, [M+H] + =444.4.

[0275] 1 H NMR(400MHz,DMSO-d6)δppm 1.18(d,J=6.6Hz,3H),1.38(s,9H),1.42(s,9H),1.92-2.11(m,2H),2.26(t,J=8.0H z,2H),2.58-2.68(m,1H),2.83(dd,J=12.2,3.7Hz,1H),3.09-3.20(m,1H),3.50(br d,J=12.0Hz,1H),3.59(br d,J=11.7Hz,1H),3.79(brd,J=13.0Hz,1H),4.08(t,J=7.3Hz,1H),4.15-4.25(m,1H),6.94(d,J=8.6Hz,2H),7.20(d,J=8.8Hz,2H). Table 10. List of intermediate B-4 compounds used in CBM-B: Step 2 Table 11. List of intermediate B-5 compounds generated by general procedure CBM-B: Step 2

[0276] Synthesis of CBM-B: Step 3 To a solution of (2S)-4-[4-(4-tert-butoxy-1-cyano-4-oxo-butyl)phenyl]-2-methyl-piperazine-1-carboxylic acid tert-butyl ester intermediate B-5a (400 mg, 0.90 mmol, 1.0 equiv) in acetic acid (4.50 mL, 0.2 M) was added concentrated sulfuric acid (0.14 mL, 2.71 mmol, 3.0 equiv). The reaction mixture was stirred at 110 ° C. After 45 min, LCMS showed complete conversion to CBM-3. The solvent was evaporated, and the residue was then purified by reverse phase flash chromatography (5% to 65% MeCN / water). The fractions were combined and concentrated to give 3-(4-((S)-3-methylpiperazin-1-yl)phenyl)piperidine-2,6-dione CBM-3 (207 mg, 76% yield) as a disulfate salt as a brown solid.

[0277] LCMS: [M+H] + =288.2.

[0278] 1 H NMR(400MHz,DMSO-d6)δppm 1.23(br d,J=6.1Hz,3H),1.81(s,1H),1.94-2.05(m,1H),2.06-2.21(m,1H),2.57-2.71(m,2H),2.79 -2.91(m,1H),3.03-3.13(m,1H),3.15(s,1H),3.26-3.35(m,3H),3.63-3.79(m,3H),4.75(br d, J=15.2Hz, 1H), 6.95 (br d, J=8.3Hz, 2H), 7.09 (br d, J=8.3Hz, 2H), 10.78 (s, 1H). Table 12. List of CBM-B compounds generated by the general procedure CBM-B: Step 3 Example S3. General Procedure for CBM-C

[0279] Synthesis of CBM-C: Step 1 Under N2, to a solution of N-bis(2-oxoethyl)carbamic acid tert-butyl ester intermediate C-2 (553.18 mg, 2.75 mmol) in DCE (20 mL), (3S)-3-(4-aminophenyl)-3-methyl-piperidine-2,6-dione intermediate C-1a (500 mg, 2.29 mmol) was added. After 5 minutes, sodium triacetoxyborohydride (1165.42 mg, 5.5 mmol) was added. The reaction was stirred at room temperature for 24 h. HPLC and LCMS showed incomplete conversion of the starting material. Therefore, sodium triacetoxyborohydride (1165.42 mg, 5.5 mmol) was added at room temperature and the reaction was continued for another 24 h. HPLC and LCMS showed complete conversion of the starting material. The reaction mixture was distributed between DCM and saturated NaHCO3. The organic phase was extracted with DCM (2 x 60 mL). The combined organics were washed with brine, dried over MgSO4 and concentrated to give the crude product, which was purified by reverse phase column chromatography (5% to 100% MeCN / water with 0.1% formic acid) to afford (S)-tert-butyl 4-(4-(3-methyl-2,6-dioxopiperidin-3-yl)phenyl)piperazine-1-carboxylate intermediate C-3a (300 mg, 33.7% yield) as an off-white solid.

[0280] LCMS: [M+H] + =388.2.

[0281] 1 H NMR(400MHz,DMSO-d6)δppm 1.41(s,9H),2.00-2.10(m,2H),2.27-2.33(m,1H),2.39-2.45(m,1H),3.05-3.12(m,3H),3 .32(s,4H),3.41-3.49(m,4H),6.94(d,J=8.8Hz,2H),7.12(d,J=8.8Hz,2H),10.85(s,1H). Table 13. List of intermediate C-1 compounds used in CBM-C: Step 1 Table 14. List of intermediate C-3 compounds generated by general procedure CBM-C: Step 1

[0282] Synthesis of (S)-3-(4-aminophenyl)-3-methylpiperidine-2,6-dione (C-1a) and (R)-3-(4-aminophenyl)-3-methylpiperidine-2,6-dione (C-1b)

[0283] Step 1: 2-(4-nitrophenyl)propionitrile: To a solution of 2-(4-nitrophenyl)acetonitrile (90 g, 555 mmol) in dry THF (2500 mL) was added LiHMDS (666 mL, 666 mmol, 1 M in THF) at -78 °C under a nitrogen atmosphere. The mixture was stirred at -78 °C for 1 h, and then iodomethane (86.7 g, 611 mmol) was added. After stirring at -78 °C for 1 h, the reaction mixture was allowed to warm to room temperature and stirred for 2 h. The reaction mixture was cooled to 0 °C and quenched dropwise with citric acid (1000 mL, 10%) at 0 °C, and then extracted with ethyl acetate (1000 mL x 2). The combined organic layers were washed with brine (1500 mL), dried over anhydrous Na2SO4, filtered, and concentrated. The residue was purified by column chromatography (petroleum ether / ethyl acetate, 1:1 to 10:1) to give 2-(4-nitrophenyl)propionitrile (36.8 g, 37.6% yield) as a yellow solid.

[0284] 1 H NMR (400MHz, CDCl3) δ: 8.25 (d, J = 8.4Hz, 2H), 7.55 (d, J = 8.4Hz, 2H), 4.05-3.99 (m, 2H), 1.68 (d, J = 4.8Hz, 3H).

[0285] Step 2: Methyl 4-cyano-4-(4-nitrophenyl)pentanoate: To a solution of 2-(4-nitrophenyl)propionitrile (65 g, 369 mmol) in toluene (650 mL) was added methyl prop-2-enoate (63.5 g, 738 mmol), KCO (102 g, 738 mmol) and BTEAC (16.8 g, 73.8 mmol). The reaction was stirred at 65 ° C for 4 h. The reaction was filtered and the filtrate was concentrated to give methyl 4-cyano-4-(4-nitrophenyl)pentanoate (75 g, crude) as a yellow solid, which was used directly without further purification.

[0286] Step 3: 3-methyl-3-(4-nitrophenyl)piperidine-2,6-dione: To a solution of methyl 4-cyano-4-(4-nitrophenyl)pentanoate (75 g, 286 mmol) in acetic acid (700 mL) was added H2SO4 (2.8 g, 28.6 mmol), and the reaction was stirred at 120°C for 12 h. The reaction mixture was cooled to room temperature and concentrated, and the residue was washed with MTBE (100 mL) and dried to give crude 3-methyl-3-(4-nitrophenyl)piperidine-2,6-dione (72 g, crude) as a gray solid, which was used directly without further purification.

[0287] Step 4: rac-3-(4-aminophenyl)-3-methylpiperidine-2,6-dione: To a solution of 3-methyl-3-(4-nitrophenyl)piperidine-2,6-dione (72 g, 290 mmol) in ethanol (720 mL) was added Fe (81 g, 1450 mmol), NHCl (155 g, 2900 mmol) and water (360 mL), and the reaction was stirred at 80 ° C for 1 h. The reaction mixture was cooled to room temperature and filtered, and the filtrate was concentrated. The residue was extracted with ethyl acetate (400 mL x 3), and the combined organic layers were washed with brine (600 mL), dried over anhydrous NaSO, filtered and concentrated to give 3-(4-aminophenyl)-3-methyl-piperidine-2,6-dione (50.2 g, 79.3% yield) as a gray solid.

[0288] 1 H NMR (400MHz, CDCl3) δ: 8.0 (br s, 1H), 7.04-7.01 (m, 1H), 6.66 (t, J = 2.0Hz, 1H), 3.73 (br s, 2H), 2.56-2.41 (m, 1H), 2.38-2.33 (m, 2H), 2.32-2.08 (m 1H),1.53(s,3H).

[0289] SFC: (S)-3-(4-aminophenyl)-3-methylpiperidine-2,6-dione (intermediate C-1a) and (R)-3-(4-aminophenyl)-3-methylpiperidine-2,6-dione (intermediate C-1b): Separation conditions: instrument – ​​Thar SFC350 preparative SFC; column – Daicel Chiralcel OJ, 250x 50mm, id10μm; mobile phase – A represents CO2, B represents IPA containing 0.1% ammonium hydroxide; gradient – ​​B% = 60%; flow rate – 200g / min; wavelength – 220nM; column temperature – 40℃; system back pressure – 100 bar. 55 g of racemic-3-(4-aminophenyl)-3-methylpiperidine-2,6-dione was separated into 25.9 g of (S)-3-(4-aminophenyl)-3-methylpiperidine-2,6-dione (Intermediate C-1a) [peak 1] and 26.5 g of (R)-3-(4-aminophenyl)-3-methylpiperidine-2,6-dione (Intermediate C-1b) [peak 2].

[0290] Synthesis of 1-(6-amino-1-methyl-1H-indazol-3-yl)dihydropyrimidine-2,4(1H,3H)-dione (C-1c)

[0291] Step 1: 1-Methyl-6-nitro-1H-indazol-3-amine: To a solution of 2-fluoro-4-nitro-benzonitrile (5.0 g, 30.1 mmol, 1.0 equiv) in dimethylacetamide (25.0 mL) was added methylhydrazine (10.4 g, 90.3 mmol, 11.9 mL, 3.0 equiv), N-ethyl-N-isopropylpropan-2-amine (4.28 g, 33.1 mmol, 5.8 mL, 1.1 equiv) and dimethylacetamide (25.0 mL). The mixture was stirred at 150 ° C for 0.5 h. The mixture was diluted with petroleum ether (300 mL) and ethyl acetate (30 mL). The mixture solution was stirred for 1 h and filtered to obtain a residue. 1-Methyl-6-nitro-indazol-3-amine (7.0 g, crude material) was obtained as a brown solid and used directly without further purification.

[0292] LCMS: [M+H] + :193.1.

[0293] 1 H NMR (400MHz, DMSO-d6): δ8.35 (s, 1H), 7.90 (d, J = 8.8 Hz, 1H), 7.69 (d, J = 8.4 Hz, 1H), 5.78 (s, 2H), 3.86 (s, 3H).

[0294] Step 2: 3-((1-methyl-6-nitro-1H-indazol-3-yl)amino)propanoic acid: To 1-methyl-6-nitro-indazol-3-amine (1.2 g, 6.24 mmol, 1.0 equiv) was added aluminum oxide (1.91 g, 18.73 mmol, 3.0 equiv), dioxane (5.0 mL), and acrylic acid (0.9 g, 12.49 mmol, 0.9 mL, 2.0 equiv). The mixture was stirred at 110° C. for 12 h. The mixture was diluted with dichloromethane (100 mL), filtered, and concentrated to give 3-((1-methyl-6-nitro-1H-indazol-3-yl)amino)propanoic acid (15 g, crude) as a brown oil and used without further purification.

[0295] LCMS: [M+H] + :265.0.

[0296] Step 3: Methyl 3-((1-methyl-6-nitro-1H-indazol-3-yl)amino)propanoate: To a solution of 3-((1-methyl-6-nitro-1H-indazol-3-yl)amino)propanoic acid (15.0 g, 56.77 mmol, 1.0 equiv) in methanol (70 mL) and toluene (70 mL) was added trimethylsilyldiazomethane (2 M, 85.2 mL, 3.0 equiv) at 0°C. The mixture was stirred at 15°C for 12 h. The reaction mixture was concentrated to give a residue. The reaction mixture was concentrated to give a residue. The residue was purified by silica gel column chromatography (1% to 20% EtOAc / petroleum ether) to give methyl 3-((1-methyl-6-nitro-1H-indazol-3-yl)amino)propanoate (4.5 g, crude) as a brown solid.

[0297]

[0266] Step 4: Methyl 3-(1-(1-methyl-6-nitro-1H-indazol-3-yl)ureido)propanoate: To a solution of methyl 3-((1-methyl-6-nitro-1H-indazol-3-yl)amino)propanoate (4.5 g, 16.17 mmol, 1.00 equiv) in acetic acid (50 mL) was added potassium cyanate (6.89 g, 80.86 mmol, 5.0 equiv). The mixture was stirred at 15 °C for 12 h. The mixture was concentrated, diluted with water (150 mL) and extracted with ethyl acetate (3 x 150 mL). The combined organic layers were washed with brine (150 mL), dried over sodium sulfate, filtered and concentrated under reduced pressure to give methyl 3-(1-(1-methyl-6-nitro-1H-indazol-3-yl)ureido)propanoate (6.100 g, crude) as a brown oil which was used without further purification.

[0298] LCMS: [M+H] + :322.6.

[0299] Step 5: 1-(1-methyl-6-nitro-1H-indazol-3-yl)dihydropyrimidine-2,4(1H,3H)-dione: To methyl 3-(1-(1-methyl-6-nitro-1H-indazol-3-yl)ureido)propanoate (6.0 g, 18.67 mmol, 1.0 equiv) was added HCl (12 M, 50.0 mL, 32.13 equiv) at 0°C. The mixture was stirred at 15°C for 12 h. Water (100 mL) was added to the reaction mixture. The mixture was concentrated and lyophilized to give a residue. The residue was purified by semi-preparative reverse phase HPLC (23% to 48% MeCN / water with 0.05% ammonium hydroxide) to give 1-(1-methyl-6-nitro-indazol-3-yl)hexahydropyrimidine-2,4-dione (1.0 g, 3.46 mmol, 18.5% yield) as a yellow solid.

[0300] LCMS: [M+H] + :290.0.

[0301] 1 H NMR (400MHz, DMSO-d6): δ10.66 (s, 1H), 8.70 (d, J = 1.2 Hz, 1H), 8.05-7.88 (m, 2H), 4.14 (s, 3H), 3.98 (t, J = 6.8 Hz, 2H), 2.78 (t, J = 6.8 Hz, 2H).

[0302] Step 6: 1-(6-amino-1-methyl-1H-indazol-3-yl)dihydropyrimidine-2,4(1H,3H)-dione (Intermediate C-1c): To a solution of 1-(1-methyl-6-nitro-indazol-3-yl)hexahydropyrimidine-2,4-dione (1.0 g, 3.46 mmol, 1.0 equiv) in ethanol (40 mL) and water (20 mL) were added iron (0.965 g, 17.29 mmol, 5.0 equiv) and ammonium chloride (1.85 g, 34.57 mmol, 10.0 equiv). The mixture was stirred at 85° C. for 1 h. The mixture was filtered and concentrated to give a residue, which was purified by silica gel column chromatography (1% to 5% MeOH / DCM) to afford 1-(6-amino-1-methyl-indazol-3-yl)hexahydropyrimidine-2,4-dione (0.599 g, 2.31 mmol, 67% yield, 100% purity) as a white solid.

[0303] LCMS: [M+H] + :260.0.

[0304] 1H NMR (400MHz, DMSO-d6): δ10.48(s,1H),7.27(d,J=8.8Hz,1H),6.47(dd,J=1.6,8.8Hz,1H),6.39 (d,J=1.2Hz,1H),5.41(s,2H),3.85(t,J=6.8Hz,2H),3.80-3.72(m,3H),2.71(t,J=6.8Hz,2H).

[0305] Synthesis of CBM-C: Step 2 To a solution of tert-butyl 4-[4-(3S)-3-methyl-2,6-dioxo-3-piperidinyl]phenyl]piperazine-1-carboxylate intermediate C-3a (300 mg, 0.7700 mmol) in 1,4-dioxane (5 mL) was added 4M HCl (3.88 mL, 15.52 mmol) in dioxane at room temperature. The reaction was stirred at room temperature for 5 h. HPLC and LCMS showed that the starting material was completely converted into the desired product. The reaction mixture was concentrated to dryness by careful treatment with acetonitrile to obtain the desired (S)-3-methyl-3-(4-(piperazine-1-yl)phenyl)piperidine-2,6-dione CBM-22 (280 mg, quantitative) as a white solid.

[0306] LCMS: [M+H] + =288.2.

[0307] 1 H NMR(400MHz,DMSO-d6)δppm 1.39(s,3H),2.02-2.12(m,2H),2.29-2.37(m,1H),2.40-2.45(m,1H),3.19(br s,4H),3.32-3.40(m,4H),6.98(d,J=8.8Hz,2H),7.16(d,J=8.8Hz,2H),9.20(br s,3H),10.86(s,1H). Table 15. List of CBM-C compounds generated by the general procedure CBM-C: Step 2 Example S4. CBM-5 Procedure

[0308] Synthesis of 3-(1-oxo-5-(piperazin-1-yl)isoindolin-2-yl)piperidine-2,6-dione (CBM-5)

[0309]

[0266] Step 1: tert-Butyl 4-(2-(2,6-dioxopiperidin-3-yl)-1-oxoisoindolin-5-yl)piperazine-1-carboxylate: To a round-bottom flask was added 3-(5-bromo-1-oxoisoindolin-2-yl)piperidine-2,6-dione (5.0 g, 15.47 mmol, 1 eq), tert-butyl piperazine-1-carboxylate (3.17 g, 17.02 mmol, 1.1 eq), methanesulfonic acid (2-dicyclohexylphosphino-2',6'-di-isopropoxy-1,1'-biphenyl)(2'-amino-1,1'-biphenyl-2-yl)palladium(II) (0.647 g, 0.77 mmol, 0.05 eq) and cesium carbonate (7.56 g, 23.21 mmol, 1.5 eq) and dioxane (100.00 mL). In 4- (2- (2,6- dioxopiperidin-3-yl) -1- oxoisoindolin-5-yl) piperazine -1- tert-butyl formate (0.300g, yield: 4.52%) as white solid.

[0310] LCMS: [M+H] + =429.1.

[0311] 1 HNMR (400MHz, CDCl3): δ8.04(br s,1H),7.76(d,J=11.6Hz,1H),7.00(dd,J=11.6,2.4Hz,1H),6.89(s,1H),5.21(dd,J=17.6,6.8Hz,1H),4. 48-4.21(m,2H),3.67-3.53(m,4H),3.35-3.20(m,4H),2.98-2.76(m,2H),2.42-2.15(m,2H),1.50(s,9H).

[0312] Step 2: 3-(1-oxo-5-(piperazin-1-yl)isoindolin-2-yl)piperidine-2,6-dione (CBM-5): To a solution of tert-butyl 4-(2-(2,6-dioxopiperidin-3-yl)-1-oxoisoindolin-5-yl)piperazine-1-carboxylate (0.150 g, 0.35 mmol, 1 eq) in dichloromethane (2.00 mL) was added trifluoroacetic acid (0.798 g, 7.00 mmol, 0.52 mL, 20 eq) in one portion. The mixture was then stirred at 25° C. for approximately 1 hour. The solvent was removed in vacuo to give a residue which was partitioned between water (5.00 mL) and dichloromethane (3.00 mL). The aqueous phase was washed with dichloromethane (3.00 mL x 2), and the aqueous layer was concentrated under high pressure in a 45 °C water bath to give the product 3-(1-oxo-5-(piperazin-1-yl)isoindolin-2-yl)piperidine-2,6-dione (CBM-5) (64.43 mg, 42% yield, 98.9% purity, TFA salt) as a light red gum.

[0313] LCMS: [M+H] + =329.1.

[0314] 1 HNMR(400MHz, DMSO-d6)10.97(s,1H),8.89(brs,2H),7.58(d,J=8.4Hz,1H),7.21-7.06(m,2H),5.06(dd,J=13.6,5.2Hz,1H),4.4 3-4.17(m,2H),3.59-3.44(m,4H),3.31-3.20(m,4H),2.98-2.84(m,1H),2.61-2.56(m,1H),2.43-2.32(m,1H),2.02-1.91(m,1H). Example S5. Procedure for CBM-7

[0315] Synthesis of 3-(1-oxo-6-(piperazin-1-yl)isoindolin-2-yl)piperidine-2,6-dione (CBM-7)

[0316] Step 1: tert-Butyl 4-(3-bromo-4-formylphenyl)piperazine-1-carboxylate: To a solution of 2-bromo-4-fluoro-benzaldehyde (8 g, 39.4 mmol) and tert-butyl piperazine-1-carboxylate (8.81 g, 47.3 mmol) in DMF (80 mL) was added KCO (10.9 g, 78.8 mmol). The reaction mixture was stirred at 100 ° C for 16 hours. The reaction mixture was cooled to room temperature and diluted with EtOAc (200 mL) and water (200 mL). The aqueous layer was extracted with EtOAc (100 mL x 2). The combined organic layers were washed with brine (400 mL), dried over anhydrous NaSO, filtered and concentrated. The resulting residue was purified by silica gel column chromatography (10% to 50% EtOAc / petroleum ether) to give the product tert-butyl 4-(3-bromo-4-formyl-phenyl)piperazine-1-carboxylate (11.8 g, 81.1% yield) as an off-white solid.

[0317] 1 H NMR: (400MHz, DMSO-d6) δ: 9.96 (s, 1H), 7.68 (d, J = 8.8Hz, 1H), 7.16 (s, 1H), 7.04-7.01 (m, 1H), 3.45 (s, 8H), 1.43 (s, 9H).

[0318] Step 2: tert-Butyl 4-(3-bromo-4-(((2,6-dioxopiperidin-3-yl)amino)methyl)phenyl)piperazine-1-carboxylate: To a stirred solution of 3-aminopiperidine-2,6-dione (3.92 g, 23.8 mmol, HCl salt) in MeOH (250 mL) was added a solution of NH in MeOH until pH = 7. The reaction was stirred at 20 °C. Then, AcOH was added dropwise until pH = 6. At this time, tert-butyl 4-(3-bromo-4-formyl-phenyl)piperazine-1-carboxylate (8.8 g, 23.8 mmol) was added and stirred at 20 °C for 10 min. After that, NaBHCN (4.49 g, 71.5 mmol) was added at 0 °C. The reaction mixture was stirred at 20 °C for 16 h. The reaction mixture was concentrated, the residue was filtered, and the filter cake was washed with petroleum ether / EtOAc (5:1, 50 mL) to give tert-butyl 4-[3-bromo-4-[[(2,6-dioxo-3-piperidinyl)amino]methyl]phenyl]piperazine-1-carboxylate (7.5 g, 65.4% yield) as a white solid.

[0319] 1H NMR: (400MHz, DMSO-d6)δ: 10.74(s,1H),7.34(d,J=8.8Hz,1H),7.12(d,J=2.0Hz,1H),6.97-6.95(m,1H),3.78(d ,J=6.0Hz,1H),3.42(t,J=4.8Hz,4H),3.11(t,J=5.2Hz,4H),2.73-2.68(m,1H),2.56-2.54(m,1H),2.13-2.10(m 1H),1.42(s,9H).

[0320] Step 3: tert-Butyl 4-(2-(2,6-dioxopiperidin-3-yl)-3-oxoisoindolin-5-yl)piperazine-1-carboxylate: To a solution of tert-butyl 4-[3-bromo-4-[[(2,6-dioxo-3-piperidinyl)amino]methyl]phenyl]piperazine-1-carboxylate (5 g, 10.4 mmol) and dicyclohexyl(3-dicyclohexylphosphoniumpropyl)phosphonium bis(tetrafluoroborate) (636 mg, 1.04 mmol) in DMF (50 mL) was added diacetoxypalladium (233 mg, 1.04 mmol) and KCO (2.15 g, 15.6 mmol) under nitrogen. The suspension was degassed under vacuum and purged with CO several times. The mixture was stirred at 80° C. under CO (50 psi) for 48 h. The reaction mixture was cooled to room temperature and filtered. The filtrate was concentrated, and the residue was purified by preparative (20% to 50% MeCN / water with 0.1% TFA) to give tert-butyl 4-[2-(2,6-dioxo-3-piperidinyl)-3-oxo-isoindolin-5-yl]piperazine-1-carboxylate (3 g, 67.4% yield) as a grey solid.

[0321] LCMS: [M+H] + =429.1.

[0322] Step 4: 3-(1-oxo-6-(piperazin-1-yl)isoindolin-2-yl)piperidine-2,6-dione (CBM-7): tert-Butyl 4-[2-(2,6-dioxo-3-piperidinyl)-3-oxo-isoindolin-5-yl]piperazine-1-carboxylate (5 g, 11.7 mmol) was added to HCl (12 N, 15 mL) at 0°C. The reaction mixture was stirred at 20°C for 1 h. The reaction mixture was diluted with MeCN at 0-10°C. The resulting precipitate was filtered, and the filter cake was dried to give 3-(1-oxo-6-piperazin-1-yl-isoindolin-2-yl)piperidine-2,6-dione (4.3 g, HCl salt, 100% yield) as a gray solid.

[0323] 1H NMR: (400MHz, DMSO-d6) δ: 10.98 (s, 1H), 9.34 (s, 2H), 7.49 (d, J = 5.2Hz, 1H), 7.33-7.27 (m, 2H), 5.13-5.09 (m, 1H), 4.30 (dd, J = 17.2Hz, J =58.2Hz,2H),3.46(d,J=4.8Hz,4H),3.23(d,J=4.8Hz,1H),2.93-2.78(m,1H),2.62-2.51(m,1H),2.48-2.37(m,1H),2.01-1.98(m,1H). Example S6. Procedure for CBM-10

[0324] Synthesis of 3-(1-methyl-6-(piperazin-1-yl)-1H-indazol-3-yl)piperidine-2,6-dione (CBM-10)

[0325] Step 1: tert-Butyl 4-(3-(2,6-bis(benzyloxy)pyridin-3-yl)-1-methyl-1H-indazol-6-yl)piperazine-1-carboxylate: A mixture of 6-bromo-3-(2,6-dibenzyloxy-3-pyridinyl)-1-methyl-indazole (30.0 g, 60.0 mmol), tert-butyl piperazine-1-carboxylate (16.8 g, 89.9 mmol), RuPhos-Pd-G3 (10 g, 12.0 mmol) and Cs2CO3 (23.4 g, 71.9 mmol) in degassed 1,4-dioxane (150 mL) was warmed to 75° C. for 18 h and then cooled to room temperature. The mixture was filtered through celite and the filter cake was washed with EtOAc (3×150 mL). The filtrate was concentrated under reduced pressure and the residue was purified by column chromatography on silica gel using a 0-40% EtOAc / hexanes gradient to give the title compound tert-butyl 4-(3-(2,6-bis(benzyloxy)pyridin-3-yl)-1-methyl-1H-indazol-6-yl)piperazine-1-carboxylate (35.6 g, 98% yield) as a solid.

[0326] MS (ESI) [M+H] + 607.5.

[0327] Step 2: tert-Butyl 4-(3-(2,6-dioxopiperidin-3-yl)-1-methyl-1H-indazol-6-yl)piperazine-1-carboxylate: tert-Butyl 4-[3-(2,6-dibenzyloxy-3-pyridinyl)-1-methyl-indazol-6-yl]piperazine-1-carboxylate (35.6 g, 58.8 mmol) and Pearlman's catalyst (8.90 g, 25 wt loading) in EtOH (300 mL) and THF (300 mL) were hydrogenated (1 atm) for 10 h at 50° C. The mixture was filtered through celite, and the filter cake was washed with a 1:1 mixture of MeCN and MeOH (4×250 mL). The filtrate was concentrated under reduced pressure and the residue was purified by column chromatography on silica gel using a 0-100% EtOAc / hexanes gradient to give the title compound tert-butyl 4-(3-(2,6-dioxopiperidin-3-yl)-1-methyl-1H-indazol-6-yl)piperazine-1-carboxylate (21.0 g, 84% yield) as a solid.

[0328] MS (ESI) [M+H] + 428.3.

[0329] Step 3: 3-(1-Methyl-6-(piperazin-1-yl)-1H-indazol-3-yl)piperidine-2,6-dione (CBM-10): To a solution of tert-butyl 4-[3-(2,6-dioxo-3-piperidinyl)-1-methyl-indazol-6-yl]piperazine-1-carboxylate (21.0 g, 49.1 mmol) in 1,4-dioxane (150 mL) was added 4N HCl in 1,4-dioxane (98.2 mL, 393 mmol) and the reaction mixture was stirred at room temperature for 20 h. Et2O (250 mL) was added and the precipitate was collected by filtration, washed with Et2O (3×30 mL), then dried under vacuum and lyophilized to give the title compound 3-(1-methyl-6-(piperazin-1-yl)-1H-indazol-3-yl)piperidine-2,6-dione as a solid (17.6 g, 98% yield).

[0330] MS (ESI) [M+H] + 328.2.

[0331] 1H NMR (500MHz, DMSO-d6) δ10.85(s,1H),9.46(s,2H),7.56(d,J=8.9Hz,1H),7.04–6.90(m,2H),4.28(dd,J=9.4,5. 0Hz,1H),3.92(s,3H),3.53–3.41(m,4H),3.23(s,4H),2.73–2.55(m,2H),2.39–2.26(m,1H),2.23–2.08(m,1H). Example S7. Procedure for CBM-11

[0332] Synthesis of 3-(1-methyl-7-(piperazin-1-yl)-1H-indazol-3-yl)piperidine-2,6-dione (CBM-11)

[0333] Step 1: 7-bromo-3-iodo-1-methyl-1H-indazole. Three batches were performed: t-BuOK (334 g, 2.97 mol, 2 equiv) was added in portions to a solution of 7-bromo-3-iodo-1H-indazole (480 g, 1.49 mol, 1 equiv) in THF (2.4 L) at 0 ° C. After addition, the suspension was stirred at 0 ° C for 1 h. A solution of CH3I (422 g, 2.97 mol, 185 mL, 2 equiv) in THF (400 mL) was then added dropwise to the cold (0 ° C) reaction mixture. The suspension was then stirred at 25 ° C for 3 h. TLC (PE / EtOAc=5 / 1, Rf=0.5) showed that the reaction was complete. The three batches of reaction mixture were combined, and the resulting suspension was poured into water (10 L) and stirred for 10 min. The aqueous phase was extracted with EtOAc (5.0 L, then 3.0 L). The combined organic layers were washed with brine (3.0 L), dried over anhydrous NaSO, filtered, and concentrated in vacuo. The crude material was purified by column chromatography on silica gel (PE / EtOAc=25 / 1, 5 / 1) to afford 7-bromo-3-iodo-1-methyl-1H-indazole (900 g, 2.67 mol, 60% yield) as a yellow solid.

[0334] 1 H NMR (400MHz, DMSO-d6) δppm 7.72 (d, J = 8.4 Hz, 1H), 7.49 (d, J = 7.2 Hz, 1H), 7.11 (t, J = 7.6 Hz, 1H), 4.34 (s, 3H).

[0335] Step 2: 3-(2,6-bis(benzyloxy)pyridin-3-yl)-7-bromo-1-methyl-1H-indazole. Three batches were performed: To a solution of 7-bromo-3-iodo-1-methyl-1H-indazole (313 g, 929 mmol, 1 eq) in 1,4-dioxane (2.0 L) and H₂O (1.0 L) were added (2,6-bis(benzyloxy)pyridin-3-yl)boronic acid (389 g, 929 mmol, 80% purity, 1 eq), K₃PO₄ (493 g, 2.32 mol, 2.5 eq), and Pd(PPh₃)₄ (21.5 g, 18.6 mmol, 0.02 eq). The suspension was then purged with N₂ three times and stirred at 90°C for 12 h. The three batches were combined and the reaction mixture was then poured into water (10 L) and stirred for 10 min. The aqueous phase was extracted with EtOAc (5 L, then 3 L). The combined organic phase was washed with brine (3 L), dried over anhydrous Na2SO4, filtered and concentrated in vacuo. The crude material was purified by column chromatography on silica gel (PE / EtOAc=25 / 1, 5 / 1). The residue was wet-milled with PE / EtOAc (2 / 1) at 25 ° C for 3 h, and then the solid was collected by vacuum filtration to obtain 3-(2,6-bis(benzyloxy)pyridin-3-yl)-7-bromo-1-methyl-1H-indazole (920 g, 63% yield) as an off-white solid.

[0336] 1 H NMR(400MHz,DMSO-d6)δppm 7.86(d,J=8.4Hz,1H),7.54(dd,J=8.0,0.8Hz,1H),7.37(dd,J=7.2,0.8Hz,1H),7.33(m, 2H), 7.28 (m, 8H), 6.94 (t, J = 7.6Hz, 1H), 6.60 (d, J = 7.6Hz, 1H), 5.43 (s, 4H), 4.36 (s, 3H).

[0337] Step 3: tert-Butyl 4-(3-(2,6-bis(benzyloxy)pyridin-3-yl)-1-methyl-1H-indazol-7-yl)piperazine-1-carboxylate. To a solution of 3-(2,6-bis(benzyloxy)pyridin-3-yl)-7-bromo-1-methyl-1H-indazole (100 g, 200 mmol) and tert-butyl piperazine-1-carboxylate (55.8 g, 300 mmol) in 1,4-dioxane (700 mL) were added Cs2CO3 (130 g, 400 mmol), RuPhos (18.6 g, 40 mmol) and Pd2(dba)3 (18.3 g, 20 mmol), then the suspension was purged with N2 three times and stirred at 110°C for 12 h. TLC (PE / EtOAc=3 / 1, R f=0.6) shows that the reaction is complete. The reaction is cooled to 20 ° C and filtered through a celite pad. The filtrate is concentrated under vacuum and the residue is purified by silica gel chromatography (100-200 mesh silica gel, PE / EtOAc=20 / 1,3 / 1) to obtain the product. The product is further purified by grinding with PE / EtOAc=(2 / 1,200mL) for 1h. The solid is collected by filtration and dried under vacuum to obtain tert-butyl 4-(3-(2,6-bis(benzyloxy)pyridin-3-yl)-1-methyl-1H-indazole-7-yl)piperazine-1-carboxylate (84g, 133mmol, 67% yield, 96% purity) as a yellow solid.

[0338] 1 H NMR(400MHz DMSO-d6)δppm 7.85(d,J=8.4Hz,1H),7.27-7.45(m,12H),6.96-6.99(m,2H),6.53(d,J=8.0Hz,1H),5.47(s,2H) ,5.40(s,2H),4.41(s,3H),4.10-4.16(m,2H),3.20-3.23(m,4H),2.84-2.89(m,2H),1.51(s,9H).

[0339] Step 4: tert-Butyl 4-(3-(2,6-dioxopiperidin-3-yl)-1-methyl-1H-indazol-7-yl)piperazine-1-carboxylate. To a suspension of tert-butyl 4-(3-(2,6-bis(benzyloxy)pyridin-3-yl)-1-methyl-1H-indazol-7-yl)piperazine-1-carboxylate (42 g, 69.3 mmol) and AcOH (4.16 g, 69.3 mmol, 3.97 mL) in THF (210 mL) and EtOH (210 mL) was added 10% Pd / C (8.0 g) and 20% Pd(OH) (8.0 g, 57 mmol), then the black suspension was purged with H three times and stirred at 50° C. at 50 psi for 12 h. The suspension was filtered through a pad of celite, and the filter cake was washed with hot THF (2 L). The filtrate was concentrated under vacuum at 45°C to give the crude product. The crude material was purified by silica gel chromatography (100-200 mesh silica gel, DCM / MeOH = 0 / 1, 10 / 1) to give a solid. The solid was further triturated with MTBE (50 mL) for 1 h. The solid was collected by filtration and dried under vacuum. tert-Butyl 4-(3-(2,6-dioxopiperidin-3-yl)-1-methyl-1H-indazol-7-yl)piperazine-1-carboxylate (18.6 g, 41.7 mmol, 30% yield, 96% purity) was obtained as a blue solid.

[0340] MS (ESI) [M+H]+ 428.4.

[0341] Step 5: 3-(1-methyl-7-(piperazin-1-yl)-1H-indazol-3-yl)piperidine-2,6-dione (CBM-11). To a solution of tert-butyl 4-(3-(2,6-dioxopiperidin-3-yl)-1-methyl-1H-indazol-7-yl)piperazine-1-carboxylate (18.6 g, 43.5 mmol, 1 eq) in DCM (420 mL) was added HCl / EtOAc (4M, 93.0 mL), and the suspension was stirred at 20 ° C for 2 h. The solid was collected by filtration and dried under vacuum at 45 ° C for 2 h. The solid was suspended in MeCN (100 mL) and dried under vacuum at 45 ° C for 2 h. The operation was repeated twice more. 3-(1-Methyl-7-(piperazin-1-yl)-1H-indazol-3-yl)piperidine-2,6-dione (16.5 g, 41.2 mmol, 95% yield, 2HCl) was obtained as a light blue solid.

[0342] 1 H NMR(400MHz,DMSO-d6)δppm 10.87(s,1H),9.49-9.58(m,2H),7.45(d,J=6.8Hz,1H),7.03-7.06(m,2H),4.33-4.37(m,1H ),4.24(s,3H),3.15-3.44(m,8H),2.60-2.67(m,2H),2.31-2.50(m,1H),2.14-2.18(m,1H). Example S8. Procedure for CBM-19

[0343] Synthesis of 1-methyl-3-(4-(piperazin-1-yl)phenyl)piperidine-2,6-dione (CBM-19)

[0344] Step 1: tert-Butyl 4-(4-(1-methyl-2,6-dioxopiperidin-3-yl)phenyl)piperazine-1-carboxylate: To a solution of MeOH (0.02 mL, 0.57 mmol, 1.0 equiv) in THF (6.0 mL) was added DIAD (0.11 mL, 0.57 mmol, 1.0 equiv) followed by tert-butyl 4-[4-(2,6-dioxopiperidin-3-yl)phenyl]piperazine-1-carboxylate Intermediate A-4a (300 mg, 0.57 mmol, 1.0 equiv) at 0° C. A solution of PPh (224.4 mg, 0.86 mmol, 1.5 equiv) in THF (3.62 mL; total concentration 0.06 M) was then added dropwise and the mixture was warmed to room temperature and stirred for 18 h. The solvent was removed under reduced pressure and the crude residue was purified by reverse phase column chromatography (5% to 70% MeCN / water with 0.1% formic acid). The fractions were concentrated to a minimum volume of water, neutralized with NaHCO 3 , and extracted with DCM (3×). The combined organic layers were dried over MgSO 4 , filtered, and concentrated to yield tert-butyl 4-(4-(1-methyl-2,6-dioxopiperidin-3-yl)phenyl)piperazine-1-carboxylate (156 mg, 84%) as an off-white solid.

[0345] LCMS: [M+H] + =388.2.

[0346] 1 H NMR (400MHz, DMSO-d6): δppm 1.42(s,9H),1.95-2.05(m,1H),2.07-2.19(m,1H),2.56-2.66(m,1H),2.69-2.81(m,1H),3.02(s,3H),3.04 -3.09(m,4H),3.41-3.48(m,4H),3.84(dd,J=11.0,4.9Hz,1H),6.91(d,J=8.6Hz,2H),7.06(d,J=8.6Hz,2H).

[0347] Step 2: 1-Methyl-3-(4-(piperazin-1-yl)phenyl)piperidine-2,6-dione (CBM-19): To a solution of tert-butyl 4-[4-(1-methyl-2,6-dioxo-3-piperidinyl)phenyl]piperazine-1-carboxylate (184 mg, 0.47 mmol) in DCM (4.74 mL, 0.10 M) was added 4 M HCl in dioxane (1.78 mL, 7.12 mmol, 15.0 equiv). The reaction was stirred at room temperature for 18 h. The solvent was removed under reduced pressure to yield the HCl salt of 1-methyl-3-(4-(piperazin-1-yl)phenyl)piperidine-2,6-dione (CBM-19) as a white solid (153 mg, 99.9% yield).

[0348] LCMS: [M+H] + =288.2.

[0349] 1 H NMR (400MHz, DMSO-d6): δppm 1.95-2.04(m,1H),2.09-2.20(m,1H),2.64(s,1H),2.70-2.82(m,1H),3.02(s,3H),3.18-3.26(m ,4H),3.30-3.37(m,4H),3.82-3.91(m,1H),6.95(d,J=8.6Hz,2H),7.10(d,J=8.6Hz,2H),8.89(br s,1H). Example S9. Procedure for CBM-25

[0350] Synthesis of 3-(4-(2,6-diazaspiro[3.3]heptane-2-yl)phenyl)piperidine-2,6-dione (CBM-25)

[0351] Step 1: tert-Butyl 6-(4-(2,6-dioxopiperidin-3-yl)phenyl)-2,6-diazaspiro[3.3]heptane-2-carboxylate: To a 250 mL screw-cap flask was added DMSO (34.7 mL) and molecular sieves (~10 grains, ). The solvent was stirred at room temperature for 10 minutes with nitrogen bubbling. Then, 3-(4-bromophenyl)piperidine-2,6-dione (1.86 g, 6.94 mmol), tert-butyl 2,6-diazaspiro[3.3]heptane-2-carboxylate oxalate (2.03 g, 4.16 mmol), DMPAO (1.34 g, 6.94 mmol), CuI (660.62 mg, 3.47 mmol) and tetrabutylammonium acetate (6.28 g, 20.81 mmol) were added and nitrogen was bubbled through the solution for 10 minutes. The reaction mixture was stirred at 110 ° C until completion (ca. 16 h). Water was added and the aqueous phase was extracted three times with EtOAc. The organic phases were combined, washed with water and brine, dried over Na2SO4 and concentrated to dryness. The resulting residue was purified first by reverse phase column chromatography (5 to 100% MeCN / water with 0.1% formic acid) followed by silica gel column chromatography (0 to 100% EtOAc / heptane) to afford tert-butyl 6-(4-(2,6-dioxopiperidin-3-yl)phenyl)-2,6-diazaspiro[3.3]heptane-2-carboxylate (136 mg, 0.35 mmol, 5% yield) as an off-white solid.

[0352] LCMS: [M+H] + =386.2.

[0353] 1 H NMR (400MHz, DMSO-d6): δppm 1.38(s,9H),1.93-2.02(m,1H),2.07(s,2H),2.56-2.65(m,1H),3.69(dd,J=10.8,4.9 Hz, 1H), 3.98-4.04 (m, 5H), 6.38 (d, J = 8.6Hz, 2H), 7.00 (d, J = 8.3Hz, 2H), 10.75 (s, 1H).

[0354] Step 2: 3-(4-(2,6-diazaspiro[3.3]heptane-2-yl)phenyl)piperidine-2,6-dione (CBM-25): In a round-bottom flask, tert-butyl 6-[4-(2,6-dioxo-3-piperidinyl)phenyl]-2,6-diazaspiro[3.3]heptane-2-carboxylate 3 (152.8 mg, 0.40 mmol) was dissolved in DCM (2 mL), followed by the addition of trifluoroacetic acid (0.61 mL, 7.93 mmol) and the mixture was stirred at room temperature for 30 min. The solvent was evaporated under reduced pressure and co-evaporated with acetonitrile three times to give 3-(4-(2,6-diazaspiro[3.3]heptane-2-yl)phenyl)piperidine-2,6-dione (CBM-25) (204.1 mg, quantitative yield, TFA salt) as a grey oil, which was used in the next step without further purification.

[0355] 1 H NMR (400MHz, DMSO-d6): δppm 1.93-2.01(m,1H),2.08-2.14(m,1H),2.57-2.65(m,1H),3.70(dd,J=11.0,4.9H z,1H),4.13-4.17(m,4H),6.42(d,J=8.6Hz,2H),7.02(d,J=8.6Hz,2H),8.43(br s,2H),10.76(s,1H). Example S10. Procedure for CBM-26

[0356] Synthesis of 1-(4-(piperazin-1-yl)phenyl)pyrimidine-2,4(1H,3H)-dione (CBM-26)

[0357] Step 1: tert-Butyl 3-(4-(4-(tert-Butoxycarbonyl)piperazin-1-yl)phenyl)-2,6-dioxo-3,6-dihydropyrimidine-1(2H)-carboxylate: To a solution of tert-butyl 2,4-dioxo-1H-pyrimidine-3-carboxylate (300 mg, 1.41 mmol, 1.0 equiv) in EtOAc (10 mL) was added [4-(4-tert-Butoxycarbonylpiperazin-1-yl)phenyl]boronic acid (562 mg, 1.84 mmol, 1.3 equiv), Et3N (0.49 mL, 3.53 mmol, 2.5 equiv), followed by Cu(OAc)2 (385 mg, 2.12 mmol, 1.5 equiv). The reaction was stirred at room temperature overnight under air. The reaction mixture was diluted with EtOAc and water. The phases were separated and the organic layer was washed with NH4Cl (2x), water, brine, dried over sodium sulfate, filtered and concentrated. The mixture was purified by reverse phase column chromatography (5% to 100% MeCN / water with 0.1% formic acid) to afford tert-butyl 3-[4-(4-tert-butoxycarbonylpiperazin-1-yl)phenyl]-2,6-dioxo-pyrimidine-1-carboxylate (260 mg, 39% yield) as an off-white solid.

[0358] LCMS: [M+H] + =473.2.

[0359] 1H NMR (400MHz, DMSO-d6): δppm 1.42(s,9H),1.51(s,9H),3.11-3.20(m,4H),3.41-3.51(m,4H),5.81(d ,J=7.8Hz,1H),6.99-7.07(m,2H),7.26-7.34(m,2H),7.74-7.80(m,1H).

[0360] Step 2: 1-(4-(piperazin-1-yl)phenyl)pyrimidine-2,4(1H,3H)-dione (CBM-26): To a solution of tert-butyl 3-[4-(4-tert-butoxycarbonylpiperazin-1-yl)phenyl]-2,6-dioxo-pyrimidine-1-carboxylate (260 mg, 0.550 mmol, 1.0 equiv) in CHCl (2 mL) was added HCl (1 mL, 5.5 mmol, 10 equiv) at room temperature for 4 h. The solvent was evaporated and co-evaporated with MeCN (3x) and MTBE (2x) to give 1-(4-piperazin-1-ylphenyl)pyrimidine-2,4-dione (CBM-26) (240 mg, quantitative, bis-HCl salt) as an off-white solid.

[0361] LCMS: [M+H] + =473.2.

[0362] 1 H NMR (400MHz, DMSO-d6): δ3.22(br s,4H),3.38-3.47(m,4H),5.63(dd,J=7.8,2.2Hz,1H),7.06(d,J=9.0Hz,2H) ,7.28(d,J=9.0Hz,2H),7.63(d,J=7.8Hz,1H),9.17(brs,2H),11.38(s,1H). Example S11. Procedure for CBM-30

[0363] Synthesis of 5-fluoro-1-(4-(piperazin-1-yl)phenyl)pyrimidine-2,4(1H,3H)-dione (CBM-30)

[0364] Step 1: 5-Fluoro-1-(4-nitrophenyl)pyrimidine-2,4(1H,3H)-dione: To a solution of 5-fluoro-1H-pyrimidine-2,4-dione (500 mg, 3.84 mmol, 1.0 equiv) in DMSO (10 mL) was added 1-fluoro-4-nitro-benzene (542 mg, 3.84 mmol, 1.0 equiv) and K2CO3 (1.33 g, 9.61 mmol, 2.5 equiv) at room temperature. The reaction mixture was then heated to 80°C for 1.5 h. LCMS showed 55-60% conversion. The reaction mixture was cooled to room temperature and water (5 mL) was added. The mixture was diluted with water and EtOAc. The phases were separated and the aqueous phase was concentrated and purified by reverse phase column chromatography (5% to 50% MeCN / water with 0.1% formic acid) to give 5-fluoro-1-(4-nitrophenyl)pyrimidine-2,4(1H,3H)-dione (291 mg, 30%) as a white solid.

[0365] LCMS: [M+H] + =252.2.

[0366] 1 H NMR (400MHz, DMSO-d6): δppm 7.72-7.82(m,2H), 8.29-8.41(m,3H), 12.04-12.15(m,1H).

[0367] Step 2: 1-(4-aminophenyl)-5-fluoropyrimidine-2,4(1H,3H)-dione: To a solution of 5-fluoro-1-(4-nitrophenyl)pyrimidine-2,4(1H,3H)-dione (360 mg, 1.43 mmol, 1.0 equiv) in 1,4-dioxane (3 mL) and water (3 mL) was added Fe (640 mg, 11.47 mmol, 8.0 equiv) and NHCl (613 mg, 11.47 mmol, 8.0 equiv) at room temperature. The reaction mixture was heated at 75° C. overnight. LCMS showed complete conversion. The mixture was filtered through celite and washed with MeCN to give 1-(4-aminophenyl)-5-fluoropyrimidine-2,4(1H,3H)-dione (260 mg, 82%) as an off-white solid.

[0368] LCMS: [M+H] + =222.2.

[0369] 1 H NMR (400MHz, DMSO-d6): δppm 5.35 (s, 2H), 6.52-6.64 (m, 2H), 6.98-7.06 (m, 2H), 7.16-7.27 (m, 1H), 8.05 (d, J = 6.6Hz, 1H).

[0370] 19 F NMR (377MHz, DMSO-d6): δppm-170.47 (d, J=6.8Hz, 1F).

[0371] Step 3: tert-Butyl 4-(4-(5-fluoro-2,4-dioxo-3,4-dihydropyrimidin-1(2H)-yl)phenyl)piperazine-1-carboxylate: To a solution of 1-(4-aminophenyl)-5-fluoropyrimidine-2,4(1H,3H)-dione (150 mg, 0.68 mmol, 1.0 eq) and tert-butyl N,N-bis(2-oxoethyl)carbamate (205 mg, 1.02 mmol, 1.5 eq) in DCE (5 mL) was added NaBH(OAc) (287 mg, 1.36 mmol, 2.0 eq) at room temperature. The reaction mixture was stirred at room temperature overnight. LCMS showed the enamine intermediate with a trace of the desired product. NaBH(OAc) (287 mg, 1.36 mmol, 2.0 eq) was added and the mixture was stirred at room temperature overnight. LCMS showed approximately 50% conversion. To the 4- (4- (5-fluoro-2,4-dioxo-3,4-dihydropyrimidin-1 (2H) -yl) phenyl) piperazine-1-carboxylic acid tert-butyl ester (60 mg, 23% yield) was added NaBH (OAc) (287 mg, 1.36 mmol, 2.0 equivalents) and the mixture was stirred at room temperature for 72 h. LCMS showed almost complete conversion. Water and CH Cl were added and the phases were separated. The aqueous phase was extracted with CH Cl (2x), and the combined organic layers were dried over sodium sulfate, filtered, and concentrated. The crude product was purified by reverse phase column chromatography (5 to 100% MeCN / water, containing 0.1% FA) to obtain tert-butyl 4- (4- (5-fluoro-2,4-dioxo-3,4-dihydropyrimidin-1 (2H) -yl) phenyl) piperazine-1-carboxylate (60 mg, 23% yield) as an off-white solid.

[0372] LCMS: [M+H] + =391.2.

[0373] 1 H NMR (400MHz, DMSO-d6): δppm 1.42(s,9H),3.11-3.21(m,4H),3.42-3.50(m,4H),6.98-7.04(m,2H),7.23-7.31(m,2H),8.12(d,J=6.6Hz,1H),11.85(br s,1H).

[0374] 19 F NMR (377MHz, DMSO-d6): δppm-170.13 (br d, J=5.4Hz, 1F).

[0375] Step 4: 5-Fluoro-1-(4-(piperazin-1-yl)phenyl)pyrimidine-2,4(1H,3H)-dione (CBM-30): To a solution of tert-butyl 4-(4-(5-fluoro-2,4-dioxo-3,4-dihydropyrimidin-1(2H)-yl)phenyl)piperazine-1-carboxylate (60 mg, 0.15 mmol, 1.0 equiv) in CHCl (2 mL) was added HCl (1 mL, 3.75 mmol, 25 equiv) at room temperature and the reaction mixture was stirred at room temperature for 5 h. LCMS showed complete conversion. The solvent was evaporated and co-evaporated with MeCN (3x) and CHCl (2x) to give 5-fluoro-1-(4-(piperazin-1-yl)phenyl)pyrimidine-2,4(1H,3H)-dione (CBM-30) (60 mg, quantitative yield, bis-HCl salt) as an off-white solid.

[0376] LCMS: [M+H] + =291.2.

[0377] 1 H NMR (400MHz, DMSO-d6): δppm 3.19-3.27(m,4H),3.38-3.45(m,4H),7.02-7.10(m,2H),7.26-7.35(m,2H),8.12(d,J=6.6Hz,1H),8.93(br dd,J=2.2,1.5Hz,2H), 11.88(d,J=5.1Hz,1H).

[0378] 19 F NMR (377MHz, DMSO-d6): δppm-170.14--169.97(m,1F). Example S12. Procedure for CBM-31

[0379] Synthesis of 1-(1-(piperidin-4-yl)-1H-pyrazol-4-yl)dihydropyrimidine-2,4(1H,3H)-dione (CBM-31)

[0380] Step 1: tert-Butyl 4-(4-((2-cyanoethyl)amino)-1H-pyrazol-1-yl)piperidine-1-carboxylate: A solution of tert-butyl 4-(4-aminopyrazol-1-yl)piperidine-1-carboxylate (1.37 g, 5.14 mmol), saturated aqueous Na2CO3 (57 mL, 5.71 mmol) and prop-2-enenitrile (6.85 mL, 104.57 mmol) in THF (10 mL) was stirred at room temperature for 48 h. The aqueous solution was extracted with EtOAc, and the combined organic phases were washed with NaCl and dried over MgSO4. The mixture was concentrated to dryness and purified by silica gel column chromatography (0% to 100% EtOAc / heptane) to give tert-butyl 4-(4-((2-cyanoethyl)amino)-1H-pyrazol-1-yl)piperidine-1-carboxylate (1.6 g, 97% yield) as a brown oil.

[0381] LCMS: [M+H] + =320.2.

[0382] 1 H NMR (400MHz, CDCl3): δppm 1.41 (s, 9H), 1.71 (ddd, J=24.0, 12.0, 4.2Hz, 2H), 1.88-1.95 (m, 2H), 2.65 (t, J=6.6Hz, 2H), 2.87 (br s,2H),3.10(q,J=6.6Hz,2H),4.00(brs,2H),4.09-4.22(m,1H),4.66(t,J=6.4Hz,1H),7.00(br s,1H),7.25(br s,1H).

[0383] Step 2: tert-Butyl 4-(4-(N-(2-cyanoethyl)cyanamido)-1H-pyrazol-1-yl)piperidine-1-carboxylate: A solution of tert-butyl 4-(4-((2-cyanoethyl)amino)-1H-pyrazol-1-yl)piperidine-1-carboxylate (1.6 g, 5.01 mmol) in EtOH (20 mL) was slowly added to a 0° C. solution of cyanogen bromide (6.68 mL, 20.04 mmol) and NaOAc (1.42 mL, 12.52 mmol) in EtOH (5 mL). The reaction was stirred until completion (ca. 18 h) as shown by LCMS. The mixture was concentrated to dryness and the residue was washed with 5% aqueous citric acid solution and extracted with EtOAc. The organic phase was washed with brine and dried over MgSO4 and the crude material was purified by silica gel column chromatography (0% to 100% EtOAc / heptane) to give tert-butyl 4-(4-(N-(2-cyanoethyl)cyanamido)-1H-pyrazol-1-yl)piperidine-1-carboxylate (1.5 g, 81% yield) as an orange oil.

[0384] LCMS: [M+Na] + =367.0.

[0385] 1 H NMR (400MHz, CDCl3): δppm 1.41(s,9H),1.74(ddd,J=24.9,12.2,4.4Hz,2H),1.91-1.98(m,2H),2.79-2.99(m,J=6.4,6.4Hz,4H),3 .77(t,J=6.5Hz,2H),3.96-4.02(m,2H),4.25-4.36(m,1H),7.49(d,J=0.7Hz,1H),7.97(d,J=0.7Hz,1H).

[0386] Step 3: 1-(1-(Piperidin-4-yl)-1H-pyrazol-4-yl)dihydropyrimidine-2,4(1H,3H)-dione (CBM-31): A solution of tert-butyl 4-(4-(N-(2-cyanoethyl)cyanamido)-1H-pyrazol-1-yl)piperidine-1-carboxylate (1.5 g, 4.36 mmol) in 6N aqueous HCl (4.5 mL, 27 mmol) was heated to 100° C. until complete by LCMS (ca. 3 h). The reaction mixture was concentrated to dryness and the crude material was suspended in a mixture of 30% MeOH / DCM and neutralized with a saturated NaHCO sol (pH ˜7). The aqueous phase was concentrated to dryness and the residue was purified by reverse phase column chromatography (0% to 100% MeCN / water with 0.1% formic acid) to afford 1-(1-(piperidin-4-yl)-1H-pyrazol-4-yl)dihydropyrimidine-2,4(1H,3H)-dione (CBM-31) as a tan solid after lyophilization (764.7 mg, 67% yield).

[0387] LCMS: [M+H] + =264.2.

[0388] 1 H NMR (400MHz, DMSO-d6): δppm 1.74(ddd,J=24.0,11.7,4.2Hz,2H),1.85-1.94(m,2H),2.53-2.61(m,2H),2.68(t,J=6.8Hz,2H),2.97 -3.06(m,J=12.5Hz,2H),3.75(t,J=6.8Hz,2H),4.04-4.20(m,1H),7.59(s,1H),7.92(s,1H),10.36(br s,1H). Procedure for target binding moiety (TBM) Example S13. General Procedure for TBM-D

[0389] Synthesis of TBM-D: Step 1 To a round-bottom flask were added 5,8-dibromoimidazo[1,2-a]pyrazine intermediate D-1a (5.0 g, 18 mmol, 1 equiv), tert-butyl 4-(4-aminopyrazol-1-yl)piperidine-1-carboxylate intermediate D-2a (5.29 g, 19.8 mmol, 1.1 equiv), and citric acid (27.66 g, 271 mmol, 15 equiv). The flask was then placed in a preheated oil bath at 100°C. After 2 h, LCMS indicated complete conversion with traces of the deprotected product. The brown residue was quenched with saturated NaHCO3 solution (until pH 7-9) and extracted with MeTHF (3x). The combined organic layers were washed with brine, dried over Na2SO4, and concentrated to yield a dark red residue. The residue was then purified by reverse-phase column chromatography (5% to 100% MeOH / water containing 0.1% formic acid). The fractions were combined and concentrated to give tert-butyl 4-(4-((5-bromoimidazo[1,2-a]pyrazin-8-yl)amino)-1H-pyrazol-1-yl)piperidine-1-carboxylate intermediate D-3a as a brown solid (5.99 g, 70% yield).

[0390] LCMS: [M+H] + =462.2,464.2.

[0391] 1 H NMR (400MHz, DMSO-d6) δppm 1.42 (s, 9H), 1.76 (qd, J=12.1, 4.2Hz, 2H), 1.99 (br dd,J=12.3,2.3Hz,2H),2.78-2.99(m,2H),3.97-4.12(m,2H),4.30-4.39(m,1H),7.59(s, 1H),7.70(d,J=1.2Hz,1H),7.76(s,1H),7.98(d,J=1.0Hz,1H),8.18(s,1H),10.00(s,1H). Table 16. TBM-D: List of intermediates used in step 1

[0392] Synthesis of 5,8-dibromo-3-methylimidazo[1,2-a]pyrazine (Intermediate D-1c) To a round-bottom flask was added 2-bromo-1,1-diethoxy-propane 1 (1.59 mL, 9.9 mmol, 5 eq) in IPA (9.9 mL, 0.2 M). HBr (1.18 mL, 21.7 mmol, 11 eq) was added to the reaction mixture and the reaction mixture was heated to 95°C and stirred overnight. The resulting yellow solution was cooled to room temperature, neutralized with solid NaHCO3, filtered, and rinsed with isopropanol. The resulting solution was placed in a flask and 3,6-dibromopyrazin-2-amine (500 mg, 2.0 mmol) was added and heated to reflux at 95°C and reacted overnight. The solution was cooled to room temperature and concentrated. The residue was dissolved in EtOAc, washed with NaHCO3 (2x) and brine, dried over sodium sulfate, filtered, and concentrated. The crude product was purified by reverse phase column chromatography (5% to 100% in water with 0.1% formic acid). The fractions were combined and concentrated to give 5,8-dibromo-3-methylimidazo[1,2-a]pyrazine intermediate D-1c (212 mg, 36% yield) as a yellow solid.

[0393] LCMS: [M+H] + =292.0.

[0394] 1 H NMR (400MHz, DMSO-d6) δppm 2.82 (s, 3H), 7.72 (s, 1H), 7.84 (s, 1H).

[0395] Synthesis of 8-chloro-5,6-dimethylimidazo[1,2-a]pyrazine (Intermediate D-1d) To a pressure tube was added 3-bromo-5,6-dimethyl-pyrazin-2-amine (200 mg, 0.99 mmol), 2-chloroacetaldehyde in H2O (50% w / w, 1.34 mL, 9.9 mmol) and 1,4-dioxane (0.825 mL), purged with N2, sealed and heated to 90°C until completion by LCMS analysis (ca. 2 h). The reaction mixture was concentrated to dryness and the crude material was taken up in DMSO / 1M NaOH aqueous solution and purified by reverse phase column chromatography (5% MeCN / water with pH 10 buffer, then 5 to 100% MeCN / water with formic acid) to give 8-chloro-5,6-dimethylimidazo[1,2-a]pyrazine intermediate D-1d (128 mg, 71% yield) as an off-white solid.

[0396] LCMS: [M+H] + =182.2.

[0397] 1H NMR (400MHz, DMSO-d6) δppm 2.45 (s, 3H), 2.58 (s, 3H), 7.86 (d, J = 1.0 Hz, 1H), 8.18 (d, J = 1.0 Hz, 1H).

[0398] Synthesis of tert-butyl (2S,4S)-4-(4-amino-1H-pyrazol-1-yl)-2-methylpiperidine-1-carboxylate (Intermediate D-2b)

[0399] Step 1: To a solution of (2S, 4R)-4-hydroxy-2-methyl-piperidine-1-carboxylic acid tert-butyl ester 1 (500.0 mg, 2.32 mmol, 1 eq) in CHCl (15 mL) was added EtN (0.97 mL, 6.97 mmol, 3 eq) and MsCl (0.27 mL, 3.48 mmol, 1.5 eq) at room temperature. After 90 min, LCMS showed the expected mass. The reaction was quenched with saturated NHCl solution and diluted with CHCl. ​​The phases were separated, and the organic layer was dried over sodium sulfate, filtered, and concentrated to give (2S, 4R)-2-methyl-4-((methylsulfonyl)oxy)piperidine-1-carboxylic acid tert-butyl ester (701 mg, 95% yield) as a light yellow solid.

[0400] LCMS: [M+H] + =194.2.

[0401] 1 H NMR (400MHz, chloroform-d) δppm 1.19(d,J=7.1Hz,3H),1.44-1.49(m,9H),1.66(brdd,J=12.1,5.0Hz,1H),1.83(dt,J=12.1,6.2Hz,1H ),1.99(dt,J=12.6,2.3Hz,1H),2.07-2.21(m,1H),2.93(td,J=13.7,2.7Hz,1H),3.03(s,3H),4.10(br d,J=12.7Hz,1H),4.48-4.63(m,1H),4.89-5.02(m,1H).

[0402] Step 2: To a solution of tert-butyl (2S,4R)-2-methyl-4-((methylsulfonyl)oxy)piperidine-1-carboxylate (778 mg, 2.65 mmol, 1.2 eq) and 4-nitro-1H-pyrazole 3 (250 mg, 2.21 mmol, 1 eq) in DMF (10.2 mL) was added CsCO (1.1 g, 3.1 mmol, 1.4 eq) at room temperature. The reaction flask was placed in an oil bath pre-equilibrated to 90 ° C. After 18 h, LCMS showed complete conversion. The reaction mixture was partitioned between water and EtOAc. The phases were separated and the aqueous layer was extracted with EtOAc (3x). The combined organics were washed with brine (3x), dried over sodium sulfate, filtered, concentrated, and purified by reverse phase column chromatography (5% to 100% MeOH / water with 0.1% formic acid). Fractions were combined and concentrated to give tert-butyl (2S,4S)-2-methyl-4-(4-nitro-1H-pyrazol-1-yl)piperidine-1-carboxylate (529 mg, 75% yield) as an orange oil.

[0403] LCMS: [M-tBu+H] + =255.2.

[0404] Step 3: Pd / C (267.47 mg, 0.25 mmol, 0.15 equiv) was added to a solution of (2S, 4S)-2-methyl-4-(4-nitro-1H-pyrazol-1-yl)piperidine-1-carboxylic acid tert-butyl ester (529 mg, 1.68 mmol, 1 equiv) in ethanol (8.5 mL) under nitrogen. The atmosphere of the flask was replaced with hydrogen by bubbling hydrogen into the mixture. The reaction mixture was then stirred at room temperature under 1 atm of hydrogen. After 1.5 h, LCMS showed complete conversion. The mixture was filtered through a celite pad, washed with EtOAc, and concentrated under reduced pressure to obtain (2S, 4S)-4-(4-amino-1H-pyrazol-1-yl)-2-methylpiperidine-1-carboxylic acid tert-butyl ester intermediate D-2b (486 mg, 98% yield) as a purple oil.

[0405] LCMS: [M+H] + =281.4.

[0406] 1H NMR(400MHz,DMSO-d6)δppm 0.93(d,J=6.6Hz,3H),1.39-1.42(m,9H),1.89-1.97(m,2H),2.00-2.11(m,2H),3.23-3.30(m,1H),3 .60-3.69(m,1H),3.69-3.87(m,2H),3.87-3.98(m,1H),4.14-4.23(m,1H),6.92(s,1H),7.10(s,1H).

[0407] Synthesis of tert-butyl (2R,4R)-4-(4-amino-1H-pyrazol-1-yl)-2-methylpiperidine-1-carboxylate (Intermediate D-2c)

[0408] Step 1: To a solution of triphenylphosphine (1.50 g, 5.71 mmol 1.50 equiv), (2R,4S)-4-hydroxy-2-methyl-piperidine-1-carboxylic acid tert-butyl ester (0.82 g, 3.8 mmol, 1.00 equiv) and 4-nitro-1H-pyrazole (0.65 g, 5.71 mmol, 1.50 equiv) in dry THF was added diisopropyl azodicarboxylate (1.12 mL, 5.71 mmol) in THF (19.00 mL, total concentration 0.10 M) at 0° C. under argon (19.00 mL) and the reaction was allowed to warm to room temperature over a period of 18 h. The reaction was concentrated under reduced pressure and purified by reverse phase column chromatography (5% to 90% MeOH / water with 0.1% formic acid). Fractions were combined and concentrated to give tert-butyl (2R,4R)-2-methyl-4-(4-nitro-1H-pyrazol-1-yl)piperidine-1-carboxylate (1.14 g, 91% yield) as a colorless semisolid.

[0409] LCMS: [M+H] + =211.2.

[0410] 1 H NMR(400MHz,DMSO-d6)δppm 0.98(d,J=6.6Hz,3H),1.41(s,9H),2.03-2.18(m,4H),3.31(ddd,J=14.2,9.4,5.5Hz,1 H),3.60-3.74(m,1H),3.90-4.03(m,1H),4.47-4.57(m,1H),8.30(s,1H),8.99(s,1H).

[0411] Step 2: To a solution of (2R, 4R)-2-methyl-4-(4-nitro-1H-pyrazol-1-yl)piperidine-1-carboxylic acid tert-butyl ester (1.14 g, 3.49 mmol, 1.00 equiv) in ethanol (34.89 mL, 0.10 M) was added 10% w / w Pd / C (371 mg, 0.35 mmol, 0.10 equiv). H2 gas was bubbled into the mixture using a rubber balloon and a stainless steel needle. After bubbling for 10 min, the needle was removed and the reaction was stirred at room temperature under an H2 atmosphere for 18 h. LCMS showed complete conversion to the desired product. The crude mixture was filtered over a 2-inch Celite pad, which was further washed with MeOH. The mixture was concentrated to give tert-butyl (2R,4R)-4-(4-amino-1H-pyrazol-1-yl)-2-methylpiperidine-1-carboxylate intermediate D-2c (1.05 g, quantitative yield) as a purple solid, which was used without purification.

[0412] LCMS: [M+H] + =281.4.

[0413] 1 H NMR(400MHz,DMSO-d6)δppm 0.93(d,J=6.8Hz,3H),1.40(s,9H),1.87-2.10(m,4H),3.22-3.31(m,1H),3.60-3.69(m,1H ),3.75-3.86(m,2H),3.93(sxt,J=6.6Hz,1H),4.13-4.25(m,1H),6.92(s,1H),7.10(s,1H).

[0414] Synthesis of tert-butyl (3R,4R)-4-(4-amino-1H-pyrazol-1-yl)-3-fluoropiperidine-1-carboxylate (Intermediate D-2e)

[0415] Step 1: To a solution of 4-nitro-1H-pyrazole (618.9 mg, 5.47 mmol, 2 equiv), diisopropyl azodicarboxylate (1.1 mL, 5.47 mmol, 2 equiv) and triphenylphosphine polymer bond (1.82 g, 5.47 mmol, 2 equiv) in dry THF (7 mL, 0.2 M) was added (3R,4S)-3-fluoro-4-hydroxy-piperidine-1-carboxylic acid tert-butyl ester (600 mg, 2.74 mmol, 1 equiv) in dry THF (7 mL, 0.2 M) under nitrogen at 0° C. After 19 h at room temperature, the reaction mixture was filtered and concentrated in vacuo, and the crude residue was purified by reverse phase column chromatography (5% to 100% MeOH / water with 0.1% formic acid). Fractions were combined and concentrated to give tert-butyl (3R,4R)-3-fluoro-4-(4-nitro-1H-pyrazol-1-yl)piperidine-1-carboxylate (404 mg, 47% yield) as a yellow oil.

[0416] LCMS: [M-tBu+H] + =259.2.

[0417] 1 H NMR (400MHz, DMSO-d6) δppm 1.37(d,J=6.4Hz,2H),1.43(s,9H),1.95(qd,J=12.4,4.4Hz,1H),2.04-2.14(m,1H),3.98(br d,J=13.2Hz,1H),4.22-4.39(m,1H),4.68(qd,J=10.7,4.6Hz,1H),4.74-4.95(m,1H),8.35(s,1H),9.09(s,1H).

[0418] 19 F NMR (377MHz, DMSO-d6) δppm-188.01 (br dd, J=50.4, 5.4Hz, 1F).

[0419] Step 2: A solution of (3R, 4R)-3-fluoro-4-(4-nitro-1H-pyrazol-1-yl)piperidine-1-carboxylic acid tert-butyl ester (404 mg, 1.29 mmol, 1 equivalent) in ethanol (23 mL, 0.06 M) was sparged with nitrogen three times, and Pd / C (137 mg, 0.13 mmol, 0.1 equivalent) was added. The mixture was then purged with hydrogen (balloon) three times and then stirred at room temperature under a hydrogen atmosphere for 19 h. The mixture was filtered on a celite pad and rinsed with EtOAc and EtOH. The filtrate was concentrated in vacuo to obtain (3R, 4R)-4-(4-amino-1H-pyrazol-1-yl)-3-fluoropiperidine-1-carboxylic acid tert-butyl ester intermediate D-2e (370 mg, 65% yield) as a purple oil, which was used without further purification.

[0420] LCMS: [Mt-Bu+H] + =229.2.

[0421] 1 H NMR(400MHz,DMSO-d6)δppm 1.18(d,J=6.1Hz,2H),1.42(s,9H),1.80-1.97(m,2H),3.77-3.97(m,2H),4 .22-4.37(m,2H),4.57-4.83(m,2H),6.95-7.02(m,1H),7.11-7.20(m,1H).

[0422] 19 F NMR (377MHz, DMSO-d6) δppm-186.81--186.56(m,1F).

[0423] Synthesis of tert-butyl (3R,4S)-4-(4-amino-1H-pyrazol-1-yl)-3-fluoropiperidine-1-carboxylate (Intermediate D-2f)

[0424] Step 1: To a solution of 4-nitro-1H-pyrazole (645 mg, 5.71 mmol), diisopropyl azodicarboxylate (1.1 mL, 5.49 mmol), and triphenylphosphine polymer bond (1.98 g, 5.94 mmol) in dry THF (18 mL) was slowly added a solution of (3R,4R)-3-fluoro-4-hydroxy-piperidine-1-carboxylic acid tert-butyl ester (607 mg, 2.77 mmol) in dry THF (18 mL) at 0°C under nitrogen. After 18 h, the reaction mixture was filtered at room temperature and concentrated in vacuo. The residue was purified by reverse phase column chromatography (5% to 100% MeOH / water with 0.1% formic acid). Collected fractions gave tert-butyl (3R,4S)-3-fluoro-4-(4-nitro-1H-pyrazol-1-yl)piperidine-1-carboxylate (511 mg, 1.63 mmol, 59% yield) as a white solid.

[0425] LCMS: [M-tBu+H] + =259.2.

[0426] 1 H NMR(400MHz,DMSO-d6)δppm 1.19(br d,J=6.1Hz,1H),1.42(s,9H),1.99–2.08(m,1H),2.24–2.36(m,1H),2.84–3.10(m,1H),4.09–4 .22(m,1H),4.24–4.43(m,1H),4.68–4.83(m,1H),4.99–5.19(m,1H),8.35(s,1H),8.95(s,1H).

[0427] 19 F NMR (377MHz, DMSO-d6) δppm-203.26--202.52(m,1F).

[0428] In 40mL 4-(4-amino-1H-pyrazoles-1-yl)-3-fluoropiperidine-1-t-butyl formate intermediate D-2f (453mg, 1.05mmol, 66% productive rate) is added in the 4-amino-1H-pyrazoles-1-yl)-3-fluoropiperidine-1-t-butyl formate intermediate D-2f (50mL).Then mixture is purged three times with hydrogen (balloon) and then in room temperature under hydrogen (balloon), stirred 19h.Mixture is flushed with nitrogen and filtered on diatomite pad and rinsed with MeOH (50mL).Filtrate is concentrated in a vacuum to obtain (3R, 4S)-4-(4-amino-1H-pyrazoles-1-yl)-3-fluoropiperidine-1-t-butyl formate intermediate D-2f (453mg, 1.05mmol, 66% productive rate) as purple oil, it is not used without further purification.

[0429] LCMS: [M+H] + =285.2.

[0430] 1H NMR (400MHz, DMSO-d6) δppm 1.14–1.27(m,1H),1.41(s,9H),1.81–1.93(m,1H),1.99–2.18(m,1H),2.79–3.01(m,1H),4.12(br d,J=2.9Hz,1H),4.18–4.31(m,1H),4.36–4.48(m,1H),4.67–4.82(m,2H),4.84–5.07(m,1H),7.02(s,1H),7.12–7.17(m,1H).

[0431] 19 F NMR (377MHz, DMSO-d6) δppm-202.38--201.24(m,1F).

[0432] Synthesis of tert-butyl (3S,4R)-4-(4-amino-1H-pyrazol-1-yl)-3-fluoropiperidine-1-carboxylate (Intermediate D-2h)

[0433] Step 1: Prepare a solution of 4-nitro-1H-pyrazole (643 mg, 5.69 mmol), (3S,4S)-3-fluoro-4-hydroxy-piperidine-1-carboxylic acid tert-butyl ester (604 mg, 2.76 mmol), and triphenylphosphine (1.91 g, 5.74 mmol) in dry THF (18 mL) and cool to 0°C under nitrogen. Then, slowly add a solution of diisopropyl azodicarboxylate (1.1 mL, 5.59 mmol) in dry THF (18 mL). After 17 h at room temperature, filter the reaction mixture and purify by reverse phase column chromatography (5% to 100% MeOH / water with 0.1% formic acid). Collected fractions gave tert-butyl (3S,4R)-3-fluoro-4-(4-nitro-1H-pyrazol-1-yl)piperidine-1-carboxylate (511 mg, 1.62 mmol, 59% yield) as a white solid.

[0434] LCMS: [M-tBu+H] + =259.2.

[0435] 1H NMR (400MHz, DMSO-d6) δppm 1.19–1.24(m,1H),1.41(s,9H),2.01(br dd,J=12.7,3.2Hz,1H),2.22–2.37(m,1H),3.15–3.36(m,1H),4.15(br s,1H),4.22–4.38(m,1H),4.68–4.81(m,1H),5.00–5.18(m,1H),8.34(s,1H),8.94(s,1H).

[0436] 19 F NMR (377MHz, DMSO-d6) δppm-205.91--197.72(m,1F).

[0437] Step 2: A solution of (3S, 4R) -3- fluoro- 4- (4- nitro -1H- pyrazol -1- bases) piperidine -1- carboxylic acid tert-butyl ester (511 mg, 1.62 mmol) in ethanol (16 mL) was purged with nitrogen three times and Pd / C (187.0 mg, 0.18 mmol) was added. The mixture was then purged with hydrogen (balloon) three times and then stirred at room temperature under hydrogen (balloon) for 22 h. The mixture was flushed with nitrogen and filtered on a celite pad and rinsed with MeOH (50 mL). The filtrate was concentrated in vacuo to obtain tert-butyl (3S, 4R) -4- (4- amino -1H- pyrazol -1- bases) -3- fluoropiperidine -1- carboxylate (427 mg, 1.05 mmol, 64% yield) as a red oil, which was used without further purification.

[0438] LCMS: [M+H] + =285.2.

[0439] 1 H NMR (400 MHz, DMSO-d6) δ ppm 1.21–1.28 (m, 1H), 1.41 (s, 9H), 1.85 (br dd, J = 12.8, 3.1 Hz, 1H), 2.07 (qd, J = 12.7, 4.7 Hz, 1H), 3.05–3.25 (m, 1H), 4.05–4.13 (m, 1H), 4.20–4.28 (m, 1H), 4.30–4.48 (m, 1H), 4.81–5.02 (m, 1H), 6.95 (s, 1H), 7.04 (s, 1H). Two protons were not observed.

[0440] 19 F NMR (377MHz, DMSO-d6) δppm-202.48--200.97(m,1F).

[0441] Synthesis of tert-butyl (3S,4S)-4-(4-amino-1H-pyrazol-1-yl)-3-fluoropiperidine-1-carboxylate (Intermediate D-2i)

[0442] Step 1: Prepare a solution of 4-nitro-1H-pyrazole (414 mg, 3.66 mmol), (3S,4R)-3-fluoro-4-hydroxy-piperidine-1-carboxylic acid tert-butyl ester (375 mg, 1.71 mmol) and triphenylphosphine polymer bond (1.31 g, 3.93 mmol) in dry THF (18 mL) and cool to 0° C. under nitrogen. Slowly add a solution of diisopropyl azodicarboxylate (0.66 mL, 3.34 mmol) in dry THF (18 mL). After 18 h at room temperature, the reaction mixture was filtered, concentrated, and purified by reverse phase column chromatography (5% to 100% MeOH / water with 0.1% formic acid) to give tert-butyl (3S,4S)-3-fluoro-4-(4-nitro-1H-pyrazol-1-yl)piperidine-1-carboxylate (452 ​​mg, 1.42 mmol, 83% yield) as a yellow solid.

[0443] LCMS: [M-tBu+H] + =259.2.

[0444] 1 H NMR (400MHz, DMSO-d6) δppm 1.21–1.25(m,1H),1.43(s,9H),1.88–2.04(m,1H),2.08–2.13(m,1H),3.98(br d,J=12.6Hz,1H),4.22–4.39(m,1H),4.62–4.74(m,1H),4.75–4.83(m,1H),4.84–4.96(m,1H),8.35(s,1H),9.09(s,1H).

[0445] 19 F NMR (377MHz, DMSO-d6) δppm-188.02 (br dd, J=49.0, 5.5Hz, 1F).

[0446] Step 2: a solution of (3S, 4S)-3-fluoro-4-(4-nitro-1H-pyrazol-1-yl)piperidine-1-carboxylic acid tert-butyl ester (441 mg, 1.40 mmol) in ethanol (12 mL) was purged with nitrogen three times and Pd / C (20.2 mg, 0.20 mmol) was added. The mixture was purged with hydrogen (balloon) three times and then stirred at room temperature under hydrogen (balloon) for 18 h. Due to no conversion being observed, the reaction mixture was purged with nitrogen three times and more Pd / C (54.6 mg, 0.5100 mmol) was added. The mixture was then purged with hydrogen (balloon) three times and then stirred at room temperature under hydrogen (balloon) for 17 h. The mixture was rinsed with nitrogen and filtered on a celite pad and rinsed with MeOH (100 mL). The filtrate was concentrated to give tert-butyl (3S,4S)-4-(4-amino-1H-pyrazol-1-yl)-3-fluoropiperidine-1-carboxylate (425 mg, 1.50 mmol, quantitative yield) as a red oil, which was used without further purification.

[0447] LCMS: [M-tBu+H] + =229.2.

[0448] 1 H NMR (400MHz, DMSO-d6) δppm 1.14–1.17(m,1H),1.41(s,9H),1.82–1.92(m,2H),3.81(br s,2H),3.91(br d,J=13.0Hz,2H),4.29–4.38(m,1H),4.54–4.66(m,1H),4.68–4.78(m,1H),6.96(s,1H),7.12(s,1H).

[0449] 19 F NMR (377MHz, DMSO-d6) δppm-186.83--186.56(m,1F).

[0450] Synthesis of (S)-tert-butyl 4-(4-amino-1H-pyrazol-1-yl)azepane-1-carboxylate (Intermediate D-2j) and (R)-tert-butyl 4-(4-amino-1H-pyrazol-1-yl)azepane-1-carboxylate (Intermediate D-2k)

[0451] Step 1: Triethylamine (2.59 mL, 18.58 mmol, 4 eq) was added to a solution of tert-butyl 4-hydroxyazepane-1-carboxylate 1 (1.0 g, 4.64 mmol, 1 eq) in DCM (15.48 mL). The flask was cooled to 0 ° C and methanesulfonyl chloride (1.08 mL, 13.93 mmol, 3 eq) was added dropwise. The reaction mixture was stirred at room temperature under a nitrogen atmosphere for 18 h. After stirring for 18 h, the reaction was quenched with water (50 mL) and the organic layer was extracted three times with DCM and washed with brine, dried over Na2SO4 and concentrated under vacuum to give tert-butyl 4-((methylsulfonyl)oxy)azepane-1-carboxylate (1.35 g, 99% yield) as an orange oil, which was used without further purification.

[0452] 1 H NMR (400MHz, CDCl3) δppm 1.46(s,9H),1.60–1.75(m,2H),1.94–2.10(m,4H),3.02(s,3H),3.32–3.56(m,4H),4.84–4.97(m,1H).

[0453] Step 2: To a round-bottom flask to which 4-nitro-1H-pyrazole (0.51 g, 4.51 mmol, 1 eq) was added tert-butyl 4-((methylsulfonyl)oxy)azepane-1-carboxylate (1.32 g, 4.51 mmol, 1 eq) in DMF (10.25 mL). CS2CO3 (2.94 g, 9.02 mmol, 2 eq) was then added and the reaction mixture was stirred at 90 ° C under a nitrogen atmosphere for 18 h. After heating for 18 h, LCMS showed complete conversion. The reaction was cooled to room temperature and water was added. The aqueous layer was extracted with EtOAc (3x), the combined organic layers were washed with brine, dried over Na2SO4 and concentrated to dryness to give an orange oil, which was purified by silica gel column chromatography (0% to 100% EtOAc / heptane). The fractions were combined and concentrated to give tert-butyl (rac)-4-(4-nitro-1H-pyrazol-1-yl)azepane-1-carboxylate (0.873 g) as a yellow oil. The material was then separated by chiral SFC to give two enantiomers, ent-1 (designated (S), 0.381 g, 27% yield) as a pale yellow oil and ent-2 (designated (R), 0.393 g, 28% yield) as a pale yellow oil. Note: Absolute stereochemistry is arbitrarily assigned from the first and second eluting enantiomers of the chiral separation.

[0454] ent-1: LCMS: [M-Boc+H] + =211.2. 1H NMR(400MHz,DMSO-d6)δppm 1.42(s,9H),1.58–1.73(m,1H),1.79–1.90(m,1H),1.90–2.03(m,2H),2.07(s,2H),3.20–3.2 9(m,1H),3.34–3.42(m,2H),3.50–3.66(m,1H),4.35–4.48(m,1H),8.26(s,1H),8.92(s,1H).

[0455] ent-2: LCMS: [M-boc+H] + =211.2. 1 H NMR(400MHz,DMSO-d6)δppm 1.42(s,9H),1.58–1.73(m,1H),1.79–1.90(m,1H),1.90–2.03(m,2H),2.07(s,2H),3.20–3.2 9(m,1H),3.34–3.42(m,2H),3.50–3.66(m,1H),4.35–4.48(m,1H),8.26(s,1H),8.92(s,1H).

[0456] Step 3: To a solution of (S)-4-(4-nitro-1H-pyrazol-1-yl)azepane-1-carboxylic acid tert-butyl ester ent-1 (0.33 g, 1.05 mmol, 1.0 equiv) in ethanol (10.47 mL, 0.1 M) was added Pd / C (167 mg, 0.16 mmol, 0.2 equiv) under nitrogen. The atmosphere in the flask was replaced with hydrogen by bubbling into the mixture for 10 min. The reaction mixture was stirred at room temperature under 1 atm of hydrogen for 48 h. The mixture was filtered through a celite pad and washed with EtOAc. The filtrate was then dried in vacuo and purified by silica gel column chromatography (0% to 100% EtOAc / heptane). The fractions were combined and concentrated to give (S)-tert-butyl 4-(4-amino-1H-pyrazol-1-yl)azepane-1-carboxylate intermediate D-2j as a red solid (185 mg, 57% yield).

[0457] LCMS: [M+H] + =281.2.

[0458] 1H NMR (400 MHz, DMSO-d6) δ ppm 1.41 (s, 9H), 1.53–1.70 (m, 1H), 1.72–1.86 (m, 3H), 1.91–2.04 (m, 3H), 3.14–3.26 (m, 2H), 3.46–3.58 (m, 1H), 3.72–3.82 (m, 1H), 4.02–4.11 (m, 1H), 6.88 (s, 1H), 7.00 (d, J = 0.7 Hz, 1H). One proton in 1 Not obvious in the H NMR spectrum.

[0459] Step 3 ': Pd / C (394 mg, 0.37 mmol) was added to a solution of (R)-4-(4-nitro-1H-pyrazol-1-yl)azepane-1-carboxylic acid tert-butyl ester ent-2 (0.38 g, 1.23 mmol) in ethanol (12.3 mL) under nitrogen. The atmosphere in the flask was replaced with hydrogen by bubbling hydrogen into the mixture. The reaction mixture was stirred at room temperature overnight under 1 atm of hydrogen. The mixture was filtered through a celite pad, and the pad was rinsed with EtOAc. The filtrate was then passed through a silica gel pad. The filtrate was then concentrated to give (R)-4-(4-amino-1H-pyrazol-1-yl)azepane-1-carboxylic acid tert-butyl ester intermediate D-2k (285 mg 67% yield) as a red solid, which was used without further purification.

[0460] LCMS: [M+H] + =281.2.

[0461] 1 H NMR(400MHz,DMSO-d6)δppm 1.42(s,9H),1.58–1.68(m,2H),1.72–1.89(m,4H),3.17–3.26(m,2H), 3.47–3.59(m,2H),4.04–4.13(m,1H),6.88(s,1H),7.01–7.02(m,1H). Table 17. List of intermediate D-3 compounds generated by general procedure TBM-D: Step 1

[0462] Synthesis of TBM-D: Step 2 To a solution of 2-(2,4-difluoro-5-methyl-phenyl)-4,4,5,5-tetramethyl-1,3,2-dioxaborolane intermediate D-4b (2.37 g, 8.76 mmol, 1.5 eq) and tert-butyl 4-[4-[(5-bromoimidazo[1,2-a]pyrazin-8-yl)amino]pyrazol-1-yl]piperidine-1-carboxylate intermediate D-3a (2.7 g, 5.84 mmol, 1 eq) in 1,4-dioxane (21.2 mL, 0.2 M) in a sealed tube were added NaHCO (1.47 g, 17.52 mmol, 3 eq) and water (9.7 mL, 0.2 M), followed by N bubbling for 5 min. Then, Pd(PPh3)4 (675mg, 0.58mmol, 0.1 equivalent) was added and N2 was bubbled for 10min. The resulting solution was stirred at 90°C in a sealed tube under N2 atmosphere for 16h. The reaction mixture was cooled to room temperature, water and EtOAc were added to the solution and the phases were separated. The aqueous phase was extracted with EtOAc (3x). The combined organic layers were dried over Na2SO4, filtered and evaporated to obtain a yellow oil. The yellow oil was then purified by reverse phase column chromatography (5% to 100% MeOH / water, containing 0.1% formic acid). The fractions were combined and concentrated to obtain tert-butyl 4-(4-((5-(2,4-difluoro-5-methylphenyl)imidazo[1,2-a]pyrazine-8-yl)amino)-1H-pyrazol-1-yl)piperidine-1-carboxylate intermediate D-5b (2.65g, 88% yield) as a light orange solid.

[0463] LCMS: [M+H] + =510.2.

[0464] 1 H NMR(400MHz,DMSO-d6)δppm 1.42(s,9H),1.71–1.84(m,2H),1.97–2.04(m,2H),2.29(s,3H),3.15–3.19(m,1H),3.98–4.14(m,3H),4 .31–4.41(m,1H),7.41–7.48(m,2H),7.58–7.63(m,J=5.6Hz,3H),7.81(s,1H),8.25(s,1H),9.99(s,1H). Table 18. TBM-D: List of intermediates used in step 2

[0465] Synthesis of 7-fluoro-4-methyl-6-(4,4,5,5-tetramethyl-1,3,2-dioxaborolan-2-yl)-3,4-dihydro-2H-benzo[b][1,4]oxazine (Intermediate D-41)

[0466] Step 1: To a solution of 6-bromo-7-fluoro-3,4-dihydro-2H-1,4-benzoxazine (600 mg, 2.59 mmol, 1.0 equiv) in DMF (6 mL) was added NaH (155 mg, 3.88 mmol, 1.5 equiv) at 0 ° C. The reaction mixture was stirred at room temperature for 20 minutes and then MeI (0.32 mL, 5.17 mmol, 2.0 equiv) was added. The reaction mixture was stirred at room temperature overnight. The reaction mixture was partitioned between EtOAc and water. The phases were separated and the aqueous phase was extracted with EtOAc (2x). The combined organic layers were washed with water (3x), brine (2x), dried over sodium sulfate, filtered, and concentrated. The crude product was extracted by silica gel column chromatography (0 to 40% EtOAc / heptane) to give the product 6-bromo-7-fluoro-4-methyl-2,3-dihydro-1,4-benzoxazine (420 mg, 66% yield) as a light yellow solid.

[0467] Step 2: In a sealed tube, 6-bromo-7-fluoro-4-methyl-2,3-dihydro-1,4-benzoxazine (200 mg, 0.81 mmol), B2pin2 (310 mg, 1.22 mmol) and KOAc (239 mg, 2.44 mmol) were mixed under N2. 1,4-dioxane (4 mL) was added and N2 was bubbled through the solution for 10 minutes. Then, Pd(dppf)Cl2·CH2Cl2 (66 mg, 0.08 mmol) was added and N2 was bubbled for 5 minutes. The tube was sealed and heated at 90°C overnight. The reaction mixture was cooled to room temperature. The reaction mixture was filtered through celite, washed with EtOAc and the filtrate was evaporated. The crude product is purified by normal phase chromatography (0% to 30% EtOAc / heptane) to afford 7-fluoro-4-methyl-6-(4,4,5,5-tetramethyl-1,3,2-dioxaborolan-2-yl)-3,4-dihydro-2H-benzo[b][1,4]oxazine intermediate D-41 (120 mg, 40% corrected yield) as a light yellow semisolid.

[0468] LCMS: [M+H] + =294.2. Table 19. List of intermediate D-4 compounds generated by general procedure TBM-D: Step 2 ND = Not Determined

[0469] Synthesis of (R)-tert-butyl 4-(3-((5-(2,4-difluoro-5-methylphenyl)imidazo[1,2-a]pyrazin-8-yl)amino)pyrrolidin-1-yl)piperidine-1-carboxylate (Intermediate D-51) from Intermediate D-51'

[0470] Step 1: To a solution of (3R)-3-[[5-(2,4-difluoro-5-methyl-phenyl)imidazo[1,2-a]pyrazin-8-yl]amino]pyrrolidine-1-carboxylic acid tert-butyl ester intermediate D-51' (290 mg, 0.68 mmol, 1 eq) in MeOH (5 mL, 0.14 M) was added 4 M HCl in 1,4-dioxane (2.53 mL, 10.1 mmol, 15 eq) and the reaction was stirred at room temperature for 2 h. The solvent was evaporated and co-evaporated with MeOH and MTBE to give (R)-5-(2,4-difluoro-5-methylphenyl)-N-(pyrrolidin-3-yl)imidazo[1,2-a]pyrazin-8-amine (283 mg, 80% yield) as a bis-HCl salt as a brown solid.

[0471] LCMS: [M+H] + =330.2.

[0472] 1 H NMR(400MHz,DMSO-d6)δppm 2.16–2.21(m,1H),2.23–2.35(m,4H),3.22–3.39(m,2H),3.39–3.48(m,1H),3.48–3.56(m,1H),4.79–4.8 4(m,1H),7.43–7.51(m,2H),7.54–7.61(m,1H),7.71–7.79(m,2H),8.97–9.10(m,1H),9.20–9.33(m,1H).

[0473] Step 2: To (R)-5-(2,4-difluoro-5-methylphenyl)-N-(pyrrolidin-3-yl)imidazo[1,2-a]pyrazin-8-amine (280 mg, 0.77 mmol, 1 eq) in DMSO (2.56 mL, 0.3 M) was added DIPEA (0.67 mL, 3.8 mmol, 5 eq) followed by tert-butyl 4-oxopiperidine-1-carboxylate (152.51 mg, 0.77 mmol, 1 eq) and AcOH (0.03 mL, 0.54 mmol, 0.7 eq) at room temperature and the solution was stirred for 10-15 min. NaBH(Oac)3 (324 mg, 1.53 mmol, 2 eq) was then added and the reaction mixture was stirred at room temperature overnight. The solvent was evaporated and the reaction mixture was purified by reverse phase column chromatography (5% to 100% MeOH / water with 0.1% formic acid). The fractions were combined and concentrated to give (R)-tert-butyl 4-(3-((5-(2,4-difluoro-5-methylphenyl)imidazo[1,2-a]pyrazin-8-yl)amino)pyrrolidin-1-yl)piperidine-1-carboxylate intermediate D-51 (320 mg, 82% yield) as the formate salt as an orange semi-solid.

[0474] LCMS: [M+H] + =513.2.

[0475] 1 H NMR(400MHz,DMSO-d6)δppm 1.20-1.33(m,2H),1.39(s,9H),1.74-1.86(m,2H),1.86-1.99(m,1H), 2.15-2.26(m,1H),2.28(s,3H),2.29-2.37(m,1H),2.59-2.70(m,2H), 2.75-2.91(m,3H),2.96-3.03(m,1H),3.75-3.90(m,2H),4.55-4.69(m ,1H),7.31(s,1H),7.38-7.50(m,2H),7.51-7.60(m,3H),8.16(s,1H).

[0476] 19 F NMR (377MHz, DMSO-d6) δppm -112.17 (q, J = 9.1Hz, 1F), -111.44 (q, J = 8.2Hz, 1F).

[0477] Synthesis of (S)-tert-Butyl 4-(3-((5-(2,4-difluoro-5-methylphenyl)imidazo[1,2-a]pyrazin-8-yl)amino)pyrrolidin-1-yl)piperidine-1-carboxylate (Intermediate D-5p) from Intermediate D-5p'

[0478] Step 1: To a solution of (S)-tert-butyl 3-((5-(2,4-difluoro-5-methylphenyl)imidazo[1,2-a]pyrazin-8-yl)amino)pyrrolidine-1-carboxylate (273 mg, 0.64 mmol, 1 eq) in MeOH (5 mL, 0.13 M) was added 4 M HCl in 1,4-dioxane (2.38 mL, 9.54 mmol, 15 eq) and the reaction was stirred at room temperature for 48 h. The solvent was evaporated and co-evaporated with MeOH and MTBE to give (S)-5-(2,4-difluoro-5-methylphenyl)-N-(pyrrolidin-3-yl)imidazo[1,2-a]pyrazin-8-amine (310 mg, quantitative) as the bis-HCl salt as a brown solid.

[0479] LCMS: [M+H] + =330.2.

[0480] 1 H NMR(400MHz,DMSO-d6)δppm 2.13-2.23(m,1H),2.24-2.37(m,4H),3.24-3.48(m,3H),3.51-3.61(m ,1H),4.76-4.89(m,1H),7.43-7.54(m,2H),7.55-7.62(m,1H),7.82(br s,1H),7.88(br s,1H),8.95-9.34(m,2H),9.49(br s,1H).

[0481] Step 2: To (S)-5-(2,4-difluoro-5-methylphenyl)-N-(pyrrolidin-3-yl)imidazo[1,2-a]pyrazin-8-amine (308 mg, 0.74 mmol, 1 eq) in CHCl (3 mL, 0.2 M) and DMSO (0.50 mL, 0.2 M) was added DIPEA (0.65 mL, 3.7 mmol, 5 eq) followed by tert-butyl 4-oxopiperidine-1-carboxylate (148 mg, 0.74 mmol, 1 eq) and AcOH (0.04 mL, 0.74 mmol, 1 eq) at room temperature and the solution was stirred for 10-15 min. NaBH(OAc) (316 mg, 1.49 mmol, 2 eq) was then added and the reaction mixture was stirred at room temperature overnight. The solvent was evaporated and the reaction mixture was purified by reverse phase column chromatography (5% to 100% MeOH / water with 0.1% formic acid). The fractions were combined and concentrated to give (S)-tert-butyl 4-(3-((5-(2,4-difluoro-5-methylphenyl)imidazo[1,2-a]pyrazin-8-yl)amino)pyrrolidin-1-yl)piperidine-1-carboxylate intermediate D-5p (344 mg, 79% yield) as the formate salt as an orange semi-solid.

[0482] LCMS: [M+H] + =513.2.

[0483] 1 H NMR(400MHz,DMSO-d6)δppm 1.21-1.34(m,2H),1.39(s,9H),1.75-1.85(m,2H),1.88-1.98(m,1H),2.15-2 .37(m,5H),2.59-2.71(m,2H),2.74-2.89(m,3H),2.97-3.04(m,1H),3.83(br d,J=12.0Hz,2H),4.57-4.67(m,1H),7.31(s,1H),7.43(t,J=9.9Hz,1H),7.49(d,J=7.1Hz,1H),7.52-7.59(m,3H),8.16(s,1H).

[0484] Synthesis of TBM-D: Step 3 To a solution of tert-butyl 4-(4-((5-(2,4-difluoro-5-methylphenyl)imidazo[1,2-a]pyrazin-8-yl)amino)-1H-pyrazol-1-yl)piperidine-1-carboxylate intermediate D-5b (2.65 g, 5.2 mmol, 1 eq) in MeOH (14 mL, 0.37 M) was added 4 M HCl in 1,4-dioxane (19.5 mL, 78 mmol, 15 eq). The reaction mixture was stirred at room temperature overnight. The reaction mixture was concentrated under reduced pressure and the residue was co-evaporated with MeOH (2x) and MTBE (2x) to give 5-(2,4-difluoro-5-methylphenyl)-N-(1-(piperidin-4-yl)-1H-pyrazol-4-yl)imidazo[1,2-a]pyrazin-8-amine TBM-2 as a bis-HCl salt (2.62 g, quantitative yield) as a tan solid.

[0485] LCMS: [M+H] + =410.2.

[0486] 1 H NMR (400MHz, DMSO-d6) δppm 2.16-2.24(m,4H),2.27-2.31(m,3H),2.99-3.12(m,2H),3.34-3.43(m,2H),4.49-4.59(m,1H),7.49(br t,J=9.9Hz,1H),7.57-7.63(m,2H),7.81(s,1H),7.86(br s,1H),7.96(s,1H),8.23(s,1H),8.95-9.09(m,1H),9.19-9.32(m,1H),10.77-10.90(m,1H).

[0487] 19 F NMR (377MHz, DMSO-d6) δppm -111.93 (s, 1F), -110.28 (s, 1F). Table 20. List of TBM-D compounds generated by the general procedure TBM-D: Step 3 Example S14. General Procedure for TBM-E

[0488] Synthesis of TBM-E: Step 1 A solution of 5-bromo-N-(1-(1-(2,2-dimethoxyethyl)piperidin-4-yl)-1H-pyrazol-4-yl)imidazo[1,2-a]pyrazin-8-amine intermediate E-2 (150 mg, 0.33 mmol), p-tolueneboronic acid intermediate E-3d (75 mg, 0.55 mmol), Xphos (61 mg, 0.05 mmol) and NaHCO (85 mg, 1.01 mmol) in 1,4-dioxane (3 mL) and water (1 mL) was degassed by bubbling nitrogen for 10 minutes. Xphos-Pd-G (44 mg, 0.05 mmol) was added and the mixture was further bubbled with nitrogen for 10 minutes. The reaction mixture was then heated to 90 ° C for 18 h. The reaction mixture was then cooled to room temperature, filtered on a pad of celite, rinsed with EtOAc and concentrated. The residue was purified by reverse phase column chromatography (5% to 100% MeCN / water with 0.1% formic acid). The fractions were collected and N-(1-(1-(2,2-dimethoxyethyl)piperidin-4-yl)-1H-pyrazol-4-yl)-5-(p-tolyl)imidazo[1,2-a]pyrazin-8-amine TBM-31 (126 mg, 0.27 mmol, 82% yield) was obtained as a white semisolid.

[0489] LCMS: [M+H] + =462.3.

[0490] 1 H NMR (400MHz, DMSO-d6) δppm 1.88-2.01(m,4H),2.16-2.25(m,2H),2.40(s,3H),2.47(br d,J=5.3Hz,2H),2.99(br d,J=11.6Hz,2H),3.27(s,6H),4.11(tat,J=10.5,5.2Hz,1H),4.49(t,J=5.1Hz,1H),7.38(d,J=7.8Hz,2H),7.41( s,1H),7.57(d,J=7.9Hz,2H),7.63(d,J=1.0Hz,1H),7.79(s,1H),7.86(d,J=1.0Hz,1H),8.24(s,1H),9.86(s,1H). Table 21. List of intermediates used in TBM-E: Step 1

[0491] Synthesis of 5-bromo-N-(1-(1-(2,2-dimethoxyethyl)piperidin-4-yl)-1H-pyrazol-4-yl)imidazo[1,2-a]pyrazin-8-amine (Intermediate E-2)

[0492] Step 1: To a solution of tert-butyl 4-(4-((5-bromoimidazo[1,2-a]pyrazin-8-yl)amino)-1H-pyrazol-1-yl)piperidine-1-carboxylate intermediate D-3a (7.15 g, 15.45 mmol) in DCM (150 mL) was added 4 M HCl in 1,4-dioxane (38.0 mL, 152 mmol). After 17 h at room temperature, the reaction mixture was concentrated in vacuo to give 5-bromo-N-(1-(piperidin-4-yl)-1H-pyrazol-4-yl)imidazo[1,2-a]pyrazin-8-amine intermediate E-1 (7.26 g, 19.86 mmol, quantitative) as a brown oil. The product was used in the next step without further purification.

[0493] LCMS: [M+H] + =362.0.

[0494] 1 H NMR (400 MHz, DMSO-d6) δ ppm 2.11-2.22 (m, 4H), 2.97-3.10 (m, 2H), 3.37 (br d, J = 12.6 Hz, 2H), 4.51 (br t, J = 7.3 Hz, 1H), 7.68 (s, 1H), 7.79 (s, 1H), 7.87 (d, J = 0.9 Hz, 1H), 8.09 (d, J = 1.2 Hz, 1H), 8.17 (s, 1H), 10.38 (s, 1H). One proton was not observed.

[0495] Step 2: To a solution of 5-bromo-N-(1-(piperidin-4-yl)-1H-pyrazol-4-yl)imidazo[1,2-a]pyrazin-8-amine intermediate E-1 (5.6 g, 15.45 mmol), DIPEA (10.0 mL, 57.41 mmol), and 60% wt. 2,2-dimethoxyacetaldehyde in DCM (150 mL) in H2O (0.77 mL, 53 mmol) was added NaBH(OAc)3 (6.64 g, 31.3 mmol). After 19 h at room temperature, the reaction mixture was concentrated and purified by reverse phase column chromatography (5% to 100% MeOH / water with 0.1% formic acid). The fractions were collected and gave 5-bromo-N-(1-(1-(2,2-dimethoxyethyl)piperidin-4-yl)-1H-pyrazol-4-yl)imidazo[1,2-a]pyrazin-8-amine intermediate E-2 as a brown oil (6.60 g, 14.40 mmol, 93% yield).

[0496] LCMS: [M+H] + =516.2.

[0497] 1 H NMR (400 MHz, DMSO-d6) δ ppm 1.81-1.99 (m, 4H), 2.18-2.31 (m, 2H), 2.50-2.53 (m, 1H), 3.01 (br d, J = 11.6 Hz, 2H), 3.27 (s, 6H), 4.11 (tt, J = 10.4, 5.1 Hz, 1H), 4.50 (t, J = 5.1 Hz, 1H), 7.59 (s, 1H), 7.70 (d, J = 1.0 Hz, 1H), 7.75 (s, 1H), 7.98 (d, J = 1.0 Hz, 1H), 8.13-8.17 (m, 1H), 9.96-10.06 (m, 1H). One proton was not observed. Table 22. List of TBM-E compounds generated by the representative procedure TBM-E: Step 1

[0498] Synthesis of N-(1-(1-(2,2-dimethoxyethyl)piperidin-4-yl)-1H-pyrazol-4-yl)-5-ethylimidazo[1,2-a]pyrazin-8-amine (TBM-28) from TBM-28' To a solution of N-(1-(1-(2,2-dimethoxyethyl)piperidin-4-yl)-1H-pyrazol-4-yl)-5-vinylimidazo[1,2-a]pyrazin-8-amine TBM-28' (115 mg, 0.17 mmol, 1 eq) in methanol (2.9 mL) was added Pd / C (17 mg, 0.02 mmol, 0.1 eq) at room temperature under N2, followed by bubbling H2 into the mixture and stirring it under an H2 atmosphere. The mixture was filtered through celite, rinsed with MeOH, and concentrated. The residue was then purified by reverse phase column chromatography (5% to 100% MeOH / water, containing 0.1% formic acid). The fractions were combined and concentrated to give N-(1-(1-(2,2-dimethoxyethyl)piperidin-4-yl)-1H-pyrazol-4-yl)-5-ethylimidazo[1,2-a]pyrazin-8-amine TBM-28 (30 mg, 40% yield) as a light yellow oil.

[0499] 1 H NMR(400MHz, DMSO-d6)δppm 1.29(t,J=7.5Hz,3H),1.79–2.09(m,6H),2.13–2.25(m,2H),2.83(q,J=7.3Hz,2H),2.98(br d,J=11.2Hz,2H),3.27–3.27(m,6H),4.03–4.16(m,1H),4.49(t,J=5.1Hz,1H),7.24(s, 1H),7.63–7.66(m,1H),7.74(s,1H),7.95–7.98(m,1H),8.17–8.19(m,1H),9.59(s,1H). Example S15. General Procedure for TBM-F

[0500] Synthesis of TBM-F: Step 1 To a round-bottom flask was added 1-((trans)-4-(2-((tert-butyldiphenylsilyl)oxy)ethyl)cyclohexyl)-1H-pyrazol-4-amine intermediate F-1a (1.64 g, 3.36 mmol, 1 eq), 5,8-dibromoimidazo[1,2-a]pyrazine 6 (0.93 g, 3.36 mmol, 1 eq), and acetic acid (4.68 g, 45.8 mmol, 13.6 eq). The reaction was placed in an oil bath preheated to 100°C. After 2 h, the reaction was quenched with saturated NaHCO3 solution (until pH 7-9) and extracted with MeTHF (3x). The combined organic layers were washed with brine, dried over Na2SO4, and concentrated to give a dark red residue, which was then purified by reverse phase column chromatography (5% to 100% MeOH / water with 0.1% formic acid). The fractions were combined and concentrated to give 5-bromo-N-(1-((trans)-4-(2-((tert-butyldiphenylsilyl)oxy)ethyl)cyclohexyl)-1H-pyrazol-4-yl)imidazo[1,2-a]pyrazin-8-amine intermediate F-2a as a light orange solid (1.56 g, 67% yield).

[0501] LCMS: [M+H] + =643.2,645.2.

[0502] 1 H NMR(400MHz, DMSO-d6)δppm 1.01(s,9H),1.03–1.13(m,2H),1.42–1.53(m,3H),1.60–1.72(m,2H),1.72–1.80(m,2H),1.94–2.02(m,2H),3.71(br t,J=5.9Hz,2H),4.00–4.08(m,1H),7.42–7.50(m,6H),7.59(s,1H),7.60–7.65(m,4H) ,7.70(d,J=1.0Hz,1H),7.73(s,1H),7.98(d,J=1.0Hz,1H),8.13(s,1H),9.97(s,1H). Table 23. List of intermediates used in TBM-F: Step 1

[0503] Synthesis of 1-((trans)-4-(2-((tert-butyldiphenylsilyl)oxy)ethyl)cyclohexyl)-1H-pyrazol-4-amine (Intermediate F-1a) and 1-((cis)-4-(2-((tert-butyldiphenylsilyl)oxy)ethyl)cyclohexyl)-1H-pyrazol-4-amine (Intermediate F-1b)

[0504] Step 1: To a solution of 4-(2-hydroxyethyl)cyclohexanol (5g, 34.7mmol, 1 equivalent) in DCM (63mL) was added N-methyl-imidazole (8.29mL, 104mmol, 3 equivalents), iodine (17.6g, 69mmol, 2 equivalents) and tert-butylchlorodiphenylsilane (11.72mL, 45mmol, 1.3 equivalents). After stirring at room temperature under a nitrogen atmosphere for 18h, the reaction was washed with 10% sodium thiosulfate solution (2x 80mL) and brine (1x 80mL). The organic phase was then dried over sodium sulfate, filtered and concentrated to give a viscous orange oil. The residue was then purified by silica gel column chromatography (0% to 30% EtOAc / heptane). After purification twice by silica gel column chromatography, the fractions were combined and concentrated to give (cis)-4-(2-((tert-butyldiphenylsilyl)oxy)ethyl)cyclohexan-1-ol cis (4.62 g, 35% yield) as a colorless oil and (trans)-4-(2-((tert-butyldiphenylsilyl)oxy)ethyl)cyclohexan-1-ol trans (2.6 g, 19% yield) as a colorless oil; the combined yield was 54%.

[0505] Cis: LCMS: [M+Na+H] + =405.4. 1 H NMR(400MHz,DMSO-d6)δppm 0.99(s,9H),1.25–1.40(m,6H),1.42–1.54(m,5H),3.64–3.71(m,3H),4.20(d,J=3.4Hz,1H),7.40–7.48(m,6H),7.58–7.63(m,4H).

[0506] Trans: LCMS: [M+Na+H] + =405.4. 1 H NMR(400MHz,DMSO-d6)δppm 0.78–0.91(m,2H),0.99(s,9H),1.01–1.11(m,2H),1.24–1.36(m,1H),1.40(q,J=6.4Hz,2H),1.53–1.62(m,2H),1.72 –1.79(m,2H),3.22–3.31(m,1H),3.66(t,J=6.5Hz,2H),4.41(d,J=4.6Hz,1H),7.40–7.48(m,6H),7.57–7.63(m,4H).

[0507] Step 2: To a solution of (cis)-4-(2-((tert-butyldiphenylsilyl)oxy)ethyl)cyclohexan-1-ol cis (2.2 g, 5.23 mmol, 1 eq), triphenylphosphine (2058 mg, 7.85 mmol, 1.5 eq), and 4-nitro-1H-pyrazole (887 mg, 7.9 mmol, 1.5 eq) in anhydrous THF (10 mL) at 0° C. was slowly added diisopropyl azodicarboxylate (1.54 mL, 7.6 mmol, 1.5 eq) as a solution in anhydrous THF (15 mL). The reaction was warmed to room temperature and stirred for 18 h. The reaction was concentrated and purified by reverse phase column chromatography (5% to 100% MeOH / water with 0.1% formic acid). Fractions were combined and concentrated to give 1-((trans)-4-(2-((tert-butyldiphenylsilyl)oxy)ethyl)cyclohexyl)-4-nitro-1H-pyrazole (960 mg, 38% yield) as a colorless oil.

[0508] LCMS: [M+H] + =478.4.

[0509] 1 H NMR(400MHz,DMSO-d6)δppm 1.00(s,9H),1.02-1.12(m,2H),1.43-1.54(m,3H),1.67-1.81(m,4H),2.01(br d,J=11.0Hz,2H),3.70(t,J=5.7Hz,2H),4.15-4.24(m,1H),7.42-7.49(m,6H),7.59-7.65(m,4H),8.24(s,1H),8.91(s,1H).

[0510] Step 3: To a solution of 1-((trans)-4-(2-((tert-butyldiphenylsilyl)oxy)ethyl)cyclohexyl)-4-nitro-1H-pyrazole (329 mg, 0.69 mmol, 1 eq) in ethanol (8 mL, 0.08 M) was added Pd / C (110 mg, 0.1 mmol, 0.15 eq) under nitrogen. The atmosphere of the flask was replaced with hydrogen (5 purges). The reaction mixture was stirred at room temperature for 3 h under 1 atm of hydrogen. The reaction mixture was filtered through a pad of celite and rinsed with EtOAc (3 x 20 mL). The solution was concentrated to give 1-((trans)-4-(2-((tert-butyldiphenylsilyl)oxy)ethyl)cyclohexyl)-1H-pyrazole-4-amine intermediate F-1a (339 mg, quantitative yield) as a purple oil which was used without further purification.

[0511] LCMS: [M+H] + =448.2.

[0512] 1 H NMR(400MHz,DMSO-d6)δppm 1.00(s,9H),1.01-1.05(m,1H),1.38-1.50(m,3H),1.50-1.63(m,2H),1.71(br d,J=12.0Hz,2H),1.84-1.94(m,2H),3.69(t,J=6.2Hz,2H),3.73-3.89(m,2H),6. 87(d,J=0.7Hz,1H),7.01(d,J=0.7Hz,1H),7.41-7.49(m,6H),7.57-7.66(m,4H).

[0513] Step 2': To a solution of triphenylphosphine (617 mg, 2.35 mmol), (trans)-4-(2-((tert-butyldiphenylsilyl)oxy)ethyl)cyclohexan-1-ol trans (600 mg, 1.57 mmol, 1.0 eq), and 4-nitro-1H-pyrazole (266 mg, 2.35 mmol, 1.5 eq) in dry THF (6 mL, 0.2 M) was slowly added diisopropyl azodicarboxylate (0.46 mL, 2.35 mmol) at 0°C. The reaction was allowed to warm to room temperature and stirred for 18 h. The solvent was evaporated, and the residue was then purified by reverse phase column chromatography (5% to 100% MeOH / water with 0.1% formic acid). Fractions were combined and concentrated to give 1-((cis)-4-(2-((tert-butyldiphenylsilyl)oxy)ethyl)cyclohexyl)-4-nitro-1H-pyrazole (520 mg, 1.087 mmol, 69% yield) as a yellow solid.

[0514] LCMS: [M+H] + =478.2.

[0515] 1 H NMR(400MHz,DMSO-d6)δppm 0.99(s,9H),1.36-1.48(m,2H),1.50-1.56(m,2H),1.59(q,J=6.8Hz,2H),1.72-1.86(m,3H),1.96-2.06(m,2H),3 .68(t,J=6.4Hz,2H),4.21-4.35(m,1H),7.39-7.50(m,6H),7.57-7.66(m,4H),8.26(d,J=0.5Hz,1H),8.93(s,1H).

[0516] Step 3': Nitrogen was bubbled through a solution of 1-((cis)-4-(2-((tert-butyldiphenylsilyl)oxy)ethyl)cyclohexyl)-4-nitro-1H-pyrazole (520 mg, 1.09 mmol, 1.0 eq) in methanol (2.18 mL, 0.5 M) for 5 minutes. Pd / C 10% (232 mg, 0.22 mmol, 0.2 eq) was then added and nitrogen was bubbled for an additional 5 minutes. H2 (1 atm) was then bubbled through the solution for 5 minutes and the reaction mixture was stirred at room temperature under a hydrogen atmosphere for 18 hours. The reaction mixture was filtered through a pad of celite and washed with EtOAc. The residue was concentrated under vacuum to give 1-((cis)-4-(2-((tert-butyldiphenylsilyl)oxy)ethyl)cyclohexyl)-1H-pyrazol-4-amine intermediate F-1b (465 mg, 1.039 mmol, 95% yield) as a purple oil, which was used in the next step without further purification.

[0517] LCMS: [M+H] + =448.4.

[0518] 1 H NMR (400 MHz, DMSO-d6) δ ppm 0.99 (s, 9H), 1.32-1.44 (m, 2H), 1.44-1.51 (m, 2H), 1.55 (q, J = 6.8 Hz, 2H), 1.60-1.70 (m, 2H), 1.70-1.79 (m, 1H), 1.83-1.96 (m, 2H), 3.68 (t, J = 6.5 Hz, 2H), 3.72-3.80 (m, 1H), 3.90-4.00 (m, 1H), 6.88 (s, 1H), 7.04 (s, 1H), 7.39-7.50 (m, 6H), 7.57-7.66 (m, 4H). One proton in 1 Not observed in the H NMR spectrum. Table 24. List of intermediate F-2 compounds generated by representative procedure TBM-F: Step 1

[0519] Synthesis of TBM-F: Step 2

[0520] To a solution of 2-(2,4-difluoro-5-methyl-phenyl)-4,4,5,5-tetramethyl-1,3,2-dioxaborolane intermediate D-4b (0.69 g, 2.54 mmol, 1.5 eq) and 5-bromo-N-(1-((trans)-4-(2-((tert-butyldiphenylsilyl)oxy)ethyl)cyclohexyl)-1H-pyrazol-4-yl)imidazo[1,2-a]pyrazin-8-amine intermediate F-2a (1.09 g, 1.69 mmol) in 1,4-dioxane (15 mL) was added NaHCO (0.43 g, 5.1 mmol, 3 eq) and water (5 mL). Nitrogen was bubbled into the reaction mixture for 10 min. Next, Pd(PPh3)4 (0.2 g, 0.17 mmol, 0.1 equiv) was added and nitrogen was bubbled for another 10 min. The resulting solution was stirred at 90 ° C in a sealed tube under a nitrogen atmosphere for 18 h. The reaction mixture was filtered through celite, rinsed with EtOAc and concentrated. The resulting residue was purified by reverse phase column chromatography (5% to 100% MeOH / water, containing 0.1% formic acid). The fractions were combined and concentrated to give N-(1-((trans)-4-(2-((tert-butyldiphenylsilyl)oxy)ethyl)cyclohexyl)-1H-pyrazol-4-yl)-5-(2,4-difluoro-5-methylphenyl)imidazo[1,2-a]pyrazine-8-amine intermediate F-4a (1.07 g, 89% yield) as a light orange oil.

[0521] LCMS: [M-TBDPS+H] + =453.2.

[0522] 1 H NMR (400MHz, DMSO-d6) δppm 1.00-1.03(m,9H),1.04-1.18(m,2H),1.45-1.53(m,3H),1.61-1.80(m,4H),1.95-2.04(m,2H),2.29(s,3H),3.72(br t,J=6.0Hz,2H),4.02-4.13(m,1H),7.41-7.49(m,8H),7.58-7.65(m,7H),7.78(s,1H),8.19(s,1H),9.96(s,1H).

[0523] 19 F NMR (377MHz, DMSO-d6) δppm-112.22--112.10(m,1F),-111.33--111.21(m,1F). Table 25. List of intermediates used in TBM-F: Step 2

[0524] Synthesis of tert-butyl 7-fluoro-6-(4,4,5,5-tetramethyl-1,3,2-dioxaborolan-2-yl)-2,3-dihydro-4H-benzo[b][1,4]oxazine-4-carboxylate (Intermediate F-3)

[0525] Step 1: To a solution of 6-bromo-7-fluoro-3,4-dihydro-2H-1,4-benzoxazine (200 mg, 0.86 mmol, 1.0 equiv) in CHCl (2 mL, 0.4 M) was added tert-butyl tert-butyloxycarbonyl carbonate (207 mg, 0.95 mmol, 1.1 equiv) and EtN (0.26 mL, 1.9 mmol, 2.2 equiv). The resulting solution was stirred at room temperature for 2 h. LCMS showed traces of the desired product. DMAP (105 mg, 0.86 mmol, 1.0 equiv) was added and the reaction mixture was stirred overnight. The reaction mixture was diluted with CHCl and saturated NaHCO and extracted with CHCl. ​​The organic layer was then washed with saturated NaHCO, dried over sodium sulfate, filtered and concentrated. The crude residue was purified by reverse phase column chromatography (5% to 100% MeOH / water, containing 0.1% formic acid). Fractions were combined and concentrated to give tert-butyl 6-bromo-7-fluoro-2,3-dihydro-4H-benzo[b][1,4]oxazine-4-carboxylate (180 mg, 63% yield) as a white solid.

[0526] LCMS: [M-Boc+H] + =232.0.

[0527] 1 H NMR (400MHz, DMSO-d6) δppm 1.49 (s, 9H), 3.78 (t, J = 4.5Hz, 2H), 4.24 (t, J = 4.5Hz, 2H), 6.99 (d, J = 9.8Hz, 1H), 8.08 (br s, 1H).

[0528] 19 F NMR (377MHz, DMSO-d6) δppm-113.57 (s, 1F).

[0529] Step 2: A mixture of tert-butyl 6-bromo-7-fluoro-2,3-dihydro-4H-benzo[b][1,4]oxazine-4-carboxylate (190 mg, 0.57 mmol, 1.0 eq), B2pin2 (218 mg, 0.86 mmol, 1.5 eq) and KOAc (168 mg, 1.72 mmol, 3.0 eq) in 1,4-dioxane (2.3 mL, 0.25 M) was bubbled with nitrogen for 10 min. Then, Pd(dppf)Cl2·CHCl2 (47 mg, 0.06 mmol, 0.1 eq) was added and nitrogen was bubbled for another 5 min. The resulting solution was stirred at 90 ° C in a sealed tube overnight. The reaction mixture was filtered through celite, rinsed with EtOAc and concentrated. The resulting residue was purified by silica gel column chromatography (0 to 30% EtOAc / heptane). The fractions were combined and concentrated to give tert-butyl 7-fluoro-6-(4,4,5,5-tetramethyl-1,3,2-dioxaborolan-2-yl)-2,3-dihydro-4H-benzo[b][1,4]oxazine-4-carboxylate intermediate F-3 as a light yellow solid (170 mg, 65% yield).

[0530] LCMS: [M+H] + =380.2.

[0531] 1 H NMR (400MHz, DMSO-d6) δppm 1.27 (s, 12H), 1.49 (s, 9H), 3.78 (t, J = 4.3Hz, 2H), 4.25 (t, J = 4.4Hz, 2H), 6.67 (d, J = 9.8Hz, 1H), 8.04 (br d, J = 5.1Hz, 1H). 19 F NMR (377MHz, DMSO-d6) δppm-107.09 (s, 1F). Table 26. List of intermediate F-4 compounds generated by the representative procedure TBM-F: Step 2

[0532] Synthesis of TBM-F: Step 3 To a stirred solution of N-(1-((trans)-4-(2-((tert-butyldiphenylsilyl)oxy)ethyl)cyclohexyl)-1H-pyrazol-4-yl)-5-(2,4-difluoro-5-methylphenyl)imidazo[1,2-a]pyrazin-8-amine intermediate F-4a (1.07 g, 1.55 mmol, 1 eq) in THF (16 mL) was added a 1 M TBAF solution in THF (3.5 mL, 3.5 mmol, 2.2 eq) at room temperature for 3 h. The solvent was removed under reduced pressure and the residue was purified by silica gel column chromatography (0% to 30% MeOH / DCM). The fractions were combined, concentrated, and the residue was then purified by reverse phase column chromatography (5% MeOH to 100% MeOH / water with 0.1% formic acid). Fractions were combined and concentrated to give 2-((trans)-4-(4-((5-(2,4-difluoro-5-methylphenyl)imidazo[1,2-a]pyrazin-8-yl)amino)-1H-pyrazol-1-yl)cyclohexyl)ethan-1-ol TBM-43 as a light yellow solid (425 mg, 59% yield).

[0533] LCMS: [M+H] + =453.2.

[0534] 1 H NMR(400MHz,DMSO-d6)δppm 1.04-1.17(m,2H),1.37(q,J=6.5Hz,2H),1.41-1.50(m,1H),1.71(qd, J=12.5,2.9Hz,2H),1.80-1.88(m,2H),1.99-2.07(m,2H),2.27-2.30( m,3H),3.43-3.49(m,2H),4.04-4.14(m,1H),4.31-4.38(m,1H),7.40- 7.48(m,2H),7.58-7.64(m,3H),7.78(s,1H),8.20(s,1H),9.96(s,1H). Table 27. List of TBM-F compounds generated by the representative procedure TBM-F: Step 3 Example S16. General Procedure for TBM-G

[0535] Synthesis of TBM-G: Step 1 To a solution of ethyl 2-((trans)-3-(4-nitro-1H-pyrazol-1-yl)cyclobutyl)acetate intermediate G-1a (414 mg, 1.6 mmol, 1 equiv) in ethanol (8 mL, 0.2 M) was added Pd / C (261 mg, 0.25 mmol, 0.15 equiv) under nitrogen. The atmosphere of the flask was replaced with hydrogen (5 purges). The reaction mixture was stirred at room temperature for 20 h under 1 atm of hydrogen. The reaction mixture was filtered through a celite pad and rinsed with EtOAc (3 x 50 mL). The resulting filtrate was concentrated to obtain ethyl 2-((trans)-3-(4-amino-1H-pyrazol-1-yl)cyclobutyl)acetate intermediate G-2a (391 mg, 94% yield) as a purple oil, which was used without further purification.

[0536] LCMS: [M+H] + =224.2.

[0537] 1 H NMR(400MHz,DMSO-d6)δppm 1.15-1.23(m,4H),2.05-2.16(m,2H),2.52-2.59(m,4H),3.79(br s, 2H), 4.05 (q, J = 7.1Hz, 2H), 4.76 (quint, J = 7.6Hz, 1H), 6.93 (s, 1H), 7.07 (s, 1H). Table 28. List of intermediates used in TBM-G: Step 1

[0538] Synthesis of ethyl 2-((trans)-3-(4-nitro-1H-pyrazol-1-yl)cyclobutyl)acetate (Intermediate G-1a) and ethyl 2-((cis)-3-(4-nitro-1H-pyrazol-1-yl)cyclobutyl)acetate (Intermediate G-1d) To a solution of ethyl 2-(3-hydroxycyclobutyl)acetate (850 mg, 5.37 mmol, 1 eq), triphenylphosphine polymer linkage (3.58 g, 10.75 mmol, 2 eq), and 4-nitro-1H-pyrazole 2 (1.22 g, 10.75 mmol, 2 eq) in anhydrous THF (13 mL, 0.2 M) at 0°C was slowly added diisopropyl azodicarboxylate (2.11 mL, 10.75 mmol, 2 eq) as a solution in anhydrous THF (13 mL, 0.2 M) and the reaction was allowed to warm to room temperature. After stirring under nitrogen for 20 h, the solvent was removed under reduced pressure and the residue was purified by silica gel column chromatography (0 to 10% EtOAc / DCM). Fractions were combined and concentrated to give 747 mg of a mixture of cis and trans isomers. The isomers were separated by chiral SFC to give ethyl 2-((trans)-3-(4-nitro-1H-pyrazol-1-yl)cyclobutyl)acetate Intermediate G-1a (414 mg, 30% yield) as a white solid and ethyl 2-((cis)-3-(4-nitro-1H-pyrazol-1-yl)cyclobutyl)acetate Intermediate G-1d (239 mg, 18% yield) as a colorless oil. Intermediate G-1a:

[0539] LCMS: [M+H] + =254.2.

[0540] 1 H NMR (400 MHz, CHLOROFORM-d) δ ppm 1.28 (t, J = 7.1 Hz, 3H), 2.36-2.45 (m, 2H), 2.59 (d, J = 7.6 Hz, 2H), 2.73-2.83 (m, 2H), 2.85-2.97 (m, 1H), 4.16 (q, J = 7.1 Hz, 2H), 4.89 (quin, J = 7.5 Hz, 1H), 8.12 (s, 1H), 8.18 (s, 1H).

[0541] Synthesis of (trans)-4-(4-nitro-1H-pyrazol-1-yl)cyclohexane-1-carboxylic acid methyl ester (Intermediate G-1b)

[0542] Step 1: To a solution of cis-4-hydroxycyclohexanecarboxylic acid methyl ester (3 g, 19 mmol) in CHCl (65 mL) was added EtN (7.66 mL, 56.9 mmol) and MsCl (1.91 mL, 24.7 mmol). After 3 h at room temperature, the reaction mixture was quenched with saturated NHCl aqueous solution and diluted with CHCl. ​​The organic phase was separated, dried over NaSO, filtered, and concentrated in vacuo to give cis-4-methylsulfonyloxycyclohexanecarboxylic acid methyl ester (4.5 g, 19 mmol, quantitative) as a light yellow solid, which was used in the next step without further purification.

[0543] 1 H NMR (400MHz, CDCl3) δppm 1.67-1.83(m,4H),1.87-1.98(m,2H),1.99-2.09(m,2H),2.35-2.45(m,1H),3.01(s,3H),3.68(s,3H),4.87-4.96(m,1H).

[0544] Step 2: Cs is added to a solution of cis-4-methylsulfonyloxycyclohexanecarboxylic acid methyl ester (4.26g, 18mmol) and 4-nitro-1H-pyrazole (1.7g, 15.03mmol) in DMF (20mL) CO (6.86g, 21.05mmol). After 4h at 90°C, the reaction mixture is cooled to room temperature and distributed between water (10mL) and EtOAc (10mL). The aqueous phase is separated and extracted with EtOAc (3x 10mL). The organic phase is merged, washed with water (3x) and salt water, through Na SO Drying, filtered and concentrated in vacuo. The residue was purified by reverse phase column chromatography (5% to 100% MeCN / water with 0.1% formic acid) to give (trans)-methyl 4-(4-nitro-1H-pyrazol-1-yl)cyclohexane-1-carboxylate (Intermediate G-1b) (1.24 g, 4.90 mmol, 33%) as a yellow solid.

[0545] 1 H NMR (400MHz, CDCl3) δppm 1.60-1.72(m,2H),1.81(qd,J=12.6,2.7Hz,2H),2.19-2.33(m,4H),2.37-2.4 6(m,1H),3.71(s,3H),4.15(tt,J=11.7,3.8Hz,1H),8.08(s,1H),8.16(s,1H).

[0546] Synthesis of (cis)-4-(4-nitro-1H-pyrazol-1-yl)cyclohexane-1-carboxylic acid methyl ester (Intermediate G-1c)

[0547] Step 1: Under nitrogen, a solution of trans-4-hydroxycyclohexanecarboxylic acid methyl ester (100 mg, 0.63 mmol, 1 eq) in CHCl (2 mL, 0.32 M) was cooled to 0 ° C, followed by the addition of methanesulfonyl chloride (64 μL, 0.82 mmol, 1.3 eq) and triethylamine (0.13 mL, 0.95 mmol, 1.5 eq). After stirring at 0 ° C for 2 h, the reaction was quenched by the addition of water. The phases were then separated and the aqueous phase was extracted 3 times with CHCl. ​​The combined organic phases were washed with brine, dried over magnesium sulfate, filtered and concentrated to give trans-4-methylsulfonyloxycyclohexanecarboxylic acid methyl ester (150 g, quantitative yield) as a light yellow solid.

[0548] 1 H NMR (400MHz, DMSO-d6) δppm 1.43-1.58(m,4H), 1.88-2.08(m,4H), 2.30-2.41(m,1H), 3.17(s,3H), 3.59(s,3H), 4.53-4.62(m,1H).

[0549] Step 2: Under nitrogen, a solution of methyl trans-4-methylsulfonyloxycyclohexanecarboxylate (150 mg, 0.64 mmol, 1 eq), 4-nitro-1H-pyrazole (72 mg, 0.64 mmol, 1 eq) and DMF (3 mL, 0.21 M) was stirred at room temperature for 5 minutes in an oven-dried round-bottom flask, followed by the addition of CsCO (415 mg, 1.27 mmol, 2.0 eq). The resulting mixture was stirred at 90°C for 16 h. The reaction was quenched by the addition of water. Ethyl acetate was added and the phases separated. The aqueous phase was extracted three times with ethyl acetate, and the combined organic phases were washed once with water and once with brine, dried over magnesium sulfate, filtered and concentrated. The crude product was purified by reverse phase column chromatography (5% to 100% MeOH / water containing 0.1% formic acid). The fractions were combined and concentrated to give (cis)-methyl 4-(4-nitro-1H-pyrazol-1-yl)cyclohexane-1-carboxylate intermediate G-1c (68.1 mg, 42% yield) as a tan solid.

[0550] LCMS: [M+H] + =254.4.

[0551] 1H NMR(400MHz,DMSO-d6)δppm 1.61-1.73(m,2H),1.89-1.96(m,4H),2.00-2.07(m,2H),2.70(quin,J=4.5Hz,1H),3.64(s,3H),4.25-4.38(m,1H),8.26(s,1H),8.91(s,1H).

[0552] Synthesis of methyl (trans)-3-(4-nitro-1H-pyrazol-1-yl)cyclobutane-1-carboxylate (Intermediate G-1e)

[0553] To a solution of 4-nitro-1H-pyrazole (1.04 g, 9.22 mmol), cis-3-hydroxycyclobutanecarboxylic acid methyl ester (800 mg, 6.15 mmol) and triphenylphosphine (8.22 g, 12.29 mmol) in dry THF (15 mL) was slowly added a solution of diisopropyl azodicarboxylate (1.82 mL, 9.22 mmol) in THF (5 mL) under nitrogen at 0°C. The next morning, the reaction mixture was filtered, concentrated in vacuo and purified by reverse phase column chromatography (5% to 100% MeCN / water with 0.1% formic acid). The fractions were collected and concentrated to give (trans)-3-(4-nitro-1H-pyrazol-1-yl)cyclobutane-1-carboxylic acid methyl ester (Intermediate G-1e) (963 mg containing 30% triphenylphosphine oxide, corresponding to 675 mg of the desired compound, 49% yield).

[0554] LCMS: [M+H] + =226.2.

[0555] 1 H NMR(400MHz,DMSO-d6)δppm 2.63-2.71(m,2H),2.76-2.85(m,2H),3.19-3.29(m,1H),3.66-3.70(m,3H),5.06-5.16(m,1H),8.34(s,1H),9.01(s,1H). Table 29. List of intermediate G-2 compounds generated by representative procedure TBM-G: Step 1

[0556] Synthesis of TBM-G: Step 2 To a round-bottom flask was added ethyl 2-((trans)-3-(4-amino-1H-pyrazol-1-yl)cyclobutyl)acetate Intermediate G-2a (391 mg, 1.75 mmol, 1 eq), 5,8-dibromoimidazo[1,2-a]pyrazine 5 (485 mg, 1.75 mmol, 1 eq), and thiazoline (2.68 g, 26.3 mmol, 15 eq). The reaction was placed in a preheated 100°C oil bath for 45 min. The brown residue was quenched with a saturated solution of NaHCO₃ (until pH 7-9) and extracted with MeTHF (3x). The combined organic layers were washed with brine, dried over Na₂SO₄, and concentrated to give a dark red residue, which was then purified by reverse phase column chromatography (5% to 100% MeOH / water with 0.1% formic acid). The fractions were combined and concentrated to give ethyl 2-((trans)-3-(4-((5-bromoimidazo[1,2-a]pyrazin-8-yl)amino)-1H-pyrazol-1-yl)cyclobutyl)acetate intermediate G-3a as a brown solid (533 mg, 70% yield).

[0557] LCMS: [M+H] + =419.0,421.0.

[0558] 1 H NMR (400MHz, DMSO-d6) δppm 1.19 (t, J = 7.1Hz, 3H), 2.19 (br t,J=8.7Hz,2H),2.56-2.66(m,5H),4.07(q,J=7.3Hz,2H),4.94-5.03(m,1H),7.59(s,1H ),7.70(d,J=1.0Hz,1H),7.79(s,1H),7.99(d,J=1.2Hz,1H),8.17(s,1H),10.00(s,1H). Table 30. List of intermediate G-3 compounds generated by representative procedure TBM-G: Step 2

[0559] Synthesis of TBM-G: Step 3 In a sealed tube, 2-(2,4-difluoro-5-methyl-phenyl)-4,4,5,5-tetramethyl-1,3,2-dioxaborolane intermediate D-4b (485 mg, 1.9 mmol, 1.5 eq), ethyl 2-((trans)-3-(4-((5-bromoimidazo[1,2-a]pyrazin-8-yl)amino)-1H-pyrazol-1-yl)cyclobutyl)acetate intermediate G-3a (533 mg, 1.3 mmol, 1 eq) and NaHCO (320 mg, 3.8 mmol, 3 eq) were added to 1,4-dioxane (3 mL, 0.32 M) and water (1 mL, 0.32 M). Nitrogen was bubbled into the reaction for 10 min. Then, Pd(PPh 3 ) 4 (220 mg, 0.19 mmol, 0.15 equiv) was added and nitrogen was bubbled for another 10 min. The resulting solution was stirred at 90 ° C in a sealed tube under a nitrogen atmosphere for 18 h. The reaction mixture was filtered through celite, rinsed with EtOAc and concentrated. The resulting residue was purified by reverse phase column chromatography (5% to 100% MeOH / water, containing 0.1% formic acid). The fractions were combined and concentrated to obtain ethyl 2-((trans)-3-(4-((5-(2,4-difluoro-5-methylphenyl)imidazo[1,2-a]pyrazine-8-yl)amino)-1H-pyrazol-1-yl)cyclobutyl)acetate intermediate G-4a (562 mg, 91% yield) as a yellow semisolid.

[0560] LCMS: [M+H] + =467.2.

[0561] 1 H NMR (400MHz, DMSO-d6) δppm 1.19 (t, J = 7.1Hz, 3H), 2.20 (br t,J=8.7Hz,2H),2.29(s,3H),2.55-2.69(m,5H),4.07(q,J=7.1Hz,2H),4.95-5.05(m ,1H),7.42-7.50(m,2H),7.58-7.64(m,3H),7.84(s,1H),8.23(s,1H),10.00(s,1H). Table 31. List of intermediate G-4 compounds generated by representative procedure TBM-G: Step 3

[0562] Synthesis of TBM-G: Step 4 To a stirred solution of ethyl 2-((trans)-3-(4-((5-(2,4-difluoro-5-methylphenyl)imidazo[1,2-a]pyrazin-8-yl)amino)-1H-pyrazol-1-yl)cyclobutyl)acetate Intermediate G-4a (562 mg, 1.2 mmol, 1 eq) in THF (8 mL) was added LiAlH4 (137 mg, 3.61 mmol, 3 eq) overnight at 0° C. The reaction was quenched with water (0.5 mL) and 1 N NaOH (0.5 mL) at 0° C. and stirred at room temperature for 20 min. The precipitate was filtered off and the filtrate was concentrated to give 2-((trans)-3-(4-((5-(2,4-difluoro-5-methylphenyl)imidazo[1,2-a]pyrazin-8-yl)amino)-1H-pyrazol-1-yl)cyclobutyl)ethan-1-ol TBM-51 (451 mg, 65% yield) as a light yellow oil which was used without further purification.

[0563] LCMS: [M+H] + =425.2. Table 32. List of TBM-G compounds generated by the representative procedure TBM-G: Step 4 Example S17. General Procedure for TBM-H

[0564] Synthesis of TBM-H: Step 1 In a sealed tube, a mixture of benzyl 3-(4,4,5,5-tetramethyl-1,3,2-dioxaborolan-2-yl)-4,5,6,7-tetrahydropyrazolo[1,5-a]pyridine intermediate D-4m (146.93 mg, 0.59 mmol), 2-((trans)-4-(4-((5-bromoimidazo[1,2-a]pyrazin-8-yl)amino)-1H-pyrazol-1-yl)cyclohexyl)ethan-1-ol intermediate H-2 (200 mg, 0.49 mmol) and K3PO4 (313 mg, 1.48 mmol) in THF (3.7 mL) and water (1.3 mL) was purged with nitrogen for 10 min; then Pd(dtbpf)Cl2 (32 mg, 0.05 mmol) was added and the mixture was purged with nitrogen again for 10 minutes. In the 4-thiazolyl-1H-pyridine-2-yl) imidazo [1,2-a] pyrazine-8-yl) amino) -1H-pyrazoles-1-yl) cyclohexyl) 2- ((trans) -4- (4- ((5- (4,5,6,7- tetrahydropyrazolo [1,5-a] pyridin-3-yl) imidazo [1,2-a] pyrazine-8-yl) amino) -1H-pyrazoles-1-yl) cyclohexyl) 2- (137.3 mg, 63% yield) of orange oil.

[0565] LCMS: [M+H] + =447.2.

[0566] 1 H NMR(400MHz,DMSO-d6)δppm 1.38(q,J=6.4Hz,2H),1.64-1.76(m,2H),1.79-1.88(m,4H),1.99-2.06(m,4H), 2.75(t,J=6.2Hz,2H),3.17(d,J=3.7Hz,2H),3.43-3.48(m,2H),4.04-4.13(m,2H ),4.16(t,J=6.0Hz,2H),4.33-4.38(m,1H),7.30(s,1H),7.61(d,J=1.2Hz,1H), 7.74(s,1H),7.78(d,J=1.2Hz,1H),7.80(s,1H),8.19-8.20(m,1H),9.76(s,1H). Table 33. List of intermediates used in TBM-H: Step 1

[0567] Synthesis of 2-((trans)-4-(4-((5-bromoimidazo[1,2-a]pyrazin-8-yl)amino)-1H-pyrazol-1-yl)cyclohexyl)ethan-1-ol (Intermediate H-2)

[0568] Step 1: (trans)-4-(4-nitro-1H-pyrazol-1-yl)cyclohexane-1-carboxylic acid methyl ester intermediate G-1b (7.38 g, 29.1 mmol, 1.0 equiv) was dissolved in 2:1 MeOH / THF (150 mL) and cooled to 0 ° C., followed by the sequential addition of calcium chloride (6.47 g, 58.3 mmol, 2.0 equiv) and sodium borohydride (4.41 g, 117 mmol, 4.0 equiv). The reaction mixture was stirred at room temperature overnight. Water (50 mL) and brine (50 mL) were added to the mixture while stirring vigorously. The aqueous phase was extracted with EtOAc (3×100 mL) and the combined organic layers were dried over MgSO 4 , filtered, and evaporated under reduced pressure. The residue was purified by normal phase column chromatography (0% to 100% EtOAc / heptane). The fractions were collected and concentrated to give ((trans)-4-(4-nitro-1H-pyrazol-1-yl)cyclohexyl)methanol (3.3 g, 14.6 mmol, 50% yield) as a yellow solid.

[0569] LCMS: [M+H] + =276.2.

[0570] 1 H NMR (400MHz, DMSO-d6) δppm 1.08 (qd, J=12.8, 3.2Hz, 2H), 1.34-1.50 (m, 1H), 1.75 (qd, J=12.5, 3.5Hz, 2H), 1.86 (br d,J=11.6Hz,2H),2.00-2.12(m,2H),3.25(t,J=5.7Hz,2H),4.20(tt,J=11.9,3.9Hz,1H),4.47(t,J=5.3Hz,1H),8.25(s,1H),8.90(s,1H).

[0571] Step 2: To a cooled (0 °C) solution of ((trans)-4-(4-nitro-1H-pyrazol-1-yl)cyclohexyl)methanol (3.3 g, 14.6 mmol, 1.0 eq) and triethylamine (2.66 mL, 19.0 mmol, 1.3 eq) in DCM (73 mL) was added methanesulfonyl chloride (1.25 mL, 16.1 mmol, 1.1 eq) dropwise. The reaction mixture was stirred at room temperature for 2 hours. Water (100 mL) was added to the reaction mixture, the phases were separated, and the aqueous layer was extracted with DCM (3 x 50 mL). The organic layers were combined and washed with 1.0 M HCl. (aq) The mixture was washed with 4-nitro-1H-pyrazol-1-yl)cyclohexylmethyl methanesulfonate (3 x 50 mL) and brine (50 mL), then dried over MgSO4, filtered and evaporated under reduced pressure to give ((trans)-4-(4-nitro-1H-pyrazol-1-yl)cyclohexyl)methyl methanesulfonate (4.25 g, 14.0 mmol, 96% yield) as a yellow solid which was used without further purification.

[0572] LCMS: [M+H] + =304.2.

[0573] 1 H NMR (400MHz, DMSO-d6) δppm 1.16-1.29(m,2H),1.66-1.83(m,3H),1.88(br d,J=12.0Hz,2H),2.09(br dd,J=12.8,3.5Hz,2H),3.18(s,3H),4.08(d,J=6.2Hz,2H),4.24(tt,J=11.8,3.9Hz,1H),8.26(s,1H),8.92(s,1H).

[0574] Step 3: A solution of ((trans)-4-(4-nitro-1H-pyrazol-1-yl)cyclohexyl)methyl methanesulfonate (4.25 g, 14.0 mmol, 1.0 equiv) and sodium cyanide (1.58 g, 32.2 mmol, 2.3 equiv) in DMSO (45 mL) was stirred at 50 ° C overnight. The reaction mixture was cooled to room temperature and then water (100 mL) and EtOAc (50 mL) were added. The phases were separated and the aqueous layer was extracted with EtOAc (3×50 mL). The organic layers were combined and washed with saturated NaHCO 3(aq) (50 mL) and brine (50 mL), dried over MgSO4, filtered and evaporated under reduced pressure to give 2-((trans)-4-(4-nitro-1H-pyrazol-1-yl)cyclohexyl)acetonitrile (3.1 g, 13.2 mmol, 94% yield) as an orange oil which was used without further purification.

[0575] LCMS: [M+H]+ =285.2.

[0576] 1 H NMR (400MHz, DMSO-d6) δppm 1.26 (qd, J=12.7, 3.3Hz, 2H), 1.63-1.75 (m, 1H), 1.75-1.86 (m, 2H), 1.89 (br d,J=12.0Hz,2H),2.03-2.13(m,2H),2.51-2.55(m,2H),4.24(tt,J=11.8,3.9Hz,1H),8.26(s,1H),8.90(s,1H).

[0577] Step 4: To a solution of 2-((trans)-4-(4-nitro-1H-pyrazol-1-yl)cyclohexyl)acetonitrile (3.1 g, 13.2 mmol, 1.0 equiv) in DCM (66 mL) cooled to -78 °C (dry ice / acetone bath) under nitrogen atmosphere was added 1.0 M DIBAL-H in DCM (39.7 mL, 39.7 mmol, 3.0 equiv) dropwise. The reaction mixture was stirred at -78 °C for 2.5 hours. Aqueous Rochelle salt (50 mL) was carefully added to the reaction mixture, which was stirred at room temperature overnight. EtOAc (150 mL) was added, and the organic phase was then washed with 1.0 M HCl. (aq) The mixture was washed with 4% paraformaldehyde (3×20 mL), dried over MgSO 4 , filtered and concentrated under reduced pressure to give 2-((trans)-4-(4-nitro-1H-pyrazol-1-yl)cyclohexyl)acetaldehyde (3.42 g, 12.6 mmol, 95% yield) as an orange oil which was used without further purification.

[0578] LCMS: [M+H] + =238.2.

[0579] 1 H NMR(400MHz,DMSO-d6)δppm 1.12-1.20(m,2H),1.74-1.84(m,5H),2.02-2.06(m,2H),2.37(dd,J=6.6,1. 8Hz, 2H), 4.19-4.26 (m, 1H), 8.25 (s, 1H), 8.91 (s, 1H), 9.68 (t, J = 1.9Hz, 1H).

[0580] Step 5: To a cooled suspension (0 °C) of 2-((trans)-4-(4-nitro-1H-pyrazol-1-yl)cyclohexyl)acetaldehyde (3.13 g, 13.2 mmol, 1.0 equiv) in methanol (66 mL) was added sodium borohydride (998 mg, 26.4 mmol, 2.0 equiv) in one portion. The reaction mixture was stirred at room temperature overnight. Water (80 mL) was added to the reaction mixture, and the aqueous layer was then extracted with EtOAc (3 x 50 mL). The organic layers were combined, washed with brine (50 mL), dried over MgSO4, filtered, and evaporated under reduced pressure. The crude residue was purified by silica gel column chromatography (0% to 100% EtOAc / heptane) to give 2-((trans)-4-(4-nitro-1H-pyrazol-1-yl)cyclohexyl)ethan-1-ol (1.81 g, 7.56 mmol, 57% yield) as a light yellow solid.

[0581] LCMS: [M+H] + =240.2.

[0582] 1 H NMR (400MHz, DMSO-d6) δppm 1.02-1.14(m,2H),1.36(q,J=6.6Hz,2H),1.39-1.53(m,1H),1.67-1.80(m,2H),1.84(br d,J=12.0Hz,2H),1.98-2.11(m,2H),3.38-3.53(m,2H),4.21(tt,J=11.9,3.9Hz,1H),4.35(t,J=5.1Hz,1H),8.25(s,1H),8.90(s,1H).

[0583] Step 6: 2-((trans)-4-(4-nitro-1H-pyrazol-1-yl)cyclohexyl)ethan-1-ol (1.81 g, 7.56 mmol, 1.0 eq) was dissolved in ethyl acetate (38 mL), and the solution was degassed by bubbling nitrogen under ultrasound for 20 minutes. Pd / C 10% w / w (1.61 g, 1.51 mmol, 0.2 eq) was added, and the mixture was further degassed by bubbling nitrogen under ultrasound for 20 minutes. The nitrogen balloon was replaced with a hydrogen-filled balloon, which was bubbled through the reaction mixture for 10 minutes, and the reaction mixture was then stirred under a static hydrogen atmosphere overnight. The reaction mixture was filtered through celite and the celite was thoroughly washed with EtOAc and MeOH. The solvent was evaporated under reduced pressure to give 2-((trans)-4-(4-amino-1H-pyrazol-1-yl)cyclohexyl)ethan-1-ol intermediate H-1 (1.45 g, 6.92 mmol, 91% yield) as a purple solid, which was used without further purification.

[0584] LCMS: [M+H] + =210.2.

[0585] 1 H NMR (400MHz, DMSO-d6) δppm 0.98-1.12(m,2H),1.31-1.37(m,2H),1.38-1.47(m,1H),1.60(br dd,J=12.3,3.3Hz,2H),1.79(br d,J=12.7Hz,2H),1.88-1.98(m,2H),3.38-3.52(m,2H),3.58-4.19(br s, 2H), 3.87 (tt, J = 11.9, 3.8Hz, 1H), 4.33 (t, J = 5.1Hz, 1H), 6.87 (s, 1H), 7.02 (s, 1H).

[0586] Step 7: To a solution of 2-((trans)-4-(4-amino-1H-pyrazol-1-yl)cyclohexyl)ethan-1-ol intermediate H-1 (2.0 g, 9.56 mmol) in 1,4-dioxane (43.44 mL) was successfully added 5,8-dibromoimidazo[1,2-a]pyrazine intermediate D-1a (2.41 g, 8.69 mmol) and vinyl alcohol (2.13 g, 20.85 mmol). The mixture was heated to 95 ° C in an oil bath overnight. After cooling to room temperature, the reaction was partitioned between EtOAc and H2O and the aqueous layer was adjusted to pH 9 with 2M NaOH. The aqueous layer was extracted with EtOAc (3x), and the combined organics were washed with brine, dried over MgSO4, filtered and concentrated. The crude residue was purified by reverse phase column chromatography (5% to 100% MeCN / water, containing 0.1% formic acid). The fractions were collected and concentrated to give 2-((trans)-4-(4-((5-bromoimidazo[1,2-a]pyrazin-8-yl)amino)-1H-pyrazol-1-yl)cyclohexyl)ethan-1-ol intermediate H-2 as a brown solid (1.28 g, 51% yield).

[0587] LCMS: [M+H] + =405.0.

[0588] 1H NMR (400MHz, DMSO-d6) δppm 1.11(br dd,J=14.3,11.4Hz,2H),1.37(q,J=6.8Hz,2H),1.41-1.51(m,1H),1.70(qd,J=12.5,3.3Hz,2H),1.83(br d,J=13.7Hz,2H),2.02(br d,J=12.2Hz,2H),3.39-3.52(m,2H),4.08(tt,J=11.9,3.9Hz,1H),4.34(t,J=5.1Hz,1H),7.5 9(s,1H),7.70(d,J=1.0Hz,1H),7.73(s,1H),7.98(d,J=1.0Hz,1H),8.14(s,1H),9.97(s,1H). Table 34. List of TBM-H compounds generated by the representative procedure TBM-H: Step 1 Example S18. Procedure for TBM-59

[0589] Synthesis of 2-((trans)-4-(4-((5-(2,4-difluoro-5-methylphenyl)imidazo[1,2-a]pyridin-8-yl)amino)-1H-pyrazol-1-yl)cyclohexyl)ethan-1-ol (TBM-59)

[0590] Step 1: In a sealed tube, 2-((trans)-4-(4-amino-1H-pyrazol-1-yl)cyclohexyl)ethan-1-ol intermediate H-1 (250 mg, 1.19 mmol, 1.0 eq), 5-chloro-8-iodoimidazo[1,2-a]pyridine intermediate D-1b (332.6 mg, 1.19 mmol, 1.0 eq), rac-BINAP (74.6 mg, 0.120 mmol, 0.1 eq), and cesium carbonate (1.95 g, 5.97 mmol, 5.0 eq) were combined with toluene (9.5 mL), and nitrogen was bubbled through the mixture under ultrasound for 30 minutes. Pd(OAc)2 (26.9 mg, 0.120 mmol, 0.1 eq) was then added quickly and the nitrogen bubble was continued for an additional 10 minutes. The tube was sealed and the reaction mixture was stirred at 120°C for 4 hours. The reaction mixture was cooled to room temperature and the volatile components were evaporated. The crude material was purified by reverse phase column chromatography (5% to 100% MeOH / water with 0.1% formic acid). The fractions were collected and concentrated to give 2-((trans)-4-(4-((5-chloroimidazo[1,2-a]pyridin-8-yl)amino)-1H-pyrazol-1-yl)cyclohexyl)ethanol-1-ol (275 mg, 0.739 mmol, 62% yield) as a yellow semisolid.

[0591] LCMS: [M+H] + =360.2.

[0592] 1 H NMR (400MHz, CDCl3) δppm 1.10-1.24(m,2H),1.56(br t,J=4.6Hz,3H),1.78(qd,J=12.6,3.1Hz,2H),1.97(br d,J=12.5Hz,2H),2.01(s,1H),2.24(br d,J=11.6Hz,2H),3.74(br t,J=6.3Hz,1H),3.77-3.98(m,2H),4.10(tt,J=12.0,3.7Hz,1H),6.53(d,J=8.2Hz,1H) ,6.79(d,J=8.1Hz,1H),7.49(s,1H),7.52-7.67(m,3H),7.75(d,J=1.1Hz,1H),8.19(br s,1H).

[0593] Step 2: In a sealed tube, 2-((trans)-4-(4-((5-chloroimidazo[1,2-a]pyridin-8-yl)amino)-1H-pyrazol-1-yl)cyclohexyl)ethan-1-ol (275. mg, 0.764 mmol, 1.0 equiv), 2-(2,4-difluoro-5-methyl-phenyl)-4,4,5,5-tetramethyl-1,3,2-dioxaborolane (Intermediate D-4b) (233 mg, 0.917 mmol, 1.2 equiv) and sodium bicarbonate (232 mg, 2.29 mmol, 3.0 equiv) were mixed with 1,4-dioxane (0.6 mL) and water (0.2 mL), and nitrogen was bubbled through the mixture under sonication for 30 min. Then quickly add Pd (PPh ) (177 mg, 0.153 mmol, 0.2 equivalent) and continue to make nitrogen bubble for another 10 minutes. The tube is sealed and the reaction mixture is stirred at 110 ° C for 18h. The reaction mixture is cooled to room temperature, and then the volatile component is evaporated. The crude residue is purified by reverse phase column chromatography (5% to 100% MeOH / water, containing 0.1% formic acid). The fractions are collected and concentrated to obtain 2- ((trans) -4- (4- ((5- (2,4- bis-fluoro-5-methylphenyl) imidazo [1,2-a] pyridin-8-yl) amino) -1H- pyrazole -1- bases) cyclohexyl) ethanol -1- alcohol TBM-59 (135 mg, 0.282 mmol, 37% yield) as a brown semi-solid.

[0594] LCMS: [M+H] + =452.2.

[0595] 1 H NMR (400MHz, DMSO-d6) δppm 1.03-1.17(m,2H),1.38(q,J=6.6Hz,2H),1.41-1.55(m,1H),1.75(qd,J=12.5,3.2Hz,2H),1.85(br d,J=11.9Hz,2H),2.05(br d,J=10.6Hz,2H),2.28(s,3H),3.40-3.54(m,2H),4.08(tt,J=11.9,3.9Hz,1H),4.35(t,J=5.0Hz,1H),6.51(d,J=7.8Hz ,1H),6.74(d,J=7.7Hz,1H),7.41(t,J=10.0Hz,1H),7.45(d,J=1.8Hz,1H),7.49-7.58(m,3H),7.87(s,1H),8.10(s,1H).

[0596] 19F NMR (377MHz, DMSO-d6) δppm -112.35 (q, J = 9.1Hz, 1F), -111.89 (q, J = 8.2Hz, 1F). Example S19. TBM-60 Procedure

[0597] Synthesis of 5-(2,4-difluoro-5-methylphenyl)-N-(1-(1-(2,2-dimethoxyethyl)piperidin-4-yl)-1H-pyrazol-4-yl)-N-methylimidazo[1,2-a]pyrazin-8-amine (TBM-60)

[0598] Step 1: To a solution of 5-(2,4-difluoro-5-methylphenyl)-N-(1-(piperidin-4-yl)-1H-pyrazol-4-yl)imidazo[1,2-a]pyrazin-8-amine TBM-2 (bis-HCl salt, 1.0 g, 2.07 mmol, 1 eq) in DCM (4.5 mL, 0.42 M) and DMSO (0.40 mL, 0.42 M) was added 60% w / w 2,2-dimethoxyacetaldehyde in HO (0.63 mL, 4.14 mmol, 2 eq) and NaBH(OAc) (878.76 mg, 4.15 mmol, 2 eq). The reaction mixture was stirred at room temperature for 2 h. The solvent was evaporated and the residue was then purified by reverse phase column chromatography (5% to 40% MeOH / water with 0.1% formic acid). The fractions were collected and concentrated to afford 5-(2,4-difluoro-5-methylphenyl)-N-(1-(1-(2,2-dimethoxyethyl)piperidin-4-yl)-1H-pyrazol-4-yl)imidazo[1,2-a]pyrazin-8-amine (936 mg, 91% yield) as a tan solid.

[0599] LCMS: [M+H] + =498.2.

[0600] 1 H NMR(400MHz,DMSO-d6)δppm 1.87-2.02(m,4H),2.18-2.27(m,2H),2.29(s,3H),3.00(br d,J=11.7Hz,2H),3.27(s,6H),4.08-4.17(m,1H),4.50(t,J=5.0Hz,1H),7.42-7. 48(m,2H),7.58-7.65(m,3H),7.80(s,1H),8.14(s,1H),8.23(s,1H),9.98(s,1H).

[0601] 19F NMR (377MHz, DMSO-d6) δppm -112.17 (q, J = 9.5Hz, 1F), -111.25 (q, J = 8.2Hz, 1F).

[0602] Step 2: To a solution of 5-(2,4-difluoro-5-methylphenyl)-N-(1-(1-(2,2-dimethoxyethyl)piperidin-4-yl)-1H-pyrazol-4-yl)imidazo[1,2-a]pyrazin-8-amine (250 mg, 0.5000 mmol, 1.0 eq) in THF (5 mL) was added NaH (24.12 mg, 0.6000 mmol, 1.2 eq) at 0 °C. The reaction was stirred at room temperature for 15-20 min. Then, MeI (0.03 mL, 0.5300 mmol, 1.05 eq) was added. The reaction mixture was stirred at room temperature overnight. LCMS showed a mixture of starting material and 3 products, two of which had the expected mass. MeI (0.03 mL, 0.5300 mmol, 1.05 eq) was added and after 2 h the reaction was quenched with water and extracted with EtOAc (3x). The combined organic layers were dried over sodium sulfate, filtered and concentrated. The crude product was purified by reverse phase column chromatography (5 to 100% MeCN / water, containing 0.1% formic acid) to give the product (80% purity by LCMS at high pH). The mixture was further purified by reverse phase column chromatography (0% to 100% MeCN / water, containing pH 10 buffer) to give 5-(2,4-difluoro-5-methylphenyl)-N-(1-(1-(2,2-dimethoxyethyl)piperidin-4-yl)-1H-pyrazol-4-yl)-N-methylimidazo[1,2-a]pyrazin-8-amine TBM-60 (40 mg, 16%) as a white solid.

[0603] LCMS: [M+H] + =512.3.

[0604] 1 H NMR(400MHz,DMSO-d6)δppm 1.97(br s,4H),2.16-2.24(m,2H),2.28(br s,3H),2.43-2.47(m,2H),2.94-3.04(m,2H),3.27(br s,6H),3.97(br s,3H),4.03-4.15(m,1H),4.44-4.54(m,1H),7.38-7.50(m,2H),7.55-7.68(m,4H),8.04-8.12(m,1H).

[0605] 19F NMR (377MHz, DMSO-d6) δppm-112.15--111.82(m,1F),-111.14--110.99(m,1F). Procedure for Compound Nos. 1-120 Example S20. Representative Procedure LDD-I

[0606] Step 1: TBM-1 (68.0 mg, 0.16 mmol) was dissolved in a mixture of DMF (0.80 mL) and DIPEA (113 μL, 0.65 mmol), followed by the addition of 2-bromoacetic acid (14.0 μL, 0.19 mmol) and the solution was stirred at room temperature. HPLC showed complete conversion to product and a small amount of impurities. The mixture was purified by reverse phase column chromatography (5% to 100% MeOH / water, containing 0.1% formic acid). The fractions were combined and evaporated to give 2-(4-(4-((5-(m-tolyl)imidazo[1,2-a]pyrazin-8-yl)amino)-1H-pyrazol-1-yl)piperidin-1-yl)acetic acid (54 mg, 77% yield) as a white solid.

[0607] LCMS: [M+H] + =433.2.

[0608] 1 H NMR(400MHz,DMSO-d6)δppm 1.99-2.08(m,4H),2.41(s,3H),2.53-2.59(m,2H),3.10(br d,J=11.5Hz,3H),4.16-4.24(m,1H),7.31-7.34(m,1H),7.43(s,1H),7.45-7.52(m,3H ),7.64(d,J=1.0Hz,1H),7.81(s,1H),7.89(d,J=1.0Hz,1H),8.25(s,1H),9.90(s,1H).

[0609] Step 2: To a solution of CBM-1 (53.4 mg, 0.12 mmol), DIPEA (102 μL, 0.59 mmol) and 2-(4-(4-((5-(m-tolyl)imidazo[1,2-a]pyrazin-8-yl)amino)-1H-pyrazol-1-yl)piperidin-1-yl)acetic acid (53 mg, 0.12 mmol) in DMF (1 mL) was added PyAOP (61 mg, 0.12 mmol) at 0°C. The mixture was stirred at room temperature for 3 hours. The mixture was purified by reverse phase column chromatography (5% to 100% MeCN / water with 0.1% formic acid). The fractions were collected and evaporated to give 2-(2,6-dioxopiperidin-3-yl)-5-(4-(2-(4-(4-((5-(m-tolyl)imidazo[1,2-a]pyrazin-8-yl)amino)-1H-pyrazol-1-yl)piperidin-1-yl)acetyl)piperazin-1-yl)isoindoline-1,3-dione as a light yellow solid (22.4 mg, 25% yield, formate salt).

[0610] LCMS: [M+H] + =756.2.

[0611] 1 H NMR(400MHz,DMSO-d6)δppm 1.90-2.06(m,5H),2.23(br.t,J=10.4Hz,2H),2.40(s,3H),2.53-2.62(m,2H),2.83-2.90(m,1H),2.92-2.99(m, 2H),3.25(s,2H),3.46-3.51(m,2H),3.53-3.59(m,2H),3.59-3.65(m,2H),3.74-3.80(m,2H),4.09-4.19(m,1H), 5.07(dd,J=12.8,5.5Hz,1H),7.25-7.34(m,2H),7.38(br.d,J=1.0Hz,1H),7.42(s,1H),7.44-7.47(m,2H),7.50( br.s,1H),7.64(s,1H),7.70(d,J=8.3Hz,1H),7.80(s,1H),7.89(s,1H),8.23(s,1H),9.88(s,1H),11.08(s,1H). Table 35. List of Examples Prepared by Procedure LDD-I Example S21. General Procedure for LDD-J

[0612] Step 1: To a solution of 5-(2,4-difluoro-5-methylphenyl)-N-(1-(piperidin-4-yl)-1H-pyrazol-4-yl)imidazo[1,2-a]pyrazin-8-amine dihydrochloride TBM-2 (1.0 g, 2.07 mmol, 1 eq) in DCM (4.5 mL, 0.42 M) and DMSO (0.40 mL, 0.42 M) was added 60% w / w 2,2-dimethoxyacetaldehyde in H2O8 (0.63 mL, 4.14 mmol, 2 eq) and NaBH(OAc)3 (879 mg, 4.15 mmol, 2 eq). The reaction mixture was stirred at room temperature for 2 h. The solvent was evaporated and the residue was then purified by reverse phase column chromatography (5% to 100% MeOH / water with 0.1% formic acid). Fractions were combined and concentrated to give 5-(2,4-difluoro-5-methylphenyl)-N-(1-(1-(2,2-dimethoxyethyl)piperidin-4-yl)-1H-pyrazol-4-yl)imidazo[1,2-a]pyrazin-8-amine (936 mg, 91% yield) as a tan solid.

[0613] LCMS: [M+H] + =498.2.

[0614] 1 H NMR(400MHz,DMSO-d6)δppm 1.87-2.02(m,4H),2.18-2.27(m,2H),2.29(s,3H),3.00(br d,J=11.7Hz,2H),3.27(s,6H),4.08-4.17(m,1H),4.50(t,J=5.0Hz,1H),7.42-7. 48(m,2H),7.58-7.65(m,3H),7.80(s,1H),8.14(s,1H),8.23(s,1H),9.98(s,1H).

[0615] 19 F NMR (377MHz, DMSO-d6) δppm -112.17 (q, J = 9.5Hz, 1F), -111.25 (q, J = 8.2Hz, 1F).

[0616] Step 2: To a round-bottom flask was added 5-(2,4-difluoro-5-methylphenyl)-N-(1-(1-(2,2-dimethoxyethyl)piperidin-4-yl)-1H-pyrazol-4-yl)imidazo[1,2-a]pyrazin-8-amine (871 mg, 1.75 mmol, 1 eq), 4M HCl in dioxane (13.14 mL, 52.57 mmol, 30 eq), and water (0.48 mL, 4.1 M). The reaction was stirred at room temperature for 16 h. The solvent was removed under reduced pressure and the residue was co-evaporated with MeCN (3x) to give 2-(4-(4-((5-(2,4-difluoro-5-methylphenyl)imidazo[1,2-a]pyrazin-8-yl)amino)-1H-pyrazol-1-yl)piperidin-1-yl)acetaldehyde (880 mg) as a yellow solid as the HCl salt.

[0617] LCMS: [M+H2O+H] + =470.2.

[0618] 1 H NMR(400MHz,DMSO-d6)δppm 2.20-2.41(m,7H),3.29-3.42(m,4H),3.60-3.76(m,2H),4.35-4.62(m,1H),4.87-5.03(m,1H),7.45-7.54 (m,2H),7.61(t,J=8.2Hz,1H),7.74-7.80(m,1H),7.82(d,J=4.6Hz,2H),8.24(s,1H),9.92-10.11(m,1H).

[0619] 19 F NMR (377MHz, DMSO-d6) δppm -112.07 (s, 1F), -110.60 (s, 1F).

[0620] Step 3: To a solution of 2-(4-(4-((5-(2,4-difluoro-5-methylphenyl)imidazo[1,2-a]pyrazin-8-yl)amino)-1H-pyrazol-1-yl)piperidin-1-yl)acetaldehyde hydrochloride (100 mg, 0.20 mmol, 1 eq), 3-(4-piperazin-1-ylphenyl)piperidine-2,6-dione CBM-2 (56 mg, 0.20 mmol, 1 eq) and DIPEA (0.25 mL, 1.43 mmol, 7 eq) in DCM (0.73 mL, 0.14 M) and DMSO (0.73 mL, 0.14 M) was added NaBH(OAc) (87 mg, 0.4100 mmol, 2 eq) under N atmosphere. The reaction mixture was stirred at room temperature for 40 h. The solvent was evaporated and the residue was then purified by reverse phase column chromatography (5% to 95% MeCN / water with 0.1% formic acid). Fractions were combined and concentrated to give 3-(4-(4-(2-(4-(4-((5-(2,4-difluoro-5-methylphenyl)imidazo[1,2-a]pyrazin-8-yl)amino)-1H-pyrazol-1-yl)piperidin-1-yl)ethyl)piperazin-1-yl)phenyl)piperidine-2,6-dione Ex. 8 as a white solid (37 mg, 25% yield).

[0621] LCMS: [M+H] + =709.3.

[0622] 1 H NMR (400MHz, DMSO-d6) δppm 1.86-2.05(m,5H),2.09-2.18(m,3H),2.29(s,3H),2.42-2.48(m,1H),2.54-2.69(m,6H),3.01(br d,J=11.2Hz,2H),3.11(br s,4H),3.72(dd,J=11.2,5.1Hz,1H),4.08-4.18(m,1H),6.89(d,J=8.6Hz,2H),7.04(d,J=8.6Hz, 2H),7.41-7.48(m,2H),7.58-7.65(m,3H),7.80(s,1H),8.23(s,1H),9.98(s,1H),10.77(s,1H).

[0623] 19 F NMR (377MHz, DMSO-d6) δppm -112.16 (q, J = 8.6Hz, 1F), -111.26 (q, J = 10.0Hz, 1F). Table 36. List of Examples Prepared by Procedure LDD-J Example S22. Representative Procedure for LDD-K

[0624] Step 1: To a round-bottom flask containing N-[1-[1-(2,2-dimethoxyethyl)-4-piperidinyl]pyrazol-4-yl]-5-ethyl-imidazo[1,2-a]pyrazin-8-amine TBM-28 (30.0 mg, 0.07 mmol) was added 4M HCl in 1,4-dioxane (254 μL, 1.01 mmol, 15 equiv) and water (0.10 mL) at room temperature. The reaction mixture was heated to 60° C. and stirred for 3 h. The solvent was removed under reduced pressure and excess HCl was added, followed by MeCN (4×) to give 2-(4-(4-((5-ethylimidazo[1,2-a]pyrazin-8-yl)amino)-1H-pyrazol-1-yl)piperidin-1-yl)acetaldehyde (26 mg, quantitative) as a yellow solid, which was used directly in the next step.

[0625] Step 2: To a solution of 2-(4-(4-((5-ethylimidazo[1,2-a]pyrazin-8-yl)amino)-1H-pyrazol-1-yl)piperidin-1-yl)acetaldehyde (27. mg, 0.07 mmol, 1 eq) and DIPEA (0.06 mL, 0.35 mmol, 5 eq) in CHCl (0.69 mL) was added 3-(4-piperazin-1-ylphenyl)piperidine-2,6-dione dihydrochloride (29 mg, 0.08 mmol, 1.2 eq) and NaBH(OAc) (29 mg, 0.14 mmol, 2 eq). The reaction mixture was stirred at room temperature for 18 h. The solvent was evaporated and the residue was purified by reverse phase column chromatography (5% to 100% MeCN / water with 0.1% formic acid). The fractions were combined, concentrated, and purified again by reverse phase column chromatography (5% to 100% MeCN / water). The fractions were combined, concentrated, and lyophilized to give 3-(4-(4-(2-(4-(4-((5-ethylimidazo[1,2-a]pyrazin-8-yl)amino)-1H-pyrazol-1-yl)piperidin-1-yl)ethyl)piperazin-1-yl)phenyl)piperidine-2,6-dione Example 70 as an off-white solid (5 mg, 11% yield).

[0626] LCMS: [M+H] + =611.4.

[0627] 1H NMR(400MHz,DMSO-d6)δppm 1.29(t,J=7.3Hz,3H),1.84-2.05(m,6H),2.07-2.19(m,3H),2.41-2.47(m,3H),2.52-2.65(m,6H),2.84(q,J=7.6Hz,2H),3.00(br d,J=11.7Hz,2H),3.09-3.13(m,4H),3.72(dd,J=10.8,4.6Hz,1H),4.04-4.16(m,1H),6.89(d,J=8.8Hz,2H),7.05(d,J=8.8Hz,2H), 7.24(s,1H),7.64(d,J=1.2Hz,1H),7.74-7.75(m,1H),7.96(d,J=1.0Hz,1H),8.18(s,1H),8.19(s,1H),9.58(s,1H),10.77(s,1H). Table 37. List of Examples Prepared by Procedure LDD-K Example S23. Representative Procedure for LDD-L

[0628] Step 1: To a suspension of 2-((trans)-4-(4-((5-(2,4-difluoro-5-methylphenyl)imidazo[1,2-a]pyrazin-8-yl)amino)-1H-pyrazol-1-yl)cyclohexyl)ethan-1-ol TBM-43 (425 mg, 0.94 mmol, 1 eq) in ethyl acetate (10 mL) was added IBX (526.0 mg, 1.88 mmol, 2 eq) at room temperature. The reaction was then stirred at 80 °C for 5 h. The reaction was cooled to room temperature, filtered through celite, rinsed with EtOAc, and concentrated. The residue was purified by reverse phase column chromatography (5% to 100% MeCN / water). Fractions were combined and concentrated to give 2-((trans)-4-(4-((5-(2,4-difluoro-5-methylphenyl)imidazo[1,2-a]pyrazin-8-yl)amino)-1H-pyrazol-1-yl)cyclohexyl)acetaldehyde as a tan solid (375 mg, 78% yield).

[0629] LCMS: [M+H] + =451.2.

[0630] Step 2: To a solution of 2-((trans)-4-(4-((5-(2,4-difluoro-5-methylphenyl)imidazo[1,2-a]pyrazin-8-yl)amino)-1H-pyrazol-1-yl)cyclohexyl)acetaldehyde (330 mg, 0.73 mmol, 1 eq) in DCM (7.5 mL) was added 3-(4-piperazin-1-ylphenyl)piperidine-2,6-dione CBM-2 (380 mg, 1.1 mmol, 1.5 eq, bis-HCl salt) and DIPEA (0.38 mL, 2.2 mmol, 3 eq) at room temperature. NaBH(OAc)3 (310 mg, 1.47 mmol, 3 eq) was then added and the mixture was stirred at room temperature under nitrogen for 20 h. DCM was removed under reduced pressure and the residue was purified by reverse phase column chromatography (5% to 100% MeCN / water with 0.1% formic acid). Fractions were combined, concentrated and lyophilized to give 3-(4-(4-(2-((trans)-4-(4-((5-(2,4-difluoro-5-methylphenyl)imidazo[1,2-a]pyrazin-8-yl)amino)-1H-pyrazol-1-yl)cyclohexyl)ethyl)piperazin-1-yl)phenyl)piperidine-2,6-dione (Compound 11) (427 mg, 80% yield) as a white solid as the complete formate salt.

[0631] LCMS: [M+H] + =708.3.

[0632] 1H NMR(400MHz,DMSO-d6)δppm 1.07-1.21(m,2H),1.32-1.46(m,3H),1.73(qd,J=12.3,3.1Hz,2H),1.84-1.92(m,2H),1.95-2.19(m,4H),2.29(s,3H),2.39(br t,J=7.2Hz,2H),2.41-2.49(m,2H),2.52-2.54(m,3H),2.58-2.67(m,1H),3 .08-3.15(m,4H),3.72(dd,J=10.9,5.0Hz,1H),4.10(tt,J=11.9,3.8Hz,1H) ,6.89(d,J=8.8Hz,2H),7.05(d,J=8.8Hz,2H),7.42-7.48(m,2H),7.58-7.6 3(m,3H),7.78(s,1H),8.14(s,1H),8.21(s,1H),9.96(s,1H),10.77(s,1H).

[0633] 19 F NMR (377MHz, DMSO-d6) δppm -112.17 (q, J = 8.2Hz, 1F), -111.27 (q, J = 9.1Hz, 1F). Table 38. List of Examples Prepared by Procedure LDD-L Example S24. Representative Procedure for LDD-M

[0634] Step 1: To a solution of 3-(4-piperazin-1-ylphenyl)piperidine-2,6-dione hydrochloride CBM-2 (200 mg, 0.65 mmol, 1 eq) in DMF (0.80 mL) was added 2-bromoacetic acid (99 mg, 0.71 mmol, 1.1 eq) and DIPEA (0.56 mL, 3.23 mmol, 5 eq) at room temperature. The mixture was stirred at room temperature for 19 h. The solution was then purified directly by reverse-phase column chromatography (5% to 100% MeCN / water with 0.1% formic acid). Fractions were combined and concentrated to afford 2-(4-(4-(2,6-dioxopiperidin-3-yl)phenyl)piperazin-1-yl)acetic acid (45 mg, 21% yield) as a yellow solid. 1 HNMR showed about 15% w / w of bis-alkylated product.

[0635] LCMS: [M+H] + =332.2.

[0636] 1 H NMR(400MHz,DMSO-d6)δppm 1.95-2.05(m,1H),2.06-2.20(m,1H),2.41-2.47(m,1H),2.58-2.67(m,1H),2.68-2.75(m,4H),3.12-3.17(m ,4H),3.21(s,2H),3.73(dd,J=11.0,4.9Hz,1H),6.89(d,J=8.6Hz,2H),7.05(d,J=8.6Hz,2H),10.77(s,1H).

[0637] Step 2: To a solution of 2-(4-(4-(2,6-dioxopiperidin-3-yl)phenyl)piperazin-1-yl)acetic acid (45 mg, 0.14 mmol, 1 eq) in DMF (0.68 mL) was added DIPEA (0.24 mL, 1.36 mmol, 10 eq) and 5-(2,4-difluoro-5-methylphenyl)-N-(1-(piperidin-4-yl)-1H-pyrazol-4-yl)imidazo[1,2-a]pyrazin-8-amine dihydrochloride TBM-2 (92 mg, 0.19 mmol, 1.4 eq) at room temperature. The resulting solution was stirred at room temperature for 10 min. PyAOP (92 mg, 0.18 mmol, 1.3 eq) was then added in one portion. The reaction mixture was stirred at room temperature for 2.5 h. The crude mixture was purified by reverse phase column chromatography (5% to 100% MeCN / water with 0.1% formic acid). The fractions were combined, collected, and purified by preparative-LCMS to give 3-(4-(4-(2-(4-(4-((5-(2,4-difluoro-5-methylphenyl)imidazo[1,2-a]pyrazin-8-yl)amino)-1H-pyrazol-1-yl)piperidin-1-yl)-2-oxoethyl)piperazin-1-yl)phenyl)piperidine-2,6-dione (Compound 23) (25 mg, 24% yield) as a white solid.

[0638] LCMS: [M+H] + =723.4.

[0639] 1 H NMR(400MHz,DMSO-d6)δppm 1.64-1.78(m,1H),1.89-2.17(m,5H),2.29(s,3H),2.40-2.48(m,1H),2.54-2.66(m,5H),2. 70-2.81(m,1H),3.11-3.22(m,6H),3.34-3.41(m,1H),3.70(dd,J=11.0,4.9Hz,1H),4.20(br d,J=12.7Hz,1H),4.41-4.51(m,2H),6.89(d,J=8.8Hz,2H),7.01(d,J=8.6Hz,2H),7.42 -7.48(m,2H),7.58-7.63(m,3H),7.80(s,1H),8.24(s,1H),10.00(s,1H),10.76(s,1H).

[0640] 19 F NMR (377MHz, DMSO-d6) δppm-112.23--112.05(m,1F),-111.29--111.14(m,1F). Table 39. List of Examples Prepared by Procedure LDD-M Example S25. Representative Procedures for LDD-N

[0641] Step 1: In a flask under N2 at 0°C, 2-((cis)-3-(4-((5-(2,4-difluoro-5-methylphenyl)imidazo[1,2-a]pyrazin-8-yl)amino)-1H-pyrazol-1-yl)cyclobutyl)ethan-1-ol TBM-54 (43.0 mg, 0.10 mmol) was dissolved in DCM (3 mL). Then, NEt3 (0.04 mL, 0.30 mmol) was added, followed by MsCl (0.02 mL, 0.30 mmol). The reaction was stirred at room temperature overnight. The reaction was quenched with water and extracted with DCM. The organic layers were combined, dried over MgSO4, filtered and concentrated. The residue was purified by reverse phase column chromatography (5% to 100% MeCN / water with 0.1% formic acid). The fractions were collected and concentrated to give 2-((cis)-3-(4-((5-(2,4-difluoro-5-methylphenyl)imidazo[1,2-a]pyrazin-8-yl)amino)-1H-pyrazol-1-yl)cyclobutyl)ethyl methanesulfonate (42 mg, 82% yield) as a light orange semisolid.

[0642] LCMS: [M+H] + =503.2.

[0643] 1 H NMR(400MHz,DMSO-d6)δppm 1.85-1.93(m,2H),2.12-2.22(m,3H),2.30(s,3H),2.54-2.61(m,2H),3.19(s,3H),4.20(t,J=6.4Hz,2H),4.66 -4.79(m,1H),7.42-7.50(m,2H),7.59-7.66(m,3H),7.80-7.86(m,1H),8.22-8.26(m,1H),9.96-10.03(m,1H).

[0644] Step 2: In a flask under N2 atmosphere, 2-((cis)-3-(4-((5-(2,4-difluoro-5-methylphenyl)imidazo[1,2-a]pyrazin-8-yl)amino)-1H-pyrazol-1-yl)cyclobutyl)ethyl methanesulfonate (42 mg, 0.08 mmol), 3-(4-piperazin-1-ylphenyl)piperidine-2,6-dione CBM-2 (39 mg, 0.13 mmol), 18-crown-6 (11 mg, 0.04 mmol) and K2CO3 (24 mg, 0.17 mmol) were dissolved in DMF (2 mL). The reaction was then stirred at 70 °C for 3 hours, with LCMS indicating 15% conversion. The reaction was quenched, concentrated and purified by reverse phase column chromatography (5% to 50% MeCN / 0.02 M aqueous HCl). The fractions were collected and further purified by preparative-HPLC (C18, using MeCN / HCl 0.02 M as eluent) to give 3-(4-(4-(2-((cis)-3-(4-((5-(2,4-difluoro-5-methylphenyl)imidazo[1,2-a]pyrazin-8-yl)amino)-1H-pyrazol-1-yl)cyclobutyl)ethyl)piperazin-1-yl)phenyl)piperidine-2,6-dione (Compound 96) (8.6 mg, 15% yield) as a light yellow semisolid.

[0645] LCMS: [M+H] + =680.4,[M+2H] 2+ =340.7.

[0646] 1 H NMR(400MHz,DMSO-d6)δppm 1.91-2.12(m,5H),2.12-2.23(m,3H),2.26-2.36(m,4H),2.55-2.70(m,4H),3.04-3.19(m,6H),3.54-3.61(m,3H), 4.70-4.80(m,1H),6.97(d,J=8.8Hz,2H),7.11(d,J=8.6Hz,2H),7.45-7.50(m,2H),7.60(t,J=8.3Hz,1H),7.72(br s,1H),7.76(s,1H),7.82(s,1H),8.23(s,1H),10.33-10.47(m,2H),10.80(s,1H). Table 40. List of Examples Prepared by Procedure LDD-N Example S26. Representative procedures for LDD-O

[0647] Step 1: To a solution of 3-(4-piperazin-1-ylphenyl)piperidine-2,6-dione dihydrochloride CBM-2 (500 mg, 1.44 mmol) in DCM (14.4 mL) was added 60% w / w 2,2-dimethoxyacetaldehyde and NaBH(OAc) (0.61 g, 2.89 mmol) in H2O (0.33 mL, 2.17 mmol). The reaction mixture was stirred at room temperature for 18 h. The suspension was then concentrated to dryness and purified by reverse-phase column chromatography (5% to 100% MeCN / water with 0.1% formic acid). Fractions were collected and concentrated to give 3-(4-(4-(2,2-dimethoxyethyl)piperazin-1-yl)phenyl)piperidine-2,6-dione (408 mg, 73% yield) as a tan solid.

[0648] LCMS: [M+H] + =362.2.

[0649] 1 H NMR(400MHz,DMSO-d6)δppm 1.92-2.06(m,1H),2.06-2.21(m,1H),2.42-2.49(m,3H),2.55-2.60(m,4H),2.60-2.70(m,1H),3.04-3.14(m,4H),3.27 (s, 6H), 3.72 (dd, J = 11.0, 4.9Hz, 1H), 4.52 (t, J = 5.1Hz, 1H), 6.88 (d, J = 8.8Hz, 2H), 7.05 (d, J = 8.8Hz, 2H), 10.77 (s, 1H).

[0650] Step 2: To a flask charged with 3-(4-(4-(2,2-dimethoxyethyl)piperazin-1-yl)phenyl)piperidine-2,6-dione (147 mg, 0.410 mmol) was added 4.0 M HCl in 1,4-dioxane (3.05 mL, 12.2 mmol), followed by water (0.050 mL). The reaction was stirred at room temperature for 5 h. The mixture was then concentrated to dryness, washed with MeCN (3x) and dried in vacuo to afford 2-(4-(4-(2,6-dioxopiperidin-3-yl)phenyl)piperazin-1-yl)acetaldehyde dihydrochloride (189 mg, 99% yield) as a tan solid.

[0651] LCMS: [M+H] + =316.2

[0652] Step 3: To a suspension of 5,6-dimethyl-N-(1-(piperidin-4-yl)-1H-pyrazol-4-yl)imidazo[1,2-a]pyrazin-8-amine TBM-21 (95 mg, 0.31 mmol) and 2-(4-(4-(2,6-dioxopiperidin-3-yl)phenyl)piperazin-1-yl)acetaldehyde dihydrochloride (181 mg, 0.40 mmol) in DCM (4.07 mL) was added DIPEA (0.21 mL, 1.22 mmol). The mixture was stirred for 10 min, followed by the addition of NaBH(OAc)3 (84 mg, 0.40 mmol) and continued stirring for 18 h, after which additional acetaldehyde (95 mg, 0.310 mmol) and NaBH(OAc)3 (84. mg, 0.40 mmol) were added. After an additional 18 h, the reaction mixture was concentrated and purified by reverse phase column chromatography (5% to 100% MeCN / water with 0.1% formic acid) followed by preparative HPLC to afford 3-(4-(4-(2-(4-(4-((5,6-dimethylimidazo[1,2-a]pyrazin-8-yl)amino)-1H-pyrazol-1-yl)piperidin-1-yl)ethyl)piperazin-1-yl)phenyl)piperidine-2,6-dione (Compound 84) (6.9 mg, 3% yield) as an off-white solid.

[0653] LCMS: [M+H] + =611.4.

[0654] 1 H NMR (400MHz, DMSO-d6) δppm 1.86-2.04(m,5H),2.07-2.18(m,3H),2.37(s,3H),2.44(s,3H),2.53-2.58(m,4H),2.58-2.65(m,1H),3.00(br d,J=12.2Hz,2H),3.09-3.13(m,4H),3.72(dd,J=11.0,4.9Hz,1H),4.04-4.17(m,1H),6.89(d,J=8.8Hz,2H),7.05(d,J=8.6Hz ,2H),7.58(d,J=1.0Hz,1H),7.78(s,1H),7.84(d,J=1.0Hz,1H),8.22(s,1H),8.43(s,2H),9.52(s,1H),10.77(s,1H),7H is not obvious. Table 41. List of Examples Prepared by Procedure LDD-O Biological Examples Example B1. IRAK3-ePL overexpression degradation assay

[0655] Stable cell lines were generated by the following protocol. 3 × 10 5 Lenti-X 293T cells (Clonetech) were seeded in 0.8 mL of culture medium in 12-well plates and incubated overnight at 37°C / 5% CO2. The packaging plasmid (0.4 μg, pMD), envelope plasmid (0.4 μg, pSP), and lentiviral transfer IRAK3-ePL plasmid (0.8 μg, IRAK3 sequence NM_007199.3) were mixed in 0.1 mL of Opti-MEM and incubated for 5 minutes. Simultaneously, 2.4 μL of Lipofectamine 2000 (Invitrogen) was added to 0.1 mL of Opti-MEM (Gibco) and incubated for 5 minutes. The plasmid DNA and liposomes were combined, and the mixture was incubated for 20 minutes. The DNA:Lipofectamine Opti-MEM mixture was then added dropwise to the previously seeded cells and incubated at 37°C / 5% CO2 for ~16 hours. Following incubation, the culture medium was removed and 1.2 mL of fresh culture medium was added to each well. Lenti-X 293T cells were incubated at 37°C / 5% CO2 for 30 h. 6 293T CRBN OE / GSPT1 G575N KI Cells were seeded in 0.5 mL of culture medium per well of a 12-well plate and incubated at 37°C / 5% CO2 for ~16 h. After incubation, the culture medium was removed from the Lenti-X 293T wells and filtered through a 0.45 μM filter. A portion of the viral supernatant was used to transfect the cells and the remainder was stored at -80°C. The virus was then added separately (0.5 mL of virus) to each well of the 293T. CRBN OE / GSPT1 G575N KIThe cells were then incubated at 37°C / 5% CO2 for 24 h. After the culture medium was aspirated from the plate, the cells were washed with DPBS, digested with trypsin and planted in 15 mL of culture medium and 1 μg / mL puromycin in a 10 cm dish. After the cells were incubated at 37°C / 5% CO2 for 72 h, the culture medium was aspirated from the plate and the cells were washed with DPBS and digested with trypsin. The cells were planted in 40 mL of culture medium with 1.0 μg / mL puromycin (Gibco) in a 15 cm dish and incubated at 37°C / 5% CO2 for 72 h. After incubation, the culture medium was removed and the cells were washed with DPBS and digested with trypsin. Most of the cells were resuspended and stored in Invitrogen freezing medium (~6-8 × 10 6 cells / bottle).

[0656] The IRAK3-ePL cell dose response curve degradation assay was performed according to the following protocol. The test compound was dispensed into a white 384-well tissue culture treated plate using an ultrasonic liquid processor. Based on a 25 μL assay volume, aliquots were prepared by repeated 10.3-fold serial dilutions starting from a 10 μM dose. Negative control wells containing only 0.2% DMSO were included to calculate the 100% signal. Positive control wells containing 30 μM Ataluren (luciferase inhibitor) were included to calculate the background signal level. All wells were backfilled to a final DMSO concentration of 0.2% to ensure consistency of DMSO between wells. Cells expressing IRAK3-ePL (IRAK3-ePL Lenti-X 293T CRBN / GSPT1 G575N) were washed, trypsinized, counted, and resuspended in fresh DMEM (Gibco) to a cell concentration of 200,000 cells / mL. 25 μL of cells (5,000 cells / well) were distributed into the wells of a 384-well plate pre-spotted with compounds in the previous step and incubated overnight at 37°C / 5% CO2. After incubation, the 384-well plate was removed from the incubator and allowed to stand at room temperature for 30 minutes. InCELL capture reagent was prepared according to the manufacturer's instructions (EA reagent, lysis buffer, and substrate reagent in a 1:1:4 ratio, Cat#96-0002, DiscoverX) and added to the 384-well plate at 25 μL per well. After incubating the plate at room temperature for 1 hour, the fluorescence signal was read using a ViewLux plate reader. The data were processed and analyzed in ActivityBase software. Briefly, the average luminescence value of the positive control wells was subtracted from the rest of the wells for background correction, and all luminescence values ​​were normalized to the DMSO control wells. The average value of the DMSO control wells was set equal to 100% of the relative IRAK3-ePL protein level. The normalized luminescence values ​​were plotted as a function of compound concentration. The x-axis represents the compound concentration and the y-axis represents the corresponding relative IRAK3-ePL protein level. The EC values ​​of the compounds used to degrade IRAK3-ePL were calculated. 50 The values ​​were calculated using a four-parameter logistic model (half-maximal effective concentration) (sigmoidal dose-response model) (FIT = (A + {(B A) / 1 + [(C / x) D ]}))(where C is the inflection point (EC 50 ), D is the correlation coefficient, and A and B are the lower and upper limits of the fit, respectively).

[0657] D max Calculated by determining the maximum percentage loss of target protein after compound treatment. min The percentage of target protein remaining after compound treatment was calculated (% D max =100-Y min) . Example B2. IRAK endogenous HTRF degradation assay

[0658] Cells (~50k) were seeded in Cisbio 96-well low volume white plates (Cisbio: cat#66PL96005). The compounds were dissolved in DMSO and serially diluted 3-fold using TECAN D300E. The cells were incubated with the compounds overnight. The total-IRAK3 HTRF kit from Cisbio was used for degradation analysis (Cisbio: 63ADK101PEH). Cryptate and D2 antibodies were diluted in detection buffer according to the manufacturer's recommendations. 2 μL of each solution was then added to 16 μL of lysate. Buffer control (lysis buffer detection buffer), Cryptate control (lysis buffer + cryptate antibody + detection buffer) and negative control (lysis buffer + cryptate antibody + D2 antibody) were prepared according to the manufacturer's recommendations. After incubation with the antibody, the HTRF signal was measured using a Perkin Elmer Envision reader and the HTRF signal was calculated using the formula: (665 nm emission / 615 nm emission) * 10,000. All HTRF values ​​were normalized to the mean of DMSO. The mean of the DMSO control wells was set equal to 100% of the relative IRAK3 protein level. The normalized luminescence values ​​were plotted as a function of compound concentration. The x-axis represents the compound concentration and the y-axis represents the corresponding normalized IRAK3-ePL protein level. The EC values ​​of the compounds used to degrade IRAK3 were calculated. 50 The values ​​(half-maximal effective concentration) were calculated using a four-parameter logistic model (sigmoidal dose-response model) (FIT = (A + {(B-A) / 1 + [(C / x)D]})) (where C is the inflection point (EC 50 ), D is the correlation coefficient, and A and B are the lower and upper limits of the fit, respectively). min Calculated by determining the lowest percentage of target protein remaining after compound treatment. max By Y min To calculate (%D max =100-Y min ). Example B3. IRAK3 biochemical binding assay

[0659] Eu kinase binding assay was performed as described by the supplier (ThermoFisher Scientific Waltham, MA). In short, 100X compound solution was prepared in DMSO by serially diluting 10mM stock solution at 3-fold intervals in a 384-well reagent plate to reach a final concentration. 1 μL of compound serial dilution was added to the corresponding wells of a 384-well reagent plate containing 32.3uL 1x buffer (50mM HEPES pH 7.4, 10nM MgCl2, 1mM EGTA, 0.01% Brij-35). 5 μL of buffer-diluted compound was transferred to the wells of the corresponding 384-well assay plate. 5 μL of 3X tracer was transferred to each well of the assay plate to a final tracer concentration of 10nM. Finally, 5 μL of 3X Eu-Anti-GST and IRAK3 mixture was transferred to each well to a final concentration of 2nM and 10nM, respectively. The reaction was incubated at room temperature for 1 hour. The TR-FRET signal of the interaction was read at room temperature using an Envision plate reader (λex 340 / λem 665 / λem 615) with a delay time of 100 μs and an integration time of 200 μs. The background-corrected emission signal ratio at each compound concentration was used to calculate the percentage of inhibition (% inhibition). The plot of % inhibition versus inhibitor concentration was fitted to the dose-response equation (Eq. 1) using Dotmatics software (Dotmatics, Bishops Stortford, Hertfordshire, England) to generate the IC 50 and Hillslope value. Using these assays, the ICs of the following compounds were determined 50 、D max 、EC 50 and DC 50 Value. D max Defined as the maximum degradation percentage achieved and DC 50 The data are summarized in Table 2. Table 42. ND = Not Determined

[0660] Although the present invention has been described in some detail by way of illustration and example for the purpose of clarity of understanding, these descriptions and examples should not be construed as limiting the scope of the invention. The disclosures of all patents and scientific literature cited herein are expressly incorporated herein by reference in their entirety.

Claims

1. A compound of formula (I) or a pharmaceutically acceptable salt thereof, in: A is a C1-C6 alkyl group, a phenyl group, a C3-C6 cycloalkyl group, a 5- to 6-membered heteroaryl group, or a 6- to 10-membered heterocyclic group, wherein the phenyl group, the cycloalkyl group, the heteroaryl group, and the heterocyclic group are substituted by x R 1 Group substituted, and wherein the heteroaryl and heterocyclic groups contain 1-3 heteroatoms selected from N and O; Each R 1 independently halo, C1-C6 alkyl, C3-C6 cycloalkyl, C1-C6 alkoxy, C1-C6 haloalkyl, or -SO2(C1-C6 alkyl); or two R on adjacent carbon atoms 1 The groups together form a fused C3-C6 cycloalkyl or a fused group; R a and R b Each is H or together form an oxo group; R c is H or C1-C6 alkyl; x is 0-5; R 2 is H or C1-C6 alkyl; R 3 is H or C1-C6 alkyl; R 4 is H or C1-C6 alkyl; X 1 is CH or N; X 2 N or CH2; Ring B is a C3-C6 cycloalkylene group or a 5- to 7-membered heterocyclylene group containing 1 or 2 nitrogen atoms; Each R 5 are independently halo, C1-C6 alkyl or C1-C6 haloalkyl; w is 0-5; L 1 -C(O)(CH2) n -、-(CH2) n -or-(CH2) n C(O)-; n is 1-6; Ring C is a 5- to 10-membered heterocyclylene group containing 1 or 2 nitrogen atoms; Each R 6 are independently halo, C1-C6 haloalkyl or C1-C6 alkyl; y is 0-5; Ring D is R 7a and R 7b Each is H or together form an oxo group; Each R 8 are independently halo, C1-C6 alkyl, C1-C6 haloalkyl or C1-C6 alkoxy; z is 0-4; X 3 N or CR 9 ; R 9 is H or C1-C6 alkyl; R 10 is H or C1-C6 alkyl; Each R 11 are independently halo, C1-C6 alkyl or C1-C6 haloalkyl; v is 0-4; and Each are independently a single bond or a double bond.

2. The compound according to claim 1 or a pharmaceutically acceptable salt thereof, wherein: A is C1-C3 alkyl, phenyl, C3-C5 cycloalkyl, 5- to 6-membered heteroaryl, or 8- to 10-membered heterocyclyl, wherein the phenyl, cycloalkyl, heteroaryl, and heterocyclyl groups are substituted by x R 1 group substitution; x is 0-3; and Each R 1 independently halo, C1-C3 alkyl, C3-C5 cycloalkyl, C1-C3 alkoxy, C1-C3 haloalkyl or -SO2(C1-C3 alkyl); or two R on adjacent carbon atoms 1 The groups together form a fused C3-C5 cycloalkyl or a fused group; R a and R b are each H or together form an oxo group; and R c is H or C1-C3 alkyl.

3. The compound according to claim 1 or 2, or a pharmaceutically acceptable salt thereof, wherein: A is -CH3 or -CH2CH3.

4. The compound according to any one of claims 1 to 3 or a pharmaceutically acceptable salt thereof, wherein: R 2 is H or C1-C3 alkyl; R 3 is H or C1-C3 alkyl; and R 4 is H or C1-C3 alkyl.

5. The compound according to any one of claims 1 to 4 or a pharmaceutically acceptable salt thereof, wherein: X 1 is N.

6. The compound according to any one of claims 1 to 5, or a pharmaceutically acceptable salt thereof, wherein: for 7. The compound according to any one of claims 1 to 6, or a pharmaceutically acceptable salt thereof, wherein: Ring B is a C4-C6 cycloalkylene group or a 6- to 7-membered heterocyclylene group containing one nitrogen atom; w is 0-2; and Each R 5 are independently halo, C1-C3 alkyl or C1-C3 haloalkyl.

8. The compound according to any one of claims 1 to 7, or a pharmaceutically acceptable salt thereof, wherein: for 9. The compound according to any one of claims 1 to 8, or a pharmaceutically acceptable salt thereof, wherein: L 1 -C(O)CH2-, -(CH2) n - or -CH2C(O)-; and n is 1-5.

10. The compound according to any one of claims 1 to 9, or a pharmaceutically acceptable salt thereof, wherein: Ring C is a 6- to 8-membered heterocyclylene group containing 1 or 2 nitrogen atoms; y is 0-3; and Each R 6 are independently halo, C1-C3 haloalkyl or C1-C3 alkyl.

11. The compound according to any one of claims 1 to 10, or a pharmaceutically acceptable salt thereof, wherein: for 12. The compound according to any one of claims 1 to 11, or a pharmaceutically acceptable salt thereof, wherein: Ring D is 13. The compound according to any one of claims 1 to 12, or a pharmaceutically acceptable salt thereof, wherein: X 3 CR 9 ; R 9 is H or C1-C3 alkyl; R 10 is H or C1-C3 alkyl; v is 0-2; and Each R 11 are independently halo, C1-C3 alkyl or C1-C3 haloalkyl.

14. The compound according to any one of claims 1 to 13, or a pharmaceutically acceptable salt thereof, wherein: for 15. The compound according to any one of claims 1 to 14, or a pharmaceutically acceptable salt thereof, wherein the compound is of formula (IIIa), (IIIb) or (IIIc):

16. The compound according to any one of claims 1 to 15, or a pharmaceutically acceptable salt thereof, wherein the compound is of formula (IVa) or (IVb):

17. A compound or a pharmaceutically acceptable salt thereof, wherein the compound is selected from the compounds in Table 1.

18. A pharmaceutical composition comprising the compound according to any one of claims 1 to 17 or a pharmaceutically acceptable salt thereof, and a pharmaceutically acceptable excipient.

19. A method for regulating interleukin-1 receptor associated kinase 3 (IRAK3), the method comprising contacting IRAK3 with an effective amount of a compound according to any one of claims 1 to 17 or a pharmaceutically acceptable salt thereof, or a pharmaceutical composition according to claim 18.

20. A method of (i) treating cancer in a subject in need thereof, comprising administering to the subject an effective amount of a compound according to any one of claims 1-17, or a pharmaceutically acceptable salt thereof, or a pharmaceutical composition according to claim 18, optionally wherein the cancer is selected from bladder cancer, breast cancer, esophageal cancer, colon cancer, head and neck cancer, kidney cancer, lung cancer, pancreatic cancer, prostate cancer, melanoma, and gastric cancer; or (ii) enhancing the immunity of a vaccinated subject, comprising administering to the subject an effective amount of a compound according to any one of claims 1-17, or a pharmaceutically acceptable salt thereof, or a pharmaceutical composition according to claim 18.

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