Substituted pyrazolyl-pyridinyl compounds as ligand-directed degradation molecules of IRAK3

Abstract

The present disclosure provides compounds and compositions thereof for modulating IRAK3. In some embodiments, the compounds and compositions are provided for the treatment of cancer.

Description

[Technical Field]

[0001] (CROSS-REFERENCE TO RELATED APPLICATIONS) This application claims priority to U.S. Provisional Application No. 63 / 391,975, filed July 25, 2022, and U.S. Provisional Application No. 63 / 467,137, filed May 17, 2023, the disclosures of each of which are incorporated herein by reference in their entirety for all purposes.

[0002] (Field) FIELD OF THE DISCLOSURE The present disclosure relates generally to compounds, compositions, and methods of making and using such compounds and compositions for treating cancer. [Background technology]

[0003] The recruitment of immune cells to the site of injury involves the coordinated interaction of multiple soluble mediators. Several cytokines, including interleukin-1 (IL-1), appear to play important roles in these processes. IL-1 promotes the inflammatory response and contributes to the tissue degeneration seen in chronic inflammatory conditions. IL-1 is also involved in the processes of bone resorption and adipose tissue regulation. Therefore, IL-1 plays an important role in numerous pathological conditions, including rheumatoid arthritis, inflammatory bowel disease, multiple sclerosis, diabetes, obesity, cancer, and sepsis.

[0004] Treatment of cells with IL-1 induces the formation of a complex consisting of two IL-1 receptor chains, IL-1R1 and IL-1RAcP, and the resulting heterodimer recruits an adaptor molecule called MyD88, which binds to 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 and a centrally located kinase domain that mediates interaction with MyD88 family adaptor proteins. Of the four members of the mammalian IRAK family, IRAK2 and IRAK3 are thought to be catalytically inactive pseudokinases (Wesche et al., J. Biol. Chem. 1999, 274, 19403-19410), but the precise roles of these two kinases remain largely unknown (Lagne et al., Structure 2021, 29, 238-251). However, a link between IRAK3 and the negative regulation of Toll-like receptor (TLR) signaling, which is involved in microbial detection and protection of multicellular organisms from infection, has been suggested (Kobayashi et al., Cell 2002, 110, 191-202). Furthermore, recent studies have shown that mutations or high expression levels of IRAK3 are associated with 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).This suggests that IRAK3 has potential as a drug target and the need for small molecules that bind to IRAK3.

[0005] Protein degradation is a highly regulated and essential process that maintains cellular homeostasis. Damaged, misfolded, or excess proteins are selectively identified and removed by the ubiquitin-proteasome pathway (UPP). The UPP plays a central role in regulating almost all cellular processes. Protein ubiquitination occurs when an E3 ubiquitin ligase binds to a protein and attaches a ubiquitin molecule to it, marking it for degradation by the proteasome.

[0006] There has been considerable interest in harnessing the UPP for therapeutic purposes (Zhou et al., Mol. Cell 2000, 6, 751-756). One promising therapeutic approach is the use of proteolytic chimeric molecules, commonly referred to as PROTACs, to remove unwanted proteins through proteolysis (Scheepstra et al., Comp. Struct. Biotech. J. 2019, 17, 160-176). PROTACs are ligand-directed degradation molecules that link an E3 ligase with a target protein for degradation. These bivalent molecules typically consist of an E3 ligase ligand connected to a small molecule that binds to the target protein via a linker. PROTACs position the E3 ligase at the appropriate distance and orientation relative to the target protein, allowing it to ubiquitinate. The ubiquitinated target protein is then recognized and degraded by the proteasome.

[0007] This provides, in some embodiments, compounds that are intended to degrade IRAK3. Summary of the Invention

[0008] In certain embodiments, the present invention provides compounds and compositions thereof for degrading IRAK3. In various embodiments, the compounds and compositions thereof may be used to treat cancer.

[0009] The present embodiments may be more fully understood by reference to the detailed description and examples, which are intended to exemplify non-limiting embodiments.

[0010] Embodiment A1. Formula (IA): [ka] [In the formula, L 1 is -(CH2CH2O) n -or in conjunction; n is 1 to 10; L 2 is a C1-C6 alkylene, -(C1-C6 alkylene)N(H)-, a 6- to 12-membered spiroheterocyclylene, or a bond, wherein the heterocyclylene contains 1 to 3 heteroatoms selected from N and O; D is [ka] and; R 1a and R 1b are each H or together form oxo; and L 3 is -CH2O- or a bond] or a pharmaceutically acceptable salt thereof.

[0011] Embodiment A2. The compound has the formula (I): [ka] [In the formula, L 1 is -(CH2CH2O) n -or in conjunction; n is 1 to 10; L 2 is a C1-C6 alkylene, -(C1-C6 alkylene)N(H)-, a 6- to 12-membered spiroheterocyclylene, or a bond, wherein the heterocyclylene contains 1 to 3 heteroatoms selected from N and O; D is [ka] and; R 1a and R 1b are each H or together form oxo; and L 3 is -CH2O- or a bond] A compound of embodiment A1, or a pharmaceutically acceptable salt thereof.

[0012] Embodiment A3. During the ceremony, L 1 -(CH2CH2O) n - and; and The compound of embodiment A1 or A2, wherein n is 1 to 7, or a pharmaceutically acceptable salt thereof.

[0013] Embodiment A4. In the formula, L 1 The compound of embodiment A1 or A2, or a pharmaceutically acceptable salt thereof, wherein is a bond.

[0014] Embodiment A5. In the formula, L 2 is C1-C3 alkylene, -(C1-C3 alkylene)N(H)-, a 10-12 membered spiroheterocyclylene, or a bond, wherein the heterocyclylene contains 1 to 2 nitrogen atoms; A compound of any one of embodiments A1 to A4, or a pharmaceutically acceptable salt thereof.

[0015] Embodiment A6. In the formula, -L 1 -L 2 -but, [ka] The compound of any one of Embodiments A1 through A5, wherein:

[0016] Embodiment A7. In the formula, D is [ka] The compound of any one of Embodiments A1 through A6, wherein:

[0017] Embodiment A8. In the formula, D is [ka] The compound of any one of Embodiments A1 through A6, wherein:

[0018] Embodiment A9. In the formula, R 1a and R 1b and R are each H, or a pharmaceutically acceptable salt thereof.

[0019] Embodiment A10. In the formula, R 1a and R 1b The compound of embodiment A8, or a pharmaceutically acceptable salt thereof, wherein: taken together form oxo.

[0020] Embodiment A11. In the formula, L 3 The compound of any one of Embodiments A8 through A10, or a pharmaceutically acceptable salt thereof, wherein is —CH 2 O—.

[0021] Embodiment A12. In the formula, L 3 The compound of any one of embodiments A8 through A10, or a pharmaceutically acceptable salt thereof, wherein is a bond.

[0022] Embodiment A13. During the ceremony, [ka] but, [ka] The compound of any one of Embodiments A8 through A12, wherein:

[0023] Embodiment A14. In the formula, D is [ka] The compound of any one of Embodiments A1 through A6, wherein:

[0024] Embodiment A15. wherein the compound has formula (II), (III), or (VII): [ka] [In the formula, L 1 is -(CH2CH2O) n -or in conjunction; n is 1 to 10; and L 2 is C1-C6 alkylene], [ka] [In the formula, L 1 is -(CH2CH2O) n -or in conjunction; n is 1 to 10; and L 2 is -(C1-C6 alkylene)-N(H)-, a 6- to 12-membered spiroheterocyclylene, or a bond, wherein the heterocyclylene contains 1 to 3 heteroatoms selected from N and O; [ka] [In the formula, L 1 is a bond; and L 2 is a 6- to 12-membered spiroheterocyclylene containing 1 to 3 heteroatoms selected from N and O. The compound of any one of embodiments A1 to A14, or a pharmaceutically acceptable salt thereof.

[0025] Embodiment A16. A compound selected from the compounds of Table 1 and pharmaceutically acceptable salts thereof.

[0026] Embodiment A17. A pharmaceutical composition comprising a compound of any one of Embodiments A1-A16, or a pharmaceutically acceptable salt thereof, and a pharmaceutically acceptable excipient.

[0027] Embodiment A18. A method for modulating interleukin-1 receptor-associated kinase 3 (IRAK3), comprising contacting IRAK3 with an effective amount of a compound of any one of embodiments A1 to A16, or a pharmaceutically acceptable salt thereof, or a pharmaceutical composition of embodiment A17.

[0028] Embodiment A19. A method of treating cancer in a patient in need of such treatment, comprising administering to the subject an effective amount of a compound of any one of embodiments A1 to A16, or a pharmaceutically acceptable salt thereof, or the pharmaceutical composition of embodiment A17, wherein the cancer is suitably 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.

[0029] Embodiment A20. A method for enhancing the immunity of a subject receiving a vaccine, comprising administering to the subject an effective amount of a compound of any one of embodiments A1 to A16, or a pharmaceutically acceptable salt thereof, or a pharmaceutical composition of embodiment A17. DETAILED DESCRIPTION OF THE INVENTION

[0030] (definition) As used herein, the terms "characterized by" and "comprising" may be used interchangeably. The terms "characterized by" and "comprising" are to be interpreted as specifying the presence of a described feature or referenced component, but do not exclude the presence or addition of one or more of that feature, component, or substituent. Furthermore, the terms "characterized by" and "comprising" are intended to include examples encompassed by the term "consisting of." Thus, the term "consisting of" may be used in place of the terms "characterized by" 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 recited features or components. In other embodiments, the term "consisting of" excludes other features or components from any succeeding recitation, except those that are not essential to the resulting technical effect.

[0032] The term "or" as used herein should be interpreted as an inclusive term meaning any one or any combination. Thus, "A, B, or C" means any of "A, B, C, A and B, A and C, B and C, A and B and C." Exceptions to this definition occur only when combinations of elements, features, steps, or acts are, for some reason, inherently mutually exclusive.

[0033] As used herein, any concentration range, percentage range, ratio range, or integer range is understood to include any integer value in the stated range, and fractions thereof, as appropriate (e.g., integer tenths and hundredths), unless otherwise indicated. Also, unless otherwise indicated, any numerical range for any physical characteristic described herein (e.g., polymer subunits, size, or thickness) is understood to include any integer within the stated range. As used herein, the terms "about" and "approximately" mean ±20%, ±10%, ±5%, or ±1% of the stated range, value, or structure, unless otherwise indicated.

[0034] An "alkyl" group is a saturated, partially saturated, or unsaturated, straight- or branched-chain acyclic hydrocarbon group having from 1 to 10 carbon atoms (C1-C 10alkyl), typically groups having 1 to 8 carbon atoms (C1-C8 alkyl), or in some embodiments, groups having 1 to 6 carbon atoms (C1-C6 alkyl), 1 to 4 carbon atoms (C1-C4 alkyl), 1 to 3 carbon atoms (C1-C3 alkyl), or 2 to 6 carbon atoms (C2-C6 alkyl). In some embodiments, the alkyl group is saturated. Representative saturated alkyl groups include -methyl, -ethyl, -n-propyl, -n-butyl, -n-pentyl, and -n-hexyl, while saturated branched alkyls include -isopropyl, -sec-butyl, -isobutyl, -tert-butyl, -isopentyl, -neopentyl, tert-pentyl, -2-methylpentyl, -3-methylpentyl, -4-methylpentyl, -2,3-dimethylbutyl, and the like. In some embodiments, the alkyl group is unsaturated, also referred to as an alkenyl or alkynyl group. An "alkenyl" group is an alkyl group containing one or more carbon-carbon double bonds. An "alkynyl" group is an alkyl group containing one or more carbon-carbon triple bonds. Examples of unsaturated alkyl groups include, but are not limited to, vinyl, allyl, -CH=CH(CH), -CH=C(CH), -C(CH)=CH, -C(CH)=CH, -C(CHCH)=CH, -C≡CH, -C≡C(CH), -C≡C(CHCH), -CHC≡CH, -CHC≡C(CH), and -CHC≡C(CHCH), among others. Alkyl groups can be substituted or unsubstituted.When an alkyl group described herein is described as "substituted," it is understood that any substituent or substituents described in the example compounds and embodiments of the present disclosure, as well as halogen; hydroxy; alkoxy; cycloalkyloxy, aryloxy, heterocyclyloxy, heteroaryloxy, heterocycloalkyloxy, cycloalkylalkyloxy, aralkyloxy, heterocyclylalkyloxy, heteroarylalkyloxy, heterocycloalkylalkyloxy; oxo (=O); amino, alkylamino, cycloalkylamino, arylamino, heterocyclylamino, heteroarylamino, heterocycloalkylamino, cycloalkylalkyl It may be substituted with amino, aralkylamino, heterocyclylalkylamino, heteroaralkylamino, heterocycloalkylalkylamino; imino; imido; amidino; guanidino; enamino; acylamino; sulfonylamino; urea, nitrourea; oxime; hydroxylamino; alkoxyamino; aralkoxyamino; hydrazino; hydrazide; hydrazono; azide; nitro; thio (-SH), alkylthio; =S; sulfinyl; sulfonyl; aminosulfonyl; phosphonate; phosphinyl; acyl; formyl; carboxy; ester; carbamate; amido; cyano; isocyanato; isothiocyanato; cyanato; thiocyanato; or -B(OH)2.In certain embodiments, when an alkyl group described herein is described as "substituted," it can be substituted with any of the substituents or substituents described in the example compounds and embodiments of the present disclosure, as well as halogen (chloro, iodo, bromo, or fluoro); alkyl; hydroxyl; alkoxy; alkoxyalkyl; amino; alkylamino; carboxy; nitro; cyano; thiol; thioether; imine; imide; amidine; guanidine; enamine; aminocarbonyl; acylamino; phosphonate; phosphine; thiocarbonyl; sulfinyl; sulfone; sulfonamide; ketone; aldehyde; ester; urea; urethane; oxime; hydroxylamine; alkoxyamine; aralkoxyamine; N-oxide; hydrazine; hydrazide; hydrazone; azide; isocyanate; isothiocyanate; cyanate; thiocyanate; B(OH)2, or O(alkyl)aminocarbonyl.

[0035] An "alkylene" group refers to a divalent alkyl residue. Particular alkylene groups are those having 1 to 10 carbon atoms (C-C 10 alkylene), typically a group having 1 to 8 carbon atoms (C1-C8 alkylene), or in some embodiments, 1 to 6 carbon atoms (C1-C6 alkylene), or 1 to 3 carbon atoms (C1-C3 alkylene). Examples of alkylene include, but are not limited to, groups such as methylene (-CH2-), ethylene (-CH2CH2-), propylene (-CH2CH2CH2-), isopropylene (-CH2CH(CH3)-), butylene (-CH2(CH2)2CH2-), isobutylene (-CH2CH(CH3)CH2-), pentylene (-CH2(CH2)3CH2-), hexylene (-CH2(CH2)4CH2-), heptylene (-CH2(CH2)5CH2-), octylene (-CH2(CH2)6CH2-), and the like.

[0036] A "cycloalkyl" group refers to a saturated or partially saturated cyclic alkyl group of 3 to 10 carbon atoms (C3-C6), including optionally substituted monocyclic or polycyclic fused or bridged rings. 10In some embodiments, a cycloalkyl group contains 3 to 8 ring carbon atoms (C3-C8 cycloalkyl), while in other embodiments, the number of ring carbon atoms ranges from 3 to 5 (C3-C5 cycloalkyl), 3 to 6 (C3-C6 cycloalkyl), or 3 to 7 (C3-C7 cycloalkyl). In some embodiments, a cycloalkyl group is a saturated cycloalkyl group. Such saturated cycloalkyl groups include, by way of example, single ring structures (e.g., cyclopropyl, cyclobutyl, cyclopentyl, cyclohexyl, cycloheptyl, cyclooctyl, 1-methylcyclopropyl, 2-methylcyclopentyl, 2-methylcyclooctyl, etc.) or multiple ring or bridged ring structures (e.g., 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, a cycloalkyl group is an unsaturated cycloalkyl group. Examples of unsaturated cycloalkyl groups include, among others, cyclohexenyl, cyclopentenyl, cyclohexadienyl, butadienyl, pentadienyl, and hexadienyl. The cycloalkyl group may be substituted or unsubstituted. Examples of such substituted cycloalkyl groups include cyclohexanol and the like.

[0037] "Heterocyclyl" refers to a non-aromatic cycloalkyl in which 1 to 4 ring carbon atoms are independently replaced with heteroatoms selected from O, S, and N. In some embodiments, heterocyclyl groups contain 3 to 10 ring atoms, while other such groups contain 3 to 5, 3 to 6, or 3 to 8 ring atoms. A heterocyclyl may be attached to another group at any ring atom (i.e., any carbon atom or heteroatom of the heterocycle). A heterocyclyl group may be substituted or unsubstituted. Heterocyclyl groups include saturated and partially saturated ring systems. Furthermore, the term heterocyclyl includes any non-aromatic ring containing at least one heteroatom, which ring may be fused to an aryl or heteroaryl ring, whether or not attached to the rest of the molecule. The term also includes bridged polycyclic ring systems containing heteroatoms. Representative examples of heterocyclyl groups include, but are not limited to, aziridinyl, azetidinyl, azepanyl, pyrrolidyl, imidazolidinyl (e.g., imidazolidin-4-onyl or imidazolidin-2,4-dionyl), pyrazolidinyl, thiazolidinyl, tetrahydrothiophenyl, tetrahydrofuranyl, piperidyl, piperazinyl (e.g., piperazin-2-onyl), morpholinyl, thiomorpholinyl, tetrahydropyranyl (e.g., tetrahydro-2H-pyranyl), tetrahydrothiopyranyl, oxathianyl, dithianyl, 1,4-dioxaspiro[4.5]decanyl, homopiperazinyl, quinuclidinyl, or tetrahydropyrimidin-2(1H)-one. Representative substituted heterocyclyl groups may be mono- or di- or more substituted, for example, but not limited to, pyridyl or morpholinyl groups may be di-, tri-, tetra-, penta-, or hexa-substituted, or di-substituted, for example, with various substituents described below.

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

[0039] An "aryl" group is an alkyl group having 6 to 14 carbon atoms (C6-C8) containing a single ring (e.g., phenyl) or multiple condensed rings (e.g., naphthyl or anthryl). 14In some embodiments, an aryl group has 6 to 14 carbon atoms (C6-C8) in the ring portion. 14 aryl), and in other embodiments, aryls having 6 to 12 carbon atoms (C-C 12 aryl) or 6 to 10 carbon atoms (C6-C 10 Specific aryl groups include phenyl, biphenyl, naphthyl, and the like. Aryl groups can be substituted or unsubstituted. The term "aryl group" also includes groups having fused rings (e.g., fused aromatic-aliphatic ring systems such as indanyl, tetrahydronaphthyl, and the like).

[0040] A "heteroaryl" group is an aromatic ring system having 1 to 4 heteroatoms as ring atoms, with the remaining atoms being carbon atoms. In some embodiments, heteroaryl groups contain 3 to 6 ring atoms in the ring portion, and in other embodiments, 6 to 9, or even 6 to 10 atoms. Suitable heteroatoms include oxygen, sulfur, and nitrogen. In certain embodiments, the heteroaryl ring system is monocyclic or bicyclic. Examples include, but are not limited to, pyrrolyl, pyrazolyl, imidazolyl, triazolyl, tetrazolyl, oxazolyl, isoxazolyl, benzisoxazolyl (e.g., benzo[d]isoxazolyl), thiazolyl, pyrrolyl, pyridazinyl, pyrimidyl, pyrazinyl, thiophenyl, benzothiophenyl, furanyl, benzofuranyl, indolyl (e.g., indolyl-2-onyl or isoindolin-1-onyl), azaindolyl (pyrrolopyridyl or 1H-pyrrolo[2,3-b]pyridyl), indazolyl, benzimidazolyl (e.g., 1H-benzo[d]imidazolyl), imidazolyl, Examples of heteroaryl groups include pyridyl (e.g., azabenzimidazolyl or 1H-imidazo[4,5-b]pyridyl), pyrazolopyridyl, triazolopyridyl, benzotriazolyl (e.g., 1H-benzo[d][1,2,3]triazolyl), benzoxazolyl (e.g., benzo[d]oxazolyl), benzothiazolyl, benzothiadiazolyl, isoxazolopyridyl, thianaphthalenyl, purinyl, xanthinyl, adenyl, guanyl, quinolinyl, isoquinolinyl (e.g., 3,4-dihydroisoquinolin-1(2H)-onyl), tetrahydroquinolinyl, quinoxalinyl, and quinazolinyl. Heteroaryl groups may be substituted or unsubstituted.

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

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

[0043] "Haloalkyl" refers to an alkyl, as defined above, substituted with one or more halogens, as defined above (e.g., trifluoromethyl, difluoromethyl, trichloromethyl, 2,2,2-trifluoroethyl, 1,2-difluoroethyl, 3-bromo-2-fluoropropyl, 1,2-dibromoethyl, etc.). In some embodiments, the haloalkyl group contains 1 to 6 carbon atoms and is substituted with one or more halogens (C-C haloalkyl), or the haloalkyl group has 1 to 3 carbon atoms and is substituted with one or more halogens (C-C haloalkyl). The halogens may all be the same or different. Unless otherwise specified, haloalkyl groups may be optionally substituted.

[0044] When a group other than an alkyl group as described herein is "substituted," the group may be substituted with one or more suitable optional substituents. In the exemplary compounds and embodiments of the present disclosure, exemplary substituents include, in addition to halogen (chloro, iodo, bromo, or fluoro), alkyl, hydroxyl, alkoxy, alkoxyalkyl, amino, alkylamino, carboxy, nitro, cyano, thiol, thioether, imine, imide, amidine, guanidine, enamine, aminocarbonyl, acylamino, phosphonate, phosphine, thiocarbonyl, sulfinyl, sulfone, sulfonamide, ketone, aldehyde, ester, urea, urethane, oxime, hydroxylamine, alkoxyamine, aralkoxyamine, N-oxide, hydrazine, hydrazide, hydrazone, azide, isocyanate, isothiocyanate, cyanate, thiocyanate, oxygen (=O), B(OH), O(alkyl)aminocarbonyl, monocyclic or fused polycyclic or non-fused. cycloalkyl, which may be polycyclic (e.g., cyclopropyl, cyclobutyl, cyclopentyl, or cyclohexyl), or heterocyclyl, which may be monocyclic, polyfused rings, or polynon-fused rings (e.g., pyrrolidyl, piperidyl, piperazinyl, morpholinyl, or thiazinyl); aryl or heteroaryl, which may be monocyclic, polyfused rings, or polynon-fused rings (e.g., phenyl, naphthyl, pyrrolyl, indolyl, furanyl, thiophenyl, imidazolyl, oxazolyl, isoxazolyl, thiazolyl, triazolyl, tetrazolyl, pyrazolyl, pyridyl, quinolinyl, isoquinolinyl, acridinyl, pyrazinyl, pyridazinyl, pyrimidyl, benzimidazolyl, benzothiophenyl, or benzofuranyl), aryloxy; aralkyloxy; heterocyclyloxy; and heterocyclylalkoxy.

[0045] Embodiments of the present disclosure are intended to include pharmaceutically acceptable salts, tautomers, isotopomers, and stereoisomers of the compounds disclosed herein (e.g., compounds of Formula (IA) or Formula (I)).

[0046] As used herein, the term "pharmaceutically acceptable salts" refers to salts prepared from pharmaceutically acceptable non-toxic acids or bases, including inorganic acids and bases and organic acids and bases. Suitable pharmaceutically acceptable base addition salts of compounds of Formula (IA) or Formula (I) include, but are not limited to, metallic salts formed from aluminum, calcium, lithium, magnesium, potassium, sodium, and zinc, or organic salts formed from lysine, N,N'-dibenzylethylenediamine, chloroprocaine, choline, diethanolamine, ethylenediamine, meglumine (N-methyl-glucamine), and procaine. Suitable non-toxic acids include, but are not limited to, inorganic and organic acids (e.g., acetic acid, alginic acid, anthranilic acid, benzenesulfonic acid, benzoic acid, camphorsulfonic acid, citric acid, ethenesulfonic acid, formic acid, fumaric acid, furoic acid, galacturonic acid, gluconic acid, glucuronic acid, glutamic acid, glycolic acid, hydrobromic acid, hydrochloric acid, isethionic acid, lactic acid, maleic acid, malic acid, mandelic acid, methanesulfonic acid, mucic acid, nitric acid, pamoic acid, pantothenic acid, phenylacetic acid, phosphoric acid, propionic acid, salicylic acid, stearic acid, succinic acid, sulfanilic acid, sulfuric acid, tartaric acid, and p-toluenesulfonic acid). Specific non-toxic acids include hydrochloric acid, hydrobromic acid, maleic acid, phosphoric acid, sulfuric acid, and methanesulfonic acid. Examples of specific salts include hydrochloride, formate, and mesylate salts. Others will be known to those skilled in the art. See, e.g., Remington's Pharmaceutical Sciences, 18th eds., Mack Publishing, Easton PA (1990) or Remington: The Science and Practice of Pharmacy, 19th eds., Mack Publishing, Easton PA (1995).

[0047] Unless otherwise specified, the terms "stereoisomer" or "stereomerically pure," as used herein, mean that one stereoisomer of a particular compound is substantially free of other stereoisomers of that compound. For example, a stereomerically pure compound having one chiral center is substantially free of the other enantiomer of that compound. A stereomerically pure compound having two chiral centers is substantially free of other diastereomers of that compound. Stereomerically pure compounds typically include those that contain greater than about 80% by weight of one stereoisomer of the compound and less than about 20% by weight of the other stereoisomer of the compound, greater than about 90% by weight of one stereoisomer of the compound and less than about 10% by weight of the other stereoisomer of the compound, greater than about 95% by weight of one stereoisomer of the compound and less than about 5% by weight of the other stereoisomer of the compound, or greater than about 97% by weight of one stereoisomer of the compound and less than about 3% by weight of the other stereoisomer of the compound. The compounds of the present disclosure may contain chiral centers and may occur as racemates, individual enantiomers or diastereomers, and mixtures thereof, and all such isomers, including mixtures thereof, are included within the embodiments of the present disclosure.

[0048] The use of pure stereoisomers of the disclosed compounds, as well as mixtures of these isomers, are included in the embodiments of the present disclosure.For example, mixtures containing equal or unequal amounts of the enantiomers of a particular compound can be used in the methods and compositions of the present disclosure.These isomers can be synthesized asymmetrically or resolved by conventional methods (e.g., chiral columns or chiral resolving agents). For example, Jacques, J., et al., Enantiomers, Racemates and Resolutions (Wiley Interscience, New York, 1981); Wilen, SH, et al., Tetrahedron 33:2725 (1977); Eliel, EL, Stereochemistry of Carbon Compounds (McGraw Hill, NY, 1962); Wilen, SH, Tables of Resolving Agents and Optical Resolutions p. 268 (EL Eliel, Ed., Univ. of Notre Dame Press, Notre Dame, IN, 1972); Todd, M., Separation Of Enantiomers : Synthetic Methods (Wiley-VCH Verlag GmbH & Co. KGaA, Weinheim, Germany, 2014); Toda, F., Enantiomer Separation: Fundamentals and Practical Methods (Springer Science & Business Media, 2007); Subramanian, G. See Chiral Separation Techniques: A Practical Approach (John Wiley & Sons, 2008); Ahuja, S., Chiral Separation Methods for Pharmaceutical and Biotechnological Products (John Wiley & Sons, 2011).

[0049] It should be noted that the compounds of the present disclosure can include E and Z isomers or mixtures thereof, and cis and trans isomers or mixtures thereof.In certain embodiments, the compounds are isolated as either E or Z isomers.In other embodiments, the compounds are a mixture of E and Z isomers.

[0050] "Tautomers" refer to isomers of a compound that are in equilibrium with each other. The concentration of isomers may vary depending on the environment in which the compound is present, for example, whether the compound is a solid, in an organic solvent, or in an aqueous solution. For example, in an aqueous solution, pyrazole may exist in the following isomers: [ka] These are considered to be tautomers of each other.

[0051] As one of ordinary skill in the art would readily appreciate, various functional groups and other structures may exhibit tautomerism, and all tautomers of the compounds of Formula (IA) or Formula (I) are within the scope of the present disclosure.

[0052] It should be noted that compounds of the present disclosure may contain unnatural proportions of isotopes at one or more atoms. For example, compounds may contain radioactive isotopes (e.g., tritium ( 3 H), iodine-125( 125 I), sulfur-35( 35 S), or carbon-14 ( 14 C)) or may be labeled with, for example, deuterium ( 2 H), carbon-13( 13 C), or nitrogen-15( 15The compound may be isotopically enriched in an isotopic composition of at least one atom (N). As used herein, an "isotopically enriched" species refers to an isotopically enriched compound. The term "isotopically enriched" refers to an atom having an isotopic composition other than the naturally occurring isotopic composition of that atom. "Isotopically enriched" may refer to a compound containing at least one atom having an isotopic composition other than the naturally occurring isotopic composition of that atom. The term "isotopic composition" refers to the amount of each isotope present in an atom. Radiolabeled, isotopically enriched compounds are useful as therapeutic agents (e.g., cancer therapeutic agents), research reagents (e.g., binding assay reagents), and diagnostic agents (e.g., in vivo imaging agents). All isotopes of the compounds described herein, whether radioactive or not, are intended to be included within the scope of the embodiments provided herein. In some embodiments, isotopically enriched species of the disclosed compounds (e.g., compounds enriched in deuterium, carbon-13, and / or nitrogen-15) are provided. As used herein, "deuterated" refers to a compound in which at least one hydrogen (H) is replaced by an isotopic composition (D or 2 H) means a compound that is deuterium-substituted, i.e., enriched with deuterium at at least one position of the compound.

[0053] It is understood that each compound of the present disclosure can be provided in the form of any pharmaceutically acceptable salt disclosed herein, regardless of stereoisomeric or isotopic composition.Similarly, it is also understood that isotopic composition can vary regardless of the stereoisomeric composition of each compound mentioned herein.Furthermore, isotopic composition is limited to the elements present in each compound or its salt disclosed herein, but can also vary regardless of the choice of pharmaceutically acceptable salt of each compound.

[0054] It should be noted that if there is a discrepancy between the described structure and the name of the structure, the described structure shall prevail.

[0055] As used herein, "treatment" means alleviating, in whole or in part, a disorder, disease, or condition, or one or more symptoms associated with the disorder, disease, or condition, or slowing or halting further progression or worsening of those symptoms, or reducing or eradicating the cause of the disorder, disease, or condition itself. In certain embodiments, the disorder is a neurodegenerative disease described herein, or a symptom thereof.

[0056] As used herein, "prevention" refers to a method of delaying and / or preventing the onset, recurrence, or spread of a disorder, disease, or condition, in whole or in part; a method of inhibiting a subject from acquiring a disorder, disease, or condition; or a method of reducing a subject's risk of acquiring a disorder, disease, or condition. In certain embodiments, the disorder is a neurodegenerative disease, or symptom thereof, as described herein.

[0057] The term "effective amount" in reference to a compound of the present disclosure means an amount capable of treating or preventing a disorder, disease or condition of the present disclosure, or a symptom thereof.

[0058] As used herein, the term "subject" or "patient" includes, but is not limited to, animals such as cows, monkeys, horses, sheep, pigs, chickens, turkeys, quail, cats, dogs, mice, rats, rabbits, or guinea pigs, in certain embodiments, mammals, and in other embodiments, humans. In certain embodiments, the subject is a human suffering from, at risk for, or suffering from an IRAK3-mediated disease.

[0059] Although various features of the invention may be described in the context of a single embodiment, the features may also be provided separately or in any suitable combination. Conversely, the invention may also be practiced in a single embodiment that may for clarity be described separately herein.

[0060] compound In some embodiments, the present application provides a compound of formula (IA): [ka] [In the formula, L 1 is -(CH2CH2O) n -or in conjunction; n is 1 to 10; L 2 is a C1-C6 alkylene, -(C1-C6 alkylene)N(H)-, a 6- to 12-membered spiroheterocyclylene, or a bond, wherein the heterocyclylene contains 1 to 3 heteroatoms selected from N and O; D is [ka] and; R 1a and R 1b are each H or together form oxo; and L 3 is -CH2O- or a bond] or a pharmaceutically acceptable salt thereof.

[0061] In a further aspect, the present application provides a compound of formula (I): [ka] [In the formula, L 1 is -(CH2CH2O) n -or in conjunction; n is 1 to 10; L 2 is a C1-C6 alkylene, -(C1-C6 alkylene)N(H)-, a 6- to 12-membered spiroheterocyclylene, or a bond, wherein the heterocyclylene contains 1 to 3 heteroatoms selected from N and O; D is [ka] and; R 1a and R 1b are each H or together form oxo; and L 3 is -CH2O- or a bond] or a pharmaceutically acceptable salt thereof.

[0062] In some embodiments, L 1 is -(CH2CH2O) n - or a bond. 1 is -(CH2CH2O) n In some embodiments, L 1 is a bond.

[0063] In some embodiments, n is 1 to 10. In some embodiments, n is 1 to 9. In some embodiments, n is 1 to 8. In some embodiments, n is 1 to 7. In some embodiments, n is 1 to 6. In some embodiments, n is 1 to 5. In some embodiments, n is 1 to 4. In some embodiments, n is 1 to 3. In some embodiments, n is 1 to 2. In some embodiments, n is 1. In some embodiments, n is 2. In some embodiments, n is 3. In some embodiments, n is 4. In some embodiments, n is 5. In some embodiments, n is 6. In some embodiments, n is 7. In some embodiments, n is 8. In some embodiments, n is 9. In some embodiments, n is 10.

[0064] In some embodiments, L 2 is a C1-C6 alkylene, -(C1-C6 alkylene)N(H)-, a 6- to 12-membered spiroheterocyclylene, or a bond, wherein the heterocyclylene contains 1 to 3 heteroatoms selected from N and O. 2 is a C1-C3 alkylene, -(C1-C3 alkylene)N(H)-, a 10- to 12-membered spiroheterocyclylene, or a bond, wherein the heterocyclylene contains 1 to 2 nitrogen atoms.

[0065] In some embodiments, L 2 is C1-C6 alkylene. In some embodiments, L 2 is C1-C3 alkylene. In some embodiments, L 2 is -CH2-, -CH2CH2-, or -CH2CH2CH2-. In some embodiments, L 2 is -CH2-. In some embodiments, L 2 is -CH2CH2-. In some embodiments, L 2 is -CH2CH2CH2-.

[0066] In some embodiments, L 2 is -(C1-C6 alkylene)N(H)-. In some embodiments, L 2 is -(C1-C3 alkylene)N(H)-. In some embodiments, L 2 is —CHN(H)—, —CHCHN(H), or —CHCHCHN(H)—. In some embodiments, L 2 is —CHN(H)—. In some embodiments, L 2 is —CHCHN(H)—. In some embodiments, L 2 is -CH2CH2CH2N(H)-.

[0067] In some embodiments, L 2 is a 6-12 membered spiroheterocyclylene, wherein the heterocyclylene contains 1-3 heteroatoms selected from N and O. In some embodiments, L 2 is a 6-12 membered spiroheterocyclylene, wherein the heterocyclylene contains 1-2 heteroatoms selected from N and O. In some embodiments, L 2 is a 10-12 membered spiroheterocyclylene, wherein the heterocyclylene contains 1-2 nitrogen atoms. 2 is a 10-12 membered spiroheterocyclylene, wherein the heterocyclylene contains one nitrogen atom.2 is a 10-12 membered spiroheterocyclylene, wherein the heterocyclylene contains two nitrogen atoms. 2 is a 10-membered spiroheterocyclylene, wherein the heterocyclylene contains one nitrogen atom. 2 is an 11-membered spiroheterocyclylene, wherein the heterocyclylene contains one nitrogen atom.

[0068] In some embodiments, L 2 is a bond.

[0069] In some embodiments, L 2 -CH2-, -CH2CH2CH2-, -CH2CH2N(H)-, [ka] , or a bond.

[0070] In some embodiments, -L 1 -L 2 -teeth, [ka] is.

[0071] In some embodiments, -L 1 -L 2 -teeth [ka] is.

[0072] In some embodiments, D is [ka] In some embodiments, D is [ka] In some embodiments, D is [ka] In some embodiments, D is [ka] In some embodiments, D is [ka] is.

[0073] In some embodiments, R 1a and R 1b are each H or together form oxo. In some embodiments, R 1a and R 1b are each H. In some embodiments, R 1a and R 1b come together to form oxo.

[0074] In some embodiments, D is [ka] In some embodiments, D is [ka] is.

[0075] In some embodiments, D is [ka] is.

[0076] In some embodiments, L 3 is —CH2O— or a bond. 3 is —CH2O—. In some embodiments, L 3 is a bond.

[0077] In some embodiments, [ka] teeth, [ka] is.

[0078] In some embodiments, the compound of Formula (IA) or Formula (I) has the formula (Ia): [ka] (In the formula, L 1 , L 2 and D is as described in formula (IA) or formula (I). is a compound of

[0079] In some embodiments, the compound of Formula (IA) or Formula (I) has Formula (II): [ka] [In the formula, L 1 Ha-(CH2CH2O) n - or a bond; n is 1 to 10; and L 2 is C1-C6 alkylene] is a compound of

[0080] In some embodiments, the compound of Formula (IA) or Formula (I) has the formula (IIa): [ka] [In the formula, L 1 Ha-(CH2CH2O) n - or a bond; n is 1 to 10; and L 2 is a C1-C6 alkylene.

[0081] In some embodiments, the compound of Formula (IA) or Formula (I) has Formula (III): [ka] [In the formula, L 1 Ha-(CH2CH2O) n - or a bond; n is 1 to 10; and L 2 is -(C1-C6 alkylene)-N(H)-, a 6- to 12-membered spiroheterocyclylene, or a bond, wherein the heterocyclylene contains 1 to 3 heteroatoms selected from N and O. is a compound of

[0082] In some embodiments, the compound of Formula (IA) or Formula (I) has the formula (IIIa): [ka] [In the formula, L 1 Ha-(CH2CH2O) n - or a bond; n is 1 to 10; and L 2 is -(C1-C6 alkylene)-N(H)-, a 6- to 12-membered spiroheterocyclylene, or a bond, wherein the heterocyclylene contains 1 to 3 heteroatoms selected from N and O. is a compound of

[0083] In some embodiments, the compound of Formula (IA) or Formula (I) has Formula (IV) or (IVa): [ka] wherein n is as defined in Formula (IA) or Formula (I). In some embodiments, n is 2-7.

[0084] In some embodiments, the compound of Formula (IA) or Formula (I) has Formula (V), (Va), (Vb), or (Vc): [ka] [ka] wherein x is 0 to 10; and R 1a and R1b is as described in Formula (IA) or Formula (I). In some embodiments, x is n as described in Formula (IA) or Formula (I). In some embodiments, x is 1 to 10. In some embodiments, x is 1 to 4. In some embodiments, x is 0.

[0085] In some embodiments, the compound of Formula (IA) or Formula (I) has Formula (VI) or (VIa): [ka] (In the formula, R 1a and R 1b is a compound of formula (IA) or formula (I).

[0086] In some embodiments, the compound of Formula (IA) or Formula (I) has Formula (VII) or (VIIa): [ka] (In the formula, L 1 is a bond; and L 2 is a 6-12 membered spiroheterocyclylene containing 1-3 heteroatoms selected from N and O. In some embodiments, L 2 is an 11-membered spiroheterocyclylene containing 1 to 2 nitrogen atoms.

[0087] It is understood herein that any description, variation, embodiment, or aspect of each part may be combined with each description, variation, embodiment, or aspect of any other part, as if all combinations of descriptions were specifically and individually recited. For example, L of Formula (IA) or Formula (I) 1 Each description, variation, embodiment, or aspect herein relating to L 2 , L 3 , D, R 1a , R 1b, and n may be combined with each description, variation, embodiment, or aspect thereof. It is also understood that all descriptions, variations, embodiments, or aspects of Formula (IA) or Formula (I), if applicable, apply equally to other formulas detailed herein, and that all formulas would be equally described as if each description, variation, embodiment, or aspect were individually listed. For example, all descriptions, variations, embodiments, or aspects of Formula (IA) or Formula (I), if applicable, apply equally to any formula detailed herein (e.g., Formulas (Ia), (II), (IIa), (III), (IIIa), (IV), (IVa), (V), (Va), (Vb), (Vc), (VI), (VIa), (VII), and (VIIa)), and all formulas would be equally described as if each description, variation, embodiment, or aspect were individually listed.

[0088] In some embodiments, provided is a compound selected from the compounds in Table 1, or a pharmaceutically acceptable salt thereof. Any compound described in this disclosure, including Table 1, may be depicted in a particular stereoisomeric form and / or non-stereochemically, but it is understood that all stereochemical configurations, including any enantiomers or diastereomers, and any tautomers or other configurations, of any compound of this disclosure, including Table 1, are described herein. [Table 1] [Table 2] [Table 3] [Table 4] or a pharmaceutically acceptable salt thereof.

[0089] It is understood that in this disclosure, combinations of substituents and / or variables of the depicted formulae are permissible only if such combinations result in stable compounds.

[0090] Additionally, all compounds of Formula (IA) or Formula (I) that exist in a free base or free acid form can be converted to a pharmaceutically acceptable salt by treatment with an appropriate inorganic or organic base or acid in a manner known to those skilled in the art. Salts of compounds of Formula (IA) or Formula (I) can be converted to the free base or free acid by standard techniques.

[0091] Synthesis method The compounds described herein can be synthesized using conventional organic synthesis and commercially available starting materials, or the methods described herein. Compounds of formula (IA) or formula (I) can be prepared as outlined in scheme 1 and the following examples herein, but are by way of example only and not by way of limitation. It should be noted that those skilled in the art will understand how to modify the procedures shown in the illustrated schemes and examples to obtain the desired product.

[0092] [ka] Scheme 1 shows the synthetic procedure for exemplary compound C. Intermediates A and B can be coupled using various amide coupling agents. For example, simply treating B with HATU and DIPEA in DMF, followed by addition of A, gives exemplary compound C.

[0093] How to use An embodiment of the present disclosure provides a method for modulating IRAK3 in a patient, comprising administering to the subject an effective amount of a compound of Formula (IA) or Formula (I). Modulation (e.g., inhibition or activation) of IRAK3 can be assessed and demonstrated by various methods known in the art. Kits and commercially available assays may be used to determine whether and to what extent IRAK3 is modulated (e.g., inhibited or activated).

[0094] In some embodiments, the present application provides a method for modulating IRAK3, comprising contacting IRAK3 with an effective amount of a compound of Formula (IA) or Formula (I), or any embodiment or variation thereof. In some embodiments, the compound of Formula (IA) or Formula (I) inhibits IRAK3. In other embodiments, the compound of Formula (IA) or Formula (I) activates IRAK3. In some embodiments, the compound of Formula (IA) or Formula (I) causes degradation of IRAK3.

[0095] In some embodiments, the present application provides a method for targeting degradation of IRAK3, comprising contacting IRAK3 with an effective amount of a compound of Formula (IA) or Formula (I), or any embodiment or variation thereof.

[0096] In some embodiments, a compound of Formula (IA) or 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, a compound of Formula (IA) or Formula (I) may be administered to a subject in need thereof, which compound is capable of inhibiting IRAK3 activity by about 1 to 100%, 5 to 100%, 10 to 100%, 15 to 100%, 20 to 100%, 25 to 100%, 30 to 100%, 35 to 100%, 40 to 100%, 45 to 100%, 50 to 100%, 55 to 100%, 60 to 100%, 65 to 100%, 70 to 100%, 75 to 100%, 80 to 100%, 90 to 100%, 100 to 100%, 110 to 100%, 120 to 100%, 130 to 100%, 140 to 100%, 150 to 100%, 160 to 100%, 170 to 100%, 180 to 100%, 190 to 100%, 210 to 210%, 220 to 220%, 230 to 230%, 240 to 240%, 250 to 250%, 260 to 260%, 270 to 270%, 280 to 280%, 290 to 290%, 300 to 300%, 310 to 310%, 320 to 320%, 330 to 330%, 340 to 340%, 350 to 350%, 360 to 360%, 370 to 370%, 380 to 380%, 390 to 390%, 400 to 400%, 410 to 410%, Adjust from 00%, 85-100%, 90-100%, 95-100%, 5-95%, 5-90%, 5-85%, 5-80%, 5-75%, 5-70%, 5-65%, 5-60%, 5-55%, 5-50%, 5-45%, 5-40%, 5-35%, 5-30%, 5-25%, 5-20%, 5-15%, 5-10%, 10-90%, 20-80%, 30-70%, or 40-60%.

[0097] In addition, in certain embodiments of the present disclosure, there is also provided a method for degrading IRAK3 in a patient, characterized by administering an effective amount of a compound of formula (IA) or formula (I) to the subject.The degradation of IRAK3 can be evaluated and demonstrated by various methods known in the art.To determine whether and to what extent IRAK3 is degraded, kits and commercially available assays (such as cell-based assays) can be used.

[0098] In some embodiments, the present application provides a method of degrading IRAK3, comprising contacting IRAK3 with an effective amount of a compound of Formula (IA) or Formula (I), or any embodiment or variation thereof. In some embodiments, the compound of Formula (IA) or Formula (I) partially degrades IRAK3. In some embodiments, the compound of Formula (IA) or Formula (I) completely degrades IRAK3.

[0099] In some embodiments, a compound of Formula (IA) or 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, a compound of Formula (IA) or Formula (I) inhibits IRAK3 by about 1 to 100%, 5 to 100%, 10 to 100%, 15 to 100%, 20 to 100%, 25 to 100%, 30 to 100%, 35 to 100%, 40 to 100%, 45 to 100%, 50 to 100%, 55 to 100%, 60 to 100%, 65 to 100%, 70 to 100%, 75 to 100%, 80 to 100%, 90 to 100%, 90 to 100%, 100 to 100%, 110 to 100%, 120 to 100%, 130 to 100%, 140 to 100%, 150 to 100%, 160 to 100%, 170 to 100%, 180 to 100%, 190 to 100%, 210 to 210%, 220 to 220%, 230 to 230%, 240 to 240%, 250 to 250%, 260 to 260%, 270 to 270%, 280 to 280%, 290 to 300%, 310 to 310%, 320 to 320%, 330 to 330%, 340 to 340%, 350 to 350%, 360 to 360%, 370 to 370%, 380 to 380%, 390 to 400%, 410 to 410%, 420 to 420%, 430 to 430%, 440 to 44 %, 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% decomposition.

[0100] In another aspect, the present application provides a method of treating cancer in a patient in need thereof, comprising administering to the subject an effective amount of a compound of Formula (IA) or Formula (I). In some embodiments, the present application provides a method of inhibiting cancer in a patient, comprising administering to the subject an effective amount of a compound of Formula (IA) or Formula (I). Examples of cancers include, but are not limited to, bladder cancer, breast cancer, esophageal cancer, colon cancer, head and neck cancer, kidney cancer, lung cancer, pancreatic cancer, prostate cancer, melanoma, and gastric cancer.

[0101] In some embodiments, administering a compound of Formula (IA) or Formula (I) to a subject susceptible to cancer prevents the subject from developing any symptoms of cancer (e.g., tumor growth or metastasis). In some embodiments, administering a compound of Formula (IA) or Formula (I) to a subject who does not yet exhibit symptoms of cancer prevents the subject from developing any symptoms of cancer. In some embodiments, administering a compound of Formula (IA) or Formula (I) to a patient in need thereof alleviates the severity of the cancer in the subject. In some embodiments, administering a compound of Formula (IA) or Formula (I) to a patient in need thereof stabilizes the cancer (prevents or slows the progression of the cancer). In some embodiments, administering a compound of Formula (IA) or Formula (I) to a patient in need thereof delays the onset or recurrence of cancer. In some embodiments, administering a compound of Formula (IA) or Formula (I) to a patient in need thereof slows the progression of the cancer. In some embodiments, administering a compound of Formula (IA) or Formula (I) to a patient in need thereof results in partial remission of the cancer. In some embodiments, administering a compound of Formula (IA) or Formula (I) to a patient in need thereof results in overall remission of the cancer. In some embodiments, administering a compound of Formula (IA) or Formula (I) to a patient in need thereof reduces the dosage of one or more other drugs required to treat the cancer. In some embodiments, administering a compound of Formula (IA) or Formula (I) to a patient in need thereof enhances the effectiveness of another drug used to treat the cancer. In some embodiments, administering a compound of Formula (IA) or Formula (I) to a patient in need thereof slows the progression of the cancer. In some embodiments, administering a compound of Formula (IA) or Formula (I) to a patient in need thereof improves the quality of life of a subject suffering from cancer. In some embodiments, administering a compound of Formula (IA) or Formula (I) to a patient in need thereof prolongs the survival of a subject suffering from cancer.

[0102] In certain aspects, provided herein is a method for preventing a subject susceptible to cancer from developing cancer, comprising administering to the subject a compound of Formula (IA) or Formula (I).

[0103] In some embodiments, the present application provides a method of alleviating the severity of cancer in a subject, comprising administering to the subject a compound of Formula (IA) or Formula (I). In some embodiments, the present application provides a method of stabilizing cancer in a subject, comprising administering to the subject a compound of Formula (IA) or Formula (I). In some embodiments, the method prevents the cancer from worsening.

[0104] In another aspect, the present application provides a method for delaying the onset or recurrence of cancer in a subject, comprising administering to the subject a compound of Formula (IA) or Formula (I).

[0105] In some embodiments, the present application provides a method of delaying the progression of cancer in a subject, comprising administering to the subject a compound of Formula (IA) or Formula (I). In some embodiments, the method results in partial remission of the cancer. In some embodiments, the method results in complete remission of the cancer.

[0106] In a further aspect, the present application provides a method for reducing the dosage of one or more other drugs required to treat cancer in a subject, the method comprising administering to the subject a compound of Formula (IA) or Formula (I). In some embodiments, the present application provides a method for enhancing the effect of another drug used to treat cancer in a subject, the method comprising administering to the subject a compound of Formula (IA) or Formula (I).

[0107] Also provided herein are methods for delaying the progression of cancer in a subject, comprising administering to the subject a compound of Formula (IA) or Formula (I). In some embodiments, the methods improve the quality of life of a subject suffering from cancer. In some embodiments, the methods extend the survival of a subject suffering from cancer.

[0108] In some embodiments, the compounds of Formula (IA) or Formula (I) are useful for 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.

[0109] In some embodiments, the present application provides a method for enhancing immunity in a subject receiving a vaccine, comprising administering an effective amount of a compound of Formula (IA) or Formula (I) to the subject. In some embodiments, the compound of Formula (IA) or Formula (I) is administered to the subject before administration of the vaccine. In some embodiments, the compound of Formula (IA) or Formula (I) is administered to the subject simultaneously with administration of the vaccine. In some embodiments, the compound of Formula (IA) or Formula (I) is administered to the subject after administration of the vaccine. In some embodiments, the compound of Formula (IA) or Formula (I) is formulated as a component of a vaccine. In some embodiments, the compound of Formula (IA) or Formula (I) is formulated separately from the vaccine.

[0110] Pharmaceutical Compositions and Routes of Administration The compounds may be administered orally, topically, or parenterally to a subject in conventional formulations (e.g., capsules, microcapsules, tablets, granules, powders, troches, pills, suppositories, injections, suspensions, syrups, patches, creams, lotions, ointments, gels, sprays, solutions, and emulsions).

[0111] The compounds of the present disclosure may be administered orally, topically, or parenterally to a subject in conventional formulations (e.g., capsules, microcapsules, tablets, granules, powders, troches, pills, suppositories, injections, suspensions, syrups, patches, creams, lotions, ointments, gels, sprays, solutions, and emulsions). Suitable formulations may be prepared by common methods using conventional organic or inorganic additives. Such additives include, for example, excipients (e.g., sucrose, starch, mannitol, sorbitol, lactose, glucose, cellulose, talc, calcium phosphate, or calcium carbonate), binders (e.g., cellulose, methylcellulose, hydroxymethylcellulose, polypropylpyrrolidone, polyvinylpyrrolidone, gelatin, gum arabic, polyethylene glycol, sucrose, or starch), disintegrants (e.g., starch, carboxymethylcellulose, hydroxypropyl starch, low-substituted hydroxypropyl cellulose, sodium bicarbonate, calcium phosphate, or calcium citrate), lubricants (e.g., cellulose, methylcellulose, hydroxypropyl starch, low-substituted hydroxypropyl cellulose, sodium bicarbonate, calcium phosphate, or calcium citrate), and the like. For example, magnesium stearate, light anhydrous silicic acid, talc, or sodium lauryl sulfate), flavorings (e.g., citric acid, menthol, glycine, or powdered orange), preservatives (e.g., sodium benzoate, sodium bisulfite, methylparaben, or propylparaben), stabilizers (e.g., citric acid, sodium citrate, or acetic acid), suspending agents (e.g., methylcellulose, polyvinylpyrrolidone, or aluminum stearate), dispersing agents (e.g., hydroxypropylmethylcellulose), diluents (e.g., water), and base waxes (e.g., cocoa butter, white petrolatum, or polyethylene glycol). The effective amount of the compound of formula (IA) or formula (I) in the pharmaceutical composition can be a level that exerts the desired effect. For example, the unit dosage for both oral and parenteral administration is about 0.005 mg to about 10 mg per kg of subject body weight.

[0112] The dose of a compound of Formula (IA) or Formula (I) to be administered to a subject can vary widely and is subject to the judgment of a medical professional. Generally, the compounds of the present disclosure can be administered at a dose of about 0.001 mg to about 10 mg per kg of subject body weight, once to four times daily, although this dose may be appropriately modified depending on the subject's age, body weight, and condition, as well as the method of administration. In certain embodiments, the dose is about 0.001 mg to about 5 mg per kg of subject body weight, about 0.01 mg to about 5 mg per kg of subject body weight, about 0.05 mg to about 1 mg per kg of subject body weight, about 0.1 mg to about 0.75 mg per kg of subject body weight, or about 0.25 mg to about 0.5 mg per kg of subject body weight. In certain embodiments, administration is once daily. In any case, the amount of a compound of Formula (IA) or Formula (I) administered will vary depending on factors such as the solubility of the active ingredient, the formulation used, and the route of administration.

[0113] In some embodiments, the compound of Formula (IA) or 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.

[0114] In another embodiment, provided herein is a single dosage form 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 (IA) or Formula (I).

[0115] In certain embodiments, provided herein are single dosage forms containing about 0.1 mg or 100 mg of a compound of Formula (IA) or Formula (I).

[0116] In another embodiment, provided herein is a single dosage form 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 (IA) or Formula (I).

[0117] The compound of Formula (IA) or Formula (I) can be administered once, twice, three times, four or more times per day. In certain embodiments, doses of 100 mg or less are administered once per day, and doses of more than 100 mg are administered twice per day in an amount equal to half the total daily dose.

[0118] The compound of formula (IA) or formula (I) may be conveniently administered orally. In some embodiments, when the compound of formula (IA) or formula (I) is administered orally, it is administered with food and water. In other embodiments, the compound of formula (IA) or formula (I) is dispersed in water or juice (e.g., apple juice or orange juice), or any other liquid, and is orally administered as a solution or suspension.

[0119] The compounds of the present disclosure may be administered intradermally, intramuscularly, intraperitoneally, transdermally, intravenously, subcutaneously, intranasally, epidurally, sublingually, intracerebrally, intravaginally, transdermally, rectally, transmucosally, by inhalation, or topically to the ear, nose, eye, or skin. The mode of administration is left to the discretion of the health care practitioner and will depend in part on the site of the disease.

[0120] In certain embodiments, provided herein are capsules comprising a compound of Formula (IA) or Formula (I) without other carriers, excipients, or vehicles.

[0121] In another embodiment, provided herein is a composition comprising an effective amount of a compound of Formula (IA) or 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 certain embodiments, the composition is a pharmaceutical composition.

[0122] The compositions may be in the form of tablets, chewable tablets, capsules, solutions, parenteral solutions, lozenges, suppositories, suspensions, and the like. The compositions are formulated into dosage forms containing a daily dose or a manageable portion of the daily dose, which may be a single tablet or capsule or a manageable volume of solution. In some embodiments, solutions are prepared from water-soluble salts (e.g., hydrochlorides). Generally, all compositions are prepared according to methods known in medicinal chemistry. Capsules can be prepared by mixing a compound of Formula (IA) or Formula (I) with a suitable carrier or diluent and filling the appropriate amount of the mixture into a capsule. Typical carriers and diluents include, but are not limited to, inert powdered substances such as various types of starch, powdered cellulose (e.g., crystalline cellulose and microcrystalline cellulose), sugars (e.g., fructose, mannitol, and sucrose), grain flours, and similar edible powders.

[0123] Tablets can be manufactured by direct compression, wet granulation compression, or dry granulation compression. These formulations typically contain diluents, binders, lubricants, and disintegrants, as well as compounds. Typical diluents include, for example, various types of starch, lactose, mannitol, kaolin, calcium phosphate or sulfate, inorganic salts (e.g., sodium chloride), and powdered sugar. Powdered cellulose derivatives are also useful. Typical tablet binders include substances such as starch, gelatin, and sugars (e.g., lactose, fructose, glucose). Convenient natural and synthetic thickeners include gum arabic, alginates, methylcellulose, polyvinylpyrrolidine, and the like. Polyethylene glycol, ethylcellulose, and waxes also serve as binders.

[0124] In tablet formulations, lubricants may be necessary to prevent coloring agents from adhering to the tablet and punch. Lubricants may be selected from lubricating solids such as talc, magnesium and calcium stearate, stearic acid, and hydrogenated vegetable oil. Tablet disintegrants are substances that swell when wet, causing the tablet to disintegrate and release the compound. These include starch, clay, cellulose, algin, and gums. More specifically, corn starch, potato starch, methylcellulose, agar, bentonite, wood cellulose, powdered natural sponge, cation exchange resin, alginic acid, guar gum, citrus pulp, and carboxymethylcellulose may be used, as well as sodium lauryl sulfate. Tablets may be coated with sugar as a flavoring agent or with a film-forming protecting agent to modify the tablet's dissolution characteristics. The composition may also be formulated as a chewable tablet, for example, by using substances such as mannitol in the formulation.

[0125] When it is desired to administer the compound of formula (IA) or formula (I) as a suppository, a common base can be used. Cocoa butter is a conventional suppository base, and waxes can be added to slightly raise the melting point. In particular, water-miscible suppository bases, including polyethylene glycols of various molecular weights, are widely used.

[0126] By appropriate formulation, it is possible to delay or prolong the effect of the compound of formula (IA) or formula (I). For example, slowly dissolving pellets of the compound of formula (IA) or formula (I) can be prepared and placed in tablets or capsules, or incorporated as a sustained-release implant device. This technique also includes preparing pellets with different dissolution rates and filling a capsule with a mixture of the pellets. The tablet or capsule can be coated with a film that is difficult to dissolve for a predictable period of time. The compound of formula (IA) or formula (I) can be dissolved or suspended in an oily or emulsified vehicle that allows it to disperse slowly in serum, thereby extending the duration of action even in parenteral formulations.

[0127] Example of implementation The present disclosure is further illustrated by the following embodiments, the features of each embodiment may be combined with any other embodiment where appropriate and practical.

[0128] Embodiment 1. Formula (IA): [ka] [In the formula, L 1 is -(CH2CH2O) n -or in conjunction; n is 1 to 10; L 2 is a C1-C6 alkylene, -(C1-C6 alkylene)N(H)-, a 6- to 12-membered spiroheterocyclylene, or a bond, wherein the heterocyclylene contains 1 to 3 heteroatoms selected from N and O; D is [ka] and; R 1a and R 1b are each H or together form oxo; and L 3 is -CH2O- or a bond] or a pharmaceutically acceptable salt thereof.

[0129] Embodiment 2. The compound has the formula (I): [ka] [In the formula, L 1 is -(CH2CH2O) n -or in conjunction; n is 1 to 10; L 2is a C1-C6 alkylene, -(C1-C6 alkylene)N(H)-, a 6- to 12-membered spiroheterocyclylene, or a bond, wherein the heterocyclylene contains 1 to 3 heteroatoms selected from N and O; D is [ka] and; R 1a and R 1b are each H or together form oxo; and L 3 is -CH2O- or a bond] 2. The compound of embodiment 1, or a pharmaceutically acceptable salt thereof.

[0130] Embodiment 3. In the formula, L 1 -(CH2CH2O) n or a pharmaceutically acceptable salt thereof.

[0131] Embodiment 4. The compound of embodiment 3, wherein n is 1 to 7, or a pharmaceutically acceptable salt thereof.

[0132] Embodiment 5. In the formula, L 1 The compound of embodiment 1 or 2, or a pharmaceutically acceptable salt thereof, wherein is a bond.

[0133] Embodiment 6. In the formula, L 2 is C1-C3 alkylene, —(C1-C3 alkylene)N(H)—, a 10- to 12-membered spiroheterocyclylene, or a bond, wherein the heterocyclylene contains 1 to 2 nitrogen atoms; or a pharmaceutically acceptable salt thereof.

[0134] Embodiment 7. In the formula, L 2 is -CH2-, -CH2CH2CH2-, -CH2CH2N(H)-, [ka] or a bond, or a pharmaceutically acceptable salt thereof.

[0135] Embodiment 8. In the formula, -L 1 -L 2 -but, [ka] 8. The compound of any one of embodiments 1 to 7, wherein:

[0136] Embodiment 9. In the formula, -L 1 -L 2 -but, [ka] 8. The compound of any one of embodiments 1 to 7, wherein:

[0137] Embodiment 10. In the formula, D is [ka] 10. The compound of any one of embodiments 1-9, wherein:

[0138] Embodiment 11. In the formula, D is [ka] 10. The compound of any one of embodiments 1-9, wherein:

[0139] Embodiment 12. In the formula, R 1a and R 1b and R are each H, or a pharmaceutically acceptable salt thereof.

[0140] Embodiment 13. In the formula, R 1a and R 1b taken together to form oxo, or a pharmaceutically acceptable salt thereof.

[0141] Embodiment 14. In the formula, L 3 14. The compound of any one of embodiments 11-13, or a pharmaceutically acceptable salt thereof, wherein is —CH2O—.

[0142] Embodiment 15. In the formula, L 3 14. The compound of any one of embodiments 11-13, or a pharmaceutically acceptable salt thereof, wherein is a bond.

[0143] Embodiment 16. During the ceremony, [ka] but [ka] 16. The compound of any one of embodiments 11-15, wherein:

[0144] Embodiment 17. In the formula, D is [ka] 10. The compound of any one of embodiments 1-9, wherein:

[0145] Embodiment 18. The compound has the formula (II): [ka] [In the formula, L 1 is -(CH2CH2O) n -or in conjunction; n is 1 to 10; and L 2 is C1-C6 alkylene] 10. The compound of any one of embodiments 1 to 9, or a pharmaceutically acceptable salt thereof.

[0146] Embodiment 19. The compound has the formula (III): [ka] [In the formula, L 1 is -(CH2CH2O) n -or in conjunction; n is 1 to 10; and L 2 is -(C1-C6 alkylene)-N(H)-, a 6- to 12-membered spiroheterocyclylene, or a bond, wherein the heterocyclylene contains 1 to 3 heteroatoms selected from N and O. 17. The compound of any one of embodiments 1 to 9 and 11 to 16, or a pharmaceutically acceptable salt thereof.

[0147] Embodiment 20. 3. The compound of formula (VII): [ka] [In the formula, L 1 is a bond; and L 2 is a 6- to 12-membered spiroheterocyclylene containing 1 to 3 heteroatoms selected from N and O. 18. The compound of any one of embodiments 1 to 9 and 17, or a pharmaceutically acceptable salt thereof.

[0148] Embodiment 21. A compound selected from the compounds of Table 1 and pharmaceutically acceptable salts thereof.

[0149] Embodiment 22. 22. A pharmaceutical composition comprising a compound of any one of embodiments 1-21 or a pharmaceutically acceptable salt thereof, and a pharmaceutically acceptable excipient.

[0150] Embodiment 23. A method for modulating interleukin-1 receptor-associated kinase 3 (IRAK3), comprising contacting IRAK3 with an effective amount of a compound of any one of embodiments 1 to 21, or a pharmaceutically acceptable salt thereof, or a pharmaceutical composition of embodiment 22.

[0151] Embodiment 24. A method for treating cancer in a patient in need thereof, comprising administering to the subject an effective amount of a compound of any one of embodiments 1 to 21, or a pharmaceutically acceptable salt thereof, or a pharmaceutical composition of embodiment 22.

[0152] Embodiment 25. 25. The method of embodiment 24, 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.

[0153] Embodiment 26. A method for enhancing immunity in a subject receiving a vaccine, comprising administering to the subject an effective amount of a compound of any one of embodiments 1 to 21 or a pharmaceutically acceptable salt thereof, or a pharmaceutical composition of embodiment 22.

[0154] Embodiment 27. The method of embodiment 26, wherein a vaccine is administered to the subject before, simultaneously with, or after administration of the compound of any one of embodiments 1 to 21 or a pharmaceutically acceptable salt thereof, or the pharmaceutical composition of embodiment 22.

[0155] Example The following examples are provided for illustrative purposes, but are not limiting. Compounds were named using the automated naming tool provided in ChemBiodraw Ultra (CambridgeSoft), which generates systematic names for chemical structures incorporating the Cahn-Ingold-Prelog rules of stereochemistry. Those skilled in the art can modify the procedures of the described examples to obtain the desired products.

[0156] Salts of the compounds described herein can be prepared by standard methods, such as adding an acid (e.g., TFA, formic acid, or HCl) to the mobile phase during chromatographic purification, or by stirring the product after chromatographic purification with an acid solution (e.g., dilute hydrochloric acid).

[0157] The following abbreviations may be relevant to this application: [Table 5]

[0158] Synthesis Examples Analysis method LC / MS Method 1. Column: Luna C18(2) 50x3mm, 3um; Temperature: 45°C, Flow Rate: 2mL / min, Retention Time: 2min; Mobile Phase Conditions: Initially 95% HO (0.1% FA) / 5% MeCN (0.1% FA) with a linear gradient to 95% MeCN (0.1% FA) over 1min, followed by 95% CH3CN (0.1% FA) over 1min.

[0159] LC / MS Method 2. Waters Acquity UPLC System; Column: ACQUIty UPLC BEH C18 1.7 μm (2.1 x 50 mm); Added Acid: Formic Acid; Mobile Phase: Water (containing 0.1% formic acid) (A) and Acetonitrile (containing 0.1% formic acid) (B); Flow Rate: 0.8 mL / min; Gradient: 5% B to 95% B for 1.5 min, followed by 95% B for 0.5 min, and then 95% B to 5% B for 0.1 min; Detection: UV (214 nm and 254 nm).

[0160] LC / MS Method 3. SunFire C18 75x4.6mm, 3.5um; Temperature: 45°C, Flow rate: 1.5mL / min, Retention time: 6min; Mobile phase conditions: Initially eluted with 95% HO + 0.1% FA / 5% MeCN + 0.1% FA, then linear gradient to 95% MeCN for 4min, then 95% MeCN for 2min.

[0161] LC / MS Method 4. Luna C18(2) 50x3mm, 3um; Temperature: 45°C, Flow Rate: 1.5mL / min, Retention Time: 2.5min; Mobile Phase Conditions: Initially 95% HO (0.1% FA) / 5% MeCN (0.1% FA), linear gradient to 95% MeCN (0.1% FA) over 1.3min, then 95% MeCN (0.1% FA) for 1.2min.

[0162] LC / MS Method 5. Luna C18 50x3 mm, 3 μm; Temperature: 45 °C, Flow rate: 1.2 mL / min, Retention time: 5 min; Mobile phase conditions: Initial elution with 95% H2O (0.05% TFA) / 5% CH3CN, followed by a linear gradient to 5% H2O (0.05% TFA) / 95% CH3CN for 3.5 min, then 95% CH3CN for 1.5 min.

[0163] LC / MS Method 6. Gemini C18 4.6x50mm, 3µm; Temperature: 45°C, Flow rate: 1.5mL / min, Retention time: 6min; Mobile phase conditions: Initial equilibration with 95% NH4HCO3 / 5% MeCN for 1min, followed by a linear gradient to 95% MeCN for 3.5min, then elution for 2.5min.

[0164] Example I-1. Preparation of (R)-4-(4-(4-(6-amino-5-(1-(2,6-dichloro-3-fluorophenyl)ethoxy)pyridin-3-yl)-1H-pyrazol-1-yl)piperidin-1-yl)butanoic acid (Intermediate I-1) [ka]

[0165] Step 1. To a solution of 5-[1-(4-piperidyl)pyrazol-4-yl]-3-[(1R)-1-(2,6-dichloro-3-fluorophenyl)ethoxy]pyridin-2-amine (125 mg, 0.2800 mmol) and ethyl 4-bromobutyrate (56.85 mg, 0.2900 mmol) in DMF (0.56 mL) and DMSO (0.1481 mL) was added DIPEA (120.87 μL, 0.6900 mmol) at room temperature and stirred overnight. LCMS showed 91% conversion. This mixture was loaded directly onto a C18 column and purified by reverse-phase chromatography (elution: 5% to 100% MeOH / EtOAc) (83% yield). LCMS method 1: Retention time: 1.040 min, purity 99.9% (215 nm), [M+H] + =564.2

[0166] Step 2. To a solution of ethyl (R)-4-(4-(4-(6-amino-5-(1-(2,6-dichloro-3-fluorophenyl)ethoxy)pyridin-3-yl)-1H-pyrazol-1-yl)piperidin-1-yl)butyrate (130 mg, 0.2300 mmol) in THF (1 mL) and water (1 mL) was added LiOH (55.2 mg, 2.3 mmol) and stirred at room temperature overnight. LCMS showed complete conversion. The reaction mixture was concentrated and purified by reverse-phase column chromatography to give the desired product in 85% yield (105 mg). LCMS method 5: Retention time: 1.936 min, purity 98.3% (215 nm), [M+H] + =536.2, [M+2H] / 2 + =269.6

[0167] Example I-2. Preparation of 2-(2-(2-(((S)-1-((2S,4R)-4-hydroxy-2-((4-(4-methylthiazol-5-yl)benzyl)carbamoyl)pyrrolidin-1-yl)-3,3-dimethyl-1-oxobutan-2-yl)amino)-2-oxoethoxy)ethoxy)ethyl 4-methylbenzenesulfonate (Intermediate I-2) [ka]

[0168] Step 1. To a solution of tert-butyl 2-(2-(2-hydroxyethoxy)ethoxy)acetate (1 g, 4.54 mmol) in dry DCM (18.16 mL) was added triethylamine (1265 μL, 9.08 mmol), 4-methylbenzenesulfonyl chloride (1.3 g, 6.81 mmol), and DMAP (55.47 mg, 0.4500 mmol) at 0 °C and stirred overnight. The ice bath was allowed to warm to room temperature during this time. HPLC showed complete conversion. The reaction was quenched with aqueous NH4Cl and extracted three times with EtOAc. The organic layer was washed with brine, dried over Na2SO4, concentrated under reduced pressure, and purified by normal phase column chromatography to give the desired product (1.56 g, 88% yield) as a pale yellow solid.

[0169] Step 2. To a solution of tert-butyl 2-(2-(2-(tosyloxy)ethoxy)ethoxy)acetate (1.56 g, 4.17 mmol) in DCM (7.7 mL) was added TFA (3.8 mL, 47 mmol) dropwise at room temperature and stirred at room temperature for 2 hours. HPLC analysis showed complete conversion. The solvent was concentrated to dryness to give the crude acid as a pale yellow oil. To a solution of the crude acid in DMF (8.3 mL) was added DIPEA (2.2 mL, 12.5 mmol) and HATU (1.82 g, 4.79 mmol) at room temperature and stirred at room temperature for 10 minutes. (2S,4R)-1-((S)-2-amino-3,3-dimethylbutanoyl)-4-hydroxy-N-(4-(4-methylthiazol-5-yl)benzyl)pyrrolidine-2-carboxamide hydrochloride (2.05 g, 4.39 mmol) was then added in one portion. The reaction mixture was stirred at room temperature overnight. HPLC analysis showed complete conversion. The reaction mixture was purified by reverse-phase column chromatography to give the desired product as a white solid (2 g, 61.6% yield). 1 H NMR (400MHz, DMSO-d6) δ ppm 0.92(s, 9H), 1.88-1.96(m, 1H), 2.01-2.07(m, 1H), 2.40(s, 3H), 2.43(s, 3H), 3.48-3.56(m, 4H), 3.59-3.68(m, 4H), 3.93(s, 2H), 4.11-4.14(m, 2H), 4.28(d, J=5.3Hz, 1H), 4.34-4.39(m, 2H), 4.41-4.46(m, 1H), 4.56(d, J=9.6Hz, 1H), 5.15(d, J=3.5Hz, 1H), 7.38-7.41(m, 5H), 7.46(d, J=7.8Hz, 2H), 7.77(d, J=8.3Hz, 2H), 8.55-8.61(m, 1H), 8.98(s, 1H) LCMS method 3: Retention time: 3.660 min, purity 93.2% (215 nm), [M+H] + =731.2

[0170] Example I-3. Preparation of (R)-20-(4-(4-(6-amino-5-(1-(2,6-dichloro-3-fluorophenyl)ethoxy)pyridin-3-yl)-1H-pyrazol-1-yl)piperidin-1-yl)-3,6,9,12,15,18-hexaoxaarachidic acid (intermediate I-3) [ka]

[0171] Step 1. To a solution of (R)-3-(1-(2,6-dichloro-3-fluorophenyl)ethoxy)-5-(1-(piperidin-4-yl)-1H-pyrazol-4-yl)pyridin-2-amine (100 mg, 0.2200 mmol) and DIPEA (58 μL, 0.33 mmol) in DMF (0.29 mL) was added tert-butyl 2-[2-[2-[2-[2-[2-[2-(p-tolylsulfonyloxy)ethoxy]ethoxy]ethoxy]ethoxy]ethoxy]ethoxy]ethoxy]ethoxy]acetate (146.79 mg, 0.27 mmol) at room temperature. DMSO (0.29 mL) was added to completely dissolve the resulting yellow solution at room temperature for 4 days, after which LCMS analysis indicated that the conversion had stopped at approximately 60%. The mixture was purified by reverse phase column chromatography and the fractions were combined and concentrated to give the desired product (110 mg, 59.8% yield) as a pale yellow solid. LCMS method 5: Retention time: 1.143 min, purity 90.1% (254 nm), [M+2H] / 2 + =829.2

[0172] Step 2. To a solution of (R)-tert-butyl 20-(4-(4-(6-amino-5-(1-(2,6-dichloro-3-fluorophenyl)ethoxy)pyridin-3-yl)-1H-pyrazol-1-yl)piperidin-1-yl)-3,6,9,12,15,18-hexaoxaarachidate (110 mg, 0.1300 mmol) in water (0.74 mL) and THF (1.48 mL) was added LiOH monohydrate (27.85 mg, 0.6600 mmol) at room temperature and stirred at 40° C. for 2 hours. LCMS showed complete conversion. The reaction mixture was concentrated under reduced pressure to remove THF, and the resulting aqueous solution was directly loaded onto a GOLD C18 column (30 g) and purified by reverse-phase column chromatography. The combined fractions were concentrated to give the desired product (43 mg, 41.9% yield) as a pale yellow oil. LCMS method 5: Retention time: 0.971 min, 100% purity (254 nm), [M+H] + =772.3 [M+2H] / 2 + =387.8

[0173] Example I-4. Preparation of (R)-23-(4-(4-(6-amino-5-(1-(2,6-dichloro-3-fluorophenyl)ethoxy)pyridin-3-yl)-1H-pyrazol-1-yl)piperidin-1-yl)-3,6,9,12,15,18,21-heptaoxatricosanoic acid (intermediate I-4) [ka] The title compound was prepared using a procedure similar to that described in I-3 using ethyl 2-[2-[2-[2-[2-[2-[2-[2-(p-tolylsulfonyloxy)ethoxy]ethoxy]ethoxy]ethoxy]ethoxy]ethoxy]ethyl acetate. LCMS method 5: Retention time: 1.040 min, purity 99.0% (215 nm), [M+H] + =816.3

[0174] Example I-5. Preparation of 2-((2-(2,6-dioxopiperidin-3-yl)-1,3-dioxoisoindolin-4-yl)oxy)-N-(2-hydroxyethyl)acetamide (Intermediate I-5) [ka] To a mixture of 2-((2-(2,6-dioxopiperidin-3-yl)-1,3-dioxoisoindolin-4-yl)oxy)acetic acid (1.8 g, 5.42 mmol) and HATU (2.472 g, 6.50 mmol) in DMF (20 mL) was added DIEA (2.84 mL, 16.25 mmol) followed by 2-aminoethan-1-ol (330 mg, 5.42 mmol) at room temperature. The reaction was stirred at room temperature for 1 hour, then concentrated and purified by normal phase column chromatography (Biotage silica, 0-100% EA / hex, then 0-25% MeOH / DCM) to give the desired product. LC / MS method 2: MS(ESI)[M+H] + 376.0, rt: 0.59 minutes

[0175] Example I-6. Preparation of 2-((2-(2,6-dioxopiperidin-3-yl)-1,3-dioxoisoindolin-4-yl)oxy)-N-(2-(2-hydroxyethoxy)ethyl)acetamide (Intermediate I-6) [ka] The title compound was prepared using the same general procedure as described for I-5 using 2-(2-aminoethoxy)ethan-1-ol. LC / MS method 2: MS(ESI)[M+H] + 420.1, rt: 0.61 minutes

[0176] Example I-7. Preparation of 2-((2-(2,6-dioxopiperidin-3-yl)-1,3-dioxoisoindolin-4-yl)oxy)-N-(2-(2-(2-hydroxyethoxy)ethoxy)ethyl)acetamide (Intermediate I-7) [ka] The title compound was prepared using the same general procedure as described for I-5 using 2-(2-(2-aminoethoxy)ethoxy)ethan-1-ol. LC / MS method 2: MS(ESI)[M+H] + 464.1, rt: 0.64 minutes

[0177] Example I-8. Preparation of 2-((2-(2,6-dioxopiperidin-3-yl)-1,3-dioxoisoindolin-4-yl)oxy)-N-(2-(2-(2-(2-hydroxyethoxy)ethoxy)ethoxy)ethyl)acetamide (Intermediate I-8) [ka] The title compound was prepared using the same general procedure as described for I-5 using 2-(2-(2-(2-aminoethoxy)ethoxy)ethoxy)ethan-1-ol. LC / MS method 2: MS(ESI)[M+H] + 508.1, rt: 0.66 minutes

[0178] Example I-9. Preparation of 2-((2-(2,6-dioxopiperidin-3-yl)-1,3-dioxoisoindolin-4-yl)oxy)-N-(14-hydroxy-3,6,9,12-tetraoxatetradecyl)acetamide (Intermediate I-9) [ka] The title compound was prepared using 14-amino-3,6,9,12-tetraoxatetradecan-1-ol following the same general procedure as described for I-5. LC / MS method 2: MS(ESI)[M+H] + 552.2, rt: 0.68 minutes

[0179] Example I-10. Preparation of 2-((2-(2,6-dioxopiperidin-3-yl)-1,3-dioxoisoindolin-5-yl)oxy)-N-(2-hydroxyethyl)acetamide (Intermediate I-10) [ka] The title compound was prepared using 2-aminoethan-1-ol by the same general procedure as described for I-5. LC / MS method 2: MS(ESI)[M+H] + 376.0, rt: 0.56 minutes

[0180] Example I-11. Preparation of 2-((2-(2,6-dioxopiperidin-3-yl)-1,3-dioxoisoindolin-5-yl)oxy)-N-(2-(2-hydroxyethoxy)ethyl)acetamide (Intermediate I-11) [ka] The title compound was prepared using the same general procedure as described for I-5 using 2-(2-aminoethoxy)ethan-1-ol. LC / MS method 2: MS(ESI)[M+H] + 420.1, rt: 0.61 minutes

[0181] Example I-12. Preparation of 2-((2-(2,6-dioxopiperidin-3-yl)-1,3-dioxoisoindolin-5-yl)oxy)-N-(2-(2-(2-hydroxyethoxy)ethoxy)ethyl)acetamide (Intermediate I-12) [ka] The title compound was prepared using the same general procedure as described for I-5 using 2-(2-(2-aminoethoxy)ethoxy)ethan-1-ol. LC / MS method 2: MS(ESI)[M+H] + 464.1, rt: 0.63 minutes

[0182] Example I-13. Preparation of 2-((2-(2,6-dioxopiperidin-3-yl)-1,3-dioxoisoindolin-5-yl)oxy)-N-(2-(2-(2-(2-hydroxyethoxy)ethoxy)ethoxy)ethyl)acetamide (Intermediate I-13) [ka] The title compound was prepared using the same general procedure as described for I-5 using 2-(2-(2-(2-aminoethoxy)ethoxy)ethoxy)ethan-1-ol. LC / MS method 2: MS(ESI)[M+H] + 508.1, rt: 0.65 minutes

[0183] Example I-14. Preparation of 2-((2-(2,6-dioxopiperidin-3-yl)-1,3-dioxoisoindolin-5-yl)oxy)-N-(14-hydroxy-3,6,9,12-tetraoxatetradecyl)acetamide (Intermediate I-14) [ka] The title compound was prepared using the same general procedure as described for I-5 using 2-(2-(2-(2-aminoethoxy)ethoxy)ethoxy)ethan-1-ol. LC / MS method 2: MS(ESI)[M+H] + 552.2, rt: 0.67 minutes

[0184] Example I-15. Preparation of (R)-5-(1-(1-(3-azaspiro[5.5]undecan-9-yl)piperidin-4-yl)-1H-pyrazol-4-yl)-3-(1-(2,6-dichloro-3-fluorophenyl)ethoxy)pyridin-2-amine (Intermediate I-15) [ka] (R)-3-(1-(2,6-dichloro-3-fluorophenyl)ethoxy)-5-(1-(piperidin-4-yl)-1H-pyrazol-4-yl)pyridin-2-amine (300 mg, 0.67 mmol), tert-butyl 9-oxo-3-azaspiro[5.5]undecane-3-carboxylate (178 mg, 0.67 mmol), and sodium triacetoxyborohydride (211 mg, 1.0 mmol) were added to a vial (20 mL) containing a stir bar. The vial was then sealed with a cap and purged with nitrogen. The solids were then suspended in DCE (2.7 mL). This suspension was then treated with acetic acid (60 μL, 1.0 mmol), causing the solution to become translucent. The reaction was then stirred at 44° C. overnight. LCMS analysis indicated that the reaction mixture had been converted to the desired product. The resulting residue was then purified by reverse-phase column chromatography (15% to 100% MeCN / water, 0.1% TFA). The collected fractions were concentrated and immediately dissolved in DCM / TFA (1:1) and left with stirring for 2 hours. LCMS analysis indicated that the reaction mixture was completely deprotected. After lyophilization, the TFA salt of the product: (R)-5-(1-(1-(3-azaspiro[5.5]undecan-9-yl)piperidin-4-yl)-1H-pyrazol-4-yl)-3-(1-(2,6-dichloro-3-fluorophenyl)ethoxy)pyridin-2-amine (390 mg, 0.54 mmol, 81% yield) was obtained as an off-white salt. LC / MS method 2: MS(ESI)[M+2H] / 2 + 301.5, rt: 0.82 minutes

[0185] Example I-16. 2-(2,6-dioxopiperidin-3-yl)-3-oxoisoindoline-5-carboxylic acid (Intermediate I-16) [ka] The title compound was obtained commercially (Enamine).

[0186] Example I-17. 2-(2,6-dioxopiperidin-3-yl)-1-oxoisoindoline-5-carboxylic acid (Intermediate I-17) [ka] The title compound was obtained commercially (Enamine).

[0187] Example I-18. Preparation of 2-(2,6-dioxopiperidin-3-yl)-1-oxoisoindoline-4-carboxylic acid (Intermediate I-18) [ka] The title compound was obtained commercially (Enamine).

[0188] Example I-19. Preparation of 2-(2,6-dioxopiperidin-3-yl)-1,3-dioxoisoindoline-5-carboxylic acid (Intermediate I-19) [ka] The title compound was obtained commercially (Enamine).

[0189] Example S1. Preparation of (2S,4R)-1-((S)-2-(4-(4-(4-(6-amino-5-((R)-1-(2,6-dichloro-3-fluorophenyl)ethoxy)pyridin-3-yl)-1H-pyrazol-1-yl)piperidin-1-yl)butanamido)-3,3-dimethylbutanoyl)-4-hydroxy-N-(4-(4-methylthiazol-5-yl)benzyl)pyrrolidine-2-carboxamide (Compound 1) [ka] (R)-4-(4-(4-(6-amino-5-(1-(2,6-dichloro-3-fluorophenyl)ethoxy)pyridin-3-yl)-1H-pyrazol-1-yl)piperidin-1-yl)butanoic acid (50 mg, 0.09 mmol), (2S,4R)-1-((S)-2-amino-3,3-dimethylbutanoyl)-4-hydroxy-N-(4-(4-methylthiazol-5-yl)benzyl)pyrrolidine-2-carboxamide (52.12 mg, 0.11 mmol), and DIPEA (0.06 mL, 0.37 mmol) were dissolved in DMF (0.93 mL), and HATU (43 mg, 0.11 mmol) was added at room temperature for 3 h. LCMS showed complete conversion. The reaction mixture was purified by reverse phase chromatography (5% to 100% MeCN / water, 0.1% formic acid), washed with MeOH, and recovered with ammonium hydroxide solution (3N MeOH). The solution was concentrated under reduced pressure and lyophilized to give the desired product as a white solid (16.7% yield, 15 mg). LC / MS method 3: Purity 98.7% (215nm), [M+H] + =948.2, 950.2, [M+H] / 2 + =474.6, 475.4 1H NMR (400MHz, DMSO-d6) δ ppm 8.97(s, 1H), 8.55(t, J=5.6Hz, 1H), 7.93(s, 1H), 7.86(d, J=9.1Hz, 1H), 7.74(d, J=1.5Hz, 1H), 7.49-7.62(m, 2H), 7.29-7.47(m, 5H), 6.89(s, 1H), 6.08(q, J=6.7Hz, 1H), 5.63(s, 2H), 5.13(d, J=3.5Hz, 1H), 5.13(d, J=3.5Hz, 1H), 4.55(d, J=9.3Hz, 1H), 4.38-4.49(m, 2H), 4.35(br. s, 1H), 4.21(dd, J=15.8, 5.7Hz, 1H), 4.08(br. s, 1H), 3.60-3.73(m, 2H), 2.92(d, J=10.1Hz, 2H), 2.44(s, 3H), 2.12-2.36(m, 4H), 1.85-2.09(m, 8H), 1.80(d, J=6.6Hz, 3H), 1.57-1.74(m, 2H), 0.94(s, 9H)

[0190] Example S2. Preparation of (2S,4R)-1-((S)-2-(2-(2-(2-(4-(4-(6-amino-5-((R)-1-(2,6-dichloro-3-fluorophenyl)ethoxy)pyridin-3-yl)-1H-pyrazol-1-yl)piperidin-1-yl)ethoxy)ethoxy)acetamido)-3,3-dimethylbutanoyl)-4-hydroxy-N-(4-(4-methylthiazol-5-yl)benzyl)pyrrolidine-2-carboxamide (Compound 2) [ka] Crizotinib (R)-3-(1-(2,6-dichloro-3-fluorophenyl)ethoxy)-5-(1-(piperidin-4-yl)-1H-pyrazol-4-yl)pyridin-2-amine (40 mg, 0.09 mmol) and 2-(2-(2-(((S)-1-((2S,4R)-4-hydroxy-2-((4-(4-methylthiazol-5-yl)benzyl)carbamoyl)pyrrolidin-1-yl)-3,3-dimethyl-1-oxobutan-2-yl)amino)-2-oxoethoxy)ethoxy)ethyl 4-methylbenzenesulfonate (77.8 mg, 0.11 mmol) in DMF (400 μL) / DMSO (100 μL) was added DIPEA (50 μL, 0.29 mmol) at room temperature and stirred at room temperature for 48 hours. LCMS showed the reaction was nearly complete. The reaction mixture was purified by reverse phase column chromatography, and the combined fractions were concentrated to give the desired product (43.6 mg) as a white solid (49% yield). LC / MS method 3: [M+2H] / 2 + =504.6, [M+3H] / 3 + =336.8, rt=2.359 minutes 1H NMR (400MHz, DMSO-d6) δ ppm 8.96-8.99(m, 1H), 8.59(t, J=6.2Hz, 1H), 7.92-7.95(m, 1H), 7.75(d, J=2.0Hz, 1H), 7.56(dd, J=9.1, 5.1Hz, 1H), 7.51(s, 1H), 7.41-7.46(m, 2H), 6.88-6.92(m, 1H), 6.08(q, J=6.6Hz, 1H), 5.63(s, 2H), 5.15(d, J=3.5Hz, 1H), 4.57(d, J=9.6Hz, 1H), 4.32-4.47(m, 4H), 4.20-4.28(m, 1H), 4.00-4.11(m, 2H), 3.98(s, 2H), 3.52-3.70(m, 9H), 3.17(d, J=5.3Hz, 1H), 2.89-2.99(m, 3H), 2.43(s, 3H), 2.04-2.14(m, 3H), 1.85-1.97(m, 6H), 1.79(d, J=6.6Hz, 3H), 0.95(s, 9H)

[0191] Example S3. Preparation of (2S,4R)-1-((S)-14-(4-(4-(6-amino-5-((R)-1-(2,6-dichloro-3-fluorophenyl)ethoxy)pyridin-3-yl)-1H-pyrazol-1-yl)piperidin-1-yl)-2-(tert-butyl)-4-oxo-6,9,12-trioxa-3-azatetradecanoyl)-4-hydroxy-N-(4-(4-methylthiazol-5-yl)benzyl)pyrrolidine-2-carboxamide (Compound 3) [ka]

[0192] Step 1: (R)-2-(2-(2-(2-(4-(4-(6-amino-5-(1-(2,6-dichloro-3-fluorophenyl)ethoxy)pyridin-3-yl)-1H-pyrazol-1-yl)piperidin-1-yl)ethoxy)ethoxy)ethoxy)ethyl acetate To a solution of ethyl 2-[2-[2-[2-(p-tolylsulfonyloxy)ethoxy]ethoxy]ethoxy]acetate (208 mg, 0.53 mmol) and (R)-3-(1-(2,6-dichloro-3-fluorophenyl)ethoxy)-5-(1-(piperidin-4-yl)-1H-pyrazol-4-yl)pyridin-2-amine (200 mg, 0.44 mmol) in 580 μL of DMF at room temperature was added DIPEA (117 μL, 0.67 mmol). DMSO (580 μL) was then added to ensure complete dissolution. The resulting yellow solution was stirred overnight at room temperature, after which LCMS analysis indicated ∼75% conversion to the product. The mixture was purified by reverse-phase column chromatography (5% to 100% MeCN / water, 0.1% formic acid) to give, after concentration, the product: (R)-2-(2-(2-(2-(4-(4-(6-amino-5-(1-(2,6-dichloro-3-fluorophenyl)ethoxy)pyridin-3-yl)-1H-pyrazol-1-yl)piperidin-1-yl)ethoxy)ethoxy)ethyl acetate (225 mg, 0.34 mmol, 76% yield) as a light yellow oil. LC / MS method 1: MS(ESI)[M+2H] / 2 + 335.8, rt: 0.980 minutes

[0193] Step 2: (R)-2-(2-(2-(2-(4-(4-(6-amino-5-(1-(2,6-dichloro-3-fluorophenyl)ethoxy)pyridin-3-yl)-1H-pyrazol-1-yl)piperidin-1-yl)ethoxy)ethoxy)ethoxy)acetic acid To a solution of (R)-2-(2-(2-(2-(4-(4-(6-amino-5-(1-(2,6-dichloro-3-fluorophenyl)ethoxy)pyridin-3-yl)-1H-pyrazol-1-yl)piperidin-1-yl)ethoxy)ethoxy)ethyl acetate (225 mg, 0.34 mmol) in water (1.9 mL) was added LiOH monohydrate (70.6 mg, 1.68 mmol). After 2 h, LCMS showed the reaction was complete. The reaction mixture was concentrated and purified by reverse phase column chromatography (5% to 100% MeCN / water) to give the product (R)-2-(2-(2-(2-(4-(4-(6-amino-5-(1-(2,6-dichloro-3-fluorophenyl)ethoxy)pyridin-3-yl)-1H-pyrazol-1-yl)piperidin-1-yl)ethoxy)ethoxy)ethoxy)acetic acid (quantitatively) as a pale yellow oil after concentration. LC / MS method 1: MS(ESI)[M+2H] / 2 + 321.6, rt: 0.937 minutes

[0194] Step 3: (2S,4R)-1-((S)-14-(4-(4-(6-amino-5-((R)-1-(2,6-dichloro-3-fluorophenyl)ethoxy)pyridin-3-yl)-1H-pyrazol-1-yl)piperidin-1-yl)-2-(tert-butyl)-4-oxo-6,9,12-trioxa-3-azatetradecanoyl)-4-hydroxy-N-(4-(4-methylthiazol-5-yl)benzyl)pyrrolidine-2-carboxamide To a solution of (R)-2-(2-(2-(2-(4-(4-(6-amino-5-(1-(2,6-dichloro-3-fluorophenyl)ethoxy)pyridin-3-yl)-1H-pyrazol-1-yl)piperidin-1-yl)ethoxy)ethoxy)ethoxy)acetic acid (200 mg, 0.31 mmol) in DMF (3.2 mL) was added DIPEA (193 μL, 1.11 mmol) and (2S,4R)-1-((S)-2-amino-3,3-dimethylbutanoyl)-4-hydroxy-N-(4-(4-methylthiazol-5-yl)benzyl)pyrrolidine-2-carboxamide hydrochloride (160 mg, 0.34 mmol) at room temperature. HATU (120 mg, 0.32 mmol) was then added in one portion, and the resulting yellow solution was stirred at room temperature. After 2 hours, LCMS analysis showed the reaction was complete. The reaction mixture was purified by reverse-phase column chromatography (5% to 100% MeCN / water, 0.1% formic acid), and the fractions were combined and concentrated. The resulting oil was again purified by reverse-phase column chromatography (5% MeCN / pH 10 water) to give, after lyophilization, the product: (2S,4R)-1-((S)-14-(4-(4-(6-amino-5-((R)-1-(2,6-dichloro-3-fluorophenyl)ethoxy)pyridin-3-yl)-1H-pyrazol-1-yl)piperidin-1-yl)-2-(tert-butyl)-4-oxo-6,9,12-trioxa-3-azatetradecanoyl)-4-hydroxy-N-(4-(4-methylthiazol-5-yl)benzyl)pyrrolidine-2-carboxamide (50 mg, 0.047 mmol, 15% yield) as a white solid. 1H NMR (400MHz, DMSO-d6) δ ppm 8.98(s, 1H), 8.61(t, J=5.9Hz, 1H), 7.94(s, 1H), 7.74(s, 1H), 7.54-7.60(m, 1H), 7.52(s, 1H), 7.37-7.46(m, 6H), 6.88(s, 1H), 6.08(q, J=6.7Hz, 1H), 5.65(s, 2H), 5.16(d, J=3.3Hz, 1H), 4.56(d, J=9.9Hz, 1H), 4.32-4.48(m, 3H), 4.20-4.29(m, 1H), 4.01-4.13(m, 1H), 3.94-4.00(m, 2H), 3.42-3.73(m, 14H), 2.90-3.04(m, 2H), 2.44(s, 3H), 2.01-2.24(m, 2H), 1.84-1.99(m, 4H), 1.79(d, J=6.6Hz, 3H), 0.94(s, 9H) LC / MS method 3: MS(ESI)[M+2H] / 2 + 526.7, rt: 2.338 minutes

[0195] Example S4. Preparation of (2S,4R)-1-((S)-17-(4-(4-(6-amino-5-((R)-1-(2,6-dichloro-3-fluorophenyl)ethoxy)pyridin-3-yl)-1H-pyrazol-1-yl)piperidin-1-yl)-2-(tert-butyl)-4-oxo-6,9,12,15-tetraoxa-3-azaheptadecanoyl)-4-hydroxy-N-(4-(4-methylthiazol-5-yl)benzyl)pyrrolidine-2-carboxamide (Compound 4) [ka]

[0196] Step 1: tert-butyl (R)-14-(4-(4-(6-amino-5-(1-(2,6-dichloro-3-fluorophenyl)ethoxy)pyridin-3-yl)-1H-pyrazol-1-yl)piperidin-1-yl)-3,6,9,12-tetraoxatetradecanoate To a solution of tert-butyl 14-(tosyloxy)-3,6,9,12-tetraoxatetradecanoate (247 mg, 0.53 mmol) and (R)-3-(1-(2,6-dichloro-3-fluorophenyl)ethoxy)-5-(1-(piperidin-4-yl)-1H-pyrazol-4-yl)pyridin-2-amine (200 mg, 0.44 mmol) in DMF (580 μL) was added DIPEA (117 μL, 0.67 mmol) at room temperature. DMSO (580 μL) was then added to completely dissolve the compound. The resulting yellow solution was stirred overnight at room temperature, and LCMS analysis indicated approximately 50% conversion to the desired product. The reaction was stirred for an additional 3 hours, after which the reaction mixture was directly purified by reverse-phase column chromatography (5% to 100% MeOH / water, containing 0.1% formic acid). The fractions were combined and concentrated to give the product after concentration: tert-butyl (R)-14-(4-(4-(6-amino-5-(1-(2,6-dichloro-3-fluorophenyl)ethoxy)pyridin-3-yl)-1H-pyrazol-1-yl)piperidin-1-yl)-3,6,9,12-tetraoxatetradecanoate (127 mg, 0.17 mmol, 38% yield) as a pale yellow oil. LC / MS method 1: MS(ESI)[M+2H] / 2 + 371.6, rt: 1.038 minutes

[0197] Step 2: (R)-14-(4-(4-(6-amino-5-(1-(2,6-dichloro-3-fluorophenyl)ethoxy)pyridin-3-yl)-1H-pyrazol-1-yl)piperidin-1-yl)-3,6,9,12-tetraoxatetradecanoic acid To a solution of tert-butyl (R)-14-(4-(4-(6-amino-5-(1-(2,6-dichloro-3-fluorophenyl)ethoxy)pyridin-3-yl)-1H-pyrazol-1-yl)piperidin-1-yl)-3,6,9,12-tetraoxatetradecanoate (127 mg, 0.17 mmol) in DCM (725 μL) was added TFA (870 μL, 11 mmol) dropwise. The resulting solution was stirred at room temperature for 2 hours, after which LCMS analysis indicated the reaction was complete. The solvent was removed under reduced pressure, and the remaining TFA was removed azeotropically with toluene (2×5 mL). The resulting residue was purified by reverse-phase column chromatography (5% to 100% MeCN / water at pH 10) to give, after concentration, the product: (R)-14-(4-(4-(6-amino-5-(1-(2,6-dichloro-3-fluorophenyl)ethoxy)pyridin-3-yl)-1H-pyrazol-1-yl)piperidin-1-yl)-3,6,9,12-tetraoxatetradecanoic acid (70 mg, 0.10 mmol, 59% yield) as a pale yellow oil. LC / MS method 1: MS(ESI)[M+2H] / 2 + 342.8, rt: 0.955 minutes

[0198] Step 3: (2S,4R)-1-((S)-17-(4-(4-(6-amino-5-((R)-1-(2,6-dichloro-3-fluorophenyl)ethoxy)pyridin-3-yl)-1H-pyrazol-1-yl)piperidin-1-yl)-2-(tert-butyl)-4-oxo-6,9,12,15-tetraoxa-3-azaheptadecanoyl)-4-hydroxy-N-(4-(4-methylthiazol-5-yl)benzyl)pyrrolidine-2-carboxamide To a solution of (R)-14-(4-(4-(6-amino-5-(1-(2,6-dichloro-3-fluorophenyl)ethoxy)pyridin-3-yl)-1H-pyrazol-1-yl)piperidin-1-yl)-3,6,9,12-tetraoxatetradecanoic acid (70 mg, 0.10 mmol) in DMF (1.05 mL) was added DIPEA (63 μL, 0.36 mmol) and (2S,4R)-1-((S)-2-amino-3,3-dimethylbutanoyl)-4-hydroxy-N-(4-(4-methylthiazol-5-yl)benzyl)pyrrolidine-2-carboxamide hydrochloride (52 mg, 0.11 mmol) at room temperature. HATU (39 mg, 0.10 mmol) was then added in one portion, and the resulting yellow solution was stirred at room temperature. After 2 hours, LCMS analysis indicated the reaction was complete. The reaction mixture was purified by reverse-phase column chromatography (5% to 100% MeCN / water, 0.1% formic acid). The resulting fractions were combined, concentrated, and purified again by reverse-phase column chromatography (5% to 100% MeCN / water at pH 10). The combined fractions were concentrated to give the product in ∼85% purity. This material was then further purified again by reverse-phase column chromatography (5%–100% MeCN / water, 0.1% formic acid) and neutralized by catch-and-release purification on an SPE cartridge (tosylic acid), affording, after lyophilization, the product: (2S,4R)-1-((S)-17-(4-(4-(6-amino-5-((R)-1-(2,6-dichloro-3-fluorophenyl)ethoxy)pyridin-3-yl)-1H-pyrazol-1-yl)piperidin-1-yl)-2-(tert-butyl)-4-oxo-6,9,12,15-tetraoxa-3-azaheptadecanoyl)-4-hydroxy-N-(4-(4-methylthiazol-5-yl)benzyl)pyrrolidine-2-carboxamide (8.2 mg, 0.007 mmol, 7% yield) as an off-white solid. 1H NMR (400MHz, DMSO-d6) δ ppm 8.98(s, 1H), 8.60(br t, J=5.2Hz, 1H), 7.94(s, 1H), 7.74(s, 1H), 7.54-7.60(m, 1H), 7.52(s, 1H), 7.37-7.46(m, 6H), 6.89(s, 1H), 6.08(q, J=6.6Hz, 1H), 5.65(s, 2H), 5.16(br s, 1H), 4.56(br d, J=9.6Hz, 1H), 4.31-4.48(m, 3H), 4.17-4.31(m, 1H), 4.03-4.16(m, 1H), 3.94-3.99(m, 2H), 3.42-3.69(m, 18H), 2.95-3.10(m, 2H), 2.55-2.64(m, 2H) 2.43(s, 3H), 2.14-2.29(m, 2H), 1.85-2.11(m, 4H), 1.79(br d, J=6.6Hz, 3H), 0.93(s, 9H) LC / MS method 3: MS(ESI)[M+2H] / 2 + 548.8, rt: 2.406 minutes

[0199] Example S5. Preparation of (2S,4R)-1-((S)-20-(4-(4-(6-amino-5-((R)-1-(2,6-dichloro-3-fluorophenyl)ethoxy)pyridin-3-yl)-1H-pyrazol-1-yl)piperidin-1-yl)-2-(tert-butyl)-4-oxo-6,9,12,15,18-pentaoxa-3-azaicosanoyl)-4-hydroxy-N-(4-(4-methylthiazol-5-yl)benzyl)pyrrolidine-2-carboxamide (Compound 5) [ka]

[0200] Step 1: (R)-17-(4-(4-(6-amino-5-(1-(2,6-dichloro-3-fluorophenyl)ethoxy)pyridin-3-yl)-1H-pyrazol-1-yl)piperidin-1-yl)-3,6,9,12,15-pentaoxaheptadecanoate ethyl ester To a solution of ethyl 17-bromo-3,6,9,12,15-pentaoxaheptadecanoate (103 mg, 0.27 mmol) and (R)-3-(1-(2,6-dichloro-3-fluorophenyl)ethoxy)-5-(1-(piperidin-4-yl)-1H-pyrazol-4-yl)pyridin-2-amine (100 mg, 0.22 mmol) in DMF (800 μL) was added DIPEA (58 μL, 0.33 mmol) at room temperature. DMSO (290 μL) was then added to completely dissolve the mixture. The resulting yellow solution was stirred at room temperature for 2 days, after which LCMS analysis indicated the reaction was complete. The mixture was purified by reverse-phase column chromatography (5% to 100% MeCN / water, 0.1% formic acid) to give the product: (R)-17-(4-(4-(6-amino-5-(1-(2,6-dichloro-3-fluorophenyl)ethoxy)pyridin-3-yl)-1H-pyrazol-1-yl)piperidin-1-yl)-3,6,9,12,15-pentaoxaheptadecanoate (125 mg, 0.16 mmol, 74% yield) as a light yellow solid. LC / MS method 1: MS(ESI)[M+2H] / 2 + 379.6, rt: 1.001 minutes

[0201] Step 2: (R)-17-(4-(4-(6-amino-5-(1-(2,6-dichloro-3-fluorophenyl)ethoxy)pyridin-3-yl)-1H-pyrazol-1-yl)piperidin-1-yl)-3,6,9,12,15-pentaoxaheptadecanoic acid To a solution of LiOH monohydrate (35 mg, 0.83 mmol) in THF (1.5 mL) and water (1 mL) was added (R)-ethyl 17-(4-(4-(6-amino-5-(1-(2,6-dichloro-3-fluorophenyl)ethoxy)pyridin-3-yl)-1H-pyrazol-1-yl)piperidin-1-yl)-3,6,9,12,15-pentaoxaheptadecanoate (125 mg, 0.17 mmol). After 2 hours, LCMS showed the reaction was complete. The reaction mixture was concentrated and then purified by reverse-phase column chromatography (5% to 100% MeCN / water) to give the product: (R)-17-(4-(4-(6-amino-5-(1-(2,6-dichloro-3-fluorophenyl)ethoxy)pyridin-3-yl)-1H-pyrazol-1-yl)piperidin-1-yl)-3,6,9,12,15-pentaoxaheptadecanoic acid (82 mg, 0.11 mmol, 68% yield) as a pale yellow oil. LC / MS method 1: MS(ESI)[M+2H] / 2 + 364.8, rt: 0.977 minutes

[0202] Step 3: (2S,4R)-1-((S)-20-(4-(4-(6-amino-5-((R)-1-(2,6-dichloro-3-fluorophenyl)ethoxy)pyridin-3-yl)-1H-pyrazol-1-yl)piperidin-1-yl)-2-(tert-butyl)-4-oxo-6,9,12,15,18-pentaoxa-3-azaicosanoyl)-4-hydroxy-N-(4-(4-methylthiazol-5-yl)benzyl)pyrrolidine-2-carboxamide To a solution of (R)-17-(4-(4-(6-amino-5-(1-(2,6-dichloro-3-fluorophenyl)ethoxy)pyridin-3-yl)-1H-pyrazol-1-yl)piperidin-1-yl)-3,6,9,12,15-pentaoxaheptadecanoic acid (78 mg, 0.11 mmol) in DMF (1.1 mL) was added DIPEA (66 μL, 0.38 mmol) and (2S,4R)-1-((S)-2-amino-3,3-dimethylbutanoyl)-4-hydroxy-N-(4-(4-methylthiazol-5-yl)benzyl)pyrrolidine-2-carboxamide hydrochloride (55 mg, 0.12 mmol) at room temperature. HATU (41 mg, 0.11 mmol) was then added in one portion, and the resulting yellow solution was stirred at room temperature. After 2 hours, LCMS showed the reaction was complete, and the reaction mixture was purified by reverse-phase column chromatography (5% to 100% MeCN / water, containing 0.1% formic acid). The resulting mixed fractions were concentrated and purified again by reverse-phase column chromatography (5% to 100% MeCN / water at pH 10) to give, after lyophilization, the product: (2S,4R)-1-((S)-20-(4-(4-(6-amino-5-((R)-1-(2,6-dichloro-3-fluorophenyl)ethoxy)pyridin-3-yl)-1H-pyrazol-1-yl)piperidin-1-yl)-2-(tert-butyl)-4-oxo-6,9,12,15,18-pentaoxa-3-azaicosanoyl)-4-hydroxy-N-(4-(4-methylthiazol-5-yl)benzyl)pyrrolidine-2-carboxamide (85 mg, 0.074 mmol, 69% yield) as a white solid. 1H NMR (400MHz, DMSO-d6) δ ppm 8.98(s, 1H), 8.60(t, J=6.2Hz, 1H), 7.95(s, 1H), 7.75(d, J=1.5Hz, 1H), 7.54-7.61(m, 1H), 7.52(s, 1H), 7.37-7.46(m, 6H), 6.89(s, 1H), 6.08(q, J=6.8Hz, 1H), 5.64(s, 2H), 5.15(d, J=3.5Hz, 1H), 4.57(d, J=9.3Hz, 1H), 4.44(m, 3H), 4.21-4.30(m, 1H), 4.02-4.12 (m, 1H), 3.94-3.99 (m, 2H), 3.45-3.71 (m, 22H), 2.91-3.00 (m, 2H), 2.44 (s, 3H), 2.02-2.18 (m, 2H), 1.86-2.00 (m, 4H), 1.78-1.86 (m, 3H), 0.90-0.99 (m, 9H) (two aliphatic protons not detected). LC / MS method 3: MS(ESI)[M+2H] / 2 + 570.8, rt: 2.439 minutes

[0203] Example S6. Preparation of (2S,4R)-1-((S)-23-(4-(4-(6-amino-5-((R)-1-(2,6-dichloro-3-fluorophenyl)ethoxy)pyridin-3-yl)-1H-pyrazol-1-yl)piperidin-1-yl)-2-(tert-butyl)-4-oxo-6,9,12,15,18,21-hexaoxa-3-aza-icosanoyl)-4-hydroxy-N-(4-(4-methylthiazol-5-yl)benzyl)pyrrolidine-2-carboxamide (Compound 6) [ka] To a solution of HATU (23.28 mg, 0.0600 mmol) in DMF (0.5737 mL) was added DIPEA (33.93 μL, 0.1900 mmol) and (R)-3-(1-(2,6-dichloro-3-fluorophenyl)ethoxy)-5-(1-(piperidin-4-yl)-1H-pyrazol-4-yl)pyridin-2-amine hydrochloride (28.59 mg, 0.06 mmol) at room temperature. (R)-20-(4-(4-(6-amino-5-(1-(2,6-dichloro-3-fluorophenyl)ethoxy)pyridin-3-yl)-1H-pyrazol-1-yl)piperidin-1-yl)-3,6,9,12,15,18-hexaoxaarachidic acid (43 mg, 0.06 mmol) was then added in one portion, and the resulting yellow solution was stirred at room temperature. After 2 h, LCMS showed complete conversion. The reaction mixture was purified by reverse-phase column chromatography (5%–100% MeCN / water, 0.1% formic acid). The combined fractions were concentrated and purified by preparative HPLC. The product was subjected to catch-and-release purification on an SPE cartridge (tosylate) to give the desired product (6 mg, 9.0% yield) as a white solid. LC / MS method 3: Retention time: 2.480 min, purity 98.9% (215 nm), [M+2H] / 2 + =592.7, 593.6 1H NMR (400MHz, DMSO-d6) δ ppm 8.98(s, 1H), 8.56-8.62(m, 1H), 7.92-7.97(m, 1H), 7.74(br d, J=1.3Hz, 1H), 7.54-7.59(m, 1H), 7.52(s, 1H), 7.38-7.47(m, 6H), 6.89(s, 1H), 6.08(br d, J=6.3Hz, 1H), 5.64(s, 2H), 5.14(br d, J=3.5Hz, 1H), 4.56(br d, J=9.3Hz, 1H), 4.31-4.48(m, 4H), 4.18-4.30(m, 1H), 4.03-4.14(m, 1H), 3.96(s, 2H), 3.49(br d, J=11.1Hz, 25H), 2.88-3.05(m, 2H), 2.42-2.45(m, 3H), 2.10-2.20(m, 2H), 2.02-2.09(m, 1H), 1.85-2.01(m, 5H), 1.77-1.83(m, 3H), 0.90-0.99(m, 9H)

[0204] Example S7. Preparation of (2S,4R)-1-((S)-26-(4-(4-(6-amino-5-((R)-1-(2,6-dichloro-3-fluorophenyl)ethoxy)pyridin-3-yl)-1H-pyrazol-1-yl)piperidin-1-yl)-2-(tert-butyl)-4-oxo-6,9,12,15,18,21,24-heptaoxa-3-azahexacosanoyl)-4-hydroxy-N-(4-(4-methylthiazol-5-yl)benzyl)pyrrolidine-2-carboxamide (Compound 7) [ka] To a solution of DIPEA (211.25 μL, 1.21 mmol), HATU (131.68 mg, 0.3500 mmol), and (2S,4R)-1-((S)-2-amino-3,3-dimethylbutanoyl)-4-hydroxy-N-(4-(4-methylthiazol-5-yl)benzyl)pyrrolidine-2-carboxamide (22 mg, 0.05 mmol) in DMF (0.44 mL) was added (R)-23-(4-(4-(6-amino-5-(1-(2,6-dichloro-3-fluorophenyl)ethoxy)pyridin-3-yl)-1H-pyrazol-1-yl)piperidin-1-yl)-3,6,9,12,15,18,21-heptaoxatricosanoic acid (35 mg, 0.04 mmol) at room temperature and stirred at room temperature. After 2 hours, LCMS showed complete conversion. The reaction mixture was loaded directly onto a GOLD C18 column (30 g) and purified by reverse-phase column chromatography (5% to 100% MeCN / water, 0.1% formic acid). The combined fractions were concentrated, followed by catch-and-release purification on an SPE cartridge (tosylic acid) to give the desired product (25 mg, 66% purity). This was then purified by preparative HPLC to give the desired product (16.7 mg, 31.2% yield) as an off-white solid. LC / MS method 3: Retention time: 2.456 min, purity 98.7% (215 nm), [M+2H] / 2 + =614.8, 615.7 1H NMR (400MHz, DMSO-d6) δ ppm 8.98(s, 1H), 8.56-8.62(m, 1H), 7.92-7.97(m, 1H), 7.74-7.77(m, 1H), 7.52-7.59(m, 2H), 7.36-7.48(m, 6H), 6.87-6.91(m, 1H), 6.05-6.11(m, 1H), 5.60-5.69(m, 2H), 5.13-5.18(m, 1H), 4.54-4.59(m, 1H), 4.32-4.48(m, 3H), 4.20-4.29(m, 1H), 4.07-4.17 (m, 1H), 3.94-4.00 (m, 2H), 3.46-3.69 (m, 30H), 2.85-3.19 (m, 2H), 2.44 (s, 3H), 1.86-2.12 (m, 6H), 1.80 (d, J=6.6Hz, 3H), 0.94 (s, 9H) (two aliphatic protons are missing). 1 The H NMR spectrum shows impurity peaks at 1.09-1.23 ppm.

[0205] Example S8. Preparation of N-(2-(4-(4-(6-amino-5-((R)-1-(2,6-dichloro-3-fluorophenyl)ethoxy)pyridin-3-yl)-1H-pyrazol-1-yl)piperidin-1-yl)ethyl)-2-((2-(2,6-dioxopiperidin-3-yl)-1,3-dioxoisoindolin-4-yl)oxy)acetamide (Compound 8) [ka] To a solution of 2-((2-(2,6-dioxopiperidin-3-yl)-1,3-dioxoisoindolin-4-yl)oxy)-N-(2-hydroxyethyl)acetamide (20 mg, 0.05 mmol) in DMA (0.85 mL) was added Dess-Martin periodinane (38 mg, 0.091 mmol) in one portion at room temperature and stirred at room temperature. The solution was stirred at room temperature overnight, after which LCMS showed complete oxidation. (R)-3-(1-(2,6-dichloro-3-fluorophenyl)ethoxy)-5-(1-(piperidin-4-yl)-1H-pyrazol-4-yl)pyridin-2-amine (24 mg, 0.05 mmol) was added, followed by sodium triacetoxyborohydride (17 mg, 0.08 mmol). After stirring at room temperature for 2 hours, the mixture showed complete conversion to the desired product. The reaction mixture was purified by reverse phase column chromatography (5% to 100% MeCN / water, 0.1% formic acid) and the fractions were combined and concentrated to give the desired product (14 mg) as a yellow solid. LC / MS method 6: Retention time: 3.183 min, purity 91.7% (215 nm), [M+2H] / 2 + =404.1 1 H NMR (400MHz, DMSO-d6) δ ppm 11.10-11.15(m, 1H), 8.13(s, 1H), 7.95(br s, 1H), 7.79-7.88(m, 1H), 7.74-7.79(m, 1H), 7.55-7.61(m, 2H), 7.51(d, J=7.3Hz, 1H), 7.42-7.48(m, 2H), 6.91(s, 1H), 6.04-6.14(m, 1H), 5.65-5.79(m, 2H), 5.07-5.16(m, 1H), 4.85(br s, 2H), 3.46-3.58(m, 2H), 2.81-2.95(m, 2H), 2.09-2.29(m, 4H), 1.97-2.08(m, 2H), 1.80(d, J=6.8Hz, 3H)( 1 Seven aliphatic protons are missing in the H NMR.)

[0206] Example S9. Preparation of N-(2-(2-(4-(4-(6-amino-5-((R)-1-(2,6-dichloro-3-fluorophenyl)ethoxy)pyridin-3-yl)-1H-pyrazol-1-yl)piperidin-1-yl)ethoxy)ethyl)-2-((2-(2,6-dioxopiperidin-3-yl)-1,3-dioxoisoindolin-4-yl)oxy)acetamide (Compound 9) [ka] To a solution of (R)-3-(1-(2,6-dichloro-3-fluorophenyl)ethoxy)-5-(1-(piperidin-4-yl)-1H-pyrazol-4-yl)pyridin-2-amine (22 mg, 0.05 mmol) in DMSO (0.34 mL) was added 2-((2-(2,6-dioxopiperidin-3-yl)-1,3-dioxoisoindolin-4-yl)oxy)-N-(2-(2-hydroxyethoxy)ethyl)acetamide (20 mg, 0.05 mmol) at room temperature. The solution was stirred overnight at room temperature, and LCMS showed 50–55% conversion. The aldehyde solution was then added dropwise over 3 min to a solution of IBX (17 mg, 0.06 mmol) and sodium triacetoxyborohydride (13 mg, 0.06 mmol) in DCE (0.4 mL). After stirring the mixture at room temperature for 15 minutes, LCMS showed complete conversion of the aldehyde to the desired product. The reaction mixture was purified by reverse-phase column chromatography (5% to 100% MeCN / water, 0.1% formic acid), and the combined fractions were concentrated under reduced pressure to give the desired product (13.09 mg, 29% yield) as a pale yellow solid. LCMS method 6: Retention time: 3.175 min, purity 89.1% (215 nm), [M+H] + =851.2, 853.2 1H NMR (300MHz, DMSO-d6) δ ppm 11.07(s, 1H), 7.91(br t, J=5.6Hz, 1H), 7.86(s, 1H), 7.71-7.78(m, 1H), 7.68(d, J=1.5Hz, 1H), 7.51(dd, J=8.8, 5.0Hz, 1H), 7.44(s, 1H), 7.41(d, J=3.8Hz, 1H), 7.32-7.39(m, 2H), 6.82(d, J=1.5Hz, 1H), 6.01(q, J=6.9Hz, 1H), 5.58(s, 2H), 5.05(dd, J=12.8, 5.4Hz, 1H), 4.73(s, 2H), 3.92-4.06(m, 1H), 3.35-3.53(m, 6H), 2.75-2.94(m, 3H), 2.48-2.58(m, 2H), 1.78-2.11(m, 7H), 1.71-1.77(m, 3H)( 1 Two aliphatic protons are missing in H NMR. 1 H NMR shows traces of formic acid (1.6% w / w).

[0207] Example S10. Preparation of N-(2-(2-(2-(4-(4-(6-amino-5-((R)-1-(2,6-dichloro-3-fluorophenyl)ethoxy)pyridin-3-yl)-1H-pyrazol-1-yl)piperidin-1-yl)ethoxy)ethoxy)ethyl)-2-((2-(2,6-dioxopiperidin-3-yl)-1,3-dioxoisoindolin-4-yl)oxy)acetamide (Compound 10) [ka] To a solution of 2-((2-(2,6-dioxopiperidin-3-yl)-1,3-dioxoisoindolin-4-yl)oxy)-N-(2-(2-(2-hydroxyethoxy)ethoxy)ethyl)acetamide (20 mg, 0.04 mmol) in DMSO (0.40 mL) was added IBX (15.7 mg, 0.06 mmol) in one portion at room temperature and stirred overnight at room temperature. LCMS showed 50-55% conversion. The solution of aldehyde was added dropwise over 3 minutes to a solution of (R)-3-(1-(2,6-dichloro-3-fluorophenyl)ethoxy)-5-(1-(piperidin-4-yl)-1H-pyrazol-4-yl)pyridin-2-amine (20 mg, 0.0400 mmol) and sodium triacetoxyborohydride (11.9 mg, 0.0600 mmol) in DCE (0.44 mL). The resulting mixture was stirred at room temperature. After 15 minutes, LCMS showed that all of the aldehyde had been converted to the desired product. The reaction mixture was purified by reverse-phase column chromatography (5% to 100% MeCN / water, 0.1% formic acid), and the combined fractions were concentrated to give the desired product (7.97 mg, 20% yield) as a pale yellow solid. 1 H NMR (400MHz, DMSO-d6) δ ppm 11.13(s, 1H), 7.99(t, J=5.6Hz, 1H), 7.88-7.95(m, 1H), 7.80(dd, J=8.6, 7.3Hz, 1H), 7.74(s, 1H), 7.57(dd, J=8.9, 5.0Hz, 1H), 7.36-7.53(m, 4H), 6.89(d, J=1.5Hz, 1H), 6.03-6.12(m, 1H), 5.63(s, 2H), 5.05-5.17(m, 1H), 4.79(s, 2H), 4.00-4.12(m, 1H), 3.34-3.60(m, 12H), 2.83-3.01(m, 3H), 2.54-2.64(m, 2H), 1.86-2.15(m, 7H), 1.80(d, J=6.6Hz, 3H) LC / MS method 6: Retention time: 3.201 min, purity 97.8% (215nm), [M+H] + =895.2, 897.2

[0208] Example S11. Preparation of N-(2-(2-(2-(2-(4-(4-(6-amino-5-((R)-1-(2,6-dichloro-3-fluorophenyl)ethoxy)pyridin-3-yl)-1H-pyrazol-1-yl)piperidin-1-yl)ethoxy)ethoxy)ethoxy)ethyl)-2-((2-(2,6-dioxopiperidin-3-yl)-1,3-dioxoisoindolin-4-yl)oxy)acetamide (Compound 11) [ka] To a solution of 2-((2-(2,6-dioxopiperidin-3-yl)-1,3-dioxoisoindolin-4-yl)oxy)-N-(2-(2-(2-(2-hydroxyethoxy)ethoxy)ethoxy)ethyl)acetamide (20 mg, 0.04 mmol) in DMSO (0.37 mL) was added IBX (14.4 mg, 0.05 mmol) in one portion at room temperature, and the mixture was stirred at room temperature overnight. To a solution of (R)-3-(1-(2,6-dichloro-3-fluorophenyl)ethoxy)-5-(1-(piperidin-4-yl)-1H-pyrazol-4-yl)pyridin-2-amine (18.4 mg, 0.04 mmol) and sodium triacetoxyborohydride (10.9 mg, 0.05 mmol) in DCE (0.40 mL) was added the aldehyde solution dropwise over 3 minutes, and the mixture was stirred at room temperature. After 15 minutes, LCMS indicated that all of the aldehyde had been converted to the desired product. The reaction mixture was purified by reverse-phase column chromatography (5% to 100% MeCN / water, 0.1% formic acid), and the combined fractions were concentrated to give the desired product (9.13 mg, 23% yield) as a pale yellow solid. 1H NMR (300MHz, DMSO-d6) δ ppm 11.13(s, 1H), 8.00(br t, J=5.7Hz, 1H), 7.94(s, 1H), 7.77-7.85(m, 1H), 7.74(s, 1H), 7.57(dd, J=9.1, 5.0Hz, 1H), 7.46-7.53(m, 2H), 7.41(t, J=8.5Hz, 2H), 6.88(d, J=1.5Hz, 1H), 6.08(q, J=6.7Hz, 1H), 5.64(s, 2H), 5.11(dd, J=12.8, 5.4Hz, 1H), 4.78(s, 2H), 4.00-4.13(m, 1H), 3.42-3.57(m, 16H), 2.82-3.00(m, 3H), 2.55-2.64(m, 2H), 1.84-2.19(m, 7H), 1.80(d, J=6.5Hz, 3H) LC / MS method 6: Retention time: 3.211 min, purity 92.6% (215nm), [M+H] + =939.1, 941.0

[0209] Example S12. Preparation of N-(14-(4-(4-(6-amino-5-((R)-1-(2,6-dichloro-3-fluorophenyl)ethoxy)pyridin-3-yl)-1H-pyrazol-1-yl)piperidin-1-yl)-3,6,9,12-tetraoxatetradecyl)-2-((2-(2,6-dioxopiperidin-3-yl)-1,3-dioxoisoindolin-4-yl)oxy)acetamide (Compound 12) [ka] To a solution of 2-((2-(2,6-dioxopiperidin-3-yl)-1,3-dioxoisoindolin-4-yl)oxy)-N-(14-hydroxy-3,6,9,12-tetraoxatetradecyl)acetamide (20 mg, 0.036 mmol) in DMSO (340 μL) was added IBX (13.2 mg, 0.047 mmol) in one portion at room temperature and stirred overnight at room temperature. LCMS analysis indicated 50–55% conversion to the desired aldehyde. The resulting crude aldehyde solution was added dropwise over 3 minutes to a solution of (R)-3-(1-(2,6-dichloro-3-fluorophenyl)ethoxy)-5-(1-(piperidin-4-yl)-1H-pyrazol-4-yl)pyridin-2-amine (16.9 mg, 0.036 mmol) and sodium triacetoxyborohydride (10 mg, 0.047 mmol) in DCE (400 μL). The mixture was stirred at room temperature for 15 minutes, after which LCMS analysis indicated that all of the aldehyde had been converted to the desired product. The reaction mixture was purified by reverse-phase column chromatography (5% to 100% MeCN / water, 0.1% formic acid). The resulting material was further purified by preparative LCMS to give the product, N-(14-(4-(4-(6-amino-5-((R)-1-(2,6-dichloro-3-fluorophenyl)ethoxy)pyridin-3-yl)-1H-pyrazol-1-yl)piperidin-1-yl)-3,6,9,12-tetraoxatetradecyl)-2-((2-(2,6-dioxopiperidin-3-yl)-1,3-dioxoisoindolin-4-yl)oxy)acetamide (9 mg, 0.0087 mmol, 24% yield) as a pale yellow solid after lyophilization. 1H NMR (400MHz, DMSO-d6) δ ppm 11.12(s, 1H), 7.99(t, J=4.9Hz, 1H), 7.94(s, 1H), 7.78-7.84(m, 1H), 7.74(d, J=1.7Hz, 1H), 7.54-7.60(m, 1H), 7.37-7.52(m, 4H), 6.89(d, J=1.7Hz, 1H), 6.08(q, J=6.9Hz, 1H), 5.63(s, 2H), 5.11(dd, J=13.0, 5.4Hz, 1H), 4.78(s, 2H), 4.00-4.14(m, 1H), 3.43-3.57(m, 20H), 2.83-3.01(m, 3H), 2.54-2.68(m, 2H), 1.84-2.19(m, 7H), 1.80(d, J=6.6Hz, 3H) LC / MS method 6: MS(ESI)[M+2H] / 2 + 493.1, rt: 3.214 minutes

[0210] Example S13. Preparation of N-(2-(4-(4-(6-amino-5-((R)-1-(2,6-dichloro-3-fluorophenyl)ethoxy)pyridin-3-yl)-1H-pyrazol-1-yl)piperidin-1-yl)ethyl)-2-((2-(2,6-dioxopiperidin-3-yl)-1,3-dioxoisoindolin-5-yl)oxy)acetamide (Compound 13) [ka] To a solution of 2-((2-(2,6-dioxopiperidin-3-yl)-1,3-dioxoisoindolin-5-yl)oxy)-N-(2-hydroxyethyl)acetamide (20 mg, 0.05 mmol) in DMA (0.85 mL) was added Dess-Martin periodinane (38.4 mg, 0.091 mmol) in one portion at room temperature and stirred overnight at room temperature. LCMS showed that the oxidation was nearly complete. (R)-3-(1-(2,6-dichloro-3-fluorophenyl)ethoxy)-5-(1-(piperidin-4-yl)-1H-pyrazol-4-yl)pyridin-2-amine (24 mg, 0.05 mmol) was added, followed by sodium triacetoxyborohydride (16.9 mg, 0.08 mmol), and the mixture was stirred at room temperature for 1 hour. The reaction mixture was purified by reverse phase column chromatography (5% to 100% MeCN / water, 0.1% formic acid) and the fractions were combined and concentrated to give the desired product (7.04 mg, 16% yield) as an off-white solid. LC / MS method 6: Retention time: 3.176 min, purity 98.0% (215 nm), [M+H] + =807.1, [M+2H] / 2 + =404.8 1 H NMR (400MHz, DMSO-d6) δ ppm 11.11(s, 1H), 7.94(s, 1H), 7.88(d, J=8.3Hz, 1H), 7.75(s, 1H), 7.52-7.61(m, 2H), 7.38-7.48(m, 4H), 6.90(d, J=1.5Hz, 1H), 6.08(d, J=6.8Hz, 1H), 5.65(br. s, 2H), 5.08-5.16(m, 1H), 4.75(s, 2H), 2.92(s, 4H), 2.04(s, 6H), 1.80(d, J=6.6Hz, 3H)( 1 The H NMR spectrum shows the loss of seven aliphatic protons. 1 H NMR shows traces of formic acid (2.2% w / w).

[0211] Example S14. Preparation of N-(2-(2-(4-(4-(6-amino-5-((R)-1-(2,6-dichloro-3-fluorophenyl)ethoxy)pyridin-3-yl)-1H-pyrazol-1-yl)piperidin-1-yl)ethoxy)ethyl)-2-((2-(2,6-dioxopiperidin-3-yl)-1,3-dioxoisoindolin-5-yl)oxy)acetamide (Compound 14) [ka] IBX (17.3 mg, 0.060 mmol) was added in one portion to a solution of 2-((2-(2,6-dioxopiperidin-3-yl)-1,3-dioxoisoindolin-5-yl)oxy)-N-(2-(2-hydroxyethoxy)ethyl)acetamide (20 mg, 0.050 mmol) in DMSO (0.45 mL) at room temperature and stirred overnight at room temperature. The aldehyde solution was added dropwise over 3 minutes to a solution of (R)-3-(1-(2,6-dichloro-3-fluorophenyl)ethoxy)-5-(1-(piperidin-4-yl)-1H-pyrazol-4-yl)pyridin-2-amine (22.1 mg, 0.050 mmol) and sodium triacetoxyborohydride (13.1 mg, 0.060 mmol) in DCE (0.48 mL), and the mixture was stirred at room temperature. After 15 minutes, it was observed that all of the aldehyde had been converted to the desired product. The reaction mixture was purified by reverse-phase column chromatography (5% to 100% MeCN / water, 0.1% formic acid), and the combined fractions were concentrated to give the desired product (9.09 mg, 22% yield) as a pale yellow solid. LC / MS method 6: Retention time: 3.158 min, purity 97.6% (215 nm), [M+H] + =851.2, 853.1 1H NMR (400MHz, DMSO-d6) δ ppm 11.11(s, 1H), 8.24(br t, J=5.4Hz, 1H), 7.92-7.96(m, 1H), 7.86(d, J=8.3Hz, 1H), 7.74(s, 1H), 7.57(dd, J=9.0, 4.9Hz, 1H), 7.51(s, 1H), 7.34-7.48(m, 3H), 6.89(d, J=1.7Hz, 1H), 6.08(d, J=6.8Hz, 1H), 5.64(s, 2H), 5.05-5.17(m, 1H), 4.70-4.79(m, 2H), 3.98-4.12(m, 1H), 3.36-3.57(m, 8H), 2.84-2.99(m, 3H), 2.54-2.63(m, 2H), 1.86-2.17(m, 7H), 1.80(d, J=6.6Hz, 3H)

[0212] Example S15. Preparation of N-(2-(2-(2-(4-(4-(6-amino-5-((R)-1-(2,6-dichloro-3-fluorophenyl)ethoxy)pyridin-3-yl)-1H-pyrazol-1-yl)piperidin-1-yl)ethoxy)ethoxy)ethyl)-2-((2-(2,6-dioxopiperidin-3-yl)-1,3-dioxoisoindolin-5-yl)oxy)acetamide (Compound 15) [ka] To a solution of 2-((2-(2,6-dioxopiperidin-3-yl)-1,3-dioxoisoindolin-5-yl)oxy)-N-(2-(2-(2-hydroxyethoxy)ethoxy)ethyl)acetamide (20 mg, 0.043 mmol) in DMSO (255 μL) at room temperature was added IBX (15.7 mg, 0.056 mmol) in one portion, and the solution was stirred at room temperature overnight. LCMS analysis showed 50% conversion. The resulting crude aldehyde solution was added dropwise over 3 minutes to a solution of (R)-3-(1-(2,6-dichloro-3-fluorophenyl)ethoxy)-5-(1-(piperidin-4-yl)-1H-pyrazol-4-yl)pyridin-2-amine (20 mg, 0.045 mmol) and sodium triacetoxyborohydride (11.9 mg, 0.056 mmol) in DCE (340 μL), and the mixture was stirred at room temperature for 15 minutes. The reaction mixture was purified by reverse-phase column chromatography (5% to 100% MeCN / water, 0.1% formic acid) and the resulting material was further purified by preparative LCMS to afford, after lyophilization, the product: N-(2-(2-(2-(4-(4-(6-amino-5-((R)-1-(2,6-dichloro-3-fluorophenyl)ethoxy)pyridin-3-yl)-1H-pyrazol-1-yl)piperidin-1-yl)ethoxy)ethoxy)ethyl)-2-((2-(2,6-dioxopiperidin-3-yl)-1,3-dioxoisoindolin-5-yl)oxy)acetamide (11.1 mg, 0.012 mmol, 28% yield) as a white solid. 1H NMR (400MHz, DMSO-d6) δ ppm 11.08-11.19(m, 1H), 8.23-8.28(m, 1H), 7.94(s, 1H), 7.86(d, J=8.3Hz, 1H), 7.75(d, J=1.5Hz, 1H), 7.57(dd, J=8.9, 5.0Hz, 1H), 7.52(s, 1H), 7.42-7.47(m, 2H), 7.38(dd, J=8.3, 2.2Hz, 1H), 6.89(d, J=1.5Hz, 1H), 6.08 (q = J=6.4Hz, 1H), 5.6(s, 2H), 5.10-5.15(m, 1H), 4.74(s, 2H), 4.03-4.11(m, 1H), 3.51(s, 10H), 3.29-3.33(m, 2H), 2.83-2.96(m, 3H), 2.54-2.64(m, 2H), 2.12(d, J=2.4Hz, 3H) 1.94(s, 4H), 1.80(d, J=6.6Hz, 3H) LC / MS method 3: MS(ESI)[M+H] + 897.3, rt: 2.154 minutes

[0213] Example S16. Preparation of N-(2-(2-(2-(2-(4-(4-(6-amino-5-((R)-1-(2,6-dichloro-3-fluorophenyl)ethoxy)pyridin-3-yl)-1H-pyrazol-1-yl)piperidin-1-yl)ethoxy)ethoxy)ethoxy)ethyl)-2-((2-(2,6-dioxopiperidin-3-yl)-1,3-dioxoisoindolin-5-yl)oxy)acetamide (Compound 16) [ka] To a solution of 2-((2-(2,6-dioxopiperidin-3-yl)-1,3-dioxoisoindolin-5-yl)oxy)-N-(2-(2-(2-(2-hydroxyethoxy)ethoxy)ethoxy)ethyl)acetamide (20 mg, 0.040 mmol) in DMSO (0.30 mL) was added IBX (14.4 mg, 0.050 mmol) in one portion at room temperature, and the mixture was stirred at room temperature overnight. The solution of the aldehyde was added dropwise over 3 minutes to a solution of (R)-3-(1-(2,6-dichloro-3-fluorophenyl)ethoxy)-5-(1-(piperidin-4-yl)-1H-pyrazol-4-yl)pyridin-2-amine (18.4 mg, 0.040 mmol) and sodium triacetoxyborohydride (10.9 mg, 0.050 mmol) in DCE (0.40 mL), and the mixture was stirred at room temperature. After 15 minutes, LCMS indicated that all of the aldehyde had been converted to the desired product. The reaction mixture was purified by reverse-phase column chromatography (5% to 100% MeCN / water, 0.1% formic acid), and the combined fractions were concentrated to give the desired product (6.1 mg, 15% yield) as a pale yellow solid. 1 H NMR (300MHz, DMSO-d6) δ ppm 11.12(s, 1H), 8.20-8.30(m, 1H), 7.93-7.97(m, 1H), 7.86(d, J=8.5Hz, 1H), 7.74(d, J=1.5Hz, 1H), 7.36-7.63(m, 5H), 6.89(d, J=1.5Hz, 1H), 6.01-6.14(m, 1H), 5.64(s, 2H), 5.12(dd, J=12.8, 5.4Hz, 1H), 4.67-4.80(m, 2H), 3.98-4.16(m, 1H), 3.42-3.64(m, 16H), 2.84-3.03(m, 3H), 2.58-2.76(m, 2H), 1.86-2.20(m, 7H), 1.80(d, J=6.8Hz, 3H) LC / MS method 6: Retention time: 3.190 min, purity 91.9% (215 nm), [M+H] + =939.1, 941.0

[0214] Example 17 Preparation of N-(14-(4-(4-(6-amino-5-((R)-1-(2,6-dichloro-3-fluorophenyl)ethoxy)pyridin-3-yl)-1H-pyrazol-1-yl)piperidin-1-yl)-3,6,9,12-tetraoxatetradecyl)-2-((2-(2,6-dioxopiperidin-3-yl)-1,3-dioxoisoindolin-5-yl)oxy)acetamide (Compound 17) [ka] To a solution of 2-((2-(2,6-dioxopiperidin-3-yl)-1,3-dioxoisoindolin-5-yl)oxy)-N-(14-hydroxy-3,6,9,12-tetraoxatetradecyl)acetamide (20 mg, 0.040 mmol) in DMSO (0.30 L) was added IBX (13.2 mg, 0.050 mmol) in one portion at room temperature, and the mixture was stirred at room temperature overnight. The solution of the aldehyde was added dropwise over 3 minutes to a solution of (R)-3-(1-(2,6-dichloro-3-fluorophenyl)ethoxy)-5-(1-(piperidin-4-yl)-1H-pyrazol-4-yl)pyridin-2-amine (16.9 mg, 0.040 mmol) and sodium triacetoxyborohydride (9.99 mg, 0.050 mmol) in DCE (0.40 mL), and the mixture was stirred at room temperature. After 15 minutes, LCMS showed that all of the aldehyde had been converted to the desired product. The reaction mixture was purified by reverse-phase column chromatography (5% to 100% MeCN / water, 0.1% formic acid), and the combined fractions were concentrated to give the desired product (5.48 mg, 14.0% yield) as a pale yellow solid. 1H NMR (300MHz, DMSO-d6) δ ppm 11.12(s, 1H), 8.20-8.34(m, 1H), 7.92-7.98(m, 1H), 7.86(d, J=8.2Hz, 1H), 7.74(d, J=1.5Hz, 1H), 7.34-7.61(m, 5H), 6.89(d, J=1.5Hz, 1H), 6.08(d, J=7.0Hz, 1H), 5.64(s, 2H), 5.03-5.25(m, 1H), 4.63-4.84(m, 2H), 3.97-4.17(m, 1H), 3.39-3.68(m, 20H), 2.79-3.00(m, 3H), 2.58-2.75(m, 2H), 1.85-2.20(m, 7H), 1.80(d, J=6.8Hz, 3H) LC / MS method 6: Retention time: 3.194 min, purity 93.6% (215nm), [M+2H] 2+ =92.9

[0215] Example S18. Preparation of 5-(9-(4-(4-(6-amino-5-((R)-1-(2,6-dichloro-3-fluorophenyl)ethoxy)pyridin-3-yl)-1H-pyrazol-1-yl)piperidin-1-yl)-3-azaspiro[5.5]undecane-3-carbonyl)-2-(2,6-dioxopiperidin-3-yl)isoindoline-1,3-dione (Compound 18) [ka] A solution of (R)-5-(1-(1-(3-azaspiro[5.5]undecan-9-yl)piperidin-4-yl)-1H-pyrazol-4-yl)-3-(1-(2,6-dichloro-3-fluorophenyl)ethoxy)pyridin-2-amine (bis(hydrochloride)) (50 mg, 0.07 mmol) in DMF (750 μL) was added to 2-(2,6-dioxo-3-piperidyl)-1,3-dioxo-isoindoline-5-carboxylic acid (22 mg, 0.07 mmol), DIPEA (60 μL, 0.37 mmol), and HATU (42 mg, 0.11 mmol) and stirred under N2 for 1 h. The mixture was then purified by reverse-phase column chromatography (5-100% MeCN / water, 0.1% formic acid) to obtain mixed fractions. This was recombined and repurified by reverse phase column chromatography (5-100% MeCN / water) to give the product, after lyophilization: 5-(9-(4-(4-(6-amino-5-((R)-1-(2,6-dichloro-3-fluorophenyl)ethoxy)pyridin-3-yl)-1H-pyrazol-1-yl)piperidin-1-yl)-3-azaspiro[5.5]undecane-3-carbonyl)-2-(2,6-dioxopiperidin-3-yl)isoindoline-1,3-dione (6.3 mg, 9% yield) as a white solid. 1H NMR (400MHz, DMSO-d6) δ ppm 11.15(s, 1H), 7.97(br dd, J=15.7, 7.1Hz, 2H), 7.80-7.91(m, 2H), 7.74(br s, 1H), 7.49-7.61(m, 2H), 7.44(br t, J=8.9Hz, 1H), 6.89(br s, 1H), 6.08(br d, J=6.1Hz, 1H), 5.63(br s, 2H), 5.12-5.23(m, 1H), 3.98-4.17(m, 1H), 3.57-3.70(m, 2H), 3.24(brd, J=2.4Hz, 2H), 2.83-3.05(m, 3H), 2.55-2.70(m, 2H), 2.26-2.42(m, 3H), 1.95-2.12(m, 3H), 1.84-1.93(m, 2H), 1.76-1.83(m, 4H), 1.53-1.67(m, 3H), 1.31-1.49(m, 4H), 1.19-1.31(m, 2H), 1.04-1.19(m, 2H) LC / MS method 3: MS(ESI)[M+2H] / 2 + 444.0, rt: 2.124 minutes

[0216] Example S19. Preparation of 3-(6-(9-(4-(4-(6-amino-5-((R)-1-(2,6-dichloro-3-fluorophenyl)ethoxy)pyridin-3-yl)-1H-pyrazol-1-yl)piperidin-1-yl)-3-azaspiro[5.5]undecane-3-carbonyl)-1-oxoisoindolin-2-yl)piperidine-2,6-dione (Compound 19) [ka] HATU (11.2 mg, 0.03 mmol) and 2-(2,6-dioxo-3-piperidyl)-3-oxo-isoindoline-5-carboxylic acid (8.5 mg, 0.03 mmol) were added to a vial (1 mL) containing a stir bar, followed by DMF (350 μL) and DIPEA (11 μL, 0.11 mmol). The mixture was stirred for 5 minutes, and (R)-5-(1-(1-(3-azaspiro[5.5]undecan-9-yl)piperidin-4-yl)-1H-pyrazol-4-yl)-3-(1-(2,6-dichloro-3-fluorophenyl)ethoxy)pyridin-2-amine (20 mg, 0.03 mmol) was added. The vial was then sealed and stirred overnight at room temperature. LCMS analysis showed complete conversion of the starting material to the desired product. The reaction mixture was then purified by preparative HPLC / MS (added acid: formic acid) to give, after lyophilization, the product: 3-(6-(9-(4-(4-(6-amino-5-((R)-1-(2,6-dichloro-3-fluorophenyl)ethoxy)pyridin-3-yl)-1H-pyrazol-1-yl)piperidin-1-yl)-3-azaspiro[5.5]undecane-3-carbonyl)-1-oxoisoindolin-2-yl)piperidine-2,6-dione formate salt (10.6 mg, 0.12 mmol, 43% yield) as an off-white solid. LC / MS method 2: MS(ESI)[M+2H] / 2 + 436.6, rt: 1.00 minutes

[0217] Example S20. Preparation of 3-(5-(9-(4-(4-(6-amino-5-((R)-1-(2,6-dichloro-3-fluorophenyl)ethoxy)pyridin-3-yl)-1H-pyrazol-1-yl)piperidin-1-yl)-3-azaspiro[5.5]undecane-3-carbonyl)-1-oxoisoindolin-2-yl)piperidine-2,6-dione (Compound 20) 11.2 mg (0.03 mmol) of HATU and 8.5 mg (0.03 mmol) of 2-(2,6-dioxo-3-piperidyl)-1-oxo-isoindoline-5-carboxylic acid were added to a 1 mL vial containing a stir bar, followed by 350 μL of DMF, followed by 11 μL (0.11 mmol) of DIPEA. The reaction mixture was stirred for 5 min, and then 20 mg (0.03 mmol) of (R)-5-(1-(1-(3-azaspiro[5.5]undecan-9-yl)piperidin-4-yl)-1H-pyrazol-4-yl)-3-(1-(2,6-dichloro-3-fluorophenyl)ethoxy)pyridin-2-amine was added. The vial was then sealed with a cap and stirred overnight at room temperature. LCMS analysis showed that the starting material was completely converted to the desired product. The reaction mixture was then purified by preparative HPLC / MS (added acid: formic acid) to give the product, 3-(5-(9-(4-(4-(6-amino-5-((R)-1-(2,6-dichloro-3-fluorophenyl)ethoxy)pyridin-3-yl)-1H-pyrazol-1-yl)piperidin-1-yl)-3-azaspiro[5.5]undecane-3-carbonyl)-1-oxoisoindolin-2-yl)piperidine-2,6-dione formate salt (13 mg, 0.14 mmol, 53% yield) as an off-white solid after lyophilization. 1H NMR (400MHz, DMSO-d6) δ ppm 11.00(s, 1H), 7.95(br s, 1H), 7.78(d, J=7.8Hz, 1H), 7.74(s, 1H), 7.61(s, 1H), 7.57(dd, J=8.9, 5.0Hz, 1H), 7.47-7.53(m, 2H), 7.41-7.47(m, 1H), 6.89(s, 1H), 6.08(q, J=6.4Hz, 1H), 5.63(s, 2H), 5.13(dd, J=13.2, 5.1Hz, 1H), 4.50(d, J=18.3Hz, 1H), 4.37(d, J=18.1Hz, 1H), 4.04(br s, 1H), 3.61(br s, 2H), 3.17-3.28(m, 2H), 2.85-3.00(m, 3H), 2.55-2.65(m, 1H), 2.31(br s, 4H), 1.92-2.06(m, 3H), 1.82-1.92(m, 2H), 1.80(d, J=6.6Hz, 4H), 1.71-1.77(m, 1H), 1.50-1.64(m, 3H), 1.29-1.48(m, 4H), 1.20-1.28(m, 1H), 1.03-1.19(m, 2H) LC / MS method 2: MS(ESI)[M+2H] / 2 + 436.5, rt: 1.00 minutes

[0218] Example S21. Preparation of 3-(4-(9-(4-(4-(6-amino-5-((R)-1-(2,6-dichloro-3-fluorophenyl)ethoxy)pyridin-3-yl)-1H-pyrazol-1-yl)piperidin-1-yl)-3-azaspiro[5.5]undecane-3-carbonyl)-1-oxoisoindolin-2-yl)piperidine-2,6-dione (Compound 21) [ka] HATU (11.2 mg, 0.03 mmol) and 2-(2,6-dioxo-3-piperidyl)-1-oxo-isoindoline-4-carboxylic acid (8.5 mg, 0.03 mmol) were added to a vial (1 mL) containing a stir bar, followed by DMF (350 μL) and DIPEA (11 μL, 0.11 mmol). The reaction mixture was stirred for 5 minutes, and (R)-5-(1-(1-(3-azaspiro[5.5]undecan-9-yl)piperidin-4-yl)-1H-pyrazol-4-yl)-3-(1-(2,6-dichloro-3-fluorophenyl)ethoxy)pyridin-2-amine (20 mg, 0.03 mmol) was added. The vial was then capped and stirred at room temperature overnight. LCMS analysis showed complete conversion of the starting material to the desired product. The reaction mixture was then purified by preparative HPLC / MS (added acid: formic acid) to give, after lyophilization, the formate salt of the product: 3-(4-(9-(4-(4-(6-amino-5-((R)-1-(2,6-dichloro-3-fluorophenyl)ethoxy)pyridin-3-yl)-1H-pyrazol-1-yl)piperidin-1-yl)-3-azaspiro[5.5]undecane-3-carbonyl)-1-oxoisoindolin-2-yl)piperidine-2,6-dione (13 mg, 0.14 mmol, 52% yield) as an off-white solid. LC / MS method 2: MS(ESI)[M+2H] / 2 + 436.6, rt: 1.01 minutes

[0219] Example S22. Preparation of 3-((3-(9-(4-(4-(6-amino-5-((R)-1-(2,6-dichloro-3-fluorophenyl)ethoxy)pyridin-3-yl)-1H-pyrazol-1-yl)piperidin-1-yl)-3-azaspiro[5.5]undecane-3-carbonyl)phenyl)amino)piperidine-2,6-dione (Compound 22) [ka] To a solution of 3-[(2,6-dioxo-3-piperidyl)amino]benzoic acid (502 mg, 2.0 mmol) in DMF (8 mL) was added DIPEA (1.6 mL, 9.2 mmol) and 5-[1-[1-(3-azaspiro[5.5]undecan-9-yl)-4-piperidyl]pyrazol-4-yl]-3-[(1R)-1-(2,6-dichloro-3-fluoro-phenyl)ethoxy]pyridin-2-amine hydrochloride (1.17 g, 1.84 mmol) at room temperature and stirred for 10 minutes at room temperature. PyAOP (1.15 g, 2.21 mmol) was then added in one portion, and the mixture was stirred at room temperature. After 3 hours, LCMS showed complete conversion. The mixture was allowed to stand at room temperature overnight and then purified by reverse-phase column chromatography (5% MeCN to 100% MeCN / water, containing 0.1% HCOOH) to give 3-((3-(9-(4-(4-(6-amino-5-((R)-1-(2,6-dichloro-3-fluorophenyl)ethoxy)pyridin-3-yl)-1H-pyrazol-1-yl)piperidin-1-yl)-3-azaspiro[5.5]undecane-3-carbonyl)phenyl)amino)piperidine-2,6-dione formate salt (ratio=1:1.2, 370 mg, 0.44 mmol, 24% yield) as a pale yellow solid. LC / MS method 3: MS(ESI)[M+H] + 831.2, rt: 2.26 minutes

[0220] Biological Examples Example B1. IRAK3-ePL overexpression degradation assay Stable cell lines were generated using the following protocol: 3 × 10 5Lenti-X 293T cells (Clonetech) were plated in 0.8 mL of medium in a 12-well plate and incubated overnight at 37°C / 5% CO2. The packaging plasmid (0.4 μg, pMD), envelope plasmid (0.4 μg, pSP), and lentiviral transduction 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. At the same time, 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 Lipofectamine were combined and the mixture was incubated for 20 minutes. The DNA:Lipofectamine, Opti-MEM mixture was then added dropwise to the pre-plated cells, and the cells were incubated at 37°C / 5% CO2 for ~16 hours. After incubation, the medium in each well was removed and fresh medium (1.2 mL) was added. Lenti-X 293T cells were incubated at 37°C / 5% CO2 for ~30 hours. CRBN OE / GSPT1 G575N KI cells (0.5×10 6 The virus (0.5 mL) was plated in medium (0.5 mL / well) in a 12-well plate and incubated at 37°C / 5% CO2 for ~16 hours. After incubation, the medium was removed from the wells of the Lenti-X 293T and filtered through a 0.45 μM filter. A portion of the viral supernatant was used to transduce cells, and the remainder was stored at -80°C. The virus (0.5 mL) was then transferred to the 293T CRBN OE / GSPT1 G575N KICells were then added to each well, followed by the addition of polybrene (10 mg / mL, Millipore) to a final concentration of 5.0 μg / mL. The cells were then incubated at 37°C / 5% CO2 for ~24 hours. After the medium was aspirated from the plate, the cells were washed with DPBS, trypsinized, and plated in 10 cm Petri dishes in 15 mL of medium supplemented with puromycin (1 μg / mL). After incubation at 37°C / 5% CO2 for ~72 hours, the medium was aspirated from the plate, washed with DPBS, and trypsinized. The cells were plated in 15 cm Petri dishes in 40 mL of medium supplemented with 1.0 μg / mL puromycin (Gibco) and incubated at 37°C / 5% CO2 for ~72 hours. After incubation, the medium was removed, the cells were washed with DPBS, and trypsinized. Most of the cells were resuspended in freezing medium (Invitrogen) and stored (~6-8 × 10 6 cells / vial).

[0221] The IRAK3-ePL cell dose-response curve resolution assay was performed using the following protocol. Test compounds were dispensed into a white 384-well tissue culture-treated plate using an acoustic liquid handler. Based on the assay volume (25 μL), 10 concentrations were prepared in duplicate, with 3-fold serial dilutions starting from a 10 μM dose. To calculate 100% signal, negative control wells containing 0.2% DMSO alone were prepared, and positive control wells containing ataluren (a luciferase inhibitor, 30 μM) were prepared to calculate background signal levels. To ensure uniform DMSO concentrations across wells, all wells were adjusted to a final DMSO concentration of 0.2%. IRAK3-ePL expressing cells (IRAK3-ePL Lenti-X 293T) were cultured in 10 wells. CRBN / GSPT1 G575N) were washed, trypsinized, and the cells were 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 dispensed into wells of a 384-well plate to which compounds had been added in the previous step and incubated overnight at 37°C / 5% CO2. After incubation, the 384-well plate was removed from the incubator and left at room temperature for 30 minutes. InCELL Hunter reagent (EA reagent, lysis buffer, and substrate reagent in a 1:1:4 ratio, Cat# 96-0002, DiscoverX) was prepared according to the manufacturer's instructions and added to the 384-well plate at 25 μL / well. After incubating the plate at room temperature for 1 hour, the luminescence signal was read using a ViewLux plate reader. Data were processed and analyzed using ActivityBase software. Briefly, to correct for background, the mean luminescence value of the positive control wells was subtracted from the remaining wells, and all luminescence values were normalized to the DMSO control wells. The mean value of the DMSO control wells was set equal to 100% relative IRAK3-ePL protein level. The normalized luminescence values were plotted on a graph as a function of compound concentration. Compound concentration was plotted on the x-axis and the corresponding relative IRAK3-ePL protein level on the y-axis. The EC of compounds to degrade IRAK3-ePL was calculated. 50 The values (median effective concentrations) were calculated using a four-variable 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 bounds of the FIT, respectively. Determine the maximum rate of loss of target protein after compound treatment, D max was calculated.

[0222] Example B2. IRAK endogenous HTRF degradation assay Cells (~50k) were plated in a 96-well small-volume white plate (Cisbio: cat# 66PL96005). Compounds were dissolved in DMSO and serially diluted 3-fold using a TECAN D300E. Cells were incubated with compounds overnight. The Total-IRAK3 HTRF kit (Cisbio: 63ADK101PEH) was used for degradation analysis. Cryptate and D2 antibodies were diluted in detection buffer according to the manufacturer's recommendations. 2µL of each solution was then added to the lysate (16µL). Buffer controls (lysis buffer, detection buffer), cryptate controls (lysis buffer + cryptate antibody + detection buffer), and negative controls (lysis buffer + cryptate antibody + D2 antibody) were prepared according to the manufacturer's recommendations. After incubation with antibody, HTRF signals were measured on an Envision plate reader (PerkinElmer) and HTRF signals were calculated using the formula: (emission at 665 nm / emission at 615 nm) * 10,000. All HTRF values were normalized to the mean value of DMSO. The mean value of DMSO control wells was set equal to 100% relative IRAK3 protein levels. Normalized luminescence values were plotted on a graph as a function of compound concentration. Compound concentration was plotted on the x-axis and corresponding normalized IRAK3 protein levels on the y-axis. EC of compounds to degrade IRAK3 50 The values (median effective concentrations) were calculated using a four-variable 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 FIT, respectively. By determining the lowest percentage of target protein remaining after compound treatment, Y min was calculated. D max is Y min (%D max =100-Y min ) was calculated.

[0223] Example B3. IRAK3 Biochemical Binding Assay The LanthaScreen® Eu Kinase Binding Assay was performed as described by the vendor (ThermoFisher Scientific, Waltham, MA). Briefly, a 10 mM stock solution was serially diluted 3-fold with DMSO to create a 100x compound solution in a 384-well reagent plate to adjust the final concentration. Serial compound dilutions (1 μL) were added to corresponding wells of a 384-well reagent plate containing 32.3 μL of 1x buffer (50 mM HEPES (pH 7.4), 10 nM MgCl2, 1 mM EGTA, 0.01% Brij-35). Five μL of buffer-diluted compound was transferred to the corresponding wells of a 384-well assay plate. Five μL of 3x-diluted tracer was transferred to each well of the assay plate (final tracer concentration: 10 nM). Finally, a mixture of 3x-diluted Eu anti-GST and IRAK3 (5 μL) was transferred to each well at final concentrations of 2 nM and 10 nM, respectively. The reaction was incubated at room temperature for 1 hour. The TR-FRET signal (λ ex 340 / λ em 665 / λ em 615) were read at room temperature using an Envision plate reader with a delay time of 100 μs and an integration time of 200 μs. The background-corrected luminescence signal ratio for each compound concentration was used to calculate the percentage of inhibition (% inhibition). Plots of % inhibition versus inhibitor concentration were fitted according to the dose-response equation (Equation 1), and IC values were calculated using Dotmatics software (Dotmatics, Bishops Stortford, Hertfordshire, England). 50 and the slope of the curve was calculated. [ka]

[0224] Using these assays, the IC of the following compounds was determined: 50 , D max , E.C. 50 , and D.C. 50 The value of D was determined. max is the maximum degradation rate achieved, DC 50is defined as the concentration at which 50% degradation is achieved. The data are summarized in Table 2. [Table 6]

[0225] For purposes of clarity of understanding, the present invention has been described in some detail by way of illustration and example, but such illustrations and examples should not be construed as limiting the scope of the invention. The disclosures of all patent and scientific literature cited herein are expressly incorporated herein by reference in their entireties.

Claims

1. Equation (IA): 【Chemistry 1】 [In the formula, L 1 is, -(CH 2 CH 2 O) n -or combination; n is between 1 and 10; L 2 is C 1 -C 6 alkylene, -(C 1 -C 6 alkylene)N(H)-, a 6- to 12-membered spiroheterocyclylene, or a bond, where the heterocyclylene contains 1 to 3 heteroatoms selected from N and O; D is 【Chemistry 2】 and; R 1a and R 1b Each is either H or together forms an oxo; and L 3 is, -CH 2 [O- or bonded] Compounds of or pharmaceutically acceptable salts thereof.

2. The compound is given by formula (I): 【Transformation 3】 [In the formula, L 1 is, -(CH 2 CH 2 O) n -or combination; n is between 1 and 10; L 2 C 1 -C 6 Alkylene, -(C 1 -C 6 Alkylene)N(H)-, a 6- to 12-membered spiroheterocyclylene, or bond, where the heterocyclylene contains 1 to 3 heteroatoms selected from N and O; D is 【Chemistry 4】 and; R 1a and R 1b Each is either H or together forms an oxo; and L 3 is, -CH 2 [O- or bonded] The compound according to claim 1, or a pharmaceutically acceptable salt thereof.

3. In the formula, L 1 However, -(CH 2 CH 2 O) n - and n is 1 to 7, the compound according to claim 1, or a pharmaceutically acceptable salt thereof.

4. In the formula, L 1 The compound according to claim 1, or a pharmaceutically acceptable salt thereof, wherein the bond is a bond.

5. In the formula, L 2 However, C 1 -C 3 Alkylene, -(C 1 -C 3 The compound according to claim 1, or a pharmaceutically acceptable salt thereof, comprising an alkylene)N(H)-, a 10-12 membered spiroheterocyclylene, or a bond, wherein the heterocyclylene contains 1-2 nitrogen atoms.

6. In the formula, -L 1 -L 2 -but, 【Transformation 5】 The compound according to claim 1, or a pharmaceutically acceptable salt thereof.

7. In the formula, D 【Transformation 6】 The compound according to claim 1, or a pharmaceutically acceptable salt thereof.

8. In the formula, D 【Transformation 7】 The compound according to claim 1, or a pharmaceutically acceptable salt thereof.

9. In the formula, R 1a and R 1b The compound according to claim 8, or a pharmaceutically acceptable salt thereof, wherein each of the elements is H.

10. In the formula, R 1a and R 1b The compound according to claim 8, or a pharmaceutically acceptable salt thereof, wherein the compounds together form an oxo.

11. In the formula, L 3 ga-CH 2 The compound according to claim 8, or a pharmaceutically acceptable salt thereof, which is O-.

12. In the formula, L 3 The compound according to claim 8, or a pharmaceutically acceptable salt thereof, wherein the bond is a linkage.

13. During the ceremony, 【Transformation 8】 but, 【Chemistry 9】 The compound according to claim 8, or a pharmaceutically acceptable salt thereof.

14. In the formula, D 【Chemistry 10】 The compound according to claim 1, or a pharmaceutically acceptable salt thereof.

15. The compound is of formula (II), (III), or (VII): 【Chemistry 11】 [In the formula, L 1 is, -(CH 2 CH 2 O) n -or combination; n is between 1 and 10; and L 2 C 1 -C 6 It is alkylene. 【Chemistry 12】 [In the formula, L 1 is, -(CH 2 CH 2 O) n -or combination; n is between 1 and 10; and L 2 is, -(C 1 -C 6 [Alkylene)-N(H)-, a 6- to 12-membered spiroheterocyclylene, or bond, where the heterocyclylene contains 1 to 3 heteroatoms selected from N and O] 【Chemistry 13】 [In the formula, L 1 is a combination; and L 2 [It is a 6- to 12-membered spiroheterocyclylene containing 1 to 3 heteroatoms selected from N and O.] The compound according to claim 1, or a pharmaceutically acceptable salt thereof.

16. Compounds listed in the following table: Table 1 Table 2 Table 3 Table 4 A compound selected from or a pharmaceutically acceptable salt thereof.

17. A pharmaceutical composition comprising a compound according to any one of claims 1 to 16 or a pharmaceutically acceptable salt thereof, and a pharmaceutically acceptable excipient.

18. A pharmaceutical composition comprising a compound according to any one of claims 1 to 16 or a pharmaceutically acceptable salt thereof for modulating interleukin-1 receptor-related kinase 3 (IRAK3).

19. A pharmaceutical composition for treating cancer, comprising a compound according to any one of claims 1 to 16 or a pharmaceutically acceptable salt thereof, wherein the cancer is appropriately 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.

20. A pharmaceutical composition comprising a compound according to any one of claims 1 to 16 or a pharmaceutically acceptable salt thereof, for enhancing the immunity of a target subject to vaccination.

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