Heteroaryl compounds as ligands for IRAK4 degradation

Abstract

The present invention provides compounds and compositions for modulating IRAK4. In some embodiments, the compounds and compositions are provided for the treatment of inflammatory or autoimmune diseases.

Description

[Technical Field]

[0001] (Cross-reference of related applications) This application claims priority to U.S. Provisional Patent Application No. 63 / 465,604, filed on 11 May 2023, which is incorporated herein by reference in its entirety.

[0002] This disclosure generally relates to compounds and compositions, as well as methods for producing them and the use of said compounds and compositions for treating inflammatory or autoimmune diseases. [Background technology]

[0003] The recruitment of immune cells to the site of injury involves the coordinated interaction of numerous soluble mediators. Several cytokines, including interleukin-1 (IL-1), appear to play a crucial role in these processes. IL-1 triggers pro-inflammatory responses and is involved in tissue degeneration seen in chronic inflammatory states. Therefore, IL-1 is also involved in the process of bone resorption and the regulation of adipose tissue. Thus, IL-1 plays a vital role in many medical 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, namely IL-1R1 and IL-1RAcP. The resulting heterodimer recruits an adapter molecule called MyD88 and 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 mediate interactions with adapter proteins of the MyD88 family. Of the four members of the mammalian IRAK family, IRAK-4 is considered the "master IRAK." IRAK-4 is a serine / threonine kinase that plays a crucial role in Toll / IL-1 receptor (TIR) ​​signaling. Under overexpression conditions, all IRAKs can mediate the activation of nuclear factor κB and the stress-induced mitogen-activated protein kinase (MAPK) signaling cascade. Studies have demonstrated that IRAK4 kinase activity is essential for cytokine production, MAPK activation, and induction of NF-κB regulatory genes in response to TLR ligands (Koziczak-Holbro M. et al., J. Biol. Chem. 2007, 282, 13552-13560).Given the central role of IRAK4 in Toll-like / IL-1R signaling and immunoprotection, compounds that modulate IRAK4 function are likely to be useful in treating inflammation, cell proliferation, and immune-related symptoms and diseases associated with IRAK-mediated signaling (e.g., rheumatoid arthritis, inflammatory bowel disease, multiple sclerosis, diabetes, obesity, allergic diseases, psoriasis, asthma, graft rejection, cancer, and sepsis).

[0005] Protein degradation is a highly regulated and essential process that maintains cellular homeostasis. Through the ubiquitin-proteasome pathway (UPP), damaged, misfolded, or excess proteins are selectively identified and removed. The UPP plays a central role in regulating almost all intracellular processes. Protein ubiquitination is achieved by E3 ubiquitin ligases, which bind to proteins and attach ubiquitin molecules to them, leading to proteasomal degradation of proteins.

[0006] The therapeutic use of UPP is attracting considerable interest (Zhou et al., Mol. Cell 2000, 6, 751-756). One promising treatment method involves using proteolytic chimeric molecules (commonly called PROTACs) to remove unwanted proteins through proteolysis (Scheepstra et al., Comp. Struct. Biotech. J. 2019, 17, 160-176). PROTACs are ligand-targeted degraders that integrate an E3 ligase with the target protein to be degraded. These divalent molecules typically consist of an E3 ligase ligand and a small linker portion that binds to the target protein. PROTACs can position the E3 ligase at the appropriate distance and orientation relative to the target protein, thereby ubiquitinating the target protein. The ubiquitinated target protein is then recognized and degraded by the proteasome.

[0007] Thus, in one aspect, provided herein is a compound that targets IRAK4 for degradation.

[0008] (Summary of the Invention) Described herein are, in certain embodiments, compounds and compositions thereof for degrading IRAK4. In various embodiments, the compounds and compositions thereof can be used for the treatment of inflammatory or autoimmune diseases.

[0009] This embodiment can be more fully understood by reference to the detailed description and examples that illustrate non-limiting embodiments.

[0010] Embodiment 1 is of formula (I): [Chemical formula] [Wherein, R 1 is C1-C6 haloalkyl, C1-C6 alkyl or -CN; R 2a is H or C1-C6 alkyl; R 2b is C5-C6 cycloalkyl optionally substituted with 1 to 5 R 3 groups; or R 2a and R 2b The dotted line between represents a ring structure, and R 2a and R 2b together with the nitrogen atom to which they are attached form a 6-membered heterocyclyl optionally containing one additional heteroatom selected from N and O, and the heterocyclyl is optionally substituted with 1 to 5 R 3 groups; Each R 3 is independently -NH2, -OH, halo, C1-C6 alkyl or C1-C6 haloalkyl; X is CH or N; L 1-C(O)N(H)-, -N(H)C(O)-, -C(O)-, -(C1-C6 alkylene)N(R 4 )- or C1-C6 alkylene; R 4 is H or C1-C6 alkyl; R 5a and R 5b Each of these is either H, or they together form an oxo group; Ring A is, [ka] It is either an 8-10 membered spiroheterocyclylene group containing 1-3 nitrogen atoms, wherein the heterocyclylene group contains m R 6 It is substituted with the base; Y 1 and Y 2 It is independently CH or N; Each R 6 These are independently halo, C1-C6 alkyl, or C1-C6 haloalkyl; m is between 0 and 5; L 2 is a combination or -N(R 7 )-and; and R 7 [It is H or C1-C6 alkyl] It is a compound of or a pharmaceutically acceptable salt thereof.

[0011] Embodiment 2 is R 1 However, the compound described in Embodiment 1, or a pharmaceutically acceptable salt thereof, is a C1-C3 haloalkyl, C1-C3 alkyl, or -CN.

[0012] Embodiment 3 is R 1 However, the compound described in Embodiment 2, or a pharmaceutically acceptable salt thereof, is -CHF2, -CF3, -CH3, or -CN.

[0013] Embodiment 4 is R 2a However, it is H or C1-C3 alkyl; and R 2bHowever, 1 to 3 R 3 The compound according to any one of Embodiments 1 to 3, or a pharmaceutically acceptable salt thereof, is a cyclohexyl group which may be optionally substituted at the base.

[0014] Embodiment 5 is R 2a and R 2b However, together with the nitrogen atoms bonded to them, they form a 6-membered heterocycline which may optionally contain one additional heteroatom selected from N and O, and the heterocycline contains 1-2 R 3 The compound according to any one of Embodiments 1 to 3, or a pharmaceutically acceptable salt thereof, which may be optionally substituted with a base.

[0015] Mental 6 is each R 3 However, independently, the compound is -NH2, -OH, halo, C1-C3 alkyl, or C1-C3 haloalkyl, or a pharmaceutically acceptable salt thereof, as described in any one of Embodiments 1 to 5.

[0016] Embodiment 7 is each R 3 However, independently, these are the compounds described in Embodiment 6 or pharmaceutically acceptable salts thereof, which are -NH2, -OH, F, or -CH3.

[0017] Embodiment 8 is, [ka] but, [ka] This is a compound described in any one of Embodiments 1 to 7, or a pharmaceutically acceptable salt thereof.

[0018] Embodiment 9 is L 1 However, -C(O)N(H)-, -N(H)C(O)-, -C(O)-, -(C1-C3 alkylene)N(R 4 )- or C1-C3 alkylene; and R 4However, the compound is one of the compounds described in any one of Embodiments 1 to 8, which is H or a C1-C3 alkyl group, or a pharmaceutically acceptable salt thereof.

[0019] Embodiment 10 is L 1 The compound described in Embodiment 9, or a pharmaceutically acceptable salt thereof, is -C(O)N(H)-, -N(H)C(O)-, -C(O)-, -CH2N(CH3)-, -CH2-, or -CH2CH2-.

[0020] Embodiment 11 is in which ring A is [ka] It is either a 10-membered spiroheterocyclylene containing two nitrogen atoms, and the spiroheterocyclylene contains m R 6 The compound is substituted with a group and is one of the compounds described in any one of Embodiments 1 to 10, or a pharmaceutically acceptable salt thereof.

[0021] Embodiment 12 comprises each R 6 However, independently, the compound is a halo, C1-C3 alkyl, or C1-C3 haloalkyl, as described in any one of Embodiments 1 to 11, or a pharmaceutically acceptable salt thereof.

[0022] Embodiment 13 is a compound according to any one of Embodiments 1 to 12, or a pharmaceutically acceptable salt thereof, wherein m is 0, 1, or 2.

[0023] Embodiment 14 is, Ring A is [ka] This is a compound described in any one of Embodiments 1 to 13, or a pharmaceutically acceptable salt thereof.

[0024] Embodiment 15 is, (i) L 2 is a combination; or (ii) L 2However, -N(R 7 )-and; and R 7 However, the compound is one of any one of Embodiments 1 to 14, which is H or a C1-C3 alkyl group, or a pharmaceutically acceptable salt thereof.

[0025] Embodiment 16 is, [ka] but, [ka] This is a compound described in any one of Embodiments 1 to 15, or a pharmaceutically acceptable salt thereof.

[0026] Embodiment 17 is a compound of formula (IIa), (IIb), (IIIa), or (IIIb): [ka] This is a compound described in any one of Embodiments 1 to 16, or a pharmaceutically acceptable salt thereof.

[0027] Embodiment 18 is a compound selected from the compounds in Table 1 or their pharmaceutically acceptable salts.

[0028] Embodiment 19 is a pharmaceutical composition comprising a compound described in any one of Embodiments 1 to 18 or a pharmaceutically acceptable salt thereof, and a pharmaceutically acceptable excipient.

[0029] Embodiment 20 is a method for modulating interleukin-1 (IL1) receptor-related kinase 4 (IRAK4), characterized by (i) contacting IRAK4 with an effective amount of a compound or a pharmaceutically acceptable salt thereof described in any one of Embodiments 1 to 18 or a pharmaceutical composition described in Embodiment 19, or (ii) administering an effective amount of a compound or a pharmaceutically acceptable salt thereof described in any one of Embodiments 1 to 18 or a pharmaceutical composition described in Embodiment 19 to a subject in need of treatment, wherein the inflammatory disease or autoimmune disease may optionally be atopic dermatitis, asthma, lupus, rheumatoid arthritis, familial Mediterranean fever, psoriasis, generalized pustular psoriasis, cryopyrin-associated periodic syndromes, hidradenitis suppurativa, Beckett syndrome, or familial cold autoinflammatory syndrome. Detailed Description of the Invention

[0030] (definition) As used herein, the terms “comprising” and “including” are interchangeable. The terms “comprising” and “including” are construed to identify the presence of any described feature or component mentioned, but do not exclude the presence or addition of one or more features, components, or groups thereof. Furthermore, the terms “comprising” and “including” are also intended to include examples that are encompassed by the term “consisting of.” Consequently, the term “consisting of” may be used instead of the terms “comprising” and “including” to provide more specific embodiments of the present invention.

[0031] The term "consisting of" means that the object has at least 90%, 95%, 97%, 98%, or 99% of the described features or components that make up the object. In another embodiment, the term "consisting of" excludes other features or components from the scope of the subsequent description, except those that are not essential to the technical effect to be achieved.

[0032] As used herein, the term “or” is interpreted as an inclusive “or” meaning any one or any combination thereof. Thus, “A, B or C” means any of the following: “A; B; C; A and B; A and C; B and C; A, B and C.” Exceptions to this definition occur only if the combination of elements, functions, procedures or actions is in any way essential and mutually exclusive.

[0033] In this specification, any concentration range, percentage range, percentage range, or integer range is understood to include any integer within the range described, and, where appropriate, its decimal values ​​(such as one-tenth and one-hundredth of an integer), unless otherwise indicated. Similarly, any numerical ranges mentioned herein relating to any physical characteristics such as polymer subunits, size, or thickness are understood to include all integers within the range described, unless otherwise indicated. Where used herein, the terms “about” and “approximately” mean ±20%, ±10%, ±5%, or ±1% of the indicated range, value, or structure, unless otherwise indicated.

[0034] The "alkyl" group consists of 1 to 10 carbon atoms (C1-C 10Alkyls are saturated, partially saturated, or unsaturated linear or branched acyclic hydrocarbons, typically having 1 to 8 carbon atoms (C1-C8 alkyls), or, in some embodiments, 1 to 6 carbon atoms (C1-C6 alkyls), 1 to 3 carbon atoms (C1-C3 alkyls), or 2 to 6 carbon atoms (C2-C6 alkyls). In some embodiments, alkyls are saturated alkyls. Representative saturated alkyls include methyl, ethyl, n-propyl, n-butyl, n-pentyl, and n-hexyl; saturated branched alkyls include isopropyl, sec-butyl, isobutyl, tert-butyl, isopentyl, neopentyl, tert-pentyl, 2-methylpentyl, 3-methylpentyl, 4-methylpentyl, and 2,3-dimethylbutyl. In some embodiments, alkyls are unsaturated alkyls, also called alkenyl or alkynyl groups. 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(CH3), -CH=C(CH3)2, -C(CH3)=CH2, -C(CH3)=CH(CH3), -C(CH2CH3)=CH2, -C≡CH, -C≡C(CH3), -C≡C(CH2CH3), -CH2C≡CH, -CH2C≡C(CH3), and -CH2C≡C(CH2CH3). Alkyl groups can be substituted or unsubstituted.Where an alkyl group described herein is said to be “substituted,” it means any substituent as seen in the exemplary compounds and embodiments disclosed herein, as well as halogens; hydroxyl; alkoxy; cycloalkyloxy, aryloxy, heterocyclyloxy, heteroaryloxy, heterocycloalkyloxy, cycloalkylalkyloxy, aralkyloxy, heterocyclylalkyloxy, heteroarylalkyloxy, heterocycloalkylalkyloxy; oxo(=O); amino, alkylamino, cycloalkylamino, arylamino, heterocyclylamino, heteroarylamino, heterocycloalkylamino, cycloalkyl Alkylamino, aralkylamino, heterocyclylalkylamino, heteroaralkylamino, heterocycloalkylalkylamino; imino; imide; 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; amide; cyano; isocyanate; isothiocyanato; cyanate; thiocyanato; or may be substituted with -B(OH)2.In certain embodiments, where an alkyl group described herein is said to be “substituted,” it may be substituted with any substituent as found in the exemplary compounds and embodiments disclosed herein, as well as halogens (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] The "cycloalkyl" group is a saturated or partially saturated cyclic alkyl group (C3-C) having 3 to 10 carbon atoms and having a monocyclic or multiple fused or bridging ring, which can be optionally substituted. 10The cycloalkyl group is a cycloalkyl group. In some embodiments, the cycloalkyl group has 3 to 8 ring carbon atoms (C3-C8 cycloalkyl), but in other embodiments, the number of ring carbon atoms is in the range of 3 to 5 (C3-C5 cycloalkyl), 3 to 6 (C3-C6 cycloalkyl), or 3 to 7 (C3-C7 cycloalkyl). In some embodiments, the cycloalkyl group is a saturated cycloalkyl group. Examples of the saturated cycloalkyl group include monocyclic structures such as cyclopropyl, cyclobutyl, cyclopentyl, cyclohexyl, cycloheptyl, cyclooctyl, 1-methylcyclopropyl, 2-methylcyclopentyl, 2-methylcyclooctyl, etc., or multiple ring structures or cross-linked ring structures such as 1-bicyclo[1.1.1]pentyl, bicyclo[2.1.1]hexyl, bicyclo[2.2.1]heptyl, bicyclo[2.2.2]octyl, adamantyl, etc. In other embodiments, the cycloalkyl group is an unsaturated cycloalkyl group. Examples of unsaturated cycloalkyl groups include cyclohexenyl, cyclopentenyl, cyclohexadienyl, butadienyl, pentadienyl, and hexadienyl. Cycloalkyl groups can be substituted or unsubstituted. An example of such a substituted cycloalkyl group is cyclohexanol.

[0036] An "aryl" group is an aromatic carbocyclic group (C6-C) having 6 to 14 carbon atoms and consisting of a single ring (e.g., phenyl) or multiple fused rings (e.g., naphthyl or anthryl). 14 This refers to an aryl group. In one embodiment, the aryl group has 6 to 14 carbon atoms (C6-C) in the ring portion of the aryl group. 14 It contains aryl atoms, and in other embodiments, 6 to 12 carbon atoms (C6-C6). 12 Aryl) or 6-10 carbon atoms (C6-C 10This includes aryl groups. Certain aryl groups include phenyl, biphenyl, and naphthyl. The aryl group can be substituted or unsubstituted. The phrase "aryl group" also includes groups containing fused rings, such as fused aromatic-aliphatic ring systems (e.g., indanyl, tetrahydronaphthyl).

[0037] "Halogen" or "halo" refers to fluorine, chlorine, bromine, or iodine.

[0038] "Haloalkyl" refers to an alkyl group substituted with one or more halo groups, as defined above. Examples include trifluoromethyl, difluoromethyl, trichloromethyl, 2,2,2-trifluoroethyl, 1,2-difluoroethyl, 3-bromo-2-fluoropropyl, and 1,2-dibromoethyl. In some embodiments, the haloalkyl group has 1 to 6 carbon atoms and is substituted with one or more halo groups (C1-C6 haloalkyl), or the haloalkyl group has 1 to 3 carbon atoms and is substituted with one or more halo groups (C1-C3 haloalkyl). The halo groups may all be the same or different. Unless otherwise specified, the haloalkyl group may be substituted as desired.

[0039] A "heteroaryl" group is an aromatic ring system having 1 to 4 heteroatoms as ring atoms in a heteroaromatic ring system, with the remaining atoms being carbon atoms. In some embodiments, the heteroaryl group contains 3 to 6 ring atoms, and in other embodiments, 6 to 9 or even 6 to 10 atoms are contained in the ring portion of the group. Suitable heteroatoms include oxygen, sulfur, and nitrogen. In certain embodiments, the heteroaryl ring system is monocyclic or bicyclic. Non-limiting examples include pyrrolyl, pyrazolyl, imidazolyl, triazolyl, tetrazolyl, oxazolyl, isoxazolyl, benzisoxazolyl (e.g., benzo[d]isoxazolyl), thiazolyl, pyrrolyl, pyridadinyl, pyrimidyl, pyrazinyl, thiophenyl, benzothiophenyl, furanil, benzofuranil, indolyl (e.g., indolyl-2-onyl or isoindoline-1-onyl), azaindolyl (pyrrolopyridyl or 1H-pyrrolo[2,3-b]pyridyl), indazolyl, benzimidazolyl (e.g., 1H-benzo[d]imidazolyl), imidazopyridyl (e.g., A Examples of heteroaryl groups include, but are not limited to, zabenzimidazolyl (or 1H-imidazo[4,5-b]pyridyl), pyrazolopyridyl, triazolopyridyl, benzotriazolyl (e.g., 1H-benzo[d][1,2,3]triazolyl), benzoxazolyl (e.g., benzo[d]xazolyl), benzothiazolyl, benzothiadiazolyl, isoxazolopyridyl, thianaphthalenyl, prinyl, xanthinyl, adeninyl, guaninyl, quinolinyl, isoquinolinyl (e.g., 3,4-dihydroisoquinoline-1(2H)-onyl), tetrahydroquinolinyl, quinoxalinyl, and quinazolinyl groups. Heteroaryl groups may be substituted or unsubstituted.

[0040] A "heterocyclyl" group is a non-aromatic cycloalkyl group in which the 1 to 4 ring carbon atoms are independently substituted with heteroatoms selected from O, S, and N. In some embodiments, a heterocyclyl group contains 3 to 10 ring members, while other groups contain 3 to 5, 3 to 6, or 3 to 8 ring members. Heterocyclyls can also be bonded to another group at any ring atom (in other words, any carbon or heteroatom of the heterocycle). Heterocycloalkyl groups can be substituted or unsubstituted. Heterocyclyl groups include saturated and partially saturated ring systems. Furthermore, the term "heterocyclyl" is intended to encompass a non-aromatic ring containing at least one heteroatom, which may be fused with an aryl or heteroaryl ring, regardless of its bonding to the rest of the molecule. The term also includes bridging polycyclic ring systems containing heteroatoms. Representative examples of heterocyclyl groups include azilidinyl, azetidinyl, azepanil, pyrrolidyl, imidazolidinyl (e.g., imidazolidine-4-onyl or imidazolidine-2,4-dionyl), pyrazolidinyl, thiazolidinyl, tetrahydrothiophenyl, tetrahydrofuranil, piperidyl, piperazinyl (e.g., piperazine-2-onyl), morpholinyl, thiomorpholinyl, tetrahydropyranil (e.g., tetrahydro-2H-pyranil), tetrahydrothiopyranil, oxathianil, dithianil, 1,4-dioxaspiro[4.5]decanyl, homopiperazinyl, quinuclidyl, or tetrahydropyrimidine-2(1H)-one. Typical substituted heterocyclyl groups may be monosubstituted or bisubstituted or more. For example, a pyridyl group or morpholinyl group may be 2-, 3-, 4-, 5-, or 6-substituted, or bisubstituted with various substituents as listed below, but is not limited to these.

[0041] "Oxo" indicates a chemical group = O.

[0042] Except for alkyl groups, where a group described herein is said to be “substituted,” it may be substituted with any suitable substituent. Exemplary substituents are those found in the exemplary compounds and embodiments disclosed herein, as well as halogens (chloro, iodo, bromo or fluoro); alkyl; hydroxyl; alkoxy; alkoxyalkyl; amino; alkylamino; carboxy; nitro; cyano; thiol; thioether; imine; imide; amidine; guanidine; enamine; aminocarbonyl; acylamino; phosphonate; phosphine; thiocarbonyl; sulfinyl; sulfone; sulfonamide; ketone; aldehyde; ester; urea; urethane; oxime; hydroxylamine; alkoxyamine; aralkoxyamine; N-oxide; hydrazine; hydrazide; hydrazone; azide; isocyanate; isothiocyanate; cyanate; thiocyanate; oxygen (=O); B(OH)2, O(alkyl)aminocarbonyl; monocyclic or condensed These are cycloalkyls (e.g., cyclopropyl, cyclobutyl, cyclopentyl, or cyclohexyl), which may be non-condensed polycyclic; or heterocyclyls (e.g., pyrrolidyl, piperidyl, piperazinyl, morpholinyl, or thiadinyl), which may be monocyclic or condensed or non-condensed polycyclic; monocyclic or condensed or non-condensed polycyclic aryls or heteroaryls (e.g., phenyl, naphthyl, pyrrolyl, indolyl, furanyl, thiophenyl, imidazolyl, oxazolyl, isoxazolyl, thiazolyl, triazolyl, tetrazolyl, pyrazolyl, pyridyl, quinolinyl, isoquinolinyl, acridinyl, pyrazinyl, pyridadinyl, pyrimidyl, benzimidazolyl, benzothiophenyl, or benzofuranyl); aryloxys; aralkyloxys; heterocyclyloxys; and heterocyclylalkoxys.

[0043] Alternatively, you can use other commonly used chemical names. For example, divalent groups such as divalent alkyl, divalent phenyl, divalent heteroaryl, and divalent heterocyclyl can also be referred to as alkylene, phenylene, heteroarylene, or heterocyclylene, respectively.

[0044] Embodiments of this disclosure include pharmaceutically acceptable salts, tautomers, isotopologues, and stereoisomers of the compounds provided herein (e.g., compounds of formula (I)).

[0045] As used herein, the term “pharmaceutically acceptable salt” refers to a salt produced from a pharmaceutically acceptable, non-toxic acid or base, including inorganic acids and bases, as well as organic acids and bases. Suitable pharmaceutically acceptable base addition salts of the compound of formula (I) include, but are not limited to, metal salts derived from aluminum, calcium, lithium, magnesium, potassium, sodium, and zinc, or organic salts derived from lysine, N,N'-dibenzylethylenediamine, chloroprocaine, choline, diethanolamine, ethylenediamine, meglumine (N-methyl-glucamine), and procaine. Suitable non-toxic acids include, but are not limited to, inorganic and organic acids such as acetic acid, alginic acid, anthranilic acid, benzenesulfonic acid, benzoic acid, camphorsulfonic acid, citric acid, ethensulfonic acid, formic acid, fumaric acid, furoic acid, galacturonic acid, gluconic acid, glucuronic acid, glutamic acid, glycolic acid, hydrobromic acid, hydrochloride, isethionic acid, lactate, maleate, malate, mandelate, methanesulfonate, mucinic 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. Specific salts include hydrochloride, formate, and mesylate. Other materials are well known in the relevant technical field; see, for example, 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).

[0046] In this specification, unless otherwise stated, the terms “stereoisomer” or “stereoisomerically pure” mean one stereoisomer of a particular compound that substantially does not contain any other stereoisomers of that compound. For example, a stereoisomerically pure compound having one chiral center substantially does not contain the opposite enantiomer of that compound. A stereoisomerically pure compound having two chiral centers substantially does not contain any other diastereomer of that compound. A typical stereoisomerically pure compound is one in which one stereoisomer of the compound is present in greater proportions than about 80% by weight and the other stereoisomer of the compound in less than about 20% by weight; one stereoisomer of the compound is present in greater proportions than about 90% by weight and the other stereoisomer of the compound in less than about 10% by weight; one stereoisomer of the compound is present in greater proportions than about 95% by weight and the other stereoisomer of the compound is present in less than about 5% by weight; or one stereoisomer of the compound is present in greater proportions than about 97% by weight and the other stereoisomer of the compound is present in less than about 3% by weight. The compounds disclosed herein have a chiral center and may exist as racemates, individual enantiomers or diastereomers, and mixtures thereof. All such isomeric forms, including mixtures thereof, are included in the embodiments disclosed herein.

[0047] The use of stereoisomerically pure forms of the compounds disclosed herein and the use of mixtures of those forms are encompassed in the embodiments disclosed herein. For example, mixtures containing equimolar or unequal amounts of enantiomers of a particular compound may be used in the methods and compositions disclosed herein. These isomers can be asymmetrically synthesized or separated using standard techniques such as chiral columns or chiral resolving agents. For example, Jacques, J., et al., Enantiomers, Racemates and Resolutions (Wiley-Interscience, New York, 1981); Wilen, SH, et al., Tetrahedron 33:2725 (1977); Eliel, EL, Stereochemistry of Carbon Compounds (McGraw-Hill, NY, 1962); Wilen, SH, Tables of Resolving Agents and Optical Resolutions p. 268 (EL Eliel, Ed., Univ. of Notre Dame Press, Notre Dame, IN, 1972); Todd, M., Separation Of Enantiomers :Synthetic Methods (Wiley-VCH Verlag GmbH & Co. KGaA, Weinheim, Germany, 2014); Toda, F., Enantiomer Separation:Fundamentals and Practical Methods (Springer Science & Business Media, 2007); See Subramanian, G., Chiral Separation Techniques: A Practical Approach (John Wiley & Sons, 2008); and Ahuja, S., Chiral Separation Methods for Pharmaceutical and Biotechnological Products (John Wiley & Sons, 2011).

[0048] It should also be noted that the compounds disclosed herein may include E isomers and Z isomers or mixtures thereof, as well as cis and trans isomers or mixtures thereof. In certain embodiments, the compound is isolated as either an E isomer or a Z isomer. In other embodiments, the compound is a mixture of E and Z isomers.

[0049] A "tautomer" refers to an isomer of a compound that is in equilibrium with it. The concentration of these isomers varies depending on the environment in which the compound exists; for example, it may differ depending on whether the compound is a solid, an organic solution, or an aqueous solution. For example, in aqueous solution, pyrazoles may exhibit the following isomers, which are called tautomers: [ka]

[0050] As will be readily apparent to those skilled in the art, various functional groups and other structures may exhibit tautomerism, and all tautomers of the compound of formula (I) are within the scope of this disclosure.

[0051] Furthermore, it should be noted that the compounds disclosed herein may contain atomic isotopes in proportions not found in nature for one or more atoms. For example, a compound may contain tritium ( 3 H), Iodine-125 ( 125 I), Sulfur 35 ( 35 S) or carbon-14 ( 14 Can it be radiolabeled with radioactive isotopes such as C), or deuterium ( 2 H), carbon-13 ( 13 C) or nitrogen 15 ( 15Isotope enrichment can be achieved by means of N), etc. As used herein, “isotopologue” is an isotope-enriched compound. The term “isotopologue” refers to an atom having an isotope composition other than the natural isotope composition of that atom. The term “isotopologue” may also refer to a compound containing at least one atom having an isotope composition other than the natural isotope composition of that atom. The term “isotopologue” means the amount of each isotope present relative to a given atom. Radiolabeled compounds and isotope-enriched compounds are useful as therapeutic agents, e.g., cancer treatments, research reagents (e.g., binding assay reagents), and diagnostic agents (e.g., in vivo imaging agents). All isotopic variations of the compounds described herein, whether radioactive or not, are intended to be included within the scope of the embodiments provided herein. In some embodiments, isototopologues of the compounds disclosed herein are provided, for example, isototopologues are compounds rich in deuterium, carbon-13, and / or nitrogen-15. As used herein, “deuterated” means that at least one hydrogen (H) atom is converted into deuterium (D or 2 This refers to compounds substituted with H, i.e., compounds in which deuterium is concentrated at at least one position.

[0052] Regardless of stereoisomer or isotopic composition, it is understood that each compound disclosed herein may be provided in any form of the pharmaceutically acceptable salts discussed herein. Similarly, it is understood that the isotopic composition may vary independently of the stereoisomer composition of each compound referred to herein. Furthermore, the isotopic composition may vary, but is limited to the elements present in each compound or its salts disclosed herein, or independently of the selection of pharmaceutically acceptable salts of each compound.

[0053] Please note that in the event of any discrepancy between the described structure and its name, the described structure will take precedence.

[0054] As used herein, “treatment” means the overall or partial relief of a disorder, disease or condition, or one or more symptoms associated with a disorder, disease or condition, or the reduction or cessation of further progression or worsening of those symptoms, or the relief or eradication of the cause of the disorder, disease or condition itself. In one embodiment, a disorder is a neurodegenerative disease or its symptoms as described herein.

[0055] As used herein, “prevention” means a method of delaying and / or interfering with the onset, recurrence or progression of the whole or partial disability, disease or condition, a method of preventing a subject from acquiring a disability, disease or condition, or a method of reducing the risk of a subject acquiring a disability, disease or condition. In one embodiment, disability is a neurodegenerative disease or its symptoms as described herein.

[0056] In relation to the compounds disclosed herein, the term “effective amount” means an amount that can treat or prevent the disorder, disease or condition, or symptoms thereof, disclosed herein.

[0057] As used herein, the terms “subject” or “patient” include, but are not limited to, animals such as cattle, monkeys, horses, sheep, pigs, birds, turkeys, quail, cats, dogs, mice, rats, rabbits, or guinea pigs, and are mammals in one embodiment and humans in another embodiment. In one embodiment, the subject is a human being who has or is at risk of having a disease or symptoms mediated by S1P5.

[0058] Various features of the present invention may be described in the context of a single embodiment, but these features may also be provided individually or in any suitable combination. Conversely, the present invention may be described herein in the context of a separate embodiment for clarity, but the present invention may also be implemented in a single embodiment.

[0059] compound In one embodiment, the formula provided herein is (I): [ka] [In the formula, R 1 These are C1-C6 haloalkyl, C1-C6 alkyl, or -CN; R 2a is H or C1-C6 alkyl; R 2b This is 1 to 5 R 3 A C5-C6 cycloalkyl group which may be optionally substituted with a group; or R 2a and R 2b The dotted line between them represents a ring structure, R 2a and R 2b These, together with the nitrogen atoms bonded to them, form a six-membered heterocycline which may optionally contain one additional heteroatom selected from N and O, and the heterocycline has 1 to 5 R 3 The base may be replaced as desired; Each R 3 These are independently -NH2, -OH, halo, C1-C6 alkyl, or C1-C6 haloalkyl; X is either CH or N; L 1 -C(O)N(H)-, -N(H)C(O)-, -C(O)-, -(C1-C6 alkylene)N(R 4 )- or C1-C6 alkylene; R 4 is H or C1-C6 alkyl; R 5a and R 5b Each of these is either H, or they together form an oxo group; Ring A is, [ka] It is either an 8-10 member spiroheterocyclylene containing 1-3 nitrogen atoms, wherein the heterocyclylene contains m R6 is substituted by a base; Y 1 and Y 2 are independently CH or N; Each R 6 is independently halo, C1-C6 alkyl or C1-C6 haloalkyl; m is from 0 to 5; L 2 is a bond or -N(R 7 )-; and R 7 is H or C1-C6 alkyl) is a compound thereof or a pharmaceutically acceptable salt thereof.

[0060] In certain embodiments, R 1 is C1-C6 haloalkyl, C1-C6 alkyl or -CN. In certain embodiments, R 1 is C1-C3 haloalkyl, C1-C3 alkyl or -CN. In certain embodiments, R 1 is -CHF2, -CF3, -CH3 or -CN.

[0061] In certain embodiments, R 1 is C1-C6 haloalkyl. In certain embodiments, R 1 is C1-C6 haloalkyl containing 1 to 13 halogen atoms. In certain embodiments, R 1 is C1-C3 haloalkyl. In certain embodiments, R 1 is C1-C3 haloalkyl containing 1 to 7 halogen atoms. In certain embodiments, R 1 is -CF3, -CHF2, -CH2F, -CCl3, -CHCl2, -CH2Cl, -CF2Cl, -CFCl2, -CH2CF3, -CH2CHF2 or -CH2CCl3. In certain embodiments, R 1 is -CF3. In certain embodiments, R 1 is -CHF2.

[0062] In certain embodiments, R1 is C1-C6 alkyl. In certain embodiments, R 1 is C1-C3 alkyl. In certain embodiments, R 1 is methyl, ethyl, n-propyl or isopropyl. In certain embodiments, R 1 is methyl. In certain embodiments, R 1 is ethyl. In certain embodiments, R 1 is n-propyl. In certain embodiments, R 1 is isopropyl.

[0063] In certain embodiments, R 1 is -CN.

[0064] In certain embodiments, R 2a is H or C1-C6 alkyl. In certain embodiments, R 2a is H or C1-C3 alkyl. In certain embodiments, R 2a is H or -CH3.

[0065] In certain embodiments, R 2a is H.

[0066] In certain embodiments, R 2a is C1-C6 alkyl. In certain embodiments, R 2a is C1-C3 alkyl. In certain embodiments, R 2a is methyl, ethyl, n-propyl or isopropyl. In certain embodiments, R 2a is methyl. In certain embodiments, R 2a is ethyl. In certain embodiments, R 2a is n-propyl. In certain embodiments, R 2a is isopropyl.

[0067] In certain embodiments, R 2b is 1 to 5 R 3It is a C5-C6 cycloalkyl group which may be optionally substituted with a group. In one embodiment, R 2b This is 1 to 3 R 3 It is a cyclohexyl which may be optionally substituted with a group. In one embodiment, R 2b This is 1-2 R 3 A cyclohexyl group which may be optionally substituted at the base. In one embodiment, R 2b This is one R 3 It is a cyclohexyl compound that may be optionally substituted at the base.

[0068] One reason, R 2b This is 1 to 5 R 3 It is a C5-C6 cycloalkyl group which may be optionally substituted with a group. In one embodiment, R 2b This is 1 to 5 R 3 It is a cyclopentyl which may be optionally substituted at the base. In one embodiment, R 2b This is 1 to 5 R 3 A cyclohexyl which may be optionally substituted with R groups. In one embodiment, the cycloalkyl is 1 to 5 R groups 3 The group may be optionally substituted with R. In one embodiment, the cycloalkyl group has 1 to 3 R groups. 3 The group may be optionally substituted with R. In one embodiment, the cycloalkyl group has 1 or 2 R 3 The group may be optionally substituted with a R group. In one embodiment, the cycloalkyl group has one R group. 3 The group may be optionally substituted. In one embodiment, the cycloalkyl group is unsubstituted.

[0069] One reason, R 2a and R 2b These, together with the nitrogen atoms bonded to them, form a 6-membered heterocycline, which may optionally contain one additional heteroatom selected from N and O, and 1 to 5 R 3The base may be substituted as desired. In one embodiment, R 2a and R 2b These, together with the nitrogen atoms bonded to them, form a 6-membered heterocycline, which may optionally contain one additional heteroatom selected from N and O, and 1 to 5 R 3 The base may be substituted as desired. In one embodiment, R 2a and R 2b These, together with the nitrogen atoms bonded to them, form a 6-membered heterocycline, which may optionally contain one additional heteroatom selected from N and O, and 1 to 2 R 3 The base may be replaced as desired.

[0070] One reason, R 2a and R 2b These, together with the nitrogen atoms bonded to them, form a 6-membered heterocycline, the heterocycline containing one additional heteroatom selected from N and O, and 1 to 5 R 3 The base may be substituted as desired. In one embodiment, R 2a and R 2b These, together with the nitrogen atoms bonded to them, form a 6-membered heterocycline, the heterocycline containing one additional nitrogen atom and 1 to 5 R 3 The base may be substituted as desired. In one embodiment, R 2a and R 2b These, together with the nitrogen atoms bonded to them, form a 6-membered heterocycline, and the heterocycline contains one oxygen atom and 1 to 5 R 3 The base may be substituted as desired. In one embodiment, the heterocyclyl has 1 to 5 R 3 It is substituted with a group. In one embodiment, the heterocyclyl has 1 to 3 R groups. 3 It is substituted with a group. In one embodiment, the heterocyclyl has one or two R groups. 3It is substituted with a group. In one embodiment, the heterocyclyl has one R 3 It is substituted with a group. In one embodiment, the heterocyclyl is unsubstituted. In one embodiment, the heterocyclyl is piperidinyl, morpholinyl, or piperazinyl.

[0071] One way of doing this is for each R 3 R is independently -NH2, -OH, halo, C1-C6 alkyl, or C1-C6 haloalkyl. In one embodiment, each R 3 R is independently -NH2, -OH, halo, C1-C3 alkyl, or C1-C3 haloalkyl. In one embodiment, each R 3 These are independently -NH2, -OH, F, Cl, -CH3, or -CF3. In one embodiment, each R 3 These are independently -NH2, -OH, F, or -CH3.

[0072] One reason, R 3 It is -NH2.

[0073] One reason, R 3 It is -OH.

[0074] One reason, R 3 This is a halo. In one embodiment, R 3 is Cl, F, or Br. In one embodiment, R 3 is Cl. In one embodiment, R 3 is F. For one reason, R 3 It is Br.

[0075] One reason, R 3 is a C1-C6 alkyl group. In one embodiment, R 3 is a C1-C3 alkyl group. In one embodiment, R 3 is methyl, ethyl, n-propyl, or isopropyl. In one embodiment, R 3is methyl. In certain embodiments, R 3 is ethyl. In certain embodiments, R 3 is n-propyl. In certain embodiments, R 3 is isopropyl.

[0076] In certain embodiments, R 3 is C1-C6 haloalkyl. In certain embodiments, R 3 is C1-C6 haloalkyl containing 1 to 13 halogen atoms. In certain embodiments, R 3 is C1-C3 haloalkyl. In certain embodiments, R 3 is C1-C3 haloalkyl containing 1 to 7 halogen atoms. In certain embodiments, R 3 is -CF3, -CHF2, -CH2F, -CCl3, -CHCl2, -CH2Cl, -CF2Cl, -CFCl2, -CH2CF3, -CH2CHF2 or -CH2CCl3. In certain embodiments, R 3 is -CF3. In certain embodiments, R 3 is -CHF2.

[0077] In certain embodiments,

Chemical formula

Chemical formula

[0078] In certain embodiments,

Chemical formula

Chemical formula

[0079] In one embodiment, X is CH or N. In one embodiment, X is CH. In one embodiment, X is N.

[0080] In one statement, L 1 -C(O)N(H)-, -N(H)C(O)-, -C(O)-, -(C1-C6 alkylene)N(R 4 )- or C1-C6 alkylene. In one embodiment, L 1 -C(O)N(H)-, -N(H)C(O)-, -C(O)-, -(C1-C3 alkylene)N(R 4 )- or C1-C3 alkylene; R 4 is H or C1-C3 alkyl. In one embodiment, L 1 These are -C(O)N(H)-, -N(H)C(O)-, -C(O)-, -CH2N(CH3)-, -CH2-, or -CH2CH2-.

[0081] In one statement, L 1 is -C(O)N(H)-. In one embodiment, L 1 is -N(H)C(O)-. In one embodiment, L 1 It is -C(O)-.

[0082] In one statement, L 1 is -(C1-C6 alkylene)N(R 4 )-. In one embodiment, L 1 is -(C1-C3 alkylene)N(R 4 )-. In one embodiment, L 1 is -CH2N(R 4 )-,-CH2CH2N(R 4 )- or -CH2CH2CH2N(R 4 )-. In any of these transformations, R 4 is H or C1-C6 alkyl. In one embodiment, R 4 is H or C1-C3 alkyl. In one embodiment, R 4 H is H. In one example, R4 is a C1-C6 alkyl group. In one embodiment, R 4 is a C1-C3 alkyl group. In one embodiment, R 4 is methyl, ethyl, n-propyl, or isopropyl. In one embodiment, R 4 is methyl. In one example, R 4 is ethyl. In one embodiment, R 4 is n-propyl. In one embodiment, R 4 is isopropyl. In one embodiment, L 1 is -CH2N(H)-, -CH2CH2N(H)-, or -CH2CH2CH2N(H)-. In one embodiment, L 1 These are -CH2N(CH3)-, -CH2CH2N(CH3)-, or -CH2CH2CH2N(CH3)-.

[0083] In one statement, L 1 L is a C1-C6 alkylene. In one embodiment, L 1 L is a C1-C3 alkylene. In one embodiment, L 1 These are -CH2-, -CH2CH2-, or -CH2CH2CH2-.

[0084] One reason, R 5a and R 5b Each of them is either H, or they together form an oxo group. In one embodiment, R 5a and R 5b These are H, respectively. In one example, R 5a and R 5b These combine to form an oxo group.

[0085] In one presentation, ring A is, [ka] And here Y 1 and Y 2This is independently CH or N, or an 8-10 membered spiroheterocyclylene containing 1-3 nitrogen atoms, wherein the heterocyclylene contains m R 6 It is substituted by the base.

[0086] In one presentation, ring A is, [ka] In one embodiment, Y 1 N is Y 2 CH is CH. In one embodiment, Y 1 CH is Y 2 In one embodiment, Y 1 and Y 2 Each of these is N. In one embodiment, Y 1 and Y 2 These are each CH.

[0087] In one embodiment, ring A is an 8-10 membered spiroheterocyclylene, which contains 1-3 nitrogen atoms and m R 6 It is substituted with a group. In one embodiment, ring A is an 8-membered spiroheterocyclylene, which contains 1 to 3 nitrogen atoms and m R groups. 6 It is substituted with a group. In one embodiment, ring A is a 9-membered spiroheterocyclylene, which contains 1 to 3 nitrogen atoms and m R groups. 6 It is substituted with a group. In one embodiment, ring A is a 10-membered spiroheterocyclylene containing 1 to 3 nitrogen atoms, which is m R 6 It is substituted with a group. In one embodiment, the spiroheterocyclylene contains one nitrogen atom. In one embodiment, the spiroheterocyclylene contains two nitrogen atoms. In one embodiment, the spiroheterocyclylene contains three nitrogen atoms. In one embodiment, ring A is a 10-membered spiroheterocyclylene which contains one nitrogen atom and m R groups 6 It is substituted by the base.

[0088] One way of doing this is for each R 6 R is independently a halo, a C1-C6 alkyl, or a C1-C6 haloalkyl. In one embodiment, each R 6 R is independently a halo, a C1-C3 alkyl, or a C1-C3 haloalkyl. In one embodiment, each R 6 These are independently Cl, -CH3, or -CF3.

[0089] One reason, R 6 This is a halo. In one embodiment, R 6 is Cl, F, or Br. In one embodiment, R 6 is Cl. In one embodiment, R 6 is F. For one reason, R 6 It is Br.

[0090] One reason, R 6 is a C1-C6 alkyl group. In one embodiment, R 6 is a C1-C3 alkyl group. In one embodiment, R 6 is methyl, ethyl, n-propyl, or isopropyl. In one embodiment, R 6 is methyl. In one example, R 6 is ethyl. In one embodiment, R 6 is n-propyl. In one embodiment, R 6 It is isopropyl.

[0091] One reason, R 6 is a C1-C6 haloalkyl. In one embodiment, R 6 is a C1-C6 haloalkyl containing 1 to 13 halogen atoms. In one embodiment, R 6 is a C1-C3 haloalkyl. In one embodiment, R 6 is a C1-C3 haloalkyl containing 1 to 7 halogen atoms. In one embodiment, R6 is -CF3, -CHF2, -CH2F, -CCl3, -CHCl2, -CH2Cl, -CF2Cl, -CFCl2, -CH2CF3, -CH2CHF2, or -CH2CCl3. In one embodiment, R 6 is -CF3. In one example, R 6 It is -CHF2.

[0092] In one embodiment, m is between 0 and 5. In one embodiment, m is 0. In one embodiment, m is 1 or 2. In one embodiment, m is 1. In one embodiment, m is 2. In one embodiment, m is 3. In one embodiment, m is 4. In one embodiment, m is 5.

[0093] In one presentation, ring A is, [ka] That is the case.

[0094] In one presentation, ring A is, [ka] That is the case.

[0095] In one statement, L 2 is a combination or -N(R 7 )-. In one embodiment, L 2 L is a combination. In one example, 2 is -N(R 7 )- and R 7 is H or C1-C3 alkyl. In one embodiment, L 2 is -N(R 7 )- and R 7 This is either H or -CH3.

[0096] In one statement, L 2 It is a combination.

[0097] In one statement, L 2 is -N(R 7 )-. In one embodiment, R 7 is H or C1-C6 alkyl. In one embodiment, R 7 H is H. In one example, R 7 is a C1-C6 alkyl group. In one embodiment, R 7 is a C1-C3 alkyl group. In one embodiment, R 7 is methyl, ethyl, n-propyl, or isopropyl. In one embodiment, R 7 is methyl. In one example, R 7 is ethyl. In one embodiment, R 7 is n-propyl. In one embodiment, R 7 is isopropyl. In one embodiment, L 2 is -N(H)-. In one embodiment, L 2 is -N(CH3)-. In one embodiment, L 2 This is -N(CH2CH3)-.

[0098] One explanation, [ka] teeth, [ka] That is the case.

[0099] In one embodiment, the compound of formula (I) is formula (IIA) or (IIB): [ka] [In the formula, [ka] This is an 8-10 membered spiroheterocyclylene containing 1-3 nitrogen atoms, and the heterocyclylene contains m R 6 It is substituted by the group; here, R 1 , R 2a , R 2b , R 5a , R 5b , R 6 m, X, L 1 , L 2 , Y 1 and Y 2 This is as stated in equation (I). It is a compound of [the compound].

[0100] In one embodiment, the compound of formula (I) is formula (IIa) or (IIb): [ka] (In the formula, R 1 , R 2a , R 2b , R 6 m, X, L 1 , L 2 , Y 1 and Y 2 (This is as stated in equation (I)). It is a compound of [the compound].

[0101] In one embodiment, the compound of formula (I) is formula (IIc) or (IId): [ka] [In the formula, [ka] This is an 8-10 membered spiroheterocyclylene containing 1-3 nitrogen atoms, and the heterocyclylene contains m R 6 It is substituted by the group; here, R 1 , R 2a , R 2b , X, L 1 and L 2 This is as stated in equation (I). It is a compound of [the compound].

[0102] In one embodiment, the compound of formula (I) is formula (IIIa) or (IIIb): [ka] (In the formula, R 1 , R 2a , R 2b , R 5a , R 5b , R 7 , X, L 1 (and ring A is as shown in formula (I)) It is a compound of [the compound].

[0103] In one embodiment, the compound of formula (I) is (IVa), (IVb), (IVc), (IVd), or (IVe): [ka] (In the formula, n is an integer from 1 to 6; where R 1 , R 2a , R 2b , R 4 , R 5a , R 5b , X, ring A and L 2 (This is as stated in equation (I)). It is a compound of [the compound].

[0104] In one embodiment, the compound of formula (I) is formula (Va) or (Vb): [ka] (In the formula, R 1 , R 2a , R 2b , R 5a , R 5b , R 7 , X, L 1 (and ring A is as shown in formula (I)) It is a compound of [the compound].

[0105] In one embodiment, the compound of formula (I) is formula (VIa) or (VIb): [ka] (In the formula, R 1 , R 2a , R 2b , R 5a , R 5b , R 6 (and m are as shown in formula (I)) It is a compound of [the compound].

[0106] In this specification, any description, variation, embodiment, or aspect of any part may be combined with any description, variation, embodiment, or aspect of any other part, and each combination of descriptions is understood to be the same as if they were described specifically and individually. For example, with respect to ring A of formula (I), all descriptions, variations, embodiments, or aspects provided herein are R 1 , R 2a , R 2b , R 3 , R 4 , R 5a , R 5b , R 6 , R 7 , X, L 1 , L 2 , Y 1 , Y 2All descriptions, variations, embodiments, or aspects of and m may be combined, and each combination can be combined in the same way as if it were specifically and individually listed. Furthermore, all descriptions, variations, embodiments, or aspects of formula (I) apply equally to other formulas detailed herein where applicable, and for all formulas, each and all descriptions, variations, embodiments, or aspects are understood to be described as if they were described separately and individually. For example, all descriptions, variations, embodiments, or aspects of formula (I) apply equally to any of the formulas detailed herein where applicable, such as formulas (IIA), (IIB), (IIa), (IIb), (IIc), (IId), (IIIa), (IIIb), (IVa), (IVb), (IVc), (IVd), (IVe), (Va), (Vb), (VIa), and (VIb), and each and all descriptions, variations, embodiments, or aspects are described as if they were described separately and individually for all formulas.

[0107] In some embodiments, compounds selected from the compounds in Table 1 or their pharmaceutically acceptable salts are provided. While certain compounds described in this disclosure, including Table 1, are shown as specific stereoisomers and / or non-stereochemical forms, it will be understood that any or all stereochemical forms, including any enantiomer or diastereomer forms, as well as any tautomers or other forms of the compounds in this disclosure, including Table 1, are also described herein.

[0108] [Table 1] [Table 2] [Table 3] [Table 4] [Table 5] [Table 6] [Table 7] [Table 8] [Table 9]

[0109] In this specification, it is understood that combinations of substituents and / or variables in the depicted formulas are permissible only if such involvement results in a stable compound.

[0110] Furthermore, all compounds of formula (I) existing in the form of free bases or acids can be converted to their pharmaceutically acceptable salts by treatment with a suitable inorganic or organic base or acid using methods known to those skilled in the art. Salts of compounds of formula (I) can be converted to the form of free bases or acids by standard techniques.

[0111] Synthesis method The compounds described herein can be prepared using conventional organic synthesis methods and commercially available starting materials, or by methods provided herein. For illustrative purposes only, the compound of formula (I) may be prepared as outlined in Schemes 1 and 2, similar to the examples described herein. Those skilled in the art will understand how the methods described in the illustrative schemes and examples may be modified to obtain the desired product.

[0112] The compound of formula A can be prepared as outlined in Scheme 1. Under basic conditions, such as in the presence of DIPEA, intermediate a-1 is coupled with intermediate a-2 to form intermediate a-3, which is then deprotected under basic conditions to form intermediate a-4. Subsequently, intermediate a-4 is coupled with intermediate a-5 using TCFH / NMI to obtain intermediate a-6, which is then deprotected under basic conditions to obtain intermediate a-7. Intermediate a-7 is then coupled with intermediate a-8 to obtain the compound of formula A. [ka] [wherein X is a halo (e.g., chloro); each R is independently an alkyl (e.g., methyl or ethyl); R 1 , R a and R b This is as stated in formula (I); and [ka] teeth, [ka] This represents the base of equation (I) corresponding to [the given expression].

[0113] The compound of formula B can be prepared as outlined in Scheme 2. Intermediate a-5 is reduced to intermediate b-1 (e.g., using NaBH4), and then coupled with intermediate a-4 using TCFH / NMI to obtain intermediate b-2. Subsequent oxidation of b-2, followed by reductive amination using a-8 (e.g., using IBX and NaBH(OAc)3), yielded the compound of formula B. [ka] [wherein R is an alkyl such as methyl or ethyl; R 1 , R a and R b This is as stated in equation (I); [ka] teeth, [ka] This represents the part of equation (I) that corresponds to [the given expression].

[0114] How to use Embodiments of this disclosure provide a method for modulating IRAK4 in a subject requiring modulation, characterized by administering an effective amount of a compound of formula (I) to the subject. Modulation of IRAK4 (e.g., inhibition or activation) can be evaluated and demonstrated by various methods known in the art. Kits and commercially available assays are available to determine whether and to what extent IRAK4 has been modulated (e.g., inhibited or activated).

[0115] In one embodiment, provided herein is a method for modulating IRAK4, characterized by contacting IRAK4 with an effective amount of the compound of formula (I) or any embodiment or variation thereof. In some embodiments, the compound of formula (I) inhibits IRAK4. In another embodiment, the compound of formula (I) activates IRAK4. In some embodiments, the compound of formula (I) is an agonist of IRAK4. In some embodiments, the compound of formula (I) is an antagonist of IRAK4.

[0116] In some embodiments, provided herein are methods for targeting and degrading IRAK4, characterized by contacting IRAK4 with an effective amount of a compound of formula (I) or any embodiment or modification thereof.

[0117] In some embodiments, the compound of formula (I) adjusts the activity of IRAK4 to approximately 1%, 5%, 10%, 15%, 20%, 25%, 30%, 35%, 40%, 45%, 50%, 55%, 60%, 65%, 70%, 75%, 80%, 85%, 90%, 95%, or 100%. In some embodiments, the compound of formula (I) increases the activity of IRAK4 by approximately 1-100%, 5-100%, 10-100%, 15-100%, 20-100%, 25-100%, 30-100%, 35-100%, 40-100%, 45-100%, 50-100%, 55-100%, 60-100%, 65-100%, 70-100%, 75-100%, and 80-100%. Adjust to 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%.

[0118] Furthermore, provided in certain embodiments of this disclosure is a method for degrading IRAK4 in an object that needs to be degraded, characterized by administering an effective amount of the compound of formula (I) to the object. Degradation of IRAK4 can be evaluated and demonstrated by various methods known in the art. Kits and commercially available assays (e.g., cell line assays) can be used to determine whether and to what extent IRAK4 has been degraded.

[0119] In one embodiment, provided herein is a method for degrading IRAK4, characterized by contacting IRAK4 with an effective amount of the compound of formula (I) or any embodiment or variation thereof. In some embodiments, the compound of formula (I) partially degrades IRAK4. In some embodiments, the compound of formula (I) completely degrades IRAK4.

[0120] In some embodiments, the compound of formula (I) degrades IRAK4 by about 1%, 5%, 10%, 15%, 20%, 25%, 30%, 35%, 40%, 45%, 50%, 55%, 60%, 65%, 70%, 75%, 80%, 85%, 90%, 95%, or 100%. In some embodiments, the compound of formula (I) contains IRAK4 in amounts of approximately 1-100%, 5-100%, 10-100%, 15-100%, 20-100%, 25-100%, 30-100%, 35-100%, 40-100%, 45-100%, 50-100%, 55-100%, 60-100%, 65-100%, 70-100%, 75-100%, 80-100%, Decompose into 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%.

[0121] In another embodiment, provided herein is a method for treating an inflammatory disease or autoimmune disease in a subject requiring treatment, comprising administering an effective amount of a compound of formula (I) to the subject. In some embodiments, provided herein is a method for treating an inflammatory disease in a subject requiring treatment, comprising administering an effective amount of a compound of formula (I) to the subject. In some embodiments, provided herein is a method for treating an autoimmune disease in a subject requiring treatment, comprising administering an effective amount of a compound of formula (I) to the subject. In some embodiments, provided herein is a method for preventing an inflammatory disease or autoimmune disease in a subject requiring treatment, comprising administering an effective amount of a compound of formula (I) to the subject. In some embodiments, provided herein is a method for preventing an inflammatory disease in a subject requiring treatment, comprising administering an effective amount of a compound of formula (I) to the subject. In some embodiments, provided herein is a method for preventing an autoimmune disease in a subject requiring treatment, comprising administering an effective amount of a compound of formula (I) to the subject. Non-exclusive examples of inflammatory or autoimmune diseases include atopic dermatitis, asthma, lupus, rheumatoid arthritis, familial Mediterranean fever, psoriasis, generalized pustular psoriasis, cryopyrin-associated periodic syndromes, hidradenitis suppurativa, Beckett syndrome, or familial cold autoinflammatory syndrome.

[0122] In some embodiments, administering a compound of formula (I) to a subject predisposed to inflammatory or autoimmune diseases prevents the subject from developing any symptoms of an inflammatory or autoimmune disease (e.g., tumor growth or metastasis). In some embodiments, administering a compound of formula (I) to a subject that has not yet shown any symptoms of an inflammatory or autoimmune disease prevents the subject from developing any symptoms of an inflammatory or autoimmune disease. In some embodiments, administering a compound of formula (I) to a subject in need of treatment reduces the severity of the inflammatory or autoimmune disease in the subject. In some embodiments, administering a compound of formula (I) to a subject in need of treatment stabilizes the inflammatory or autoimmune disease (prevents or delays its worsening). In some embodiments, administering a compound of formula (I) to a subject in need of treatment delays the onset or recurrence of the inflammatory or autoimmune disease. In some embodiments, administering a compound of formula (I) to a subject in need of treatment slows the progression of the inflammatory or autoimmune disease. In some embodiments, administering a compound of formula (I) to a subject in need of treatment provides partial remission of an inflammatory or autoimmune disease. In some embodiments, administering a compound of formula (I) to a subject in need of treatment provides complete remission of an inflammatory or autoimmune disease. In some embodiments, administering a compound of formula (I) to a subject in need of treatment reduces the dose of one or more other drugs required to treat the inflammatory or autoimmune disease. In some embodiments, administering a compound of formula (I) to a subject in need of treatment enhances the effect of other drugs used to treat the inflammatory or autoimmune disease. In some embodiments, administering a compound of formula (I) to a subject in need of treatment slows the progression of the inflammatory or autoimmune disease. In some embodiments, administering a compound of formula (I) to a subject in need of treatment improves the quality of life of a subject with an inflammatory or autoimmune disease.In some embodiments, administering a compound of formula (I) to a subject in need of treatment extends the survival time of subjects with inflammatory or autoimmune diseases.

[0123] In one embodiment, the foregoing provides a method for preventing a subject predisposed to an inflammatory disease or autoimmune disease from developing any symptoms of an inflammatory disease or autoimmune disease, the method comprising administering a compound of formula (I) to the subject. In some embodiments, the foregoing provides a method for preventing a subject that has not yet shown symptoms of an inflammatory disease or autoimmune disease from developing symptoms of an inflammatory disease or autoimmune disease, the method comprising administering a compound of formula (I) to the subject.

[0124] In some embodiments, the foregoing provides a method for reducing the severity of an inflammatory or autoimmune disease in a subject, the method comprising administering a compound of formula (I) to the subject. In some embodiments, the foregoing provides a method for stabilizing an inflammatory or autoimmune disease in a subject, the method comprising administering a compound of formula (I) to the subject. In some embodiments, the method prevents the worsening of an inflammatory or autoimmune disease. In some embodiments, the method slows the worsening of an inflammatory or autoimmune disease.

[0125] In another embodiment, provided herein is a method for delaying the onset or recurrence of an inflammatory or autoimmune disease in a subject, the method comprising administering a compound of formula (I) to the subject.

[0126] In some embodiments, the methods provided herein are for slowing the progression of an inflammatory or autoimmune disease in a subject, the methods comprising administering a compound of formula (I) to the subject. In some embodiments, the methods result in partial remission of the inflammatory or autoimmune disease. In some embodiments, the methods result in complete remission of the inflammatory or autoimmune disease.

[0127] In a further embodiment, provided herein is a method for reducing the dose of one or more other agents required to treat an inflammatory or autoimmune disease in a subject, the method comprising administering a compound of formula (I) to the subject. In some embodiments, provided herein is a method for enhancing the effect of another agent used to treat an inflammatory or autoimmune disease in a subject, the method comprising administering a compound of formula (I) to the subject.

[0128] Furthermore, a method is provided for delaying the progression of inflammatory or autoimmune diseases in subjects, the method comprising administering a compound of formula (I) to the subject. In some embodiments, the method improves the quality of life of subjects with inflammatory or autoimmune diseases. In some embodiments, the method extends the survival time of subjects with inflammatory or autoimmune diseases.

[0129] In another embodiment, provided herein are methods for treating symptoms of an inflammatory or autoimmune disease caused by a disease in a subject requiring treatment, comprising administering an effective amount of the compound of formula (I) to the subject. In some embodiments, provided herein are methods for preventing symptoms of an inflammatory or autoimmune disease caused by a disease in a subject requiring treatment, comprising administering an effective amount of the compound of formula (I) to the subject. In some embodiments, administering the compound of formula (I) to a subject predisposed to a disease that causes symptoms of an inflammatory or autoimmune disease prevents the subject from developing symptoms of the inflammatory or autoimmune disease. In some embodiments, administering the compound of formula (I) to a subject that has not yet developed symptoms of an inflammatory or autoimmune disease related to a disease that causes symptoms of an inflammatory or autoimmune disease can prevent the subject from developing symptoms of the inflammatory or autoimmune disease. In some embodiments, administering the compound of formula (I) to a subject requiring treatment reduces the severity of symptoms of an inflammatory or autoimmune disease caused by a disease in the subject. In some embodiments, administering a compound of formula (I) to a subject in need of treatment stabilizes the symptoms of an inflammatory or autoimmune disease (prevents or delays the worsening of symptoms of an inflammatory or autoimmune disease). In some embodiments, administering a compound of formula (I) to a subject in need of treatment delays the onset or recurrence of symptoms of an inflammatory or autoimmune disease caused by the disease. In some embodiments, administering a compound of formula (I) to a subject in need of treatment slows the progression of symptoms of an inflammatory or autoimmune disease caused by the disease. In some embodiments, administering a compound of formula (I) to a subject in need of treatment results in partial remission of the disease causing symptoms of an inflammatory or autoimmune disease. In some embodiments, administering a compound of formula (I) to a subject in need of treatment results in complete remission of the disease causing symptoms of an inflammatory or autoimmune disease.In some embodiments, administering a compound of formula (I) to a subject in need of treatment reduces the dose of one or more other drugs required to treat a disease causing symptoms of an inflammatory or autoimmune disease. In some embodiments, administering a compound of formula (I) to a subject in need of treatment enhances the effect of other drugs used to treat symptoms of an inflammatory or autoimmune disease. In some embodiments, administering a compound of formula (I) to a subject in need of treatment delays the progression of a disease causing symptoms of an inflammatory or autoimmune disease. In some embodiments, administering a compound of formula (I) to a subject in need of treatment improves the quality of life of a subject with a disease causing symptoms of an inflammatory or autoimmune disease. In some embodiments, administering a compound of formula (I) to a subject in need of treatment extends the survival time of a subject with a disease causing symptoms of an inflammatory or autoimmune disease. In some embodiments, the disease is atopic dermatitis, asthma, lupus, rheumatoid arthritis, familial Mediterranean fever, psoriasis, generalized pustular psoriasis, cryopyrin-associated periodic syndromes, hidradenitis suppurativa, Beckett syndrome, or familial cold autoinflammatory syndrome.

[0130] In some embodiments, compounds of formula (I) are useful for treating diseases selected from atopic dermatitis, asthma, lupus, rheumatoid arthritis, familial Mediterranean fever, psoriasis, generalized pustular psoriasis, cryopyrin-associated periodic syndromes, hidradenitis suppurativa, Beckett syndrome, and familial cold autoinflammatory syndrome.

[0131] Pharmaceutical composition and route of administration The compounds provided herein may be administered orally, topically, or parenterally to a subject by conventional dosage forms, such as capsules, microcapsules, tablets, granules, powders, lozenges, tablets, suppositories, injections, suspensions, syrups, patches, creams, lotions, ointments, gels, sprays, solutions, and emulsions.

[0132] The compounds disclosed herein may be administered orally, topically, or parenterally to a subject in the form of conventional dosage forms, such as capsules, microcapsules, tablets, granules, powders, lozenges, pills, suppositories, injections, suspensions, syrups, patches, creams, lotions, ointments, gels, sprays, solutions, and emulsions. Suitable formulations may include 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 hydroxypropylcellulose, sodium bicarbonate, calcium phosphate, or calcium citrate), and lubricants (e.g., magnesium stearate, light It can be manufactured by conventional methods using conventional organic or inorganic additives such as anhydrous silicic acid, talc, sodium lauryl sulfate, flavoring agents (e.g., citric acid, menthol, glycine, or orange powder), preservatives (e.g., sodium benzoate, sodium bisulfate, methylparaben, or propylparaben), stabilizers (e.g., citric acid, sodium citrate, or acetic acid), suspending agents (e.g., methylcellulose, polyvinylpyrrolidone, or aluminum stearate), dispersants (e.g., hydroxypropyl methylcellulose), diluents (e.g., water), and base waxes (e.g., cocoa butter, white petrolatum, or polyethylene glycol). The effective amount of the compound of formula (I) in the pharmaceutical composition may be present at a level that produces the desired effect; for example, in unit doses for both oral and parenteral administration, it ranges from about 0.005 mg per kg of body weight to about 10 mg per kg of body weight of the subject.

[0133] The dose of the compound of formula (I) administered to a subject can vary over a wide range and is subject to the judgment of the healthcare professional. Generally, the compounds disclosed herein can be administered at doses of approximately 0.001 mg to approximately 10 mg per kg of body weight of the subject, 1 to 4 times a day, but the above dose can be appropriately changed depending on the subject's age, weight, medical condition, and route of administration. In any case, the amount of compound of formula (I) administered will vary depending on factors such as the solubility of the active ingredient, the formulation used, and the route of administration.

[0134] The compound of formula (I) may be administered orally for convenience. In one embodiment, when administered orally, the compound of formula (I) is administered with food and water. In another embodiment, the compound of formula (I) is administered orally as a solution or suspension, dispersed in water or juice (e.g., apple juice or orange juice) or any other liquid.

[0135] The compounds disclosed herein may also be administered intradermally, intramuscularly, intraperitoneally, percutaneously, intravenously, subcutaneously, intranasally, epidurally, sublingually, intracerebrally, intravaginally, percutaneously, rectally, mucous membrane, or by inhalation, or topically to the ear, nose, eye, or skin. The mode of administration is at the discretion of the healthcare professional and may be partially modified depending on the site of the condition.

[0136] In one embodiment, what is provided herein is a capsule containing a compound of formula (I) that does not contain additional carriers, excipients, or vehicles.

[0137] In another embodiment, provided herein is a composition comprising an effective amount of a compound of formula (I) and a pharmaceutically acceptable carrier or vehicle, wherein the pharmaceutically acceptable carrier or vehicle may include excipients, diluents, or mixtures thereof. In one embodiment, the composition is a pharmaceutical composition.

[0138] The compositions may take the form of tablets, chewable tablets, capsules, solutions, parenteral solutions, lozenges, suppositories, and suspensions. The compositions may be formulated to contain a daily dose, or a suitable portion of a daily dose, in a single tablet or capsule, or in a suitable amount of liquid, as a dosage unit. In one embodiment, the solution is prepared from a water-soluble salt such as hydrochloride. In general, all compositions are manufactured according to known methods in pharmaceutical chemistry. Capsules can be manufactured by mixing the compound of formula (I) with a suitable carrier or diluent and filling a suitable amount of the mixture into a capsule. Common carriers and diluents include, but are not limited to, a wide variety of starches, powdered cellulose, especially crystalline and microcrystalline cellulose, sugars such as fructose, mannitol and sucrose, grain flour and similar edible powders, and other inert powders.

[0139] Tablets can be manufactured by direct compression, wet granulation, or dry granulation. The formulations typically contain not only the compound but also diluents, binders, lubricants, and disintegrants. Typical diluents include, for example, various starches, lactose, mannitol, kaolin, calcium phosphate or sulfate, inorganic salts such as sodium chloride, and powdered sugars. Powdered cellulose derivatives are also useful. Typical tablet binders include substances such as starch, gelatin, and sugars (e.g., lactose, fructose, glucose). Natural and synthetic gums such as acacia, alginates, methylcellulose, and polyvinylpyrrolidine are also convenient. Polyethylene glycol, ethylcellulose, and waxes can also function as binders.

[0140] In tablets, a lubricant may be necessary to prevent dye from adhering to the tablet and the tableting machine. The lubricant can be selected from lubricating solids such as talc, magnesium stearate, calcium stearate, stearic acid, and hydrogenated vegetable oils. Tablet disintegrants are substances that swell and disintegrate the tablet upon contact with moisture, releasing compounds. These include starch, clay, cellulose, algins, and gums. More specifically, corn and potato starch, methylcellulose, agar, bentonite, wood cellulose, powdered natural sponge, cation exchange resins, alginic acid, guar gum, citrus pulp, and carboxymethylcellulose, such as sodium lauryl sulfate, can also be used. Tablets may also be coated with sugars as flavoring or odor-modifying agents, or with film-forming protective agents to alter the tablet's solubility. Furthermore, the composition can be formulated as a chewable tablet by using substances such as mannitol in the formulation.

[0141] When administering the compound of formula (I) as a suppository, typical bases can be used. Cocoa butter is a conventional suppository base and can be modified by adding wax to slightly raise its melting point. In particular, water-miscible suppository bases containing polyethylene glycol of various molecular weights are widely used.

[0142] The effects of the compound of formula (I) can be delayed or prolonged by appropriate formulation. For example, pellets of the compound of formula (I) that can dissolve gradually can be prepared and incorporated into tablets or capsules, or as a sustained-release implantable device. This technique also includes preparing pellets with several different dissolution rates and filling a capsule with a mixture of these pellets. The tablets or capsules may be coated with a film that resists dissolution for a predictable period. Even parenteral formulations can be made long-acting by dissolving or suspending the compound of formula (I) in an oily or emulsifying agent that disperses slowly in serum.

[0143] Exemplary Example This disclosure is further illustrated by the following embodiments.

[0144] Embodiment P1. Formula (I): [ka] [In the formula, R 1 These are C1-C6 haloalkyl, C1-C6 alkyl, or -CN; R 2a is H or C1-C6 alkyl; R 2b This is 1 to 5 R 3 A C5-C6 cycloalkyl group which may be optionally substituted with a group; or R 2a and R 2b The dotted line between them represents a ring structure, R 2a and R 2b These, together with the nitrogen atoms bonded to them, form a six-membered heterocycline which may optionally contain one additional heteroatom selected from N and O, and the heterocycline has 1 to 5 R 3 The base may be replaced as desired; Each R 3 These are independently -NH2, -OH, halo, C1-C6 alkyl, or C1-C6 haloalkyl; X is either CH or N; L 1 -C(O)N(H)-, -N(H)C(O)-, -C(O)-, -(C1-C6 alkylene)N(R 4 )- or C1-C6 alkylene; R 4 is H or C1-C6 alkyl; R 5a and R 5b Each of these is either H, or they together form an oxo group; Ring A is, [ka] It is either an 8-10 member spiroheterocyclylene containing 1-3 nitrogen atoms, wherein the heterocyclylene contains m R 6 It is substituted with the base; Y 1 and Y 2 It is independently CH or N; Each R 6 These are independently halo, C1-C6 alkyl, or C1-C6 haloalkyl; m is between 0 and 5; L 2 is a combination or -N(R 7 )-and; and R 7 [It is H or C1-C6 alkyl] It is a compound of or a pharmaceutically acceptable salt thereof.

[0145] Embodiment P2. R 1 However, the compound described in Embodiment P1 or a pharmaceutically acceptable salt thereof is a C1-C3 haloalkyl, C1-C3 alkyl, or -CN.

[0146] Embodiment P3. R 1 However, the compound described in Embodiment P2 or a pharmaceutically acceptable salt thereof is -CHF2, -CF3, -CH3, or -CN.

[0147] Embodiment P4. R 2a However, it is H or C1-C3 alkyl; and R 2b However, 1 to 3 R 3 The compound according to any one of embodiments P1 to P3 or a pharmaceutically acceptable salt thereof, which is a cyclohexyl group that may be optionally substituted with a base.

[0148] Embodiment P5. R 2a However, it is H or -CH3; R 2b However, 1-2 R 3 The compound according to Embodiment P4 or a pharmaceutically acceptable salt thereof, which is a cyclohexyl group that may be optionally substituted at the base.

[0149] Embodiment P6. R 2a However, H is; R 2b However, one R 3 The compound according to Embodiment P5 or a pharmaceutically acceptable salt thereof, which is a cyclohexyl group that may be optionally substituted at the base.

[0150] Embodiment P7. R 2a and R 2b However, together with the nitrogen atoms bonded to them, they form a 6-membered heterocycline which may optionally contain one additional heteroatom selected from N and O, and the heterocycline contains 1 to 5 R 3 The compound according to any one of embodiments P1 to P3 or a pharmaceutically acceptable salt thereof, which may be optionally substituted with a base.

[0151] Embodiment P8. R 2a and R 2b However, together with the nitrogen atoms bonded to them, they form a 6-membered heterocycline which may optionally contain one additional heteroatom selected from N and O, and the heterocycline contains 1 to 2 R 3 The compound described in Embodiment P7 or a pharmaceutically acceptable salt thereof, which may be optionally substituted with a base.

[0152] Embodiment P9. Each R 3 However, independently, the compound is -NH2, -OH, halo, C1-C3 alkyl, or C1-C3 haloalkyl, or a pharmaceutically acceptable salt thereof, as described in any one of Embodiments P1 to P8.

[0153] Embodiment P10. Each R 3 However, independently, these are the compounds described in Embodiment P9 or pharmaceutically acceptable salts thereof, which are -NH2, -OH, F, Cl, -CH3, or -CF3.

[0154] Embodiment P11. Each R 3However, independently, these are the compounds described in Embodiment P10 or pharmaceutically acceptable salts thereof, which are -NH2, -OH, F, or -CH3.

[0155] Embodiment P12. [ka] but, [ka] The compound is one of the compounds described in any one of embodiments P1 to P3 and P7 to P11, or a pharmaceutically acceptable salt thereof.

[0156] Embodiment P13. [ka] but, [ka] It is a compound described in any one of P1-P6 and P9-P11 or a pharmaceutically acceptable salt thereof.

[0157] Embodiment P14. X is a compound according to any one of Embodiments P1 to P13 or a pharmaceutically acceptable salt thereof, wherein X is CH.

[0158] Embodiment P15. X is a compound according to any one of Embodiments P1 to P13 or a pharmaceutically acceptable salt thereof, wherein X is N.

[0159] Embodiment P16. L 1 However, -C(O)N(H)-, -N(H)C(O)-, -C(O)-, -(C1-C3 alkylene)N(R 4 )- or C1-C3 alkylene; and R 4 The compound is one of the compounds described in any one of Embodiments P1 to P15 or a pharmaceutically acceptable salt thereof, which is H or a C1-C3 alkyl group.

[0160] Embodiment P17. L 1 The compound described in Embodiment P16 or a pharmaceutically acceptable salt thereof is -C(O)N(H)-, -N(H)C(O)-, -C(O)-, -CH2N(CH3)-, -CH2-, or -CH2CH2-.

[0161] Embodiment P18. R 5a and R 5b However, each of these is H, and is a compound or a pharmaceutically acceptable salt thereof described in any one of embodiments P1 to P17.

[0162] Embodiment P19. R 5a and R 5b However, together they form an oxo group, which is the compound described in any one of embodiments P1 to P17 or a pharmaceutically acceptable salt thereof.

[0163] Embodiment P20. Ring A is [ka] The compound is one of the embodiments P1 to P19 or a pharmaceutically acceptable salt thereof.

[0164] Embodiment P21. Y 1 However, N is; Y 2 However, the compound described in Embodiment P20 or a pharmaceutically acceptable salt thereof is CH.

[0165] Embodiment P22. Y 1 However, it is CH; Y 2 However, N is the compound described in Embodiment P20 or a pharmaceutically acceptable salt thereof.

[0166] Embodiment P23. Y 1 and Y 2 However, each of these is N, which is the compound described in Embodiment P20 or a pharmaceutically acceptable salt thereof.

[0167] Embodiment P24. Y 1and Y 2 However, each of these is a CH compound or a pharmaceutically acceptable salt thereof as described in Embodiment P20.

[0168] Embodiment P25. Ring A is an 8-10 membered spiroheterocyclylene containing two nitrogen atoms, and the heterocyclylene contains m R 6 A compound or a pharmaceutically acceptable salt thereof described in any one of embodiments P1 to P19, which is substituted with a group.

[0169] Embodiment P26. Ring A is a 10-membered spiroheterocyclylene containing two nitrogen atoms, and the heterocyclylene contains m R 6 A compound described in Embodiment P25 or a pharmaceutically acceptable salt thereof, which is substituted with a group.

[0170] Embodiment P27. Each R 6 However, independently, the compound described in any one of Embodiments P1 to P26 or a pharmaceutically acceptable salt thereof is a halo, C1-C3 alkyl, or C1-C3 haloalkyl.

[0171] Embodiment P28. Each R 6 However, independently, these are the compounds described in Embodiment P27 or pharmaceutically acceptable salts thereof, which are Cl, -CH3 or -CF3.

[0172] Embodiment P29. The compound or a pharmaceutically acceptable salt thereof described in any one of Embodiments P1 to P26, wherein m is 0.

[0173] Embodiment P30. m is a compound according to any one of Embodiments P1 to P28 or a pharmaceutically acceptable salt thereof, wherein m is 1 or 2.

[0174] Embodiment P31. Ring A is [ka] The compound is one of the compounds described in any one of embodiments P1 to P24 and P29, or a pharmaceutically acceptable salt thereof.

[0175] Embodiment P32. Ring A is [ka] The compound or a pharmaceutically acceptable salt thereof described in any one of embodiments P1 to P19, P25, P26, and P29.

[0176] Embodiment P33. L 2 However, the compound is a compound described in any one of embodiments P1 to P32 or a pharmaceutically acceptable salt thereof.

[0177] Embodiment P34. L 2 However, -N(R 7 )-and; R 7 The compound is one of the compounds described in any one of Embodiments P1 to P32 or a pharmaceutically acceptable salt thereof, which is H or a C1-C3 alkyl group.

[0178] Embodiment P35. L 2 However, -N(R 7 )-and; R 7 The compound described in Embodiment P34 or a pharmaceutically acceptable salt thereof is H or -CH3.

[0179] Embodiment P36. [ka] but, [ka] The compound is one of the compounds described in any one of embodiments P1 to P35 or a pharmaceutically acceptable salt thereof.

[0180] Embodiment P37. The compound is of formula (IIa) or (IIb): [ka] The compound or a pharmaceutically acceptable salt thereof described in any one of embodiments P1-P24, P27-P31, and P33-P35.

[0181] Embodiment P38. The compound is of formula (IIIa) or (IIIb): [ka] The compound is one of the compounds described in any one of embodiments P1 to P36 or a pharmaceutically acceptable salt thereof.

[0182] Embodiment P39. A compound selected from the compounds in Table 1 or their pharmaceutically acceptable salts.

[0183] Embodiment P40. A pharmaceutical composition comprising a compound described in any one of Embodiments P1 to P39 or a pharmaceutically acceptable salt thereof, and a pharmaceutically acceptable excipient.

[0184] Embodiment P41. A method for modulating interleukin-1 (IL1) receptor-related kinase 4 (IRAK4), characterized by contacting IRAK4 with an effective amount of a compound described in any one of Embodiments P1 to P39 or a pharmaceutically acceptable salt thereof, or a pharmaceutical composition described in Embodiment P40.

[0185] Embodiment P42. A method for treating an inflammatory disease or autoimmune disease in a subject, characterized by administering an effective amount of IRAK4, a compound described in any one of Embodiments P1 to P39 or a pharmaceutically acceptable salt thereof, or a pharmaceutical composition described in Embodiment P40, to a subject in need of treatment.

[0186] Embodiment P43. The method according to Embodiment P42, wherein the inflammatory or autoimmune disease is atopic dermatitis, asthma, lupus, rheumatoid arthritis, familial Mediterranean fever, psoriasis, generalized pustular psoriasis, cryopyrin-associated periodic syndromes, sweat gland abscess, Beckett syndrome, or familial cold autoinflammatory syndrome. [Examples]

[0187] The following examples are for illustrative purposes only and are not limiting. The compounds were named using the automated naming tool provided in ChemBiodraw Ultra (Cambridgesoft), which generates systematic names for chemical structures while adhering to the Kahn-Ingold-Prelogue order rule for stereochemistry. Those skilled in the art can modify the procedures described in the illustrated examples to arrive at the target product.

[0188] Salts of the compounds described herein can be prepared by standard methods such as mixing an acid (e.g., TFA, formic acid, or HCl) into the mobile phase during chromatographic purification, or stirring the product after chromatographic purification with an acidic solution (e.g., an aqueous solution of HCl).

[0189] The following abbreviations may be relevant to this application. Abbreviations ACN or MeCN: Acetonitrile aq: aqueous solution BCA assay: Bicinchonin acid assay n-Bu4OAc: Tetrabutylacetate CBM: Cereblon binding site CV: Column volume DABCO:1,4-Diazabicyclo[2.2.2]Octane DCE: Dichloroethane DCM: Dichloromethane DIBAL-H: Diisobutylaluminum hydride DIPEA: N,N-diisopropylethylamine DMA: Dimethylacetamide DMF: Dimethylformamide DMPAO:2-((2,6-dimethylphenyl)amino)-2-oxoacetate DMPU:N,N'-dimethylpropyleneurea DMSO: Dimethyl sulfoxide DP: Desired product equiv. or eq.: equivalent quantity ESI: Electrospray Ionization Et3N: Triethylamine æ:ethyl acetate FA: Formic acid FBS: Fetal Bovine Serum FC: Flash chromatography h: time HATU:1-[bis(dimethylamino)methylene]-1H-1,2,3-triazolo[4,5-b]pyridinium 3-oxidehexafluorophosphate HPLC: High-Performance Liquid Chromatography IBX: 2-Iodoxybenzoic acid Ir(ppy)2(dtbbpy)PF6:4,4'-bis(1,1-dimethylethyl)-2,2'-bipyridine-N1,N1']bis[2-(2-pyridinyl-N)phenyl-C]iridium(III) hexafluorophosphate i-PrOH: Isopropanol LCMS: Liquid Chromatography Mass Spectrometry MeOH: methanol MsCl: Mesyl chloride MSD: Mass Selective Detector MTBE: Methyl tert-butyl ether NaBH(OAc)3: Sodium triacetoxyborohydride Ni-dtbbpy-Br2:[4,4'-bis(1,1-dimethylethyl)-2,2'-bipyridine]nickel(II) dichloride NMI: N-methylimidazole Pd-PEPPSI-IPent: Dichloro[1,3-bis(2,6-di-3-pentylphenyl)imidazole-2-ylidene](3-chloropyridyl)palladium(II) pdt: product PhMe: Toluene Rf: Delay coefficient rt: room temperature rt: retention time sat.: saturation SM: Starting material TCFH: Chloro-N,N,N',N'-tetramethylformamidinium hexafluorophosphate TFA: Trifluoroacetic acid THF: Tetrahydrofuran THP: Tetrahydropyran TLC: Thin-layer chromatography

[0190] Synthesis Examples LCMS method LCMS method 1 Column: Luna C18 (2) 50 x 3 mm, 3 μm. Temperature: 45°C. Flow rate: 1.5 mL / min. Duration: 2.5 minutes. Mobile phase conditions: Initially 95% H2O + 0.1% FA / 5% MeCN + 0.1% FA, linear gradient to 95% MeCN + 0.1% FA over 1.3 minutes, then held at 95% MeCN + 0.1% FA for 1.2 minutes. MSD: ESI positive.

[0191] LCMS method 2 Column: Kinetex Polar C18 2.6 um, 50 x 3.0 mm. Temperature: 45°C. Flow rate: 1.2 mL / min. Duration: 3 minutes. Mobile phase conditions: Initially 95% H2O + 0.1% FA / 5% MeCN + 0.1% FA, then a linear gradient to 95% MeCN + 0.1% FA over 1.5 minutes, followed by a 1.5-minute retention at 95% MeCN + 0.1% FA. MSD: Positive.

[0192] LCMS method 3 Column: Kinetex Polar C18 2.6 um, 50 x 3.0 mm. Temperature: 40°C. Flow rate: 1.2 mL / min. Experiment duration: 6 minutes. Mobile phase conditions: Initially 95% H2O + 0.1% FA / 5% MeCN + 0.1% FA, then a linear gradient to 95% MeCN + 0.1% FA over 3.5 minutes, followed by a 2.5-minute retention at 95% MeCN + 0.1% FA. MSD: Positive.

[0193] LCMS method 4 Column: Luna C18 (2) 50 x 3 mm, 3 μm. Temperature: 45°C. Flow rate: 1.5 mL / min. Duration: 3.5 minutes. Mobile phase conditions: Initially 95% H2O + 0.1% FA / 5% MeCN 0.1% FA, then a linear gradient to 95% MeCN + 0.1% FA over 1.3 minutes, followed by a retention of 95% MeCN 0.1% FA for 2.2 minutes. MSD: ESI positive.

[0194] LCMS method 5 Column: SunFire C18 75 x 4.6 mm, 3.5 μm. Temperature: 45°C. Flow rate: 1.5 mL / min. Experiment duration: 6 minutes. Mobile phase conditions: Initially 95% H2O + 0.1% FA / 5% MeCN + 0.1% FA, then a linear gradient to 95% MeCN + 0.1% FA over 4 minutes, followed by a 2-minute hold at 95% MeCN + 0.1% FA. MSD: Positive.

[0195] LCMS method 6 Column: C18 4.6 x 100 mm. Starting gradient: 95% H2O + 0.1% FA / 5% MeCN + 0.1% FA for 10 minutes, with 1.5 minutes of equilibration. Gradient from 95% H2O to 0% over 0-8 minutes and hold for 2 minutes.

[0196] Synthesis of intermediates The designation of specific intermediate compounds with numbers such as 1 or 2 is unique to the example described. Therefore, while multiple examples may refer to the same intermediate compound number, such as 1 or 2, the chemical structure of the compound differs between different examples.

[0197] Example I-1. Synthesis of a typical intermediate T-1 [ka]

[0198] Step 1. Production of methyl 4-methylsulfonyloxycyclohexanecarboxylate (2) To a solution of methyl 4-hydroxycyclohexanecarboxylate 1 (3.0 g, 18.96 mmol) and Et3N (3.96 mL, 28.45 mmol) in CH2Cl2 (60 mL), methanesulfonyl chloride (1.91 mL, 24.65 mmol) was added dropwise at 0°C. The reaction mixture was stirred at 0°C for 65 minutes. This reaction was tracked by TLC (6 / 4: siRNA / heptane); the target compound 2 had an Rf of 0.30, and the sample was placed simultaneously with 1 (Rf = 0.25) until it was completely converted. Water was added, and the aqueous phase was extracted with CH2Cl2 (3 x 30 mL). The organic phase was dried over Na2SO4, and the solvent was evaporated to dryness to obtain 2 as a pale yellow oil (4.45 g, quantitative yield). NMR was not performed. LC-MS method 1: 99.9% purity (at 215 nm), [M-Ms+H]+ = 141.2; [M+Na]+ = 259.2.

[0199] Step 2. Preparation of methyl 4-(3-cyano-4-nitropyrazole-1-yl)cyclohexanecarboxylate (4) To a solution of methyl 4-methylsulfonyloxycyclohexanecarboxylate 2 (2.05 g, 8.69 mmol) and 4-nitro-1H-pyrazole-3-carbonitrile 3 (1.0 g, 7.24 mmol) in dry DMF (1.5 mL), cesium carbonate (3.54 g, 10.86 mmol) was added at room temperature. The resulting mixture was stirred overnight at 90°C. LC-MS showed a 50% conversion (rt = 1.70 min). Water was added to the mixture, and the aqueous phase was extracted with ELISA (3 x). The organic phase was washed once with brine and dried over sodium sulfate. The solvent was evaporated to obtain 4 as a pale yellow solid (694 mg, yield = 34%, purity 97.7% by LC-MS).

[0200] LC-MS method 1: 97.7% purity (at 215 nm), [M+H] + = 279.2. 1 H NMR (400 MHz, DMSO-d6) δ ppm 1.47 - 1.59 (m, 2 H), 1.69 - 1.73 (m, 1 H), 1.84 (qd, J = 12.5, 3.3 Hz, 2 H), 2.03 - 2.15 (m, 4 H), 2.37 - 2.47 (m, 1 H), 3.61 - 3.63 (m, 3 H), 4.42 (tt, J = 11.9, 3.9 Hz, 1 H), 9.21 (s, 1 H).

[0201] Step 3. Preparation of methyl 4-(4-amino-3-cyanopyrazole-1-yl)cyclohexanecarboxylate (T-1) Nitrogen was passed through a mixed solution of methyl 4-(3-cyano-4-nitropyrazole-1-yl)cyclohexanecarboxylate 4 (750.0 mg, 2.7 mmol) in ethyl acetate (20.2 mL) and methanol (6.7 mL) and bubbling was performed for 5 minutes. Then, 10% Pd / C (573.67 mg, 0.54 mmol) was added, and nitrogen was passed through the solution and bubbling was performed for another 5 minutes. Then, hydrogen was passed through the solution and bubbling was performed for 5 minutes, and the resulting mixture was stirred under a hydrogen atmosphere for 5 hours (1 atm). Complete conversion was shown by LC-MS. The reaction mixture was filtered through Celite and washed with siRNA. The filtrate was concentrated under reduced pressure. The crude mixture was purified by reverse-phase flash chromatography using C18 RediSep Rf Gold (eluting with DMSO-containing retention solution, MeOH / 0.1% HCOOH 5 / 95% over 4 CV, then to 60% MeOH over 12 CV). The fraction was concentrated to dryness to obtain a T-1 off-white solid (m = 603 mg, 90.1% yield).

[0202] LC-MS method 1: 99.9% purity (at 215 nm), [M+H] + = 249.4, [M+Na] + =271.2. 1 H NMR (400 MHz, DMSO-d6) δ ppm 1.44 - 1.56 (m, 2 H), 1.66 - 1.80 (m, 2 H), 2.00 (br d, J = 11.5 Hz, 4 H), 2.33 - 2.45 (m, 1 H), 3.61 (s, 3 H), 4.12 (tt, J = 11.8, 3.2 Hz, 1 H), 4.74 (s, 2 H), 7.24 (s, 1 H).

[0203] Example I-2. General intermediate T-2 [ka]

[0204] Step 1. Production of methyl 4-methylsulfonyloxycyclohexanecarboxylate (2) Methyl 4-hydroxycyclohexanecarboxylate (1) (5.0 g, 31.61 mmol, 1.0 equivalent) and Et3N (6.61 mL, 47.41 mmol, 1.5 equivalent) were dissolved in CH2Cl2 (158 mL, 0.2 M), to which methanesulfonyl chloride (3.18 mL, 41.09 mmol, 1.3 equivalent) was added dropwise at 0°C. The resulting mixture was stirred at 0°C for 30 minutes. TLC (KMnO4): SM: Rf = 0.25, Product: Rf = 0.30 (60% HCl / heptane). Water was added, and the aqueous layer was extracted with CH2Cl2 (3×). The combined organic layers were washed with 1 M HCl (1×) solution, dried over Na₂SO₄, filtered, and then concentrated to dryness under reduced pressure to obtain pure 2 (7.47 g, quantitative yield) without any further purification.

[0205] LC-MS method 1: 99.9% purity (at 215 nm); [M-MsOH+H] + = 141.2 m / z, [M+Na] + = 259.2 m / z. 1 H NMR (400 MHz, CDCl3) δ ppm 1.67 - 1.82 (m, 4 H), 1.87 - 1.97 (m, 2 H), 1.97 - 2.07 (m, 2 H), 2.36 - 2.45 (m, 1 H), 3.01 (s, 3 H), 3.68 (s, 3 H), 4.88 - 4.94 (m, 1 H).

[0206] Step 2. Preparation of methyl (1r,4r)-4-(4-nitro-3-(trifluoromethyl)-1H-pyrazole-1-yl)cyclohexane-1-carboxylate (4) To a solution of 4-nitro-3-(trifluoromethyl)-1H-pyrazole 3 (1.0 g, 5.52 mmol, 1.0 eq.) and methyl 4-methylsulfonyloxycyclohexanecarboxylate 2 (1.3 g, 5.52 mmol, 2.0 eq.) in dry DMF (18.41 mL, 0.3 M), Cs2CO3 (3.6 g, 11.05 mmol, 2.0 eq.) was added. After stirring overnight at 90°C, complete conversion to 4 was shown by LC-MS. The reaction was quenched with water, and the aqueous layer was extracted three times with siRNA. The combined organic layers were washed twice with water, once with brine, dried over Na2SO4, filtered, and concentrated under reduced pressure. The crude product was purified by flash chromatography (elution over 15 CVs with 0-35% MTBE / heptane from 80 g SiO2, followed by elution over 3 CVs with 35% MTBE / heptane). The target product was eluted with 30% MTBE / heptane (confirmed by TLC eluting with 60% MTBE / heptane, Rf=0.30, UV + KMnO4). The fractions were combined, concentrated to dryness, and 4 (671 mg, 36% yield) was obtained as a pale yellow solid.

[0207] LC-MS method 1: Retention time: 1.819 min, 99.9% purity (at 215 nm), [M+H] + = 322.2. 1 H NMR (400 MHz, DMSO-d6) δ ppm 1.52 (qd, J = 13.0, 3.0 Hz, 2 H), 1.83 (br dd, J = 12.5, 3.1 Hz, 2 H), 2.00 - 2.17 (m, 4 H), 2.42 (tt, J = 12.2, 3.5 Hz, 1 H), 3.61 (s, 3 H), 4.39 (tt, J = 11.9, 3.8 Hz, 1 H), 9.19 (s, 1 H). 19 F NMR (377 MHz, DMSO-d6) δ ppm -61.45 (s, 3 F).

[0208] Step 3. Preparation of methyl 4-[4-amino-3-(trifluoromethyl)pyrazole-1-yl]cyclohexanecarboxylate (T-2) To a solution of methyl 4-[4-nitro-3-(trifluoromethyl)pyrazole-1-yl]cyclohexanecarboxylate 4 (671 mg, 2.09 mmol, 1.0 eq.) / ethyl acetate (13.93 mL, 0.15 M), 10% Pd / C (444 mg, 0.42 mmol, 0.2 eq.) was added. Nitrogen was passed through the solution and the mixture was bubbling for 10 minutes, then the balloon was replaced with hydrogen. Hydrogen was passed through the solution and the mixture was bubbling for 5 minutes, and the resulting mixture was stirred under a hydrogen atmosphere (1 atm). After stirring at room temperature for 5 hours, complete conversion to the product was shown by LC-MS. The reaction mixture was filtered through a Celite pad and washed with siRNA. The filtrate was concentrated under reduced pressure to obtain T-2 (608 mg, quantitative yield) as a pink solid. The crude product was used in the next step without further purification.

[0209] LC-MS method 1: Retention time: 1.562 min, 99.9% purity (at 215 nm), [M+H] + = 292.2. 1 H NMR (400 MHz, DMSO-d6) δ ppm 1.41 - 1.60 (m, 2 H), 1.64 - 1.81 (m, 2 H), 1.91 - 2.06 (m, 4 H), 2.32 - 2.45 (m, 1 H), 3.60 (s, 3 H), 4.00 - 4.13 (m, 1 H), 4.21 (s, 2 H), 7.22 (s, 1 H). 19 F NMR (377 MHz, DMSO-d6) δ ppm -58.85 (s, 3 F).

[0210] Example I-3. General intermediate T-3 [ka]

[0211] Step 1. Production of methyl 4-methylsulfonyloxycyclohexanecarboxylate (2) Under nitrogen, a solution of methyl 4-hydroxycyclohexanecarboxylate 1 (5.0 g, 31.61 mmol, 1.0 eq.) / CH2Cl2 (105 mL, 0.3 M) was cooled to 0°C, and then methanesulfonyl chloride (2.69 mL, 34.77 mmol, 1.1 eq.) and triethylamine (5.28 mL, 37.93 mmol, 1.2 eq.) were added, with the latter being added dropwise. After stirring at 0°C for 2 hours, complete conversion was shown by TLC (CH2Cl2 / MeOH 5.5:0.5, KMnO4 staining). The reaction mixture was quenched by adding water, the phases were separated, and the aqueous phase was extracted three times with CH2Cl2. The combined organic phases were washed twice with saline solution, dried on magnesium sulfate, filtered, and concentrated to obtain 2 (7.45 g, 99% yield) as a yellow oily substance.

[0212] 1 H NMR (400 MHz, CDCl3) δ ppm 1.67 - 1.84 (m, 4 H), 1.87 - 1.99 (m, 2 H), 2.00 - 2.09 (m, 2 H), 2.36 - 2.46 (m, 1 H), 3.02 (s, 3 H), 3.69 (s, 3 H), 4.87 - 4.96 (m, 1 H).

[0213] Step 2. Preparation of methyl 4-[3-(difluoromethyl)-4-nitropyrazole-1-yl]cyclohexanecarboxylate (4) Under nitrogen, a solution of 2 (1.16 g, 4.91 mmol, 1.0 eq.), 3 (0.80 g, 4.91 mmol, 1.0 eq.), and DMF (12.3 mL, 0.4 M) in a heated and dried round-bottom flask was stirred at room temperature for 5 minutes, after which Cs2CO3 (3.20 g, 9.81 mmol, 2.0 eq.) was added. The resulting mixture was stirred at 90°C for 16 hours. After the incomplete conversion of 3 was confirmed by LC-MS (Method 1), a second dose of 2 (1.16 g, 4.91 mmol, 1.0 eq.) was added, and stirring was continued at 90°C for 48 hours. 90% conversion of 3 was confirmed by LC-MS. The reaction was quenched by adding water. The phases were separated by adding ethyl acetate. The aqueous phase was extracted three times with ethyl acetate, and the combined organic phase was washed once with water and once with saline solution, dried on magnesium sulfate, and concentrated. The residue was incorporated into MTBE and water, and the phases were separated. The organic phase was washed, washed five times with water, washed once with saline solution, dried on magnesium sulfate, filtered, and concentrated to obtain an orange oily substance. The residue was then purified by normal-phase flash chromatography (80 g silica column, elution: 0 to 30% MTBE / heptane over 10 CV). The fractions were combined and concentrated to obtain impurity 4 (883 mg). The residue was then purified by reverse-phase chromatography (100 g C18 RediSep Rf Gold column, holding solution (DMSO), elution: 5% MeOH / 0.1% HCOOH over 4 CV, then 5% to 100% MeOH / 0.1% HCOOH over 15 CV). The fractions were combined and concentrated to obtain 4 (618 mg, 41% yield) as a white solid.

[0214] LC-MS method 1: Retention time: 1.745 min, 99.9% purity (at 215 nm), [M+H] + = 304.2. 1H NMR (400 MHz, DMSO-d6) δ ppm 1.45 - 1.60 (m, 2 H), 1.76 - 1.89 (m, 2 H), 2.00 - 2.15 (m, 4 H), 2.36 - 2.47 (m, 1 H), 3.61 (s, 3 H), 4.28 - 4.40 (m, 1 H), 7.14 - 7.47 (m, 1 H), 9.05 (s, 1 H). 19 F NMR (377 MHz, DMSO-d6) δ ppm -117.40 (s, 2 F).

[0215] Step 3. Preparation of methyl 4-[4-amino-3-(difluoromethyl)pyrazole-1-yl]cyclohexanecarboxylate (T-3) Under nitrogen, a solution of 4 (625 mg, 2.06 mmol, 1.0 eq.) and ethyl acetate (20.6 mL, 0.1 M) was degassed by sparging with nitrogen at room temperature for 15 minutes. Pd / C (438 mg, 10% w / w, 0.41 mmol, 0.2 eq.) was added, and sparging was performed again for 15 minutes. After sparging the mixture with H2 for 15 minutes, the mixture was stirred at room temperature for 16 hours, with the needle held just above the solvent surface. Complete conversion of 4 was observed by LC-MS (Method 1). The mixture was filtered through Celite, and the filter cake was thoroughly washed with ethyl acetate. The resulting solution was concentrated to obtain T-3 (563 mg, 99% yield) as an orange solid.

[0216] LC-MS method 1: Retention time: 1.297 min, 99.9% purity (at 215 nm), [M+H] + = 274.2. 1 H NMR (400 MHz, CDCl3) δ ppm 1.53 - 1.80 (m, 5 H), 2.12 - 2.42 (m, 9 H), 3.93 - 4.03 (m, 1 H), 6.54 - 6.84 (m, 1 H), 7.07 (s, 1 H). 19F NMR (377 MHz, CDCl3) δ ppm -112.22 (s, 2 F).

[0217] Synthesis of cereblon binding groups (CBMs) Example I-4. Synthesis of a typical intermediate C-1 [ka]

[0218] Step 1. Preparation of tert-butyl 4-[2-(2,6-dioxo-3-piperidyl)-1,3-dioxo-isoindoline-5-yl]piperazine-1-carboxylate (2) In a sealed tube, 2-(2,6-dioxo-3-piperidyl)-5-fluoroisoindoline-1,3-dione 1 (2.0 g, 7.24 mmol), tert-butylpiperazine-1-carboxylate (1.4 g, 7.52 mmol), DIPEA (3.87 mL, 22.2 mmol), and DMSO (15 mL) were placed. The tube was sealed and heated at 90°C for 72 hours. The progress of the reaction was tracked by HPLC. Upon completion, the reaction mixture was directly purified by reverse-phase chromatography on C18 RediSep Rf Gold using a 5-95% MeOH / 0.1% formic acid aqueous solution as the eluent. The pure fractions were combined and concentrated under reduced pressure to obtain compound 2 (2.86 g, 89% yield) as a yellow solid.

[0219] LC-MS method 1: 99.9% purity (at 215 nm), [M-tBu+H]+ = 387.2 m / z, [M+Na] + = 466.2 m / z. 1H NMR (400 MHz, DMSO-d6) δ ppm 1.42 (s, 9 H), 1.99 - 2.09 (m, 1 H), 2.52 - 2.64 (m, 2 H), 2.82 - 2.95 (m, 1 H), 3.47 (m, 8 H), 5.07 (dd, J = 13.0, 5.4 Hz, 1 H), 7.24 (dd, J = 8.6, 2.2 Hz, 1 H), 7.35 (d, J = 2.0 Hz, 1 H), 7.70 (d, J = 8.3 Hz, 1 H), 11.08 (s, 1 H).

[0220] Step 2. Preparation of 2-(2,6-dioxo-3-piperidyl)-5-piperazine-1-ylisoindoline-1,3-dionetrifluoroacetate (C-1) TFA (10.55 mL, 129.5 mmol) was added to a solution of tert-butyl 4-[2-(2,6-dioxo-3-piperidyl)-1,3-dioxo-isoindorin-5-yl]piperazine-1-carboxylate 2 (2.87 g, 6.48 mmol) in CH2Cl2 (30 mL), and the mixture was stirred at room temperature for 2 hours. Upon completion of HPLC, volatile substances were removed under vacuum, and the residue was co-evaporated with MeCN (3x) and MTBE (2x). The residue was purified by C18 RediSep Rf Gold reverse-phase chromatography using a 5-20% MeCN / 0.05% TFA aqueous solution as the eluate. The pure fraction was concentrated under reduced pressure to obtain C-1 (2.52 g, 85% yield) as a yellow solid, which was the TFA salt.

[0221] LC-MS method 1: 99.9% purity (at 215 nm), [M+H] + = 343.2 m / z. 1H NMR (400 MHz, DMSO-d6) δ ppm 1.95 - 2.10 (m, 1 H), 2.52 - 2.64 (m, 2 H), 2.82 - 2.97 (m, 1 H), 3.18 - 3.31 (m, 4 H), 3.62 - 3.73 (m, 4 H), 5.09 (dd, J = 12.8, 5.5 Hz, 1 H), 7.33 (dd, J = 8.8, 2.2 Hz, 1 H), 7.46 (d, J = 2.0 Hz, 1 H), 7.76 (d, J = 8.3 Hz, 1 H), 11.09 (s, 1 H).

[0222] Example I-5. Synthesis of a typical intermediate C-2 [ka]

[0223] Step 1. Preparation of tert-butyl 4-[2-(2,6-dioxo-3-piperidyl)-3-oxo-isoindorin-5-yl]piperazine-1-carboxylate (3) In a 40 ml oven-dried vial, a magnetic stirrer bar was placed under atmospheric pressure. 3-(6-bromo-1-oxo-isoindolin-2-yl)piperidine-2,6-dione (1) (500 mg, 1.55 mmol), tert-butylpiperazine-1-carboxylate (2) (432 mg, 2.32 mmol), DABCO (521 mg, 4.64 mmol), and Ir(ppy)2(dtbbpy)PF6 (14.2 mg, 15.47 mmol) were added, followed by DMA (10 mL). Dibromonickel; 1,2-dimethoxyethane (23.9 mg, 77.4 mmol) was then added as a DMA solution (0.5 mL) under an N2 atmosphere. The reaction system was placed 6 cm away from a 30 W blue LED and allowed to react at 25°C for 96 hours. LC-MS confirmed that the main peak was the target product. Eight batches were combined and added dropwise to water (200 mL). The resulting solid was filtered and concentrated. The residue was purified by prep-HPLC (column: Phenomenex Luna C18 250 mm x 100 mm x 10 μm; mobile phase: [water (0.1% TFA)-ACN]; B%: 16%~46%, 25 min) to obtain tert-butyl 4-[2-(2,6-dioxo-3-piperidyl)-3-oxo-isoindorin-5-yl]piperazine-1-carboxylate (3) (2.5 g, 47.1% yield) as a gray solid. 1 H NMR:400 MHz, DMSO-d6 δ:10.97 (s, 1H), 7.45 (d, J = 8.8 Hz, 1H), 7.30-7.27 (m, 1H), 7.20 (d, J = 2.0 Hz, 1H), 5.13-5.08 (m, 1H), 4.30 (dd, J = 16.8 Hz, 51.6 Hz, 2H), 3.48 (d, J = 4.8 Hz, 4H), 3.16 (d, J = 5.2 Hz, 4H), 2.92-2.68 (m, 1H), 2.62-2.51 (m, 1H), 2.41-2.37 (m, 1H), 2.01-1.99 (m, 1st hour), 1.45 (s, 9th hour).

[0224] Step 2. Preparation of 3-(1-oxo-6-piperazine-1-yl-isoindoline-2-yl)piperidine-2,6-dione (C-2) Tert-butyl 4-[2-(2,6-dioxo-3-piperidyl)-3-oxo-isoindorin-5-yl]piperazine-1-carboxylate 3 (5 g, 11.7 mmol) was added to HCl (12 N, 15 mL) at 0°C. The reaction mixture was stirred at 20°C for 1 hour. LC-MS revealed that the major peak was the target product. The mixture was diluted with MeCN (500 mL) at 0-10°C, the resulting solid was filtered, and the cake was dried to obtain the product 3-(1-oxo-6-piperazine-1-yl-isoindorin-2-yl)piperidine-2,6-dione C-2 (4.3 g, HCl salt, 100% yield) as a gray solid.

[0225] 1 H NMR:400 MHz, DMSO-d6 δ:10.98 (s, 1H), 9.34 (s, 2H), 7.49 (d, J = 5.2 Hz, 1H), 7.33-7.27 (m, 2H), 5.13-5.09 (m, 1H), 4.30 (dd, J = 17.2 Hz, J = 58.2 Hz, 2H), 3.46 (d, J = 4.8 Hz, 4H), 3.23 (d, J = 4.8 Hz, 1H), 2.93-2.78 (m, 1H), 2.62-2.51 (m, 1H),2.48-2.37 (m, 1H), 2.01-1.98 (m, 1H).

[0226] Example I-6. Synthesis of a typical intermediate C-3 [ka]

[0227] Step 1. Preparation of tert-butyl N-[1-[2-(2,6-dioxo-3-piperidyl)-1,3-dioxo-isoindorin-5-yl]-4-piperidyl]carbamate (3) 2-(2,6-dioxo-3-piperidyl)-5-fluoroisoindoline-1,3-dione 1 (200.0 mg, 0.72 mmol) and tert-butyl N-(4-piperidyl)carbamate 2 (188.52 mg, 0.94 mmol) were dissolved in 3.62 mL of dry DMSO, to which DIPEA (252.23 μL, 1.45 mmol) was added. The resulting mixture was stirred overnight at 90°C. Complete conversion was demonstrated by LC-MS. The reaction mixture was diluted with water, and the aqueous phase was extracted with siRNA (4x). The organic layer was washed with H2O (5x), dried over Na2SO4, and concentrated to dryness. The residue was then purified by reverse-phase chromatography (30 g C18 RediSep Rf Gold column, holding solution (DMSO), elution: 5% MeCN / 0.1% HCOOH over 5 CV, followed by 5-100% MeCN / 0.1% HCOOH over 20 CV). The fraction of interest was concentrated to dryness to obtain compound 3 as a yellow solid (283 mg, 86% yield).

[0228] LC-MS method 1: 99.9% purity (at 215 nm), [M-2HCOOH+H] + = 457.2. 1H NMR (400 MHz, DMSO-d6) δ ppm 1.39 (s, 9 H), 1.40 - 1.45 (m, 2 H), 1.80 (br d, J = 11.2 Hz, 2 H), 1.97 - 2.05 (m, 1 H), 2.53 - 2.63 (m, 2 H), 2.83 - 2.95 (m, 1 H), 3.05 (br t, J = 11.9 Hz, 2 H), 3.49 - 3.60 (m, 1 H), 3.98 (br d, J = 13.4 Hz, 2 H), 5.06 (dd, J = 13.0, 5.4 Hz, 1 H), 6.87 (br d, J = 7.6 Hz, 1 H), 7.31 (d, J = 1.7 Hz, 1 H), 7.65 (d, J = 8.6 Hz, 1 H), 8.14 (s, 1 H), 11.07 (s, 1 H).

[0229] Step 2. Preparation of 5-(4-amino-1-piperidyl)-2-(2,6-dioxo-3-piperidyl)isoindoline-1,3-dionetrifluoroacetate (C-3) To a solution of tert-butyl N-[1-[2-(2,6-dioxo-3-piperidyl)-1,3-dioxo-isoindorin-5-yl]-4-piperidyl]carbamate 3 (283.0 mg, 0.62 mmol) in CH2Cl2 (6.19 mL), TFA (721.44 μL, 9.3 mmol) was added. The resulting mixture was stirred overnight at room temperature. Complete conversion was shown by LC-MS. The solvent was evaporated under high pressure. The residue was evaporated twice with MeCN / toluene and twice again with MeCN / toluene, and then dried under high vacuum to obtain C-3 as a yellow solid (291 mg, quantitative yield). The crude product was used in the next reaction without purification. LC-MS method 1: 99.9% purity (at 215 nm), [M-TFA+H] + = 357.2.

[0230] Example I-7. Synthesis of a typical intermediate C-4 [ka]

[0231] Step 1. Preparation of tert-butyl 4-[2-(2,6-dioxo-3-piperidyl)-1-oxo-isoindorin-5-yl]piperazine-1-carboxylate (3) In a sealed tube, a solution of tert-butylpiperazine-1-carboxylate 2 (74.93 mg, 0.4000 mmol) and 3-(5-bromo-1-oxo-isoindorin-2-yl)piperidine-2,6-dione 1 (100 mg, 0.3100 mmol) in 1,4-dioxane (3.0946 mL) was added. The solution was degassed, and Cs2CO3 (302.49 mg, 0.9300 mmol) and Pd-PEPPSI-IPent (12.25 mg, 0.0200 mmol) were added. The tube was sealed and stirred overnight at 90°C. After 20 hours, the reaction showed complete conversion, which was passed through Celite and washed with DCM. The filtrate was concentrated under vacuum. The residue was purified by silica gel column chromatography (40 g, DCM / MeOH, 0%~5% at 15 CV; the product eluted at 4% MeOH). The pure fractions were combined, volatile components were evaporated, and the mixture was dried under high vacuum to obtain compound 3 (82 mg, 0.1912 mmol, 61.779% yield) as a white solid.

[0232] Step 2. Preparation of 3-(1-oxo-5-piperazine-1-yl-isoindoline-2-yl)piperidine-2,6-dione (C-4) In a vial, tert-butyl 4-[2-(2,6-dioxo-3-piperidyl)-1-oxo-isoindorin-5-yl]piperazine-1-carboxylate 3 (200 mg, 0.4700 mmol) was suspended in HCl (2 mL, 8 mmol). Ethyl acetate (2 mL) was added to the mixture and stirring was continued for 30 minutes. After 30 minutes, HPLC showed that little conversion had occurred. Methanol (1 mL) was added to the reaction mixture and stirring was continued. After 30 minutes, HPLC showed incomplete conversion. The mixture was sonicated and stirred in a water bath (50°C) for 30 minutes. HPLC showed complete conversion. The mixture was concentrated to dryness and co-evaporated with MeCN (3X) to obtain the target product C-4 as an off-white solid (180 mg, 0.4687 mmol, 99% yield).

[0233] Example I-8. Synthesis of a typical intermediate C-5 [ka]

[0234] Step 1. Preparation of tert-butyl 4-[2-(2,6-dioxo-3-piperidyl)-3-oxo-isoindorin-5-yl]piperidine-1-carboxylate (3) In a Schlenk flask, a solution of 3-(6-bromo-1-oxo-isoindorin-2-yl)piperidine-2,6-dione 1 (250 mg, 0.7700 mmol), potassium (1-tert-butoxycarbonyl-4-piperidyl)-trifluoroboranoid 2 (595.02 μL, 2.32 mmol), and 2,4,6-trimethylpyridine (0.18 mL, 1.39 mmol) in 1,4-dioxane (2 mL) was added to a solution of (Ir[dF(CF3)ppy]2(dtbpy))PF6 (2.17 mg, 0.002 mmol) in 1,4-dioxane (1 mL). A solution of Ni-dtbbpy-Br2 (37.67 mg, 0.0800 mmol) in 1,4-dioxane (1 mL), which had been sonicated for 1 minute, was added. The reaction mixture was degassed three times using the freeze-pump-thaw method. The flask was placed under a dry nitrogen atmosphere and sealed with Parafilm. The reaction mixture was stirred under blue LED irradiation for 4 days. The solution was evaporated until dry. DMSO was added to the crude mixture. The crude mixture was purified by column chromatography C18 (5-100% MeCN / H2O (0.1% formic acid), from 5% MeCN to 100% at 15 CV), and the combined product 3 was concentrated to dryness. LC-MS method 1: 99.9% purity (at 215 nm), [M+H] + = 428.1.

[0235] Step 2. Preparation of 2-(2,6-dioxopiperidine-3-yl)-5-(piperidine-4-yl)isoindoline-1,3-dione (C-5) To a solution of tert-butyl 4-[2-(2,6-dioxo-3-piperidyl)-3-oxo-isoindorin-5-yl]piperidine-1-carboxylate 3 (46.0 mg, 0.1100 mmol) in DCM (1.4035 mL), TFA (0.18 mL, 2.15 mmol) was added. The reaction mixture was stirred at room temperature for 2 hours, at which point complete conversion was observed by LC-MS. The crude reaction mixture was evaporated with MeCN / PhMe (4x) to obtain product C-5 (46 mg, 0.1042 mmol, 97% yield), which was used in the next step without purification. LC-MS method 1: 99.9% purity (at 215 nm), [M+H] + = 328.2.

[0236] Example I-9. Synthesis of a common intermediate C-6 [ka]

[0237] Step 1. Preparation of tert-butyl 4-[2-(2,6-dioxo-3-piperidyl)-3-oxo-isoindorin-5-yl]piperidine-1-carboxylate (3) In a reaction vial, tert-butyl 4-bromopiperidine-1-carboxylate 2 (48.97 mg, 0.1900 mmol), 5-bromo-2-(2,6-dioxo-3-piperidyl)isoindoline-1,3-dione 1 (50 mg, 0.1500 mmol), nickel(II) iodide (23.17 mg, 0.0700 mmol), 1,10-phenanthroline (13.36 mg, 0.0700 mmol), and sodium iodide (22.25 mg, 0.1500 mmol) were added. Then DMPU (0.65 mL, 0.1500 mmol), followed by zinc (19.39 mg, 0.3000 mmol), and finally 4-ethylpyridine (16.87 μL, 0.1500 mmol) were added. The mixture was stirred overnight at 90°C. Upon completion, the mixture was purified by reverse-phase chromatography using a gradient of acetonitrile and 0.1% FA water (5% to 100% over 10 CV). The pure fractions were combined and evaporated under reduced pressure to obtain compound 2 (15.7 mg) as a pale yellow solid. LCMS method 1: Purity: 99.9% [M+Na]+:464.2; [Mt-Bu] + :386.2.

[0238] Step 2. Preparation of 2-(2,6-dioxo-3-piperidyl)-5-(4-piperidyl)isoindoline-1,3-dione(C-6) To a solution of tert-butyl 4-[2-(2,6-dioxo-3-piperidyl)-1,3-dioxo-isoindorin-5-yl]piperidine-1-carboxylate 3 (75 mg, 0.1700 mmol) in DCM (1.5856 mL), TFA (0.43 mL, 1.7 mmol) was added. The mixture was stirred at rt for 2 hours, at which point no starting material was observed. Toluene and MeCN were added to the reaction mixture, and the solvent was evaporated under reduced pressure. The crude product was co-evaporated three times with MeCN to obtain product C-6 (95 mg, 0.2063 mmol, 99% yield) as an orange solid. LC-MS method 1: 98.9% purity (at 215 nm), [M+H]+ = 342.2.

[0239] Example I-10. Synthesis of a common intermediate C-7 [ka]

[0240] Step 1. Preparation of tert-butyl N-[1-[2-(2,6-dioxo-3-piperidyl)-1-oxo-isoindorin-5-yl]-4-piperidyl]carbamate (3) In a heat-dried microwave vial, novel activated 3Å molecular sieves (100 mg) and dried DMSO (9.3 mL, 0.1 M) were added under nitrogen, and the mixture was degassed by sparging with nitrogen for 10 minutes. The reagents were then added in the following order: CuI (88.4 mg, 0.46 mmol, 0.5 eq.), DMPAO (179.4 mg, 0.93 mmol, 1.0 eq.), n-Bu4OAc (840 mg, 2.79 mmol, 3.0 eq.), 1 (300 mg, 0.93 mmol, 1.0 eq.), and 2 (223 mg, 1.11 mmol, 1.2 eq.). The heterogeneous mixture was then degassed by sparging with nitrogen for 10 minutes. The mixture was stirred and heated at 110°C for 16 hours. Complete conversion of 1 was observed by LC-MS (Method 1). The phases were separated by adding ethyl acetate and water. The aqueous phase was extracted three times with ethyl acetate. The combined organic phase was washed once with saline solution, dried over magnesium sulfate, filtered, and concentrated. The residue was purified by reverse-phase flash chromatography (50 g C18 RediSep Rf Gold column, holding solution (DMSO), elution: 5% MeCN / 0.1% HCOOH over 4 CV, then 5% to 60% MeCN / 0.1% HCOOH over 5 CV). The fractions were combined and concentrated to obtain 3 (127 mg, 30% yield) as a pale orange solid.

[0241] LC-MS method 1: Retention time: 1.555 min, 99.9% purity (at 215 nm), [M+H] + = 443.2. 1 H NMR (400 MHz, DMSO-d6) δ ppm 1.34 - 1.49 (m, 11 H), 1.75 - 1.83 (m, 2 H), 1.91 - 2.01 (m, 1 H), 2.30 - 2.43 (m, 1 H), 2.53 - 2.62 (m, 1 H), 2.83 - 2.97 (m, 3 H), 3.41 - 3.55 (m, 1 H), 3.82 (br d, J = 13.2 Hz, 2 H), 4.19 (d, J = 16.0 Hz, 1 H), 4.31 (d, J = 16.0 Hz, 1 H), 5.04 (dd, J = 13.2, 5.1 Hz, 1 H), 6.85 (br. d, J = 7.1 Hz, 1 H), 7.01 - 7.09 (m, 2 H), 7.49 (d, J = 8.3 Hz, 1 H), 10.94 (s, 1 H).

[0242] Step 2. Preparation of 3-[5-(4-amino-1-piperidyl)-1-oxo-isoindorin-2-yl]piperidine-2,6-dione dihydrochloride (C-7) Under nitrogen, a mixture of 3 (127 mg, 0.30 mmol, 1.0 eq.) and 4.0 M HCl / 1,4-dioxane (9.43 mL, 120 eq.) was stirred at room temperature for 1.25 hours. Complete conversion of 3 was observed by LC-MS (Method 1). The mixture was concentrated under reduced pressure and co-evaporated three times with acetonitrile to obtain C-7 (156 mg, quantitative yield) as a pale orange solid.

[0243] LC-MS method 1: Retention time: 0.751 min, 86.8% purity (at 215 nm), [M+H] + = 343.2. 1H NMR (400 MHz, DMSO-d6) δ ppm 1.51 - 1.64 (m, 2 H), 1.90 - 2.03 (m, 3 H), 2.30 - 2.41 (m, 1 H), 2.55 - 2.64 (m, 1 H), 2.84 - 3.00 (m, 3 H), 3.22 - 3.34 (m, 1 H), 3.44 - 3.74 (m, 1 H), 3.94 (br. d, J = 13.4 Hz, 2 H), 4.21 (d, J = 16.0 Hz, 1 H), 4.32 (d, J = 16.0 Hz, 1 H), 5.05 (dd, J = 13.4, 5.1 Hz, 1 H), 7.04 - 7.15 (m, 2 H), 7.53 (d, J = 8.6 Hz, 1 H), 7.94 - 8.07 (m, 3 H), 10.94 (s, 1 H).

[0244] Example I-11. Synthesis of a common intermediate C-8 [ka]

[0245] Step 1. Preparation of tert-butyl N-[1-[2-(2,6-dioxo-3-piperidyl)-3-oxo-isoindorin-5-yl]-4-piperidyl]carbamate (3) DMSO (4.2 mL) and molecular sieves (4 Å, beads) were added to a microwave vial and stirred at room temperature for 30 minutes while bubbling with nitrogen gas through the solvent. Then, 3-(6-bromo-1-oxo-isoindorin-2-yl)piperidine-2,6-dione 1 (150 mg, 0.46 mmol), tert-butyl N-(4-piperidyl)carbamate 2 (111.6 mg, 0.56 mmol), DMPAO (89.7 mg, 0.46 mmol), CuI (0.5 M solution in DMSO; 0.46 mL, 0.23 mmol), and tetrabutylammonium acetate (419.9 mg, 1.39 mmol) were added. Nitrogen gas was passed through the solution for 15 minutes while bubbling, and the mixture was sonicated. The mixture was capped in a microwave vial and stirred at 110°C for 24 hours. After cooling to room temperature, water and RINKAN were added. The aqueous phase was extracted three times with RINKAN. The organic phases were combined, washed with water and saline solution, dried over Na2SO4, filtered, and evaporated under reduced pressure. The oily solid was then dissolved in DMSO, and the solution was loaded onto a 50 g RediSep Rf Gold C18 chromatography column. Elution was performed with MeCN / 0.1% formic acid aqueous solution (5% over 4 CVs, then 5% to 60% over 15 CVs). The fractions of interest were combined and concentrated to obtain 3 (43.5 mg, yield 16%) as an orange solid. LC-MS method 1: 76.1% purity (at 215 nm), [M+H] + = 443.2.

[0246] Step 2. Preparation of 3-[6-(4-amino-1-piperidyl)-1-oxo-isoindorin-2-yl]piperidine-2,6-dionetrifluoroacetate (C-8) To a solution of tert-butyl N-[1-[2-(2,6-dioxo-3-piperidyl)-3-oxo-isoindorin-5-yl]-4-piperidyl]carbamate 3 (99.3 mg, 0.22 mmol) in CH2Cl2 (1.6 mL), trifluoroacetic acid (0.45 mL, 1.76 mmol) was added. The resulting mixture was stirred at room temperature for 2 hours. MeCN was added, and the solvent was removed under reduced pressure. The crude mixture was co-evaporated three times with MeCN. The residue was dissolved in water, and the solution was loaded onto a 15.5 g RediSep Rf Gold C18 chromatography column. Elution was performed using MeCN / 0.05 M TFA aqueous solution (5% for 5 CV, then 5% to 20% for 10 CV). The fractions of interest were combined and concentrated to obtain C-8 (52 mg, 51% yield) as an orange solid.

[0247] LC-MS method 1: 99.9% purity (at 215 nm), [M-TFA+H] + = 343.2. 1 H NMR (400 MHz, DMSO-d6) δ ppm 1.53 - 1.66 (m, 2 H), 1.89 - 2.02 (m, 3 H), 2.31 - 2.45 (m, 1 H), 2.56 - 2.69 (m, 1 H), 2.79 - 2.96 (m, 3 H), 3.18 - 3.30 (m, 1 H), 3.78 - 3.86 (m, 2 H), 4.15 - 4.25 (m, 1 H), 4.29 - 4.38 (m, 1 H), 5.07 (dd, J = 13.3, 5.0 Hz, 1 H), 7.20 (d, J = 2.0 Hz, 1 H), 7.28 (dd, J = 8.6, 2.2 Hz, 1 H), 7.44 (d, J = 8.3 Hz, 1 H), 7.87 (br s, 3 H), 10.96 (s, 1 H).

[0248] Example I-12. Synthesis of a common intermediate C-9 [ka]

[0249] Step 1. Preparation of tert-butyl 1-[2-(2,6-dioxo-3-piperidyl)-1,3-dioxo-isoindorin-5-yl]piperidine-4-carboxylate (3) 2-(2,6-dioxo-3-piperidyl)-5-fluoroisoindoline-1,3-dione 1 (200 mg, 0.720 mmol, 1 eq.) and tert-butylpiperidine-4-carboxylate 2 (174.4 mg, 0.940 mmol, 1.3 eq.) were dissolved in anhydrous DMSO (3.6 mL) and DIPEA (252 μL, 1.45 mmol, 2 eq.) was added. The resulting mixture was stirred at 90°C. After 18 hours, complete conversion was shown by LC-MS. The reaction mixture was diluted with water, and the aqueous phase was extracted three times with siRNA. The organic matter was washed five times with water, dried over Na₂SO₄, and concentrated to dryness. The residue was purified by reverse-phase flash chromatography (50 g C18 RediSep Rf Gold column, holding solution (DMSO), elution: 5% MeCN / 0.1% HCOOH over 3 CV, then 5 to 40% MeCN / 0.1% HCOOH over 12 CV, then 40% MeCN / 0.1% HCOOH over 3 CV, then 40 to 55% MeCN / 0.1% HCOOH over 4 CV, then 55% MeCN / 0.1% HCOOH over 7 CV). The fractions were combined and concentrated to obtain 3 (297 mg, 90% yield) as a yellow solid.

[0250] LC-MS method 1: 99.9% purity (at 215 nm), [M+H] + = 442.2. 1H NMR (400 MHz, DMSO-d6) δ ppm 1.40 (s, 9 H), 1.49 - 1.63 (m, 2 H), 1.82 - 1.91 (m, 2 H), 1.97 - 2.04 (m, 1 H), 2.52 - 2.63 (m, 3 H), 2.81 - 2.95 (m, 1 H), 3.08 (br t, J = 11.2 Hz, 2 H), 3.96 (br d, J = 13.2 Hz, 2 H), 5.06 (dd, J = 13.0, 5.4 Hz, 1 H), 7.24 (dd, J = 8.6, 2.0 Hz, 1 H), 7.32 (d, J = 1.7 Hz, 1 H), 7.66 (d, J = 8.6 Hz, 1 H), 11.07 (s, 1 H).

[0251] Step 2. Preparation of 1-[2-(2,6-dioxo-3-piperidyl)-1,3-dioxo-isoindorin-5-yl]piperidine-4-carboxylic acid (C-9) A solution of tert-butyl 1-[2-(2,6-dioxo-3-piperidyl)-1,3-dioxo-isoindorin-5-yl]piperidine-4-carboxylate 3 (297 mg, 0.650 mmol, 1 eq.) in TFA (1.51 mL, 19.52 mmol, 30 eq.) was stirred at room temperature. After 1 hour, complete conversion was shown by HPLC. The TFA was evaporated under reduced pressure, and the residue was purified by reverse-phase flash chromatography (50 g C18 RediSep Rf Gold column, holding solution (DMSO), elution: 5% MeOH / 0.1% HCOOH over 3 CV, then 5-40% MeOH / 0.1% HCOOH over 11 CV, then 40% MeCN / 0.1% HCOOH over 4 CV, then 40-60% MeCN / 0.1% HCOOH over 9 CV). The fractions were combined and concentrated to obtain C-9 (206 mg, 82% yield) as a yellow solid.

[0252] LC-MS method 1: 99.9% purity (at 215 nm), [M+H] += 386.2. 1 H NMR (400 MHz, DMSO-d6) δ ppm 1.53 - 1.67 (m, 2 H), 1.84 - 1.95 (m, 2 H), 1.97 - 2.05 (m, 1 H), 2.52 - 2.63 (m, 3 H), 2.81 - 2.94 (m, 1 H), 3.09 (br t, J = 11.4 Hz, 2 H), 3.97 (br d, J = 13.2 Hz, 2 H), 5.06 (dd, J = 12.8, 5.5 Hz, 1 H), 7.25 (dd, J = 8.6, 2.0 Hz, 1 H), 7.33 (d, J = 1.5 Hz, 1 H), 7.66 (d, J = 8.6 Hz, 1 H), 11.07 (s, 1 H), 12.29 (dd, J = 6.0, 2.1 Hz, 1 H).

[0253] Example I-13. Synthesis of a common intermediate C-10 [ka]

[0254] Step 1. Preparation of tert-butyl N-[1-[2-(2,6-dioxo-3-piperidyl)-1-oxo-isoindorin-5-yl]-4-piperidyl]-N-methyl-carbamate (3) In a heat-dried microwave vial, novel activated 3Å molecular sieves (20 mg) and dried DMSO (4.6 mL, 0.1 M) were placed under nitrogen, and the mixture was degassed by sparging with nitrogen for 20 minutes. Subsequently, the reagents were added in the following order: CuI (44.2 mg, 0.23 mmol, 0.5 eq.), 3-(5-bromo-1-oxo-isoindorin-2-yl)piperidine-2,6-dione 1 (150 mg, 0.46 mmol, 1.0 eq.), tert-butyl N-methyl-N-(4-piperidyl)carbamate 2 (119.37 mg, 0.56 mmol, 1.2 eq.), DMPAO (89.68 mg, 0.46 mmol, 1.0 eq.), and tetrabutylammonium acetate (419.88 mg, 1.39 mmol, 3.0 eq.). The heterogeneous mixture was then further degassed by sparging with nitrogen for 10 minutes. The mixture was stirred and heated at 110°C for 16 hours. Complete conversion of 1 was observed by LC-MS (Method 3). The phases were separated by adding ethyl acetate and water. The aqueous phase was extracted three times with ethyl acetate. The combined organic phases were washed once with saline solution, dried over magnesium sulfate, filtered, and concentrated. The residue was purified by normal-phase flash chromatography (40 g silica column, elution: 0 to 10% CH2Cl2 / MeOH over 15 CV, product eluted with 6.5% MeOH). The fractions were combined and concentrated to obtain tert-butyl N-[1-[2-(2,6-dioxo-3-piperidyl)-1-oxo-isoindorin-5-yl]-4-piperidyl]-N-methyl-carbamate 3 (127 mg, 47% yield) as an orange solid.

[0255] LC-MS method 2: Retention time: 1.788 min, 79.1% purity (at 215 nm), [M+H] + = 457.2. 1H NMR (400 MHz, chloroform-d) δ ppm 0.77 - 0.90 (m, 1 H), 1.48 (s, 9 H), 1.72 - 1.84 (m, 4 H), 2.19 - 2.25 (m, 1 H), 2.34 (br dd, J = 13.0, 5.1 Hz, 1 H), 2.75 (s, 3 H), 2.82 - 3.00 (m, 4 H), 3.91 (br d, J = 13.2 Hz, 2 H), 4.20 - 4.30 (m, 1 H), 4.37 - 4.45 (m, 1 H), 5.20 (dd, J = 13.3, 5.0 Hz, 1 H), 6.91 (br s, 1 H), 6.97 - 7.05 (m, 1 H), 7.74 (d, J = 8.6 Hz, 1 H), 7.96 (br s, 1 H).

[0256] Step 2. Preparation of 3-[5-[4-(methylamino)-1-piperidyl]-1-oxo-isoindorin-2-yl]piperidine-2,6-dione hydrochloride (C-10) Under nitrogen, a solution of tert-butyl N-[1-[2-(2,6-dioxo-3-piperidyl)-1-oxo-isoindorin-5-yl]-4-piperidyl]-N-methyl-carbamate 3 (125 mg, 0.27 mmol, 1.0 eq.) and 4 M HCl / 1,4-dioxane (1.37 mL, 5.48 mmol, 20.0 eq.) in MeCN (2 mL, 0.1 M) was stirred at room temperature for 1 hour. Complete conversion of 3 was confirmed by LC-MS (Method 3). The mixture was concentrated under reduced pressure and evaporated three times using acetonitrile to obtain 3-[5-[4-(methylamino)-1-piperidyl]-1-oxo-isoindorin-2-yl]piperidine-2,6-dione C-10 (124 mg, quantitative yield) as a pale orange solid.

[0257] LC-MS method 2: Retention time: 1.215 min, 85.6% purity (at 215 nm), [M-HCl+H] + = 357.2. 1 H NMR (400 MHz, DMSO-d6) δ ppm 1.50 - 1.63 (m, 2 H), 1.91 - 2.07 (m, 3 H), 2.31 - 2.40 (m, 1 H), 2.53 - 2.63 (m, 4 H), 2.82 - 2.97 (m, 3 H), 3.16 - 3.28 (m, 1 H), 3.95 - 4.03 (m, 2 H), 4.17 - 4.24 (m, 1 H), 4.29 - 4.38 (m, 1 H), 5.05 (dd, J = 13.3, 5.0 Hz, 1 H), 7.04 - 7.15 (m, 2 H), 7.53 (d, J = 8.3 Hz, 1 H), 8.67 (br s, 2 H), 10.94 (s, 1 H).

[0258] Example I-14. Synthesis of a common intermediate C-11 [ka]

[0259] Step 1. Preparation of tert-butyl 4-[[2-(2,6-dioxo-3-piperidyl)-1-oxo-isoindorin-5-yl]-methyl-amino]piperidine-1-carboxylate (3) In a sealed tube, pre-activated molecular sieves (3 Å beads) were added under high vacuum for 1.5 hours and heated with a torch every 10 minutes. Then, 3-(5-bromo-1-oxo-isoindorin-2-yl)piperidine-2,6-dione 1 (150 mg, 0.46 mmol, 1 eq.), tert-butyl 4-(methylamino)piperidine-1-carboxylate 2 (119.37 mg, 0.56 mmol, 1.2 eq.), DMPAO (89.68 mg, 0.46 mmol, 1 eq.), CuI (44.2 mg, 0.23 mmol, 0.5 eq.), and tetrabutylammonium acetate (419.88 mg, 1.39 mmol, 3 eq.) were added in a solution of dried DMSO (4.6 mL, 0.1 M). Nitrogen was passed through the solution for 15 minutes by bubbling, the tube was sealed, and the resulting mixture was stirred overnight at 110°C. After overnight stirring, LC-MS showed a conversion rate to compound 3 of 16%. Water and siRNA were added to the reaction solution, and the aqueous phase was extracted three times with siRNA. The organic phases were combined, washed three times with water and once with brine, dried over Na2SO4, filtered, and concentrated to dryness. The residue was then purified by normal-phase FC (40 g column, solid retained on silica, 0-10% DCM / MeOH over 15 CV). The fractions were combined and concentrated to obtain compound 3 (12.7 mg, 5% yield) as a yellow oily formate.

[0260] LC-MS method 2: 86.6% purity (at 215 nm), [Mt-Bu+H] + = 401.1. 1H NMR (400 MHz, DMSO-d6) δ ppm 1.40 - 1.43 (m, 9 H), 1.56 - 1.65 (m, 4 H), 2.54 (br s, 2 H), 2.77 - 2.80 (m, 3 H), 2.84 - 2.95 (m, 3 H), 3.91 - 4.11 (m, 4 H), 4.15 - 4.23 (m, 1 H), 4.27 - 4.35 (m, 1 H), 5.03 (dd, J = 13.3, 5.0 Hz, 1 H), 6.90 - 6.97 (m, 2 H), 7.47 - 7.51 (m, 1 H), 8.18 (s, 1 H), 10.93 (s, 1 H).

[0261] Step 2. Preparation of N-[3-(difluoromethyl)-1-[4-[2-[4-[2-(2,6-dioxo-3-piperidyl)-1-oxo-isoindoline-5-yl]piperazine-1-yl]ethyl]cyclohexyl]pyrazole-4-yl]-5-[(3R,5R)-3-amino-5-fluoro-1-piperidyl]pyrazolo[1,5-a]pyrimidine-3-carboxamide hydrochloride (C-11) To a solution of tert-butyl 4-[[2-(2,6-dioxo-3-piperidyl)-3-oxo-isoindorin-5-yl]-methyl-amino]piperidine-1-carboxylate 3 (62.8 mg, 0.11 mmol, 1 equivalent) in CH2Cl2 (1 mL, 0.11 M), 4 M HCl / 1,4-dioxane (0.82 mL, 3.26 mmol, 30 equivalents) was added, and the reaction mixture was stirred at room temperature. After 1 hour, LC-MS showed complete conversion to compound C-11. The solvent was evaporated, and the mixture was purified by reverse-phase FC (50 g C18 RediSep Rf Gold column, holding solution (H2O), elution: 5% MeCN / H2O + 0.02 M HCl over 4 CV, then 5 to 80% MeCN / H2O + 0.02 M HCl over 20 CV). The fractions were combined and concentrated to obtain C-11 (48.3 mg, quantitative yield) as a white solid in the form of hydrochloride.

[0262] LC-MS method 3: 99.9% purity (at 215 nm), [M-HCl+H] + = 357.2. 1 H NMR (400 MHz, DMSO-d6) δ ppm 1.73 - 1.82 (m, 2 H), 1.92 - 2.06 (m, 3 H), 2.33 - 2.43 (m, 1 H), 2.55 - 2.62 (m, 1 H), 2.82 (s, 3 H), 2.85 - 2.95 (m, 1 H), 3.00 - 3.10 (m, 2 H), 3.35 (br d, J = 13.2 Hz, 2 H), 4.13 - 4.23 (m, 2 H), 4.32 (br d, J = 16.6 Hz, 1 H), 5.04 (dd, J = 13.0, 4.9Hz, 1H), 6.96 - 7.01 (m, 2 H), 7.49 - 7.54 (m, 1 H), 8.68 - 8.96 (m, 2 H), 10.93 (s, 1 H).

[0263] Example I-15. Synthesis of a common intermediate C-12 [ka]

[0264] Step 1. Preparation of tert-butyl 8-[2-(2,6-dioxo-3-piperidyl)-1-oxo-isoindorin-5-yl]-2,8-diazaspiro[4.5]decane-2-carboxylate (3) DMSO (8.2 mL) and a 3Å molecular sieve were added to a heat-dried tube. After sparging the solvent with nitrogen for 20 minutes, tert-butyl 2,8-diazaspiro[4.5]decane-2-carboxylate 2 (357 mg, 1.49 mmol, 1.2 eq.), 3-(5-bromo-1-oxo-isoindorin-2-yl)piperidine-2,6-dione 1 (400 mg, 1.24 mmol, 1.0 eq.), CuI (118 mg, 0.62 mmol, 0.5 eq.), DMPAO (239 mg, 1.24 mmol, 1.0 eq.), and tetrabutylammonium acetate (1.12 g, 3.71 mmol, 3.0 eq.) were added. The reaction mixture was again sparged with nitrogen for 15 minutes. The tube was sealed and stirred at 110°C for 16 hours. Water and ethyl acetate were added upon completion. The aqueous phase was extracted with ethyl acetate (3x). The organic phases were combined, washed with water (3x) and saline solution (1x), dried over Na2SO4, and concentrated under reduced pressure. The residue was purified by reverse-phase chromatography (C18 column, MeCN / 0.1% HCOOH 5:95, then 5:95~50:50) to obtain title compound 3 as a pink solid (225 mg, 38%).

[0265] LC-MS method 1: 99.9% purity (at 215 nm), [M+H] + = 483.2. 1H NMR (400 MHz, DMSO-d6) δ ppm 1.40 (s, 9 H), 1.52 - 1.63 (m, 4 H), 1.70 - 1.79 (m, 2 H), 1.89 - 2.01 (m, 1 H), 2.28 - 2.44 (m, 1 H), 2.54 - 2.63 (m, 1 H), 2.82 - 2.97 (m, 1 H), 3.13 (s, 2 H), 4.15 - 4.36 (m, 2 H), 5.04 (dd, J = 13.3, 5.0 Hz, 1 H), 6.97 - 7.13 (m, 2 H), 7.50 (d, J = 8.6 Hz, 1 H), 10.94 (s, 1 H). Note: The six proton signals were partially obscured by the water peak.

[0266] Step 2. Preparation of 3-[5-(2,8-diazaspiro[4,5]decane-8-yl)-1-oxo-isoindorin-2-yl]piperidine-2,6-dione (C-12) To a solution of tert-butyl 8-[2-(2,6-dioxo-3-piperidyl)-1-oxo-isoindorin-5-yl]-2,8-diazaspiro[4.5]decane-2-carboxylate 3 (225 mg, 0.47 mmol, 1.0 eq.) in CH2Cl2 (4.7 mL), TFA (2.9 mL, 11.66 mmol, 25 eq.) was added. The reaction mixture was stirred at room temperature for 1 hour. Upon completion, the mixture was concentrated to dryness, and residual TFA was removed using toluene (2x) and MeCN (2x) to obtain the bis-TFA salt of the title compound C-12 as a pale orange semi-solid (320 mg, quantitative yield). This substance was used in the next step without purification. LC-MS method 1: 99.9% purity (at 215 nm), [M+H] + = 383.2.

[0267] Example I-16. Synthesis of a common intermediate C-13 [ka]

[0268] Step 1. Preparation of [4-[tert-butoxycarbonyl(methyl)amino]cyclohexyl]methanesulfonate (2) To a solution of tert-butyl N-(4-hydroxycyclohexyl)-N-methyl-carbamate 1 (220 mg, 0.9600 mmol) in DCM (9.5936 mL), triethylamine (0.53 mL, 3.84 mmol) and MsCl (0.22 mL, 2.88 mmol) were added dropwise at 0°C. The mixture was stirred at 0°C for 30 minutes, after which the cold bath was removed. The reaction mixture was stirred overnight at room temperature. TLC analysis showed the formation of one product (SiO2, 1:1 heptane / Depositphotos, Rf SM = 0.2, Rf DP = 0.3). The reaction solution was added to a saturated NaHCO3 solution (50 mL) to separate the organic phase. The aqueous phase was extracted with DCM (4 x 20 mL), and the combined organic layers were washed with saline solution (1 x 100 mL) and evaporated to obtain compound 2 (322 mg, 98%) as a pale yellow solid.

[0269] Step 2. Preparation of tert-butyl N-(4-azidocyclohexyl)-N-methyl-carbamate (3) [4-[tert-butoxycarbonyl(methyl)amino]cyclohexyl]methanesulfonate 2 (690 mg, 2.02 mmol) was dissolved in DMF (6.7337 mL), and then sodium azide (262.65 mg, 4.04 mmol) was added. The resulting suspension was stirred at 70°C. Subsequently, TLC analysis showed the formation of a new product (SiO2, 1:1 siRNA / heptane, Rf SM = 0.3, Rf DP = 0.6). The reaction mixture was added to water (100 mL) and extracted with siRNA (3 x 20 mL). The combined organic layers were washed with saline (1 x 50 mL) and dried to obtain product 3 (481 mg, 70%) as a pale yellow oil. This was used in the next step without further purification.

[0270] Step 3. Preparation of tert-butyl ((1r,4r)-4-aminocyclohexyl)(methyl)carbamate (4) tert-butyl N-(4-azidocyclohexyl)-N-methyl-carbamate 3 (117 mg, 0.4600 mmol) was dissolved in THF (5 mL), and then triphenylphosphine (241.32 mg, 0.9200 mmol) was added. The resulting solution was stirred at room temperature for 2 hours. Then, water (1 mL) was added and the mixture was stirred overnight. The solvent was removed by vacuum, and the residue was purified by reverse-phase chromatography (5-100% MeOH in a buffer solution at pH = 10), collected, evaporated, and then purified again to remove triphenylphosphine (5-100% MeOH / 0.1% FA) to obtain product 4 (75 mg, 59%) as a colorless semi-solid.

[0271] Step 4. Production of [4-[tert-butoxycarbonyl(methyl)amino]cyclohexyl]methanesulfonate (6) A gray suspension of tert-butyl N-(4-aminocyclohexyl)-N-methyl-carbamate 4 (92 mg, 0.4000 mmol), 2-(2,6-dioxo-3-piperidyl)-4-fluoroisoindoline-1,3-dione 5 (122.4 mg, 0.4400 mmol), and DIPEA (0.18 mL, 1.01 mmol) / DMSO (2.7 mL) was stirred at 120°C for 48 hours. Partial conversion was confirmed by HPLC, but the reaction was stopped due to the presence of some degradation products. The resulting black solution was then cooled to room temperature and directly injected onto a C18 column (50 g), and purified by reverse-phase chromatography (5-100% MeOH / 0.1% FA). The fractions were evaporated together to obtain product 6 (83 mg, 40%) as a yellow solid.

[0272] Step 5. 2-(2,6-dioxopiperidine-3-yl)-4-(((1r,4r)-4-(methylamino)cyclohexyl)amino)isoindoline-1,3-dione(C13) tert-butyl N-[4-[[2-(2,6-dioxo-3-piperidyl)-1,3-dioxo-isoindorin-4-yl]amino]cyclohexyl]-N-methyl-carbamate 6 (62 mg, 0.1200 mmol) was dissolved in 1,4-dioxane (1.2 mL), and then 4.0 M hydrogen chloride solution (0.61 mL, 2.45 mmol) was added. The resulting solution was stirred at rt. After 12 hours, the reaction was complete. The solvent was evaporated under vacuum, and the residue was co-evaporated with ACN (2 x 5 mL) to obtain product C-13 (62 mg, 99%) as a yellow solid dihydrochloride.

[0273] 1 H NMR (400 MHz, DMSO-d6) δ ppm 1.40-1.53 ​​(m, 4 H), 2.00-2.10 (m, 6 H), 2.52-2.56 (m, 4 H) 2.83-2.97 (m, 2 H), 3.53-3.57 (m, 2 H), 5.02-5.07 (dd, 1 H), 6.18 (d, 1 H), 7.05 (d, 1 H), 7.22 (d, 1 H), 7.57-7.61 (dd, 1 H), 8.75 (s, 2 H), 11.09 (s, 1 H).

[0274] Synthesis of the final compound General method for final product 1 Example S1. Synthesis of P-1 [ka]

[0275] Step 1. Preparation of ethyl 5-(4-tert-butoxycarbonylpiperazin-1-yl)pyrazolo[1,5-a]pyrimidine-3-carboxylate (3) To a solution of ethyl 5-chloropyrazolo[1,5-a]pyrimidine-3-carboxylate 1 (1.5 g, 6.65 mmol, 1.0 equivalent) and tert-butylpiperazine-1-carboxylate 2 (1.56 g, 8.38 mmol, 1.2 equivalent) in MeCN (30 mL, 0.2 M), DIPEA (2.9 mL, 16.60 mmol, 2.5 equivalent) was added. The reaction mixture was stirred overnight at 80°C. The solvent was removed under reduced pressure to obtain crude oily product 3 (2.5 g, quantitative yield), which was used in the next step without purification. LC-MS method 1: 99.9% purity (at 215 nm); [M+H] + = 376.2 m / z.

[0276] Step 2. Preparation of 5-(4-tert-butoxycarbonylpiperazin-1-yl)pyrazolo[1,5-a]pyrimidine-3-carboxylic acid (4) To a solution of ethyl 5-(4-tert-butoxycarbonylpiperazin-1-yl)pyrazolo[1,5-a]pyrimidine-3-carboxylate 3 (5.0 g, 13.3 mmol, 1.0 equivalent) in THF (22.2 mL, 0.2 M) and MeOH (22.2 mL, 0.2 M), a solution of LiOH·H2O (5.6 g, 133 mmol, 10 equivalents) / water (22.2 mL, 0.2 M) was added. The resulting mixture was stirred at 60°C for 3 hours. At completion, THF and MeOH were removed under reduced pressure. The crude mixture was diluted with water. The mixture was acidified to pH=3 (precipitate formation) with 6 M hydrochloric acid aqueous solution while vigorously stirring. The suspension was filtered through a Buchner funnel, and the solid was washed with water. The solid was co-evaporated with THF(3×) to remove water and obtain compound 4 (4.2 g, 91% yield).

[0277] LC-MS method 1: 99.9% purity (at 215 nm); [M+H] + = 348.2 m / z. 1H NMR (400 MHz, CDCl3) δ ppm 1.50 (s, 9 H), 3.60 - 3.66 (m, 4 H), 3.73 - 3.83 (m, 4 H), 6.48 (d, J = 8.1 Hz, 1 H), 8.34 (s, 1 H), 8.37 (d, J = 7.8Hz, 1H).

[0278] Step 3. Preparation of tert-butyl 4-[3-[[3-cyano-1-(4-methoxycarbonylcyclohexyl)pyrazole-4-yl]carbamoyl]pyrazolo[1,5-a]pyrimidine-5-yl]piperazine-1-carboxylate (5) 5-(4-tert-butoxycarbonylpiperazin-1-yl)pyrazolo[1,5-a]pyrimidine-3-carboxylic acid 4 (945 mg, 2.72 mmol), methyl 4-(4-amino-3-cyanopyrazole-1-yl)cyclohexanecarboxylate T-1 (675.4 mg, 2.72 mmol), and NMI (754 μL, 9.52 mmol) were dissolved in MeCN (6.8 mL) and TCFH (916 mg, 3.26 mmol) at room temperature. The resulting mixture was stirred at room temperature for 4 hours. At this point, the reaction mixture was diluted with water. The aqueous layer was extracted three times with ELISA. The combined organic layers were washed with brine and dried over Na2SO4 to concentrate to dryness. The residue was suspended in DMSO. The solid was filtered through a Buchner funnel, washed with MeCN, and then dried under high vacuum to obtain 5 (1.29 g, 82% yield) as a white solid.

[0279] LC-MS method 1: 99.9% purity (at 215 nm), [M+H] + = 578.2, [Mt-Bu+H] + = 522.2. 1H NMR (400 MHz, acetone-d6) δ ppm 1.47 (s, 9 H), 1.57 - 1.74 (m, 2 H), 1.87 - 2.01 (m, 2 H), 2.10 - 2.27 (m, 4 H), 2.48 (br t, J = 12.3 Hz, 1 H), 3.65 (s, 7 H), 3.94 (br s, 4 H), 4.37 (br t, J = 11.7 Hz, 1 H), 6.91 (d, J = 8.1 Hz, 1 H), 8.28 (s, 1 H), 8.46 (s, 1 H), 8.63 (d, J = 7.8 Hz, 1 H), 9.68 (s, 1 H).

[0280] Step 4. Preparation of (1r,4r)-4-(4-(5-(4-(tert-butoxycarbonyl)piperazin-1-yl)pyrazolo[1,5-a]pyrimidine-3-carboxamide)-3-cyano-1H-pyrazole-1-yl)cyclohexane-1-carboxylic acid (6) To a suspension of tert-butyl 4-[3-[[3-cyano-1-(4-methoxycarbonylcyclohexyl)pyrazole-4-yl]carbamoyl]pyrazolo[1,5-a]pyrimidine-5-yl]piperazine-1-carboxylate 5 (600 mg, 1.04 mmol) in methanol (1.7 mL) and THF (1.7 mL), a solution of LiOH monohydrate (174.5 mg, 4.15 mmol) / water (1.7 mL) was added at 0°C. The resulting mixture was stirred at 0°C for 10 minutes, then stirred at room temperature for 1 hour. The solvent was evaporated under reduced pressure, and the residue was suspended in nanopure water and sonicated. 6N HCl was added with vigorous stirring until the pH was 3. Then MeCN was added. The resulting suspension was sonicated and filtered through a Buchner funnel. The solid was washed with nanopure water and MeCN. Subsequently, it was suspended in nanopure water and freeze-dried to obtain 6 (569 mg, 97% yield) as a pale yellow solid.

[0281] LC-MS method 5: 97.6% purity (at 215 nm), [M+Na] + = 586.2, [M+H]+ = 564.2, [Mt-Bu+H] + = 508.2. 1 H NMR (400 MHz, DMSO-d6) δ ppm 1.43 (s, 9 H), 1.47 - 1.58 (m, 2 H), 1.76 - 1.89 (m, 2 H), 1.99 - 2.12 (m, 4 H), 2.27 - 2.36 (m, 1 H), 3.44 - 3.57 (m, 4 H), 3.71 - 4.02 (m, 4 H), 4.31 - 4.41 (m, 1 H), 6.92 (d, J = 7.8 Hz, 1 H), 8.31 (s, 1 H), 8.46 (s, 1 H), 8.84 (d, J = 8.1 Hz, 1 H), 9.61 (s, 1 H), 12.17 (s, 1 H).

[0282] Step 5. Preparation of tert-butyl 4-[3-[[3-cyano-1-[4-[4-[2-(2,6-dioxo-3-piperidyl)-1,3-dioxoisoindoline-5-yl]piperazine-1-carbonyl]cyclohexyl]pyrazole-4-yl]carbamoyl]pyrazolo[1,5-a]pyrimidine-5-yl]piperazine-1-carboxylate (7) (1r,4r)-4-(4-(5-(4-(tert-butoxycarbonyl)piperazin-1-yl)pyrazolo[1,5-a]pyrimidine-3-carboxamide)-3-cyano-1H-pyrazole-1-yl)cyclohexane-1-carboxylic acid 6 (80 mg, 0.14 mmol) and 2-(2,6-dioxo-3-piperidyl)-5-piperazin-1-ylisoindoline-1,3-dionetrifluoroacetate C-1 (90.7 mg, 0.20 mmol) / dry DMF (2 mL) were mixed with DIPEA (0.37 mL, 2.13 mmol) at room temperature. After 5 minutes, HATU (70.2 mg, 0.18 mmol) was added, and the resulting mixture was stirred at room temperature for 20 hours. The solution was then loaded directly onto a 30 g RediSep Rf Gold C18 Isco chromatography column. Elution was performed using a MeCN / 0.1% formic acid aqueous solution (5% over 4 CV, then 5-100% over 15 CV, then 100% over 2 CV). The fractions were combined and concentrated to obtain 7 (109 mg, 86% yield) as a yellow solid.

[0283] LC-MS method 1: 99.9% purity (at 215 nm), [M+H] + = 888.4, [Mt-Bu+H] + = 832.2. 1H NMR (400 MHz, DMSO-d6) δ ppm 1.53 - 1.67 (m, 3 H), 1.79 - 2.15 (m, 9 H), 2.64 (s, 2 H), 2.73 - 2.97 (m, 3 H), 3.43 - 3.57 (m, 10 H), 3.58 - 3.66 (m, 3 H), 3.68 - 3.78 (m, 3 H), 3.78 - 3.93 (m, 5 H), 4.35 - 4.47 (m, 1 H), 5.08 (dd, J = 12.3, 5.0 Hz, 1 H), 6.93 (d, J = 7.6 Hz, 1 H), 7.27 (dd, J = 8.7, 1.8 Hz, 1 H), 7.37 (d, J = 2.2 Hz, 1 H), 7.71 (d, J = 8.3 Hz, 1 H), 8.32 (s, 1 H), 8.48 (s, 1 H), 8.86 (d, J = 7.8 Hz, 1 H), 9.64 (s, 1 H), 11.08 (s, 1 H).

[0284] Step 6. Manufacturing of N-(3-cyano-1-((1r,4r)-4-(4-(2-(2,6-dioxopiperidine-3-yl)-1,3-dioxoisoindorin-5-yl)piperazine-1-carbonyl)cyclohexyl)-1H-pyrazole-4-yl)-5-(piperazine-1-yl)pyrazolo[1,5-a]pyrimidine-3-carboxamide hydrochloride (P-1) In a round-bottom flask, tert-butyl 4-[3-[[3-cyano-1-[4-[4-[2-(2,6-dioxo-3-piperidyl)-1,3-dioxo-isoindolin-5-yl]piperazine-1-carbonyl]cyclohexyl]pyrazole-4-yl]carbamoyl]pyrazolo[1,5-a]pyrimidine-5-yl]piperazine-1-carboxylate 7 (75 mg, 0.08 mmol) was dissolved in 4 M HCl / dioxane (2.1 mL, 8.40 mmol) solution. This solution was stirred at room temperature for 1.5 hours. At this point, the solvent was evaporated under reduced pressure. The remaining dioxane was co-evaporated with water (repeated 3 times). The residue was dissolved in water and freeze-dried to obtain P-1 (60.4 mg, 87% yield) as a yellow solid.

[0285] LC-MS method 5: 99.9% purity (at 215 nm), [M-HCl+Na] + = 810.2, [M-HCl+2H] 2+ = 394.7. 1 H NMR (400 MHz, DMSO-d6) δ ppm 1.53 - 1.66 (m, 2 H), 1.81 - 2.11 (m, 7 H), 2.53 - 2.63 (m, 2 H), 2.76 - 2.95 (m, 2 H), 3.22 - 3.29 (m, 4 H), 3.44 - 3.57 (m, 4 H), 3.59 - 3.76 (m, 4 H), 4.00 - 4.13 (m, 4 H), 4.37 - 4.46 (m, 1 H), 5.08 (dd, J = 12.8, 5.5 Hz, 1 H), 7.00 (d, J = 8.1 Hz, 1 H), 7.27 (dd, J = 8.7, 1.8 Hz, 1 H), 7.37 (d, J = 1.5 Hz, 1 H), 7.71 (d, J = 8.3 Hz, 1 H), 8.37 (s, 1 H), 8.48 (s, 1 H), 8.95 (d, J = 8.1 Hz, 1 H), 9.04 (br. s, 2 H), 9.59 (s, 1 H), 11.08 (s, 1 H).

[0286] Example S2. Synthesis of P-13 [ka]

[0287] Step 1. Preparation of ethyl 5-[(3R,5R)-3-(tert-butoxycarbonylamino)-5-fluoro-1-piperidyl]pyrazolo[1,5-a]pyrimidine-3-carboxylate (3) To a solution of ethyl 5-chloropyrazolo[1,5-a]pyrimidine-3-carboxylate 1 (800 mg, 3.55 mmol, 1.0 eq.) in MeCN (17.7 mL), DIPEA (1.54 mL, 8.86 mmol, 2.5 eq.) and tert-butyl N-[(3R,5R)-5-fluoro-3-piperidyl]carbamate 2 (1.01 g, 4.61 mmol, 1.3 eq.) were added. After stirring at 60°C over the weekend, complete conversion to 3 was shown by LC-MS. The solvent was removed under reduced pressure, and the residue was vacuum-dried to obtain 3 (1.44 g, quantitative yield) as a white solid. The crude product was used in the next step without further purification.

[0288] LC-MS method 1: Retention time: 1.656 min, 99.9% purity (at 215 nm), [M+H] + = 408.2. 1H NMR (400 MHz, DMSO-d6) δ ppm 1.37 - 1.45 (m, 9 H), 1.56 - 1.92 (m, 1 H), 2.09 - 2.20 (m, 1 H), 2.56 - 2.71 (m, 1 H), 2.76 - 3.03 (m, 1 H), 3.08 - 3.19 (m, 1 H), 3.29 (s, 1 H), 3.37 (br s, 1 H), 3.56 - 3.73 (m, 2 H), 4.11 - 4.25 (m, 2 H), 4.41 - 4.73 (m, 1 H), 4.99 (br s, 1 H), 5.11 (br s, 1 H), 6.85 (br d, J = 8.1 Hz, 1 H), 7.10 (br d, J = 7.8 Hz, 1 H), 8.22 (s, 1 H), 8.73 (d, J = 7.8 Hz, 1 H). 19 F NMR (377 MHz, DMSO-d6) δ ppm -184.32 (s, 1 F).

[0289] Step 2. Preparation of 5-[(3R,5R)-3-(tert-butoxycarbonylamino)-5-fluoro-1-piperidyl]pyrazolo[1,5-a]pyrimidine-3-carboxylic acid (4) To a solution of ethyl 5-[(3R,5R)-3-(tert-butoxycarbonylamino)-5-fluoro-1-piperidyl]pyrazolo[1,5-a]pyrimidine-3-carboxylate 3 (1.44 mg, 3.53 mmol, 1.0 eq) / THF (5.89 mL) and methanol (5.89 mL), a solution of LiOH·H2O (1.48 g, 35.34 mmol, 10.0 eq.) / water (5.89 mL) was added. After stirring at 60°C for 18 hours, complete conversion to the target product 4 was shown by LC-MS. The reaction mixture was concentrated under reduced pressure to remove THF / MeOH, and the crude mixture was diluted with water. Under vigorous stirring, the mixture was acidified with 6 N HCl aqueous solution to pH=3 (precipitate formation). The suspension was filtered through a Buchner funnel, and the solid was rinsed with water. The solid was dried overnight in a stove under vacuum to obtain 4 (1.40 g, quantitative yield) as a white solid. The crude product was used in the next step without further purification.

[0290] LC-MS method 1: Retention time: 1.487 min, 99.9% purity (at 215 nm), [M+H] + = 380.1. 1 H NMR (400 MHz, DMSO-d6) δ ppm 1.41 (s, 9 H), 1.66 - 1.95 (m, 1 H), 2.05 - 2.26 (m, 1 H), 2.92 - 3.11 (m, 1 H), 3.28 - 3.30 (m, 1 H), 3.34 - 3.49 (m, 1 H), 3.59 - 3.74 (m, 1 H), 4.52 - 4.75 (m, 1 H), 4.99 (br s, 1 H), 5.11 (br s, 1 H), 6.82 (br d, J = 7.6 Hz, 1 H), 7.12 (br d, J = 7.8 Hz, 1 H), 8.19 (s, 1 H), 8.73 (d, J = 7.8 Hz, 1 H). 19 F NMR (377 MHz, DMSO-d6) δ ppm-184.32 (s, 1 F).

[0291] Step 3. Preparation of methyl 4-[4-[[5-[(3R,5R)-3-(tert-butoxycarbonylamino)-5-fluoro-1-piperidyl]pyrazolo[1,5-a]pyrimidine-3-carbonyl]amino]-3-(trifluoromethyl)pyrazole-1-yl]cyclohexanecarboxylate (5) To a solution of 5-[(3R,5R)-3-(tert-butoxycarbonylamino)-5-fluoro-1-piperidyl]pyrazolo[1,5-a]pyrimidine-3-carboxylic acid 4 (330 mg, 0.87 mmol, 1.0 eq.) and methyl 4-[4-amino-3-(trifluoromethyl)pyrazole-1-yl]cyclohexanecarboxylate T-2 (255 mg, 0.87 mmol, 1.0 eq.) in MeCN (4.37 mL, 0.2 M), NMI (208 μL, 2.62 mmol, 3.0 eq.) was added, followed by TCFH (368 mg, 1.31 mmol, 1.5 eq.). The resulting mixture was stirred at room temperature. After stirring for 1 hour, complete conversion to 5 was shown by LC-MS. Nanopure water was added to the reaction mixture. The suspension was sonicated and filtered through a Buchner funnel. The solid was rinsed with nanopure water and dried under high vacuum for 2 hours to obtain 5 (279 mg, 46% yield) as a white solid. The crude product was used without further purification.

[0292] LC-MS method 1: Retention time: 1.812 min, 99.9% purity (at 215 nm), [M+H] + = 653.2. 1H NMR (400 MHz, DMSO-d6) δ ppm 1.30 (br s, 9 H), 1.49 - 1.62 (m, 3 H), 1.75 - 1.92 (m, 4 H), 2.01 - 2.11 (m, 5 H), 3.40 - 3.57 (m, 1 H), 3.63 (s, 3 H), 4.32 (br t, J = 10.9 Hz, 1 H), 4.50 - 4.64 (m, 1 H), 4.96 - 5.18 (m, 1 H), 6.92 (br d, J = 5.5 Hz, 1 H), 7.09 (br s, 1 H), 8.30 (s, 1 H), 8.41 (br s, 1 H), 8.82 (br d, J = 7.8 Hz, 1 H), 9.22 (br s, 1 H). 2H is not observed. 19 F NMR (377 MHz, DMSO-d6) δ ppm -185.73 - -184.64 (m, 1 F), -59.01 (br s, 3 F).

[0293] Step 4. Preparation of 4-[4-[[5-[(3R,5R)-3-(tert-butoxycarbonylamino)-5-fluoro-1-piperidyl]pyrazolo[1,5-a]pyrimidine-3-carbonyl]amino]-3-(trifluoromethyl)pyrazole-1-yl]cyclohexanecarboxylic acid (6) To a solution of methyl 4-[4-[[5-[(3R,5R)-3-(tert-butoxycarbonylamino)-5-fluoro-1-piperidyl]pyrazolo[1,5-a]pyrimidine-3-carbonyl]amino]-3-(trifluoromethyl)pyrazole-1-yl]cyclohexanecarboxylate 5 (279 mg, 0.43 mmol, 1.0 eq.) in THF (2.14 mL, 0.1 M), a solution of LiOH·H2O (180 mg, 4.27 mmol, 10.0 eq.) / water (2.14 mL, 0.1 M) was added, and the resulting mixture was stirred at room temperature. After stirring overnight, complete conversion to 6 was shown by LC-MS. The reaction mixture was concentrated under reduced pressure, and the residue was suspended in nanopure water. Under vigorous stirring at 0°C, aqueous 6N HCl was added until the pH became 3. The solid was filtered through a Buchner funnel, rinsed with nanopure water, and dried overnight under high vacuum to obtain 6 (232 mg, 83% yield) as a white solid. The product was used in the next step without further purification.

[0294] LC-MS method 1: Retention time: 1.667 min, 99.9% purity (at 215 nm), [M+H] + = 639.2. 1 H NMR (400 MHz, DMSO-d6) δ ppm 1.29 (br s, 9 H), 1.43 - 1.61 (m, 3 H), 1.68 - 1.93 (m, 4 H), 1.97 - 2.14 (m, 5 H), 2.25 - 2.39 (m, 1 H), 3.39 - 3.75 (m, 2 H), 4.30 (br t, J = 11.7 Hz, 1 H), 4.50 - 4.68 (m, 1 H), 4.89 - 5.19 (m, 1 H), 6.91 (br s, 1 H), 7.10 (br s, 1 H), 8.29 (br s, 1 H), 8.41 (br s, 1 H), 8.81 (br d, J = 6.1 Hz, 1 H), 9.21 (br s, 1 H), 12.15 (br s, 1 H). 19F NMR (377 MHz, DMSO-d6) δ ppm -186.50 - -181.97 (m, 1 F), -59.01 (br s, 3 F).

[0295] Step 5. Preparation of tert-butyl N-[(3R,5R)-1-[3-[[1-[4-[4-[2-(2,6-dioxo-3-piperidyl)-1,3-dioxo-isoindoline-5-yl]piperazine-1-carbonyl]cyclohexyl]-3-(trifluoromethyl)pyrazole-4-yl]carbamoyl]pyrazolo[1,5-a]pyrimidine-5-yl]-5-fluoro-3-piperidyl]carbamate (7) 4-[4-[[5-[(3R,5R)-3-(tert-butoxycarbonylamino)-5-fluoro-1-piperidyl]pyrazolo[1,5-a]pyrimidine-3-carbonyl]amino]-3-(trifluoromethyl)pyrazole-1-yl]cyclohexanecarboxylic acid 6 (75 mg, 0.12 mmol) and 2-(2,6-dioxo-3-piperidyl)-5-piperazine-1-ylisoindoline-1,3-dionetrifluoroacetate C-1 (75 mg, 0.16 mmol) were dissolved in anhydrous DMF (1 mL), to which DIPEA (0.1 mL, 0.59 mmol) was added at room temperature. After 5 minutes, HATU (53.59 mg, 0.14 mmol) was added. The resulting mixture was stirred at room temperature for 18 hours. The crude mixture was directly purified by RediSep Rf Gold reverse-phase flash chromatography using a 10-80% MeCN / 0.1% formic acid aqueous solution. The fraction containing the target product was collected, and the volatile substances were concentrated under vacuum to obtain 7 (86 mg, 90% yield) as a yellow solid.

[0296] LC-MS method 1: 99.9% purity (at 215 nm), [M+H] + = 963.2 m / z. 1H NMR (400 MHz, DMSO-d6) δ ppm 1.30 (br. s, 9 H), 1.48 - 1.68 (m, 2 H), 1.75 - 2.10 (m, 8 H), 2.53 - 2.63 (m, 2 H), 2.76 - 2.95 (m, 2 H), 3.31 (m, 3 H), 3.39 - 3.83 (m, 10 H), 4.29 - 4.42 (m, 1 H), 4.47 - 4.68 (m, 1 H), 4.96 - 5.16 (m, 2 H), 6.92 (d, J = 7.8 Hz, 1 H), 7.02 - 7.16 (m, 1 H), 7.23 - 7.31 (m, 1 H), 7.37 (s, 1 H), 7.71 (d, J = 8.6 Hz, 1 H), 8.30 (s, 1 H), 8.41 (br. s, 1 H), 8.82 (d, J = 7.8 Hz, 1 H), 9.23 (s, 1 H), 11.08 (s, 1 H). 19 F NMR (377 MHz, DMSO-d6) δ ppm -185.22 (s, 1 F), -58.91 (s, 3 F).

[0297] Step 6. Preparation of 5-[(3R,5R)-3-amino-5-fluoro-1-piperidyl]-N-[1-[4-[4-[2-(2,6-dioxo-3-piperidyl)-1,3-dioxo-isoindoline-5-yl]piperazine-1-carbonyl]cyclohexyl]-3-(trifluoromethyl)pyrazole-4-yl]pyrazolo[1,5-a]pyrimidine-3-carboxamide (P-13) 103 mg, 0.11 mmol of tert-butyl N-[(3R,5R)-1-[3-[[1-[4-[4-[2-(2,6-dioxo-3-piperidyl)-1,3-dioxo-isoindoline-5-yl]piperazine-1-carbonyl]cyclohexyl]-3-(trifluoromethyl)pyrazole-4-yl]carbamoyl]pyrazolo[1,5-a]pyrimidine-5-yl]-5-fluoro-3-piperidyl]carbamate 7 (103 mg, 0.11 mmol) was placed in a round-bottom flask, to which 4.0 M HCl / 1,4-dioxane (4 mL) was added, and the solution was stirred at rt for 30 minutes. After the reaction was completed by HPLC, the volatile components were evaporated under reduced pressure. The residue was purified by C18 RediSep Rf Gold reverse-phase chromatography using elution with 5 to 50% MeCN / 0.02 M aqueous solution. The desired fractions were combined, concentrated under reduced pressure, and lyophilized to obtain P-13 (39.8 mg, 43% yield) as a yellow solid.

[0298] LC-MS method 5: 99.9% purity (at 215 nm), [M-HCl+H] + = 863.2 m / z, [M-HCl+2H] 2+ = 432.2 m / z. 1H NMR (400 MHz, DMSO-d6) δ ppm 1.54 - 1.68 (m, 2 H), 1.81 - 2.12 (m, 8 H), 2.35 - 2.44 (m, 1 H), 2.53 - 2.64 (m, 2 H), 2.76 - 2.96 (m, 2 H),3.22 - 3.29 (m, 1 H), 3.35 - 3.78 (m, 10 H), 4.33 - 4.43 (m, 1 H), 4.44 - 4.93 (m, 2 H), 5.03 - 5.22 (m, 2 H), 6.91 (d,J = 7.6 Hz, 1 H), 7.28 (dd,J =8.6, 1.5 Hz, 1 H), 7.37 (s, 1 H), 7.71 (d,J = 8.6 Hz, 1 H), 8.13 (br s, 3 H), 8.34 (s, 1 H), 8.46 (s, 1 H), 8.94 (d,J = 8.1 Hz, 1 H), 9.25 (s, 1 H), 11.08 (s, 1 H). 19 F NMR (377 MHz, DMSO-d6) δ ppm -184.97 (s, 1 F), -58.76 (s, 3 F).

[0299] Example S3. Synthesis of P-31 [ka]

[0300] Step 1. Preparation of ethyl 5-morpholinopyrazolo[1,5-a]pyrimidine-3-carboxylate 3 In a sealed tube, ethyl 5-chloropyrazolo[1,5-a]pyrimidine-3-carboxylate 1 (1.5 g, 6.65 mmol, 1 eq.), MeCN (33.24 mL, 0.2 M), and DIPEA (2.89 mL, 16.62 mmol, 2.5 eq.) were added, followed by morpholine 2 (639.65 uL, 7.31 mmol, 1.1 eq.). The tube was sealed, and the mixture was stirred at 90°C for 16 hours. The solvent was evaporated under reduced pressure, and the mixture was dried under high vacuum to obtain ethyl 5-morpholinopyrazolo[1,5-a]pyrimidine-3-carboxylate 3 (2.96 g, 99% yield) as an orange solid.

[0301] LC-MS method 1: Retention time: 1.432 min, 99% purity (at 215 nm), [M + H] + = 277.2 1 ¹H NMR (400 MHz, chloroform-d) δ ppm 1.39 (t, J = 7.1 Hz, 3 H), 3.76 - 3.86 (m, 8 H), 4.35 (q, J = 7.1 Hz, 2 H), 6.41 (d, J = 8.1 Hz, 1 H), 8.29 - 8.34 (m, 2 H).

[0302] Step 2. Preparation of 5-morpholinopyrazolo[1,5-a]pyrimidine-3-carboxylic acid 4 A solution of ethyl 5-morpholinopyrazolo[1,5-a]pyrimidine-3-carboxylate 3 (1.84 g, 6.66 mmol, 1.0 eq.) in THF (17.8 mL, 0.1 M) and methanol (17.8 mL) was stirred for 5 minutes, and then a solution of LiOH·H2O (2.8 g, 66.6 mmol, 10 eq.) / water (17.8 mL) was added. The mixture was stirred at 60°C for 3 hours, then the oil bath was removed, and the mixture was stirred over the weekend (72 h). The reaction mixture was concentrated under reduced pressure, and the crude mixture was diluted with a small amount of water and acidified to pH 3 with 6 N HCl. The resulting precipitate was filtered and rinsed with water. The solid was dried under high vacuum to obtain 5-morpholinopyrazolo[1,5-a]pyrimidine-3-carboxylic acid 4 (1.65 g, 99% yield) as a brown solid. LC-MS method 1: Retention time: 1.192 min, 99% purity (at 215 nm), [M + H] + = 249.2 1 H NMR (400 MHz, DMSO-d6) δ ppm 3.72 (br d, J = 5.9 Hz, 8 H), 6.84 (d, J = 8.1 Hz, 1 H), 8.19 (s, 1 H), 8.74 (d, J = 7.8 Hz, 1 H), 11.73 (s, 1 H).

[0303] Step 3. Preparation of Methyl 4-[3-(difluoromethyl)-4-[(5-morpholinopyrazolo[1,5-a]pyrimidine-3-carbonyl)amino]pyrazole-1-yl]cyclohexanecarboxylate 5 In a round-bottom flask, methyl 4-[4-amino-3-(difluoromethyl)pyrazole-1-yl]cyclohexanecarboxylate T-3 (201.83 mg, 0.66 mmol, 1.1 eq.), 5-morpholinopyrazolo[1,5-a]pyrimidine-3-carboxylic acid 4 (150.0 mg, 0.60 mmol, 1.0 eq.), MeCN (7.5 mL, 0.08 M), and NMI (174.74 uL, 2.21 mmol, 3.6 eq.) were added. The mixture was stirred at 0°C for 5 minutes, and then TCFH (211.93 mg, 0.76 mmol, 1.25 eq.) was added. The mixture was stirred at 0°C for 5 minutes, and then heated to room temperature over 30 minutes. The reaction mixture was concentrated under vacuum. The solution was purified by reverse-phase chromatography (50 g C18 RediSep Rf Gold column, holding solution (DMSO), elution: 5% MeOH / 0.1% HCOOH over 4 CV, then 5% to 100% MeOH / 0.1% HCOOH over 15 CV). The pure fractions were combined, concentrated under reduced pressure, and dried under high vacuum to obtain methyl 4-[3-(difluoromethyl)-4-[(5-morpholinopyrazolo[1,5-a]pyrimidine-3-carbonyl)amino]pyrazole-1-yl]cyclohexanecarboxylate 5 (315.8 mg, 95% yield) as an orange solid.

[0304] LC-MS method 1: Retention time: 1.687 min, 92.4% purity (at 215 nm), [M + H] + = 504.0. 1H NMR (400 MHz, DMSO-d6) δ ppm 1.47 - 1.62 (m, 2 H), 1.73 - 1.88 (m, 2 H), 1.98 - 2.10 (m, 4 H), 2.37 - 2.47 (m, 1 H), 3.62 (s, 3 H), 3.69 - 3.74 (m, 4 H), 3.76 - 3.82 (m, 4 H), 4.25 (ddt, J = 11.7, 7.9, 3.9 Hz, 1 H), 6.91 (d, J = 7.8 Hz, 1 H), 6.95 - 7.25 (m, 1 H), 8.29 (s, 1 H), 8.39 (s, 1 H), 8.83 (d, J = 8.1 Hz, 1 H), 9.40 (s, 1 H).

[0305] Step 4. Preparation of 4-[3-(difluoromethyl)-4-[(5-morpholinopyrazolo[1,5-a]pyrimidine-3-carbonyl)amino]pyrazole-1-yl]cyclohexanecarboxylic acid 6 In a round-bottom flask, methyl 4-[3-(difluoromethyl)-4-[(5-morpholinopyrazolo[1,5-a]pyrimidine-3-carbonyl)amino]pyrazole-1-yl]cyclohexanecarboxylate 5 (291.8 mg, 0.58 mmol, 1.0 eq.), THF (1.9 mL, 0.1 M), and methanol (1.9 mL) were added. The solution was stirred for 5 minutes, and then a solution of LiOH·H2O (243.41 mg, 5.8 mmol, 10.0 eq.) / water (1.9 mL) was added. The mixture was stirred at 60°C for 1 hour. The reaction mixture was concentrated under reduced pressure, and the crude mixture was diluted with a small amount of water and acidified to pH 3 with 6 N HCl. The precipitate was filtered, washed with acetonitrile, and dried under high vacuum to obtain 4-[3-(difluoromethyl)-4-[(5-morpholinopyrazolo[1,5-a]pyrimidine-3-carbonyl)amino]pyrazole-1-yl]cyclohexanecarboxylic acid 6 (251.0 mg, 81% yield) as an off-white solid.

[0306] LC-MS method 1: Retention time: 1.518 min, 91.9% purity (at 215 nm), [M + H] + = 490.2. 1 H NMR (400 MHz, DMSO-d6) δ ppm 1.44 - 1.59 (m, 2 H), 1.71 - 1.86 (m, 2 H), 1.99 - 2.09 (m, 4 H), 2.23 - 2.35 (m, 1 H), 3.68 - 3.75 (m, 4 H), 3.76 - 3.84 (m, 4 H), 4.17 - 4.31 (m, 1 H), 6.91 (d, J = 8.1 Hz, 1 H), 6.95 - 7.26 (m, 1 H), 8.29 (s, 1 H), 8.39 (s, 1 H), 8.83 (d, J = 8.1 Hz, 1 H), 9.40 (s, 1 H), 12.15 (br s, 1 H).

[0307] Step 5. Manufacturing of N-[3-(difluoromethyl)-1-[4-[[1-[2-(2,6-dioxo-3-piperidyl)-1-oxo-isoindoline-5-yl]-4-piperidyl]carbamoyl]cyclohexyl]pyrazole-4-yl]-5-morpholino-pyrazolo[1,5-a]pyrimidine-3-carboxamide P-31 4-[3-(difluoromethyl)-4-[(5-morpholinopyrazolo[1,5-a]pyrimidine-3-carbonyl)amino]pyrazole-1-yl]cyclohexanecarboxylic acid 6 (45.0 mg, 0.09 mmol, 1.0 eq.) and 3-[5-(4-amino-1-piperidyl)-1-oxo-isoindorin-2-yl]piperidine-2,6-dione hydrochloride C-7 (41.8 mg, 0.11 mmol, 1.2 eq.) were dissolved in DMF (0.9 mL, 0.1 M), and then DIPEA (160.14 μL, 0.92 mmol, 10 eq.) and HATU (52.43 mg, 0.14 mmol, 1.5 eq.) were added sequentially. The reaction mixture was stirred overnight under a nitrogen atmosphere. After adding water (~7 mL) to the mixture, the solid was filtered and washed with water. The solid was collected, dissolved in DMF, and purified by reverse-phase chromatography (50 g C18 RediSep Rf Gold column, holding solution (DMF), elution: 5% MeCN / 0.1% HCOOH over 4 CV, then 5% to 50% MeCN / 0.1% HCOOH over 15 CV). The pure fractions were combined, concentrated under reduced pressure, and freeze-dried overnight to obtain N-[3-(difluoromethyl)-1-[4-[[1-[2-(2,6-dioxo-3-piperidyl)-1-oxo-isoindoline-5-yl]-4-piperidyl]carbamoyl]cyclohexyl]pyrazole-4-yl]-5-morpholino-pyrazolo[1,5-a]pyrimidine-3-carboxamide P-31 (21.8 mg, 29% yield).

[0308] LC-MS method 5: Retention time: 3.017 min, 99.8% purity (at 215 nm), [M+2H] 2+ = 407.8, [M+H] + = 814.3. 1H NMR (400 MHz, DMSO-d6) δ ppm 1.42 - 1.63 (m, 4 H), 1.67 - 1.87 (m, 6 H), 1.93 - 2.00 (m, 1 H), 2.04 - 2.11 (m, 2 H), 2.12 - 2.19 (m, 1 H), 2.35 - 2.44 (m, 1 H), 2.55 - 2.63 (m, 1 H), 2.85 - 3.02 (m, 3 H), 3.70 - 3.75 (m, 4 H), 3.77 - 3.86 (m, 7 H), 4.18 - 4.35 (m, 3 H), 5.05 (dd, J = 13.2, 5.1 Hz, 1H), 6.91 (d, J = 7.8 Hz, 1 H), 6.97 - 7.24 (m, 3 H), 7.51 (d, J = 8.6 Hz, 1 H), 7.77 (d, J = 7.8 Hz, 1 H), 8.29 (s, 1 H), 8.39 (s, 1 H), 8.83 (d, J = 7.8 Hz, 1 H), 9.40 (s, 1 H), 10.94 (s, 1 H). 19 F NMR (377 MHz, DMSO-d6) δ ppm -111.32 (d, J = 54.5 Hz, 2 F).

[0309] Example S4. The following compounds were synthesized using the same general route as described above, with modifications to amine 2 in step 1, intermediate (TX) in step 3, and CBM in step 5 (Table 2). [Table 10] [Table 11] [Table 12] [Table 13] [Table 14] [Table 15] [Table 16] [Table 17] [Table 18] [Table 19] [Table 20] [Table 21] [Table 22]

[0310] General method for the final product 2 Example S5. Synthesis of P-6 [ka]

[0311] Step 1. Preparation of 4-amino-1-[4-(hydroxymethyl)cyclohexyl]pyrazole-3-carbonitrile (2) To a solution of methyl 4-(4-amino-3-cyanopyrazole-1-yl)cyclohexanecarboxylate T-1 (25 mg, 0.100 mmol, 1 eq.) in THF (0.3 mL) and ethanol (0.6 mL), CaCl2 (22.4 mg, 0.200 mmol, 2 eq.) and then NaBH4 (15.2 mg, 0.400 mmol, 4 eq.) were added at 0°C. The resulting mixture was stirred at 0°C for 1 hour, then stirred at room temperature. After 17 hours, complete conversion was shown by LC-MS. Water was added, and the reaction mixture was stirred at room temperature. After 1 hour, the aqueous layer was extracted three times with siRNA. The organic matter was washed with brine and dried over Na2SO4 to concentrate to dryness. Crude product 2 was used in the next step without purification.

[0312] Step 2. Preparation of tert-butyl 4-[3-[[3-cyano-1-[4-(hydroxymethyl)cyclohexyl]pyrazole-4-yl]carbamoyl]pyrazolo[1,5-a]pyrimidine-5-yl]piperazine-1-carboxylate (4) 4-amino-1-[4-(hydroxymethyl)cyclohexyl]pyrazole-3-carbonitrile 2 (21.6 mg, 0.100 mmol, 1 eq.), 5-(4-tert-butoxycarbonylpiperazin-1-yl)pyrazolo[1,5-a]pyrimidine-3-carboxylic acid 3 [synthesized in general method 1; P-1 of the final product] (35 mg, 0.100 mmol, 1 eq.), and NMI (28 uL, 0.350 mmol, 3.5 eq.) were mixed in MeCN (1 mL) with TCFH (33.9 mg, 0.120 mmol, 1.2 eq.). The resulting mixture was stirred at room temperature. After 18 hours, complete conversion was shown by LC-MS. Water was added, and the reaction mixture was stirred at 0°C for 30 minutes. The solid was filtered through a Buchner funnel, rinsed with water, and dried under high vacuum to obtain 4 (37.2 mg, 67% yield) as a white solid.

[0313] LC-MS method 1: 86.9% purity (at 215 nm), [M+H] += 550.2. 1 H NMR (400 MHz, DMSO-d6) δ ppm 1.03 - 1.16 (m, 2 H), 1.43 (s, 9 H), 1.70 - 1.82 (m, 2 H), 1.82 - 1.92 (m, 2 H), 2.00 - 2.09 (m, 2 H), 3.26 (t, J = 5.6 Hz, 2 H), 3.44 (br dd, J = 4.6, 3.4 Hz, 1 H), 3.50 (br s, 4 H), 3.83 (br s, 4 H), 4.30 (tt, J = 12.0, 3.9 Hz, 1 H), 4.48 (t, J = 5.3 Hz, 1 H), 6.92 (d, J = 8.1 Hz, 1 H), 8.31 (s, 1 H), 8.46 (s, 1 H), 8.85 (d, J = 8.1 Hz, 1 H), 9.61 (s, 1 H).

[0314] Step 3. Preparation of tert-butyl 4-[3-[[3-cyano-1-[4-[[4-[2-(2,6-dioxo-3-piperidyl)-1,3-dioxoisoindoline-5-yl]piperazine-1-yl]methyl]cyclohexyl]pyrazole-4-yl]carbamoyl]pyrazolo[1,5-a]pyrimidine-5-yl]piperazine-1-carboxylate (5) Solution A: To a solution of tert-butyl 4-[3-[[3-cyano-1-[4-(hydroxymethyl)cyclohexyl]pyrazole-4-yl]carbamoyl]pyrazolo[1,5-a]pyrimidine-5-yl]piperazine-1-carboxylate 4 (37.2 mg, 0.070 mmol, 1 eq.) in anhydrous DMSO (1 mL), IBX (24.6 mg, 0.090 mmol, 1.3 eq.) was added. The resulting mixture was stirred at room temperature. After 18 hours, LC-MS showed complete conversion to tert-butyl 4-[3-[[3-cyano-1-(4-formylcyclohexyl)pyrazole-4-yl]carbamoyl]pyrazolo[1,5-a]pyrimidine-5-yl]piperazine-1-carboxylate.

[0315] 2-(2,6-dioxo-3-piperidyl)-5-piperazine-1-ylisoindoline-1,3-dione;2,2,2-trifluoroacetic acid C-1 (30.8 mg, 0.070 mmol, 1 eq.) was dissolved in anhydrous DCE (1 mL) and DIPEA (0.024 mL, 0.140 mmol, 2 eq.) was added. The resulting mixture was stirred at room temperature. After 5 minutes, NaBH(OAc)3 (18.6 mg, 0.090 mmol, 1.3 eq.) was added, followed by solution A. The resulting mixture was stirred at room temperature. After 2 hours, LC-MS showed that the conversion was almost complete. Volatile substances were removed under reduced pressure, and the residue was purified by reverse-phase flash chromatography (30 g C18 RediSep Rf Gold column, holding solution (DMSO), elution: 5% MeOH / 0.1% HCOOH over 3 CV, then 5 to 40% MeOH / 0.1% HCOOH over 2 CV, then 40 to 60% MeOH / 0.1% HCOOH over 11 CV, then 60% MeOH / 0.1% HCOOH over 6 CV). The fractions were combined and concentrated to obtain 5 (30.3 mg, 51% yield) as a yellow solid.

[0316] LC-MS method 1: 99.9% purity (at 215 nm), [M+H] + = 874.2. 1H NMR (400 MHz, DMSO-d6) δ ppm 1.01 - 1.15 (m, 2 H), 1.43 (s, 9 H), 1.62 - 1.71 (m, 1 H), 1.75 - 1.87 (m, 2 H), 1.90 - 2.12 (m, 5 H), 2.16 - 2.22 (m, 2 H), 2.53 - 2.63 (m, 2 H), 2.82 - 2.94 (m, 1 H), 3.42 - 3.55 (m, 8 H), 3.84 (br d, J = 2.7 Hz, 4 H), 4.28 - 4.39 (m, 1 H), 5.07 (dd, J = 12.8, 5.3 Hz, 1H), 6.93 (d, J = 7.8 Hz, 1 H), 7.26 (dd, J = 8.9, 1.3 Hz, 1 H), 7.35 (s, 1 H), 7.68 (d, J = 8.6 Hz, 1 H), 8.32 (s, 1 H), 8.47 (s, 1 H), 8.86 (d, J = 8.1 Hz, 1 H), 9.63 (s, 1 H), 11.08 (s, 1 H).

[0317] Step 4. Preparation of N-[3-cyano-1-[4-[[4-[2-(2,6-dioxo-3-piperidyl)-1,3-dioxo-isoindoline-5-yl]piperazine-1-yl]methyl]cyclohexyl]pyrazole-4-yl]-5-piperazine-1-yl-pyrazolo[1,5-a]pyrimidine-3-carboxamide dihydrochloride (P-6) A solution of tert-butyl 4-[3-[[3-cyano-1-[4-[[4-[2-(2,6-dioxo-3-piperidyl)-1,3-dioxo-isoindoline-5-yl]piperazine-1-yl]methyl]cyclohexyl]pyrazole-4-yl]carbamoyl]pyrazolo[1,5-a]pyrimidine-5-yl]piperazine-1-carboxylate 5 (30 mg, 0.030 mmol, 1 eq.) in 4 M HCl / 1,4-dioxane (1.29 mL, 5.15 mmol, 150 eq.) was stirred at room temperature. After 1 hour, complete conversion was shown by LC-MS. The solvent was removed under reduced pressure, and the residue was purified by reverse-phase flash chromatography (30 g C18 RediSep Rf Gold column, holding solution (DMSO), elution: 5% MeCN / 0.02 M HCl over 3 CV, then 5-20% MeCN / 0.02 M HCl over 6 CV, then 20-30% MeCN / 0.02 M HCl over 12 CV). The fractions were combined, concentrated, and lyophilized to obtain P-6 (16.2 mg, 61% yield) as a yellow solid, all as the dihydrochloride.

[0318] LC-MS method 5: 99.6% purity (at 215 nm), [M-2HCl+H] + = 774.3. 1H NMR (400 MHz, DMSO-d6) δ ppm 1.16 - 1.28 (m, 2 H), 1.78 - 1.91 (m, 2 H), 1.94 - 2.12 (m, 6 H), 2.53 - 2.64 (m, 2 H), 2.83 - 2.96 (m, 1 H), 3.03 - 3.20 (m, 4 H), 3.21 - 3.27 (m, 4 H), 3.43 - 3.53 (m, 2 H), 3.56 - 3.68 (m, 2 H), 3.99 - 4.15 (m, 4 H), 4.17 - 4.26 (m, 2 H), 4.31 - 4.44 (m, 1 H), 5.10 (dd, J = 13.0, 5.1 Hz, 1 H), 7.00 (d, J = 8.1 Hz, 1 H), 7.36 (d, J = 9.8 Hz, 1 H), 7.50 (s, 1 H), 7.77 (d, J = 8.6 Hz, 1 H), 8.37 (s, 1 H), 8.49 (s, 1 H), 8.95 (d, J = 7.8 Hz, 1 H), 9.18 (br s, 2 H), 9.59 (s, 1 H), 10.18 (br s, 1 H), 11.09 (s, 1 H).

[0319] Example S6. Synthesis of P-32 [ka] Step 1. Preparation of [4-[4-amino-3-(difluoromethyl)pyrazole-1-yl]cyclohexyl]methanol (2) To a solution of methyl 4-[4-amino-3-(difluoromethyl)pyrazole-1-yl]cyclohexanecarboxylate T-3 (1.23 g, 4.5 mmol, 1.0 eq.) in THF (15.0 mL, 0.1 M) and ethanol (30.0 mL), CaCl2 (1.0 g, 9.0 mmol, 2.0 eq.) and then NaBH4 (0.68 g, 18.0 mmol, 4.0 eq.) were added at 0°C. The resulting mixture was stirred overnight and then heated to room temperature. Water was added, and the reaction mixture was stirred at room temperature for 1 hour. The aqueous phase was extracted with SiO(3x). The organic layer was washed with saline solution, dried over Na2SO4, and concentrated to dryness to obtain [4-[4-amino-3-(difluoromethyl)pyrazole-1-yl]cyclohexyl]methanol 2 (1.1 g, 99% yield) as an orange oil.

[0320] LC-MS method 1: Retention time: 0.989 min, 99.0% purity (at 215 nm), [M+H] + = 246.2. 1 H NMR (400 MHz, DMSO-d6) δ ppm 1.04 (qd, J = 12.9, 3.2 Hz, 2 H), 1.32 - 1.43 (m, 1 H), 1.62 (qd, J = 12.6, 3.3 Hz, 2 H), 1.82 (br d, J = 11.5 Hz, 2 H), 1.93 - 1.98 (m, 2 H), 3.24 (t, J = 5.7 Hz, 2 H), 3.94 (tt, J = 11.9, 3.8 Hz, 1 H), 4.00 - 4.06 (m, 2 H), 4.44 (t, J = 5.3 Hz, 1 H), 6.72 - 7.02 (m, 1 H), 7.14 (s, 1 H). 19 F NMR (377 MHz, DMSO-d6) δ ppm -111.21 (s, 2 F).

[0321] Step 2. Preparation of tert-butyl N-[(3r,5r)-1-[3-[[3-(difluoromethyl)-1-[4-(hydroxymethyl)cyclohexyl]pyrazole-4-yl]carbamoyl]pyrazolo[1,5-a]pyrimidine-5-yl]-5-fluoro-3-piperidyl]carbamate (4) [4-[4-amino-3-(difluoromethyl)pyrazole-1-yl]cyclohexyl]methanol 2 (511.8 mg, 1.88 mmol, 1.5 eq.), 5-[(3r,5r)-3-(tert-butoxycarbonylamino)-5-fluoro-1-piperidyl]pyrazolo[1,5-a]pyrimidine-3-carboxylic acid 3 [synthesized in general method of final product 1; P-13] (475.0 mg, 1.25 mmol, 1.0 eq.), and NMI (362.72 uL, 4.58 mmol, 3.6 eq.) were mixed in MeCN (12.5 mL, 0.1 M) with TCFH (440.43 mg, 1.57 mmol, 1.2 eq.). The resulting mixture was stirred overnight at room temperature. Water was added, and the reaction mixture was stirred at room temperature for 1 hour. The solid was filtered through a Buchner funnel and rinsed with a water / MeCN mixture. It was purified by reverse-phase chromatography (50 g C18 RediSep Rf Gold column, holding solution (DMSO), elution: over 5% MeOH / 0.1% HCOOH 4 CV, then over 5% to 100% MeOH / 0.1% HCOOH 15 CV). The pure fractions were combined and concentrated under reduced pressure to obtain tert-butyl N-[(3r,5r)-1-[3-[[3-(difluoromethyl)-1-[4-(hydroxymethyl)cyclohexyl]pyrazole-4-yl]carbamoyl]pyrazolo[1,5-a]pyrimidine-5-yl]-5-fluoro-3-piperidyl]carbamate 4 (421 mg, 55% yield) as a brown solid.

[0322] LC-MS method 1: Retention time: 1.629 min, 99.9% purity (at 215 nm), [M+H] + = 607.2. 1H NMR (400 MHz, DMSO-d6) δ ppm 1.03 - 1.15 (m, 2 H), 1.34 (br s, 9 H), 1.64 - 1.80 (m, 3 H), 1.82 - 1.93 (m, 3 H), 1.99 - 2.07 (m, 2 H), 2.08 - 2.18 (m, 1 H), 2.95 - 3.14 (m, 1 H), 3.26 (t, J = 5.7 Hz, 2 H), 3.37 - 3.52 (m, 1 H), 3.62 - 3.71 (m, 1 H), 4.12 - 4.21 (m, 1 H), 4.47 (t, J = 5.3 Hz, 1 H), 4.69 (br s, 1 H), 4.95 - 5.12 (m, 1 H), 6.86 - 6.94 (m, 1 H), 7.01 - 7.22 (m, 2 H), 8.28 (s, 1 H), 8.34 (br s, 1 H), 8.82 (d, J = 7.8 Hz, 1 H), 9.31 (br s, 1 H). 19 F NMR (377 MHz, DMSO-d6) δ ppm -186.02 - -179.79 (m, 1F), -113.10 - -108.69 (m, 2F).

[0323] Step 3. Preparation of tert-butyl N-[(3r,5r)-1-[3-[[3-(difluoromethyl)-1-[4-[[[1-[2-(2,6-dioxo-3-piperidyl)-1-oxo-isoindoline-5-yl]-4-piperidyl]-methyl-amino]methyl]cyclohexyl]pyrazole-4-yl]carbamoyl]pyrazolo[1,5-a]pyrimidine-5-yl]-5-fluoro-3-piperidyl]carbamate (5) To a solution of tert-butyl N-[(3R,5R)-1-[3-[[3-(difluoromethyl)-1-[4-(hydroxymethyl)cyclohexyl]pyrazole-4-yl]carbamoyl]pyrazolo[1,5-a]pyrimidine-5-yl]-5-fluoro-3-piperidyl]carbamate 4 (75.0 mg, 0.12 mmol, 1.0 eq.) / dried DMSO (1 mL, 0.12 M), IBX (41.54 mg, 0.15 mmol, 1.2 eq.) was added. The resulting mixture was stirred at room temperature. After stirring overnight, LC-MS (Method 3) demonstrated the complete conversion to the target product, tert-butyl N-[(3R,5R)-1-[3-[[3-(difluoromethyl)-1-(4-formylcyclohexyl)pyrazole-4-yl]carbamoyl]pyrazolo[1,5-a]pyrimidine-5-yl]-5-fluoro-3-piperidyl]carbamate. To the reaction mixture, 3-[5-[4-(methylamino)-1-piperidyl]-1-oxo-isoindorin-2-yl]piperidine-2,6-dione hydrochloride (53.61 mg, 0.14 mmol, 1.2 eq.), DCE (1 mL, 0.06 M), and DIPEA (0.22 mL, 1.24 mmol, 10 eq.) were added. The mixture was stirred at room temperature for 10 minutes. Sodium triacetoxyborohydride (34.18 mg, 0.16 mmol, 1.3 eq.) was added, and the mixture was stirred at room temperature for 3 hours. The reaction mixture was concentrated under vacuum and purified by reverse-phase chromatography (50 g C18 RediSep Rf Gold column, retention solution (DMSO), elution: 5% MeCN / 0.1% HCOOH over 4 CV, then 5% to 40% MeCN / 0.1% HCOOH over 15 CV).The pure fractions were combined and concentrated to obtain tert-butyl N-[(3R,5R)-1-[3-[[3-(difluoromethyl)-1-[4-[[[1-[2-(2,6-dioxo-3-piperidyl)-1-oxo-isoindoline-5-yl]-4-piperidyl]-methyl-amino]methyl]cyclohexyl]pyrazole-4-yl]carbamoyl]pyrazolo[1,5-a]pyrimidine-5-yl]-5-fluoro-3-piperidyl]carbamate 5 (49 mg, 36% yield) as a brown solid. The target product contained IBX residue.

[0324] LC-MS method 2: Retention time: 1.685 min, 87.7% purity (at 215 nm), [M - Boc + 2H] 2+ = 423.2. 1H NMR (400 MHz, DMSO-d6) δ ppm 0.98 - 1.06 (m, 2 H), 1.31 - 1.38 (m, 7 H), 1.45 (s, 9 H), 1.48 - 1.54 (m, 2 H), 1.71 - 1.80 (m, 4 H), 1.86 - 1.94 (m, 4 H), 2.04 (dt, J = 15.2, 3.1 Hz, 2 H), 2.18 - 2.27 (m, 5 H), 2.79 - 2.88 (m, 3 H), 3.46 - 3.52 (m, 1 H), 3.62 - 3.72 (m, 2 H), 3.94 (br d, J = 11.5 Hz, 2 H), 4.20 (br d, J = 17.1 Hz, 1 H), 4.33 (br d, J = 16.9 Hz, 1 H), 4.96 - 5.11 (m, 3 H), 5.79 - 5.83 (m, 1 H), 7.06 (s, 2 H), 7.53 (dd, J = 15.9, 8.1 Hz, 2 H), 7.97 (br dd, J = 8.4, 1.3 Hz, 1 H), 8.03 (dd, J = 8.6, 1.2 Hz, 1 H), 8.29 (s, 1 H), 8.33 (br s, 1 H), 8.82 (d, J = 7.8 Hz, 1 H), 9.31 (br s, 1 H), 10.94 (s, 1 H).

[0325] Step 4. Manufacturing of N-[3-(difluoromethyl)-1-[4-[[[1-[2-(2,6-dioxo-3-piperidyl)-1-oxo-isoindoline-5-yl]-4-piperidyl]-methyl-amino]methyl]cyclohexyl]pyrazole-4-yl]-5-[(3r,5r)-3-amino-5-fluoro-1-piperidyl]pyrazolo[1,5-a]pyrimidine-3-carboxamide hydrochloride (P-32) 4.0 M HCl / dioxane (1.94 mL, 7.78 mmol, 150.0 eq.) was added to tert-butyl N-[(3r,5r)-1-[3-[[3-(difluoromethyl)-1-[4-[[[1-[[2-(2,6-dioxo-3-piperidyl)-1-oxo-isoindorin-5-yl]-4-piperidyl]-methyl-aminomethyl]cyclohexyl]pyrazole-4-yl]carbamoyl]pyrazolo[1,5-a]pyrimidine-5-yl]-5-fluoro-3-piperidyl]carbamate 5 (49 mg, 0.05 mmol, 1.0 eq.). The reaction mixture was stirred at room temperature for 2 hours. The reaction mixture was concentrated under vacuum. The solution was purified by reverse-phase chromatography (50 g C18 RediSep Rf Gold column, holding solution (H2O), elution: 5% MeCN / 0.02 M HCl over 4 CV, then 5% to 40% MeCN / 0.02 M HCl over 15 CV). The pure fractions were combined, concentrated under reduced pressure, and freeze-dried overnight to obtain N-[3-(difluoromethyl)-1-[4-[[[1-[2-(2,6-dioxo-3-piperidyl)-1-oxo-isoindorin-5-yl]-4-piperidyl]-methyl-amino]methyl]cyclohexyl]pyrazole-4-yl]-5-[(3r,5r)-3-amino-5-fluoro-1-piperidyl]pyrazolo[1,5-a]pyrimidine-3-carboxamide P-32 (34.8 mg, 79% yield).

[0326] LC-MS method 5: Retention time: 1.797 min, 99.9% purity (at 215 nm), [M-HCl+H] + = 845.5;[M-2HCl+2H] 2+ = 423.3. 1H NMR (400 MHz, DMSO-d6) δ ppm 1.12 - 1.29 (m, 2 H), 1.68 - 2.00 (m, 8 H), 2.03 - 2.19 (m, 6 H), 2.34 - 2.42 (m, 2 H), 2.54 - 2.63 (m, 2 H), 2.72 - 2.78 (m, 3 H), 2.85 - 2.96 (m, 4 H), 3.08 - 3.15 (m, 1 H), 3.28 - 3.39 (m, 2 H), 4.06 (br d, J = 12.7 Hz, 2 H), 4.20 - 4.36 (m, 3 H), 4.53 - 4.65 (m, 1 H), 4.79 - 4.91 (m, 1 H), 5.02 - 5.18 (m, 2 H), 6.90 (d, J = 8.1 Hz, 1 H), 7.00 - 7.28 (m, 3 H), 7.55 (d, J = 8.6 Hz, 1 H), 8.28 - 8.34 (m, 3 H), 8.40 (s, 1 H), 8.94 (d, J = 7.8 Hz, 1 H), 9.34 (s, 1 H), 9.56 - 9.65 (m, 1 H), 10.95 (s, 1 H). 19 F NMR (377 MHz, DMSO-d6) δ ppm -184.56 (s, 2 F), -111.32 (s, 1 F).

[0327] Example S7. The following compounds were synthesized using the same general route by combining the starting intermediate (TX) in step 1, the acid substrate 3 in step 2, and the CBM (CX) in step 3 (Table 3).

[0328] [Table 23] [Table 24] [Table 25] [Table 26] [Table 27]

[0329] Method 3 for the final product Example S8. Synthesis of P-27 [ka]

[0330] Step 1. Production of methyl 4-methylsulfonyloxycyclohexanecarboxylate (2) Under nitrogen, a solution of methyl 4-hydroxycyclohexanecarboxylate 1 (5.0 g, 31.61 mmol, 1.0 eq.) / CH2Cl2 (105 mL, 0.3 M) was cooled to 0°C, and then methanesulfonyl chloride (2.69 mL, 34.77 mmol, 1.1 eq.) and triethylamine (5.28 mL, 37.93 mmol, 1.2 eq.) were added (the latter added dropwise). After stirring at 0°C for 2 hours, complete conversion was shown by TLC (CH2Cl2 / MeOH 5:1, KMnO4 staining). The reaction mixture was quenched by adding water, the phases were separated, and the aqueous phase was extracted three times with CH2Cl2. The combined organic phases were washed twice with saline, dried on magnesium sulfate, filtered, and concentrated to obtain 2 (7.45 g, 99% yield) as a yellow oil.

[0331] 1 H NMR (400 MHz, CDCl3) δ ppm 1.67 - 1.84 (m, 4 H), 1.87 - 1.99 (m, 2 H), 2.00 - 2.09 (m, 2 H), 2.36 - 2.46 (m, 1 H), 3.02 (s, 3 H), 3.69 (s, 3 H), 4.87 - 4.96 (m, 1 H).

[0332] Step 2. Preparation of methyl 4-(3-methyl-4-nitropyrazole-1-yl)cyclohexanecarboxylate (4) To a solution of methyl 4-methylsulfonyloxycyclohexanecarboxylate 2 (976.02 mg, 4.13 mmol) and 3-methyl-4-nitro-1H-pyrazole 3 (350 mg, 2.75 mmol) in dry DMF (13.769 mL), Cs2CO3 (1.79 g, 5.51 mmol) was added. The resulting mixture was stirred overnight at 55°C. After one night, HPLC showed 51% conversion, and methyl 4-methylsulfonyloxycyclohexanecarboxylate 2 (976.02 mg, 4.13 mmol) was added, and the mixture was stirred overnight or longer. HPLC showed 66% conversion, so methyl 4-methylsulfonyloxycyclohexanecarboxylate 2 (650.68 mg, 2.75 mmol) was added, and the mixture was stirred for another night. HPLC showed 95% conversion. Water was added to the mixture, the aqueous phase was extracted three times with RINKAN, the organic phase was washed with water, dried over sodium sulfate, filtered, and concentrated. The residue was purified by normal-phase flash chromatography (elution: 0–30% MTBE / heptane over 15 CV). The pure fractions were combined and concentrated to obtain 4 (410 mg, 56% yield) as a white solid. LC-MS method 1: Retention time: 1.715 min, 99.9% purity (at 215 nm), [M+H] + = 268.2. 1 H NMR (400 MHz, DMSO-d6) δ ppm 1.45 - 1.66 (m, 3 H), 1.74 - 1.95 (m, 6 H), 2.41 - 2.45 (m, 2 H), 2.45 - 2.45 (m, 1 H), 2.65 (s, 2 H), 4.15 - 4.26 (m, 1 H), 4.30 - 4.41 (m, 1 H), 8.80 (s, 1 H).

[0333] Step 3. Preparation of methyl 4-(4-amino-3-methylpyrazole-1-yl)cyclohexanecarboxylate (5) Methyl 4-(3-methyl-4-nitropyrazole-1-yl)cyclohexanecarboxylate 4 (283 mg, 1.06 mmol) was dissolved in ethyl acetate (7.5 mL) and degassed by purging with N2 for 15 minutes. Then, Pd / C (10% w / w) (450.72 mg, 0.42 mmol) was added, and the mixture was degassed by purging with N2 for 15 minutes, followed by purging with H2 for 15 minutes. The mixture was stirred overnight at room temperature under H2 (1 atm). The reaction mixture was filtered through celite, rinsed with MeOH, and concentrated under reduced pressure to obtain 5 (220 mg, 87% yield) as a brown solid.

[0334] LC-MS method 1: 99.9% purity (at 215 nm), [M+H] + = 238.2. 1 H NMR (400 MHz, DMSO-d6) δ ppm 1.39 - 1.59 (m, 1 H), 1.60 - 1.71 (m, 1 H), 1.73 - 1.85 (m, 2 H), 1.87 - 2.04 (m, 4 H), 2.05 - 2.18 (m, 1 H), 2.28 - 2.42 (m, 1 H), 3.43 - 3.72 (m, 4 H), 3.77 - 4.02 (m, 1 H), 6.81 - 7.07 (m, 1 H).

[0335] Step 4. Preparation of methyl 4-[3-methyl-4-[[5-[(3R,5R)-3-(tert-butoxycarbonylamino)-5-fluoro-1-piperidyl]pyrazolo[1,5-a]pyrimidine-3-carbonyl]amino]pyrazole-1-yl]cyclohexanecarboxylate (7) Methyl 4-(4-amino-3-methylpyrazole-1-yl)cyclohexanecarboxylate 5 (220 mg, 0.93 mmol) and 5-[(3R,5R)-3-(tert-butoxycarbonylamino)-5-fluoro-1-piperidyl]pyrazolo[1,5-a]pyrimidine-3-carboxylic acid 6 [synthesized in general method of final product 1; P-13] (351.73 mg, 0.93 mmol) / MeCN (10.5 mL) were mixed with NMI (0.22 mL, 2.78 mmol) and TCFH (390.19 mg, 1.39 mmol). The mixture was stirred overnight at room temperature. Water was then added, and the precipitate was filtered and dried using a high vacuum pump to obtain 7 (293 mg, 52% yield) as an off-white solid.

[0336] LC-MS method 1: 68.4% purity (at 215 nm), [M+H] + = 599.3. 1 H NMR (400 MHz, DMSO-d6) δ ppm 1.17 - 1.31 (s, 9 H), 1.58 (m, 2 H), 1.95 - 2.05 (m, 4 H), 2.01 - 2.16 (m, 4 H), 2.28 (m, 2 H), 2.85 - 3.00 (m, 1 H), 3.50 (m, 8 H), 3.51 - 3.55 (m, 1 H), 4.16 - 4.44 (m, 1 H), 5.06 - 5.16 (m, 1 H), 7.27 - 7.30 (m, 1 H), 7.30 - 7.32 (m, 1 H), 7.32 - 7.35 (m, 1 H), 7.48 - 7.53 (m, 1 H), 8.64 - 8.76 (m, 2 H), 10.98 (s, 1 H).

[0337] Step 5. Preparation of 4-[3-methyl-4-[[5-[(3R,5R)-3-(tert-butoxycarbonylamino)-5-fluoro-1-piperidyl]pyrazolo[1,5-a]pyrimidine-3-carbonyl]amino]pyrazole-1-yl]cyclohexanecarboxylic acid (8) To a suspension of methyl 4-[3-methyl-4-[[5-[(3R,5R)-3-(tert-butoxycarbonylamino)-5-fluoro-1-piperidyl]pyrazolo[1,5-a]pyrimidine-3-carbonyl]amino]pyrazole-1-yl]cyclohexanecarboxylate 7 (290 mg, 0.48 mmol) in THF (2 mL) and methanol (2 mL), LiOH·H2O (81.3 mg, 1.94 mmol) / water (2 mL) was added. The resulting mixture was stirred at 60°C. After 1 hour, THF and MeOH were removed under reduced pressure. Nanopure water was added to the residue, and the pH was adjusted to 3 by adding 6 N HCl. Then, MeCN was added, and the precipitate was filtered and rinsed with water and MeCN to obtain 8 (253 mg, 89% yield) as an off-white solid.

[0338] LC-MS method 1: 99.9% purity (at 215 nm), [M+H] + = 585.4. 1 H NMR (400 MHz, DMSO-d6) δ ppm 1.34 (br s, 4 H), 1.47 - 1.58 (m, 2 H), 1.71 - 1.80 (m, 2 H), 1.82 - 1.94 (m, 2 H), 1.97 - 2.07 (m, 2 H), 2.08 (s, 1 H), 2.21 (s, 4 H), 2.66 - 2.70 (m, 1 H), 3.62 - 3.79 (m, 2 H), 4.01 - 4.17 (m, 2 H), 4.39 - 4.77 (m, 4 H), 5.00 - 5.21 (m, 3 H), 6.88 - 6.99 (m, 1H), 7.12 - 7.20 (m, 1 H), 7.68 - 7.73 (m, 1 H), 8.00 - 8.05 (m, 1 H), 8.26 (m, 1 H), 8.82 (m, 1 H), 9.03 - 9.09 (m, 1 H), 12.09 - 12.24 (m, 1 H). 19 F NMR (377 MHz, DMSO-d6) δ ppm -167.26 (s, 1 F).

[0339] Step 6. Preparation of tert-butyl N-[(3R,5R)-1-[3-[[1-[4-[[1-[2-(2,6-dioxo-3-piperidyl)-1,3-dioxo-isoindoline-5-yl]-4-piperidyl]carbamoyl]cyclohexyl]-3-methyl-pyrazole-4-yl]carbamoyl]pyrazolo[1,5-a]pyrimidine-5-yl]-5-fluoro-3-piperidyl]carbamate (9) 4-[3-methyl-4-[[5-[(3R,5R)-3-(tert-butoxycarbonylamino)-5-fluoro-1-piperidyl]pyrazolo[1,5-a]pyrimidine-3-carbonyl]amino]pyrazole-1-yl]cyclohexanecarboxylic acid 8 (250 mg, 0.43 mmol) and 5-(4-amino-1-piperidyl)-2-(2,6-dioxo-3-piperidyl)isoindoline-1,3-dione 2,2,2-trifluoroacetic acid C-3 (281.61 mg, 0.60 mmol) were dissolved in dry DMF (5 mL) and DIPEA (0.59 mL, 3.42 mmol) was added. After stirring at room temperature for 5 minutes, HATU (195.11 mg, 0.51 mmol) was added and the resulting mixture was stirred at room temperature. After 1 hour, complete conversion was shown by LC-MS. The mixture was purified by reverse-phase flash chromatography (150 g C18 RediSep Rf Gold column, retention solution (DMF), elution: 5% MeCN / 0.1% HCOOH over 5 CV, then 5 to 25% MeCN / 0.1% HCOOH over 5 CV, then 25 to 60% over 15 CV). The fractions were combined, concentrated, and lyophilized to obtain 9 (267 mg, 68% yield) as a yellow solid.

[0340] LC-MS method 1: 99.9% purity (at 215 nm), [M+H] + = 923.4. 1H NMR (400 MHz, DMSO-d6) δ ppm 1.34 (br s, 9 H), 1.40 - 1.49 (m, 2 H), 1.52 - 1.62 (m, 2 H), 1.63 - 1.73 (m, 2 H), 1.76 - 1.93 (m, 6 H), 1.98 - 2.06 (m, 2 H), 2.06 - 2.10 (m, 2 H), 2.10 - 2.18 (m, 2 H), 2.21 (s, 2 H), 2.25 - 2.30 (m, 1 H), 2.55 - 2.69 (m, 2 H), 2.82 - 2.97 (m, 2 H), 3.13 (br s, 2 H), 3.63 - 3.77 (m, 1 H), 3.81 - 3.93 (m, 1 H), 3.97 - 4.16 (m, 3 H), 5.06 (m, 2 H), 5.13 - 5.20 (m, 1 H), 6.89 - 6.98 (m, 1 H), 7.09 - 7.21 (m, 1 H), 7.23 - 7.29 (m, 1 H), 7.35 (br d, J = 1.5 Hz, 1 H), 7.68 (m, 1 H), 7.76 (br d, J = 8.1 Hz, 1 H), 8.03 (s, 1 H), 8.15 (s, 1 H), 8.23 ​​- 8.29 (m, 1 H), 8.82 (br d, J = 8.1 Hz, 1 H), 9.05 (br s, 1 H), 11.08 (s, 1 H). 19 F NMR (377 MHz, DMSO-d6) δ ppm -188.35 (s, 1 F).

[0341] Step 7. Preparation of N-[1-[4-[[1-[2-(2,6-dioxo-3-piperidyl)-1,3-dioxo-isoindoline-5-yl]-4-piperidyl]carbamoyl]cyclohexyl]-3-methyl-pyrazole-4-yl]-5-[(3R,5R)-3-amino-5-fluoro-1-piperidyl]pyrazolo[1,5-a]pyrimidine-3-carboxamide hydrochloride (P-27) tert-butyl N-[(3R,5R)-1-[3-[[1-[4-[[1-[2-(2,6-dioxo-3-piperidyl)-1,3-dioxo-isoindoline-5-yl]-4-piperidyl]carbamoyl]cyclohexyl]-3-methyl-pyrazole-4-yl]carbamoyl]pyrazolo[1,5-a]pyrimidine-5-yl]-5-fluoro-3-piperidyl]carbamate 9 (300 mg, 0.33 mmol) was dissolved in 4 M HCl / 1,4-dioxane (10 mL, 40 mmol). The solution was stirred at rt for 3 hours. The solvent was removed under reduced pressure, and the residue was purified by reverse-phase flash chromatography (150 g C18 RediSep Rf Gold column, holding solution (water), elution: 5% MeCN / 0.02 M HCl over 3 CV, then 5 to 30% MeCN / 0.02 M HCl over 15 CV). The fractions were combined, concentrated, and purified by preparative HPLC to obtain P-27 (83.97 mg, 31% yield) as a yellow solid, entirely as an HCl salt.

[0342] LC-MS method 5: 97.3% purity (at 215 nm), [M-HCl+2H] 2+ = 412.2;[M-HCl+H] + = 823.3;[M-HCl+3H] 3+ = 275.3. 1H NMR (400 MHz, DMSO-d6) δ ppm 1.37 - 1.49 (m, 2 H), 1.49 - 1.62 (m, 2 H), 1.63 - 1.76 (m, 2 H), 1.76 - 1.88 (m, 4 H), 1.90 - 2.08 (m, 4 H), 2.10 - 2.19 (m, 1 H), 2.23 (s, 3 H), 2.35 - 2.45 (m, 1 H), 2.54 - 2.64 (m, 2 H), 2.82 - 2.95 (m, 1 H), 3.12 (t, J = 11.5 Hz, 2 H), 3.25 - 3.60 (m, 4H), 3.94 - 4.12 (m, 4 H), 4.64 - 4.78 (m, 1 H), 5.03 - 5.25 (m, 2 H), 6.92 (d, J = 7.8 Hz, 1 H), 7.26 (dd, J = 8.7, 1.8 Hz, 1 H), 7.34 (d, J = 1.5 Hz, 1 H), 7.67 (d, J = 8.3 Hz, 1 H), 7.77 (d, J = 7.6 Hz, 1 H), 8.05 (s, 1 H), 8.19 - 8.43 (m, 4 H), 8.91 (d, J = 8.1 Hz, 1 H), 9.05 (s, 1 H), 11.08 (s, 1 H). 19 F NMR (377 MHz, DMSO-d6) δ ppm -184.87 (s, 1 F).

[0343] [Table 28]

[0344] Method 4 for final product Example S-9. Synthesis of P-29 [ka]

[0345] Step 1. Preparation of [4-(tert-butoxycarbonylamino)cyclohexyl]methanesulfonate (2) To a solution of tert-butyl N-(4-hydroxycyclohexyl)carbamate 1 (4.0 g, 18.6 mmol, 1 eq.) and Et3N (3.8.8 mL, 27.9 mmol, 1.5 eq.) in anhydrous DCM (93 mL), methanesulfonyl chloride (1.87 mL, 24.2 mmol, 1.3 eq.) was added dropwise at 0°C. The resulting mixture was stirred at 0°C. After 1 hour, complete conversion was shown by TLC (50% HCl / heptane, KMnO4 staining). Water was added, and the aqueous phase was extracted three times with DCM. The organic matter was washed once with 1N aqueous HCl solution, dried over Na2SO4, and concentrated to obtain 2 (5.45 g, quantitative yield) as a white solid.

[0346] LC-MS method 1: 99.9% purity (at 200 nm), [M+Na] + = 316.2. 1 H NMR (400 MHz, CDCl3) δ ppm 1.45 (s, 9 H), 1.51 - 1.64 (m, 2 H), 1.68 - 1.80 (m, 2 H), 1.80 - 1.90 (m, 2 H), 2.00 - 2.11 (m, 2 H), 3.02 (s, 3 H), 3.53 (br s, 1 H), 4.47 (br s, 1 H), 4.87 - 4.92 (m, 1 H).

[0347] Step 2. Preparation of tert-butyl N-[4-[3-(difluoromethyl)-4-nitropyrazole-1-yl]cyclohexyl]carbamate (4) 3-(difluoromethyl)-4-nitro-1H-pyrazole 3 (1.0 g, 6.13 mmol, 1 eq.) and [4-(tert-butoxycarbonylamino)cyclohexyl]methanesulfonate 2 (1.8 g, 6.13 mmol, 1 eq.) were dissolved in anhydrous DMF (20.4 mL) and Cs2CO3 (4.0 g, 12.3 mmol, 2 eq.) was added. The resulting mixture was stirred at 90°C. After 20 hours, LC-MS indicated that the conversion was not complete. Additional [4-(tert-butoxycarbonylamino)cyclohexyl]methanesulfonate 2 (0.9 g, 3.07 mmol, 0.5 eq.) was added to the reaction mixture, and the resulting mixture was stirred at 90°C. After 3 days, HPLC indicated that the conversion was not complete. An additional [4-(tert-butoxycarbonylamino)cyclohexyl]methanesulfonate 2 (1.35 g, 4.60 mmol, 0.75 eq.) was added to the reaction mixture, and the resulting mixture was stirred at 90°C. After 24 hours, HPLC indicated that the conversion was almost complete. Water was added to the reaction mixture, and the aqueous phase was extracted three times with RINKAN. The organic matter was washed three times with water, once with brine, dried over Na2SO4, and concentrated to dryness. The residue was then purified by normal-phase flash chromatography (120 g silica column, pre-adsorbed, elution: 0 to 40% MTBE / heptane over 10 CV, then 40 to 80% MTBE / heptane over 10 CV, then 80 to 100% MTBE / heptane over 10 CV). The fractions were combined and concentrated to obtain the impurity product. The crude product was then purified by reverse-phase chromatography (80 g C18 RediSep Rf Gold column, holding solution (DMSO), elution: 5% MeOH / 0.1% HCOOH over 3 CV, then 5 to 55% MeOH / 0.1% HCOOH over 0.5 CV, then 55 to 90% MeOH / 0.1% HCOOH over 15 CV). The fractions were combined and concentrated to obtain 4 (583 mg, 26% yield) as a white solid.

[0348] LC-MS method 1: 99.9% purity (at 215 nm), [M-tBu+H] + = 205.2. 1 H NMR (400 MHz, DMSO-d6) δ ppm 1.27 - 1.38 (m, 3 H), 1.38 (s, 9 H), 1.75 - 1.95 (m, 4 H), 2.04-2.07 (m, 2 H), 4.23 - 4.33 (m, 1 H), 6.83 (br d, J = 7.3 Hz, 1 H), 7.30 (t, J = 53.1 Hz, 1 H), 9.05 (s, 1 H). 19 F NMR (377 MHz, DMSO-d6) δ ppm -117.36 (s, 2 F).

[0349] Step 3. Preparation of 4-[3-(difluoromethyl)-4-nitropyrazole-1-yl]cyclohexaneamine (5) To a solution of tert-butyl N-[4-[3-(difluoromethyl)-4-nitropyrazole-1-yl]cyclohexyl]carbamate 4 (200 mg, 0.560 mmol, 1 eq.), 4 M HCl / 1,4-dioxane (4.16 mL, 16.7 mmol, 30 eq.) was added. The resulting mixture was stirred at room temperature. After 2.5 hours, complete conversion was shown by LC-MS. The solvent was evaporated under reduced pressure, and the crude product was co-evaporated twice with MeCN. The crude mixture was purified by reverse-phase flash chromatography (50 g C18 RediSep Rf Gold column, holding solution (H2O), elution: 5% MeOH / buffer solution (pH 10) over 10 CV, then 5% MeOH / H2O over 5 CV, then 5 to 100% MeOH / H2O over 12 CV). The fractions were combined and concentrated to obtain 5 (125 mg, 87% yield) as a colorless semi-solid.

[0350] LC-MS method 1: 99.9% purity (at 215 nm), [M+H] + = 261.4. 1 H NMR (400 MHz, CDCl3) δ ppm 1.27 - 1.42 (m, 4 H), 1.85 (qd, J = 12.7, 3.4 Hz, 2 H), 2.03 - 2.12 (m, 2 H), 2.20 - 2.30 (m, 2 H), 2.79 - 2.89 (m, 1 H), 4.19 (tt, J = 12.0, 3.9 Hz, 1 H), 7.12 (t, J = 53.3 Hz, 1 H), 8.21 (s, 1 H). 19 F NMR (377 MHz, CDCl3) δ ppm -117.68 (s, 2 F).

[0351] Step 4. Preparation of 1-(4-aminocyclohexyl)-3-(difluoromethyl)pyrazole-4-amine (6) A solution of 4-[3-(difluoromethyl)-4-nitropyrazole-1-yl]cyclohexaneamine 5 (150 mg, 0.580 mmol, 1 eq.) / ethyl acetate (5.8 mL) was degassed with nitrogen for 15 minutes. Then, Pd / C 10 wt% (184 mg, 0.170 mmol, 0.3 eq.) was added, and the solution was degassed for another 15 minutes. Finally, the solution was degassed with hydrogen for 15 minutes, and the mixture was stirred under hydrogen (1 atm) at room temperature. After 4 hours, complete conversion was shown by LC-MS. The solution was filtered through a Celite pad and rinsed with siRNA / MeCN. The solvent was evaporated under reduced pressure to obtain 6 (95 mg, 72% yield) as a gray oil.

[0352] LC-MS method 1: 99.9% purity (at 254 nm), [M+H] + = 231.4. 1H NMR (400 MHz, DMSO-d6) δ ppm 1.10 - 1.23 (m, 2 H), 1.60 - 1.74 (m, 2 H), 1.78 - 1.95 (m, 4 H), 2.58 (ddt, J = 11.0, 7.2, 3.7 Hz, 1 H), 3.95 (ddt, J = 11.7, 7.9, 3.8 Hz, 1 H), 4.02 (br s, 2 H), 6.86 (t, J = 54.0 Hz, 1 H), 7.13 (s, 1 H). 19 F NMR (377 MHz, CDCl3) δ ppm -111.24 (d, J = 54.5 Hz, 2 F).

[0353] Step 5. Preparation of N-[4-[4-amino-3-(difluoromethyl)pyrazole-1-yl]cyclohexyl]-1-[2-(2,6-dioxo-3-piperidyl)-1,3-dioxo-isoindorin-5-yl]piperidine-4-carboxamide (7) 1-[2-(2,6-dioxo-3-piperidyl)-1,3-dioxo-isoindorin-5-yl]piperidine-4-carboxylic acid C-9 (100 mg, 0.260 mmol, 1 eq.) and 1-(4-aminocyclohexyl)-3-(difluoromethyl)pyrazole-4-amine 6 (101.6 mg, 0.44 mmol, 1.7 eq.) were dissolved in anhydrous DMF (2.5 mL) to which DIPEA (135 μL, 0.780 mmol, 3 eq.) was added at room temperature. After 5 minutes, HATU (118.4 mg, 0.310 mmol, 1.2 eq.) was added, and the resulting mixture was stirred at room temperature. After 1.5 hours, complete conversion was shown by LC-MS. The crude mixture was directly purified by reverse-phase flash chromatography (100 g C18 gold column, retention solution (DMF), elution: 5% MeCN / 0.1% HCOOH over 5 CV, then 5 to 25% MeCN / 0.1% HCOOH over 15 CV, then 25 to 35% MeCN / 0.1% HCOOH over 10 CV). The fractions were combined and concentrated to obtain 7 (105 mg, 68% yield) as a yellow solid.

[0354] LC-MS method 1: 99.9% purity (at 215 nm), [M+H] + = 598.4. 1H NMR (400 MHz, DMSO-d6) δ ppm 1.26 - 1.39 (m, 2 H), 1.53 - 1.67 (m, 2 H), 1.68 - 1.78 (m, 3 H), 1.81 - 1.90 (m, 2 H), 1.91 - 2.06 (m, 3 H), 2.35 - 2.44 (m, 1 H), 2.53 - 2.58 (m, 1 H), 1.59 - 2.62 (m, 1 H), 2.81 - 2.93 (m, 1 H), 2.93 - 3.04 (m, 2 H), 3.51 - 3.64 (m, 1 H), 3.93 - 4.13 (m, 4 H), 5.06 (dd, J = 13.0, 5.4 Hz, 1 H), 6.87 (t, J = 54.0 Hz, 1 H), 7.16 (s, 1 H), 7.25 (dd, J = 9.0, 2.0 Hz, 1 H), 7.33 (d, J = 2.0 Hz, 1 H), 7.66 (d, J = 8.6 Hz, 1 H), 7.76 (br d, J = 7.8 Hz, 1 H), 11.07 (s, 1 H). 19 F NMR (377 MHz, DMSO-d6) δ ppm -111.30 (s, 2 F).

[0355] Step 6. Preparation of tert-butyl N-[(3R,5R)-1-[3-[[3-(difluoromethyl)-1-[4-[[1-[2-(2,6-dioxo-3-piperidyl)-1,3-dioxo-isoindoline-5-yl]piperidine-4-carbonyl]amino]cyclohexyl]pyrazole-4-yl]carbamoyl]pyrazolo[1,5-a]pyrimidine-5-yl]-5-fluoro-3-piperidyl]carbamate (9) N-[4-[4-amino-3-(difluoromethyl)pyrazole-1-yl]cyclohexyl]-1-[2-(2,6-dioxo-3-piperidyl)-1,3-dioxo-isoindorin-5-yl]piperidine-4-carboxamide 7 (100 mg, 0.170 mmol, 1 eq.) and 5-[(3R,5R)-3-(tert-butoxycarbonylamino)-5-fluoro-1-piperidyl]pyrazolo[1,5-a]pyrimidine-3-carboxylic acid 8 [Synthesized in General Method of Final Product 1; P-13] (63.5 mg, 0.170 mmol, 1 eq.) are dissolved in MeCN (1.7 mL) and NMI (0.04 mL, 0.50 mmol, 3 eq.) and TCFH (70.4 mg, 0.25 mmol, 1.5 eq.) was added at 0°C. The mixture was stirred at room temperature. After 20 hours, complete conversion was shown by LC-MS. MeCN was removed under reduced pressure, and the residue was purified by reverse-phase flash chromatography (50 g C18 RediSep Rf Gold column, holding solution (DMSO), elution: 5% MeCN / 0.1% HCOOH over 5 CV, then 5 to 40% MeCN / 0.1% HCOOH over 1 CV, then 40 to 70% MeCN / 0.1% HCOOH over 15 CV). The fractions were combined and concentrated to obtain 9 (84.3 mg, 52% yield) as a yellow solid.

[0356] LC-MS method 1: 99.9% purity (at 215 nm), [M+H] + = 959.4. 1H NMR (400 MHz, DMSO-d6) δ ppm 1.31 - 1.44 (m, 2 H), 1.54 - 1.69 (m, 2 H), 1.70 - 1.95 (m, 7 H), 1.97 - 2.06 (m, 3 H), 2.09 - 2.19 (m, 1 H), 2.35 - 2.46 (m, 2 H), 2.55 - 2.62 (m, 1 H), 2.81 - 2.94 (m, 2 H), 2.97 - 3.03 (m, 3 H), 3.35 - 3.54 (m, 2 H), 3.55 - 3.74 (m, 3 H), 4.07 (br d, J = 12.7 Hz, 2 H), 4.17 - 4.29 (m, 1 H), 4.96 - 5.11 (m, 2 H), 6.84 - 6.95 (m, 1 H), 7.05 - 7.15 (m, 1 H), 7.25 (br d, J = 8.8 Hz, 1 H), 7.33 (br s, 1 H), 7.66 (d, J = 8.6 Hz, 1 H), 7.79 (d, J = 7.8 Hz, 1 H), 8.29 (s, 1 H), 8.36 (br s, 1 H), 8.82 (d, J = 7.8 Hz, 1 H), 9.31 (br s, 1 H), 11.07 (s, 1 H). 19 F NMR (377 MHz, DMSO-d6) δ ppm -111.80 - -110.41 (m, 2 F).

[0357] Step 7. Manufacturing of 5-[(3R,5R)-3-amino-5-fluoro-1-piperidyl]-N-[3-(difluoromethyl)-1-[4-[[1-[2-(2,6-dioxo-3-piperidyl)-1,3-dioxo-isoindoline-5-yl]piperidine-4-carbonyl]amino]cyclohexyl]pyrazole-4-yl]pyrazolo[1,5-a]pyrimidine-3-carboxamide hydrochloride (P-29) tert-butyl N-[(3R,5R)-1-[3-[[3-(difluoromethyl)-1-[4-[[1-[2-(2,6-dioxo-3-piperidyl)-1,3-dioxo-isoindoline-5-yl]piperidine-4-carbonyl]amino]cyclohexyl]pyrazole-4-yl]carbamoyl]pyrazolo[1,5-a]pyrimidine-5-yl]-5-fluoro-3-piperidyl]carbamate 9 (80 mg, 0.080 mmol, 1 eq.) was mixed with 4 M HCl / 1,4-dioxane (3.13 mL, 12.5 mmol, 150 eq.). The mixture was sonicated and stirred at room temperature. After 20 hours, complete conversion was shown by HPLC. The solvent was removed under reduced pressure, and the residue was purified by reverse-phase flash chromatography (30 g C18 RediSep Rf Gold column, holding solution (DMSO), elution: 5% MeCN / 0.02 M HCl over 5 CV, then 5 to 100% MeCN / 0.02 M HCl over 10 CV). The fractions were combined, concentrated, and lyophilized to obtain P-29 (30.60 mg, 44% yield), entirely as hydrochloride, as a yellow solid.

[0358] LC-MS method 5: 99.4% purity (at 215 nm), [M-HCl+H]+ = 859.4. 1H NMR (400 MHz, DMSO-d6) δ ppm 1.31 - 1.44 (m, 2 H), 1.55 - 1.68 (m, 2 H), 1.69 - 1.94 (m, 7 H), 1.94 - 2.10 (m, 4 H), 2.37 - 2.44 (m, 1 H), 2.53 - 2.63 (m, 2 H), 2.83 - 2.94 (m, 1 H), 2.94 - 3.05 (m, 2 H), 3.22 - 3.28 (m, 1 H), 3.37 - 3.53 (m, 2 H), 3.55 - 3.67 (m, 1 H), 4.02 - 4.12 (m, 2 H), 4.20 - 4.30 (m, 1 H), 4.52 - 4.93 (m, 2 H), 5.02 - 5.19 (m, 2 H), 6.88 (d, J = 7.8 Hz, 1 H), 6.99 - 7.28 (m, 2 H), 7.33 (d, J = 2.0 Hz, 1 H), 7.67 (d, J = 8.6 Hz, 1 H), 7.80 (d, J = 7.3 Hz, 1 H), 8.14 (br s, 3 H), 8.33 (s, 1 H), 8.40 (s, 1 H), 8.93 (d, J = 7.8 Hz, 1 H), 9.33 (s, 1 H), 11.07 (s, 1 H). 19 F NMR (377 MHz, DMSO-d6) δ ppm -184.79 (s, 1 F), -111.25 (d, J = 16.3 Hz, 2 F).

[0359] Table 29

[0360] Method 5 of the final product Example S-10. Synthesis of P-30

change

[0361] Step 1. Preparation of [4-[3-(difluoromethyl)-4-nitropyrazole-1-yl]cyclohexyl]methanol (2) To a 1:2 THF (3.8 mL) / ethanol (7.5 mL) solution of methyl 4-[3-(difluoromethyl)-4-nitropyrazole-1-yl]cyclohexanecarboxylate 1 (730.0 mg, 2.26 mmol, 1 eq.) at 0°C, CaCl2 (501.2 mg, 4.52 mmol, 2 eq.) was added, followed by NaBH4 (341.6 mg, 9.03 mmol, 4 eq.). The resulting mixture was stirred at room temperature. After 16 hours at room temperature, LC-MS indicated that the reaction was complete. The reaction was quenched by dropwise addition of water, and the resulting mixture was stirred at room temperature for 1 hour. The product was then extracted three times with ELISA, the organic layers were washed once with water and once with brine, and finally dried over MgSO4. Depositphotos was removed under reduced pressure, and the residue was purified by normal-phase flash chromatography [80 g silica column, retention solution (DCM), elution: 0% Depositphotos over 3 CV, then 0 to 100% Depositphotos over 10 CV, then 100% Depositphotos over 3 CV (the target product eluted at around 50% Depositphotos)]. The fractions were combined and concentrated to obtain 2 (371.9 mg, 58% yield) as a colorless oil. LC-MS method 1: 97.5% purity (at 215 nm), [M+H] + = 276.2. 1 H NMR (400 MHz, CDCl3-d) δ ppm 1.14 - 1.30 (m, 3 H), 1.70 - 1.87 (m, 2 H), 1.97 - 2.09 (m, 2 H), 2.25 - 2.34 (m, 2 H), 3.54 (d, J = 6.1 Hz, 2 H), 4.18 (tt, J = 12.1, 3.9 Hz, 1 H), 7.10 (t, J = 53.3 Hz, 1 H), 8.22 (s, 1 H). 19F NMR (377 MHz, CDCl3-d) δ ppm -117.64 (d, J = 53.13, 2 F).

[0362] Step 2. Preparation of [4-[3-(difluoromethyl)-4-nitropyrazole-1-yl]cyclohexyl]methylmethanesulfonate (3) [4-[3-(difluoromethyl)-4-nitropyrazole-1-yl]cyclohexyl]methanol 2 (428.5 mg, 1.52 mmol) and Et3N (0.28 mL, 1.98 mmol) in DCM (7.62 mL) were mixed dropwise with MsCl (0.13 mL, 1.68 mmol) at 0°C. The reaction mixture was then slowly warmed to room temperature and stirred. After 3 hours, TLC (3:7 heptane / siRNA) indicated completion of the reaction. The reaction mixture was then partitioned with water and siRNA. The phases were separated, and the organic phase was washed three times with water, once with 1 N hydrochloric acid, and once with brine, and then dried over MgSO4. siRNA was removed under reduced pressure to obtain 3 (529 mg, 93% yield) as a pale yellow oil. The product was used in the next step without further purification. LC-MS method 1: 94.8% purity (at 215 nm), [M+H] + = 354.0. 1 H NMR (400 MHz, CDCl3-d) δ ppm 1.22 - 1.37 (m, 3 H), 1.80 - 1.90 (m, 2 H), 2.05 - 2.13 (m, 2 H), 2.27 - 2.36 (m, 2 H), 3.04 (s, 3 H), 4.08 - 4.14 (m, 2 H), 4.15 - 4.24 (m, 1 H), 7.11 (t, J = 53.4 Hz, 1 H), 8.22 (s, 1 H). 19 F NMR (377 MHz, CDCl3-d) δ ppm -117.64 (d, J = 53.13, 2 F).

[0363] Step 3. Preparation of 2-[4-[3-(difluoromethyl)-4-nitropyrazole-1-yl]cyclohexyl]acetaldehyde (4) A solution of [4-[3-(difluoromethyl)-4-nitropyrazole-1-yl]cyclohexyl]methylmethanesulfonate 3 (529.0 mg, 1.5 mmol, 1 eq.) and NaCN (183.4 mg, 3.74 mmol, 2.5 eq.) in DMSO (7.49 mL) was heated to 50°C and stirred. After 16 hours at 50°C, LC-MS indicated that the reaction was complete. The reaction mixture was cooled to rt and quenched with water while vigorously stirring. Then, ethyl acetate was added and the product was extracted twice with ethyl acetate. The organic layers were washed once with concentrated NaHCO3 aqueous solution and once with saline. Finally, the solution of the product was dried over MgSO4 to remove ethyl acetate under reduced pressure, and 4 (342 mg, 79% yield) was obtained as a yellow oil.

[0364] LC-MS method 1: 98.8% purity (at 215 nm), [M+H] + = 285.2. 1 H NMR (400 MHz, CDCl3-d) δ ppm 1.30 - 1.46 (m, 2 H), 1.76 - 1.96 (m, 3 H), 2.06 - 2.16 (m, 2 H), 2.29 - 2.37 (m, 2 H), 2.37 - 2.40 (m, 2 H), 4.20 (tt, J = 12.0, 3.9 Hz, 1 H), 7.12 (t, J = 54.2 Hz, 1 H), 8.22 (s, 1 H). 19 F NMR (377 MHz, CDCl3-d) δ ppm -117.68 (s, 2 F).

[0365] Step 4. Preparation of 2-[4-[3-(difluoromethyl)-4-nitropyrazole-1-yl]cyclohexyl]acetaldehyde (5) To a solution of 2-[4-[3-(difluoromethyl)-4-nitropyrazole-1-yl]cyclohexyl]acetonitrile 4 (342 mg, 1.18 mmol, 1 eq.) in DCM (5.88 mL), a solution of 1 M DIBAL-H / DCM (3.53 mL, 3.53 mmol, 3 eq.) was added at -78°C. The reaction mixture was then stirred at the same temperature. After 2 hours, LC-MS indicated that the reaction was complete. The reaction was slowly quenched with Rochelle salt solution at -78°C and stirred at rt for 1 hour. The product was then extracted three times with siRNA, and the combined organic layers were washed twice with 1 M hydrochloric acid and once with brine. Finally, the solution of the product was dried over MgSO4, and siRNA was removed under reduced pressure to obtain 5 (264 mg, 78% yield) as a yellow oil.

[0366] LC-MS method 1: 99.9% purity (at 215 nm), [M+H] + = 288.2. 1 H NMR (400 MHz, DMSO-d6 ) δ ppm 1.36 (q, J = 6.5 Hz, 2 H), 1.75 (qd, J = 12.5, 3.2 Hz, 2 H), 1.85 (d, J = 12.5 Hz, 2 H), 2.02 - 2.15 (m, 2 H), 3.40 - 3.50 (m, 2 H), 4.19 - 4.32 (m, 1 H), 4.32 - 4.40 (m, 1 H), 7.30 (t, J = 52.3 Hz, 1 H), 9.06 (s, 1 H). 19 F NMR (377 MHz, DMSO-d6) δ ppm -117.36 (s, 2 F).

[0367] Step 5. Production of 2-[4-[3-(difluoromethyl)-4-nitropyrazole-1-yl]cyclohexyl]ethanol (6) To a solution of 2-[4-[3-(difluoromethyl)-4-nitropyrazole-1-yl]cyclohexyl]acetaldehyde 5 (264 mg, 0.92 mmol, 1 eq.) in methanol (4.59 mL), NaBH4 (69.52 mg, 1.84 mmol) was added at 0°C. The resulting mixture was stirred at room temperature. After 16 hours, LC-MS indicated that the reaction was complete. Water was added, and the resulting mixture was stirred at room temperature for 1 hour. The product was extracted three times with siRNA, and the combined organic layers were washed twice with brine. Finally, the solution of the product was dried over MgSO4, and siRNA was removed under reduced pressure to obtain 6 (216 mg, 68% yield) as a yellow oil.

[0368] LC-MS method 1: 83.3% purity (at 215 nm), [M+H] + = 290.2. 1 H NMR (400 MHz, DMSO-d6 ) δ ppm 1.03 - 1.15 (m, 2 H), 1.36 (q, J = 6.5 Hz, 2 H), 1.41 - 1.52 (m, 1 H), 1.69 - 1.81 (m, 2 H), 1.85 (br d, J = 12.5 Hz, 2 H), 2.02 - 2.12 (m, 2 H), 3.37 - 3.52 (m, 2 H), 4.22 - 4.32 (m, 1 H), 4.33 - 4.39 (m, 1 H), 7.30 (t, J = 53.2 Hz, 1 H), 9.06 (s, 1 H). 19 F NMR (377 MHz, DMSO-d6) δ ppm -117.36 (s, 2 F).

[0369] Step 6. Production of 2-[4-[4-amino-3-(difluoromethyl)pyrazole-1-yl]cyclohexyl]ethanol (7) 2-[4-[3-(difluoromethyl)-4-nitropyrazole-1-yl]cyclohexyl]ethanol 6 (216.1 mg, 0.75 mmol, 1 eq.) was dissolved in siRNA (3.74 mL) and bubbling with N2 for 5 minutes. Then, 10% Pd / C (238.5 mg, 0.22 mmol, 0.3 eq.) was added, and bubbling was carried out again for 5 minutes with N2. Then, bubbling was carried out for 5 minutes with H2, and the resulting mixture was stirred at room temperature under 1 atm of H2. After 3 hours, LC-MS indicated that the reaction was complete. The solution was filtered through a Celite pad and thoroughly washed with siRNA. Finally, the filtrate was concentrated under reduced pressure to obtain 7 (169 mg, 81% yield) as a yellow oil. This product was used in the next step without purification. LC-MS method 1: 92.9% purity (at 215 nm), [M+H] + = 260.2.

[0370] Step 7. Preparation of tert-butyl N-[(3R,5R)-1-[3-[[3-(difluoromethyl)-1-[4-(2-hydroxyethyl)cyclohexyl]pyrazole-4-yl]carbamoyl]pyrazolo[1,5-a]pyrimidine-5-yl]-5-fluoro-3-piperidyl]carbamate (9) TCFH (168.6 mg, 0.60 mmol, 1.2 eq.) was added at 0°C to a solution of 2-[4-[4-amino-3-(difluoromethyl)pyrazole-1-yl]cyclohexyl]ethanol 7 (168.8 mg, 0.65 mmol, 1.3 eq.), 5-[(3R,5R)-3-(tert-butoxycarbonylamino)-5-fluoro-1-piperidyl]pyrazolo[1,5-a]pyrimidine-3-carboxylic acid 8 [synthesized in general method 1; P-13] (190 mg, 0.50 mmol, 1 eq.), and NMI (0.14 mL, 1.75 mmol, 3.5 eq.) in MeCN (2.50 mL). The reaction mixture was then stirred at room temperature. After 16 hours, LC-MS indicated that the reaction was complete. MeCN was removed under reduced pressure, and the residue was purified by reverse-phase flash chromatography [30 g C18 column, holding solution (DMSO), elution: 5% MeOH / 0.1% HCOOH over 3 CV, then 5-100% MeOH / 0.1% HCOOH over 15 CV, then 100% MeOH / 0.1% HCOOH over 3 CV (the target product eluted around 80% MeCN)]. The fractions were combined and concentrated to obtain 9 (155.6 mg, 50% yield) as an off-white solid.

[0371] LC-MS method 1: 99.9% purity (at 215 nm), [M+H] + = 621.2. 1H NMR (400 MHz, DMSO-d6 ) δ ppm 1.04 - 1.17 (m, 3 H), 1.29 - 1.44 (m, 12 H), 1.65 - 1.77 (m, 3 H), 1.80 - 1.93 (m, 3 H), 1.97 - 2.07 (m, 2 H), 2.10 - 2.18 (m, 1 H), 2.94 - 3.14 (m, 1 H), 3.36 - 3.53 (m, 3 H), 3.61 - 3.72 (m, 1 H), 4.11 - 4.23 (m, 1 H), 4.31 - 4.40 (m, 1 H), 4.94 - 5.13 (m, 1 H), 6.83 - 6.90 (m, 1 H), 6.91 - 7.23 (m, 2 H), 8.28 (s, 1 H), 8.33 (br s, 1 H), 8.82 (d, J = 8.1 Hz, 1 H), 9.31 (br s, 1 H). 19 F NMR (377 MHz, DMSO-d6) δ ppm -184.27 - -183.87 (m, 1 F), -112.01 - -110.10 (m, 2 F).

[0372] Step 8. Preparation of tert-butyl N-[(3R,5R)-1-[3-[[3-(difluoromethyl)-1-[4-[2-[4-[2-(2,6-dioxo-3-piperidyl)-1-oxo-isoindoline-5-yl]piperazine-1-yl]ethyl]cyclohexyl]pyrazole-4-yl]carbamoyl]pyrazolo[1,5-a]pyrimidine-5-yl]-5-fluoro-3-piperidyl]carbamate (10) IBX (44.9 mg, 0.16 mmol, 1.2 eq.) was added to a solution of tert-butyl N-[(3R,5R)-1-[3-[[3-(difluoromethyl)-1-[4-(2-hydroxyethyl)cyclohexyl]pyrazole-4-yl]carbamoyl]pyrazolo[1,5-a]pyrimidine-5-yl]-5-fluoro-3-piperidyl]carbamate 9 (81 mg, 0.13 mmol, 1 eq.) in dried DMSO (1 mL, 0.13 M). The resulting mixture was stirred overnight at room temperature. After overnight stirring, LC-MS analysis showed complete conversion to the target aldehyde. Subsequently, 3-(1-oxo-5-piperazine-1-ylisoindorin-2-yl)piperidine-2,6-dione hydrochloride C-4 (52.5 mg, 0.14 mmol, 1.1 eq.), DCE (1 mL, 0.13 M), and DIPEA (0.23 mL, 1.31 mmol, 10 eq.) were added to the reaction mixture. The mixture was stirred at rt for 10 minutes, and then sodium triacetoxyborohydride (36.1 mg, 0.17 mmol, 1.3 eq.) was added. After 90 minutes, complete conversion to compound 10 was confirmed by LC-MS. DCE was evaporated, and the reaction mixture was purified by reverse-phase FC (50 g C18 RediSep Rf Gold column, holding solution (DMSO), 5% MeCN / 0.1% HCOOH over 4 CV, then 5-95% MeCN / 0.1% HCOOH over 20 CV). The pure fractions were combined and concentrated to obtain 10 (73 mg, 48% yield) as a brown solid. The target product contained IBX residue.

[0373] LC-MS method 2: 79.9% purity (at 215 nm), [Mt-Bu+2H] 2+ = 438.2. 1H NMR (400 MHz, DMSO-d6) δ ppm 1.07 - 1.22 (m, 3 H), 1.33 (s, 9 H), 1.39 - 1.47 (m, 3 H), 1.67 - 1.82 (m, 3 H), 1.84 - 1.93 (m, 3 H), 1.93 - 1.99 (m, 1 H), 2.03 (br d, J = 11.2 Hz, 2 H), 2.09 - 2.18 (m, 1 H), 2.34 - 2.43 (m, 3 H), 2.53 (br s, 2 H), 2.55 - 2.63 (m, 1 H), 2.84 - 2.96 (m, 1 H), 2.99 - 3.14 (m, 1 H), 3.37 - 3.52 (m, 4 H), 3.66 (br s, 1 H), 4.14 - 4.24 (m, 2 H), 4.29 - 4.36 (m, 1 H), 4.50 - 4.78 (m, 1 H), 4.93 - 5.12 (m, 2 H), 6.86 - 6.93 (m, 1 H), 7.07 (s, 2 H), 7.18 - 7.48 (m, J = 1.2 Hz, 1 H), 7.52 (d, J = 8.8 Hz, 1 H), 8.14 (s, 2 H), 8.28 (s, 1 H), 8.34 (br s, 1 H), 8.82 (d, J = 7.8 Hz, 1 H), 9.31 (s, 1 H), 10.94 (s, 1 H). 19 F NMR (377 MHz, DMSO-d6) δ ppm -184.78 (br s, 1 F), -111.22 (br dd, J = 53.1, 19.1 Hz, 2 F).

[0374] Step 9. Preparation of N-[3-(difluoromethyl)-1-[4-[2-[4-[2-(2,6-dioxo-3-piperidyl)-1-oxo-isoindoline-5-yl]piperazine-1-yl]ethyl]cyclohexyl]pyrazole-4-yl]-5-[(3R,5R)-3-amino-5-fluoro-1-piperidyl]pyrazolo[1,5-a]pyrimidine-3-carboxamide dihydrochloride (P-30) In a round-bottom flask, tert-butyl N-[(3R,5R)-1-[3-[[3-(difluoromethyl)-1-[4-[2-[4-[2-(2,6-dioxo-3-piperidyl)-1-oxo-isoindolin-5-yl]piperazine-1-yl]ethyl]cyclohexyl]pyrazole-4-yl]carbamoyl]pyrazolo[1,5-a]pyrimidine-5-yl]-5-fluoro-3-piperidyl]carbamate 10 (73 mg, 0.08 mmol, 1.0 eq.) and 4.0 M HCl / dioxane (2.43 mL, 9.72 mmol, 124 eq.) were added. The reaction mixture was stirred at room temperature. After 18 hours, complete conversion was shown by LC-MS. The reaction mixture was concentrated under vacuum, and the residue was purified by reverse-phase FC (50 g C18 RediSep Rf Gold column, holding solution (H2O), elution: 5% MeCN / 0.02 M HCl over 4 CV, then 5% to 30% MeCN / 0.02 M HCl over 20 CV). The pure fractions were combined, concentrated, and lyophilized to obtain P-30 (24.2 mg, 37% yield) as a white solid dihydrochloride.

[0375] LC-MS method 3: 99.1% purity (at 215 nm), [M-2HCl+H]+ = 831.3. 1H NMR (400 MHz, DMSO-d6) δ ppm 1.11 - 1.27 (m, 2 H), 1.42 (br s, 1 H), 1.62 - 1.81 (m, 4 H), 1.84 - 1.92 (m, 2 H), 1.93 - 2.10 (m, 4 H), 2.35 - 2.43 (m, 2 H), 2.59 (br d, J = 16.6 Hz, 1 H), 2.84 - 2.98 (m, 1 H), 3.04 - 3.48 (m, 11 H), 4.02 (br d, J = 11.7 Hz, 2 H), 4.19 - 4.28 (m, 2 H), 4.36 (d, J = 15.4 Hz, 1 H), 4.58 (br s, 1 H), 4.87 (br s, 1 H), 5.02 - 5.18 (m, 2 H), 6.88 (br d, J = 7.6 Hz, 1 H), 6.98 - 7.28 (m, 3 H), 7.60 (d, J = 8.6 Hz, 1 H), 8.20 (br s, 3 H), 8.32 (s, 1 H), 8.39 (s, 1 H), 8.94 (d, J = 7.8 Hz, 1 H), 9.33 (s, 1 H), 10.27 (br s, 1 H), 10.96 (s, 1 H). 19 F NMR (377 MHz, DMSO-d6) δ ppm -184.51 (br s, 1 F), -111.23 (br dd, J = 52.5, 18.4 Hz, 2 F).

[0376] Table 30

[0377] Method 6 for the final product Example S-11. Synthesis of P-38

change

[0378] Step 1. Preparation of benzyl 4-methylsulfonyloxypiperidine-1-carboxylate (2) Methanesulfonyl chloride (0.821 mL, 10.6 mmol, 1.25 eq.) was added dropwise to a 100 mL ice-cold DCM (Digital Cholesterol) solution of 4-hydroxypiperidine-1-carboxylate 1 (2.00 g, 8.50 mmol, 1.0 eq.) and triethylamine (1.78 mL, 12.8 mmol, 1.5 eq.). The reaction mixture was stirred at room temperature. After 1 hour, LC-MS indicated complete conversion. Water (50 mL) and saturated NaHCO3 (aq) (50 mL) were added to the reaction mixture to separate the phases, and the aqueous layer was extracted with DCM (3 × 50 mL). The combined organic layers were washed with saline solution (50 mL), then dried over MgSO4 and evaporated under reduced pressure to obtain 2 (2.80 g, 8.50 mmol, quantitative yield) as a pale orange oil.

[0379] LC-MS method 1: 95.8% purity (at 215 nm), [M+H] + = 314.1. 1 H NMR (400 MHz, CDCl3) δ ppm 1.77 - 1.92 (m, 2 H), 1.93 - 2.08 (m, 2 H), 3.05 (s, 3 H), 3.43 (ddd, J = 13.7, 7.9, 3.9 Hz, 2 H), 3.70 - 3.85 (m, 2 H), 4.91 (tt, J = 7.5, 3.8 Hz, 1 H), 5.14 (s, 2 H), 7.27 - 7.43 (m, 5 H).

[0380] Step 2. Preparation of benzyl 4-[3-(difluoromethyl)-4-nitropyrazole-1-yl]piperidine-1-carboxylate (4) A mixed solution of benzyl 4-methylsulfonyloxypiperidine-1-carboxylate 2 (1.00 g, 3.19 mmol, 1.2 eq.), 3-(difluoromethyl)-4-nitro-1H-pyrazole 3 (450 mg, 2.76 mmol, 1.0 eq.), and potassium carbonate (800 mg, 5.80 mmol, 2.1 eq.) in DMF (22 mL) was stirred at 100°C under a nitrogen atmosphere. After 24 hours, complete conversion was shown by LC-MS. Water (100 mL) was added to the reaction mixture, and the aqueous phase was extracted with siRNA (3 × 50 mL). The combined organic substances were washed with 1:1 water / saline solution (3 × 50 mL) and saline solution (50 mL), dried on MgSO4, and evaporated to dryness. The crude product was purified by normal-phase flash chromatography (80 g silica column, pre-adsorbed, eluted: heptane / siRNA, 95:5 to 70:30, 12 CV) to obtain 4 (644 mg, 1.46 mmol, 53% yield) as a pale yellow oily substance.

[0381] LC-MS method 1: 86.0% purity (at 215 nm), [M+H] + = 381.1. 1 H NMR (400 MHz, CDCl3) δ ppm 1.90 - 2.01 (m, 2 H), 2.22 (br d, J = 12.1 Hz, 2 H), 2.88 - 3.04 (m, 2 H), 4.37 (tt, J = 11.7, 4.0 Hz, 3 H), 5.16 (s, 2 H), 7.12 (t, J = 53.3 Hz, 1 H), 7.32 - 7.43 (m, 5 H), 8.21 (s, 1 H). 19 F NMR (377 MHz, CDCl3) δ ppm -117.79 (s, 2 F).

[0382] Step 3. Preparation of benzyl 4-[4-amino-3-(difluoromethyl)pyrazole-1-yl]piperidine-1-carboxylate (5) Zinc (2.21 g, 33.9 mmol, 20.0 eq.; activated with 1.0 M HCl (aqueous solution)) was added to a solution of benzyl 4-[3-(difluoromethyl)-4-nitropyrazole-1-yl]piperidine-1-carboxylate 4 (644 mg, 1.46 mmol, 1.0 eq.) / i-PrOH (17 mL). Acetic acid (0.970 mL, 16.9 mmol, 10.0 eq.) was then added to the solution, and the reaction mixture was stirred at room temperature. After 1 hour, complete conversion was shown by LC-MS. The reaction mixture was filtered through a Celite pad and evaporated to dryness. The residue was taken up with ethyl acetate (50 mL), washed with saturated NaHCO3 (aq) (10 mL) and saline solution (10 mL), dried on MgSO4, and evaporated to dryness to obtain 5 (500 mg, 1.24 mmol, 73% yield) as a brown oily substance.

[0383] LC-MS method 1: 86.9% purity (at 215 nm), [M+H] + = 351.1. 1 H NMR (400 MHz, DMSO-d6) δ ppm 1.73 (qd, J = 12.2, 4.5 Hz, 2 H), 1.91 - 2.01 (m, 2 H), 2.86 - 3.10 (m, 2 H), 4.08 (br d, J = 12.8 Hz, 4 H), 4.17 - 4.30 (m, 1 H), 5.09 (s, 2 H), 6.88 (t, J = 54.2 Hz, 1 H), 7.18 (s, 1 H), 7.31 - 7.39 (m, 5 H). 19 F NMR (377 MHz, DMSO-d6) δ ppm -111.51 (s, 2 F).

[0384] Step 4. Preparation of benzyl 4-[3-(difluoromethyl)-4-[[5-[(3R,5R)-3-(tert-butoxycarbonylamino)-5-fluoro-1-piperidyl]pyrazolo[1,5-a]pyrimidine-3-carbonyl]amino]pyrazole-1-yl]piperidine-1-carboxylate (7) TCFH (614 mg, 2.19 mmol, 1.2 eq.) was added in one step to a solution of 5-[(3R,5R)-3-(tert-butoxycarbonylamino)-5-fluoro-1-piperidyl]pyrazolo[1,5-a]pyrimidine-3-carboxylic acid 6 [synthesized in General Method 1; P-13] (692 mg, 1.82 mmol, 1.0 eq.), benzyl 4-[4-amino-3-(difluoromethyl)pyrazole-1-yl]piperidine-1-carboxylate 5 (708 mg, 1.92 mmol, 1.05 eq.) and NMI (506 μL, 6.38 mmol, 3.5 eq.) in MeCN (12.1 mL). The reaction mixture was then stirred at room temperature. After 24 hours, complete conversion was shown by LC-MS. Volatile components were evaporated under reduced pressure. The residue was purified by reverse-phase flash chromatography (150 g C18 RediSep Rf Gold column, holding solution (DMSO), MeOH / 0.1% HCOOH(aq), 40%→90%, 15 CV). The fractions were combined and concentrated to obtain 7 (823 mg, 1.10 mmol, 60% yield) as an off-white solid.

[0385] LC-MS method 1: 95.0% purity (at 215 nm), [M+H] + = 712.3. 1H NMR (400 MHz, DMSO-d6) δ ppm 1.33 (br s, 9 H), 1.68 - 1.97 (m, 4 H), 2.03 (br d, J = 10.6 Hz, 2 H), 2.09 - 2.22 (m, 1 H), 2.88 - 3.18 (m, 3 H), 3.36 - 3.56 (m, 1 H), 3.67 (br d, J = 7.0 Hz, 1 H), 4.12 (br d, J = 13.2 Hz, 2 H), 4.42 - 4.56 (m, 1 H), 4.57 - 4.92 (m, 1 H), 4.97 (d, J = 45.4 Hz, 1 H), 5.10 (s, 2 H), 6.89 (br d, J = 8.1 Hz, 1 H), 6.92 - 7.26 (m, 2 H), 7.29 - 7.36 (m, 1 H), 7.38 (s, 2 H), 7.39 (s, 2 H), 8.29 (s, 1 H), 8.38 (br s, 1 H), 8.82 (d, J = 7.8 Hz, 1 H), 9.31 (br s, 1 H). 19 F NMR (377 MHz, DMSO-d6) δ ppm -186.27 - -182.64 (m, 1 F), -114.40 - -109.21 (m, 2 F).

[0386] Step 5. Preparation of tert-butyl N-[(3R,5R)-1-[3-[[3-(difluoromethyl)-1-(4-piperidyl)pyrazole-4-yl]carbamoyl]pyrazolo[1,5-a]pyrimidine-5-yl]-5-fluoro-3-piperidyl]carbamate (8) Benzyl 4-[3-(difluoromethyl)-4-[[5-[(3R,5R)-3-(tert-butoxycarbonylamino)-5-fluoro-1-piperidyl]pyrazolo[1,5-a]pyrimidine-3-carbonyl]amino]pyrazole-1-yl]piperidine-1-carboxylate 7 (739 mg, 1.04 mmol, 1.0 eq.) was dissolved in methanol (4.0 mL), and this solution was sparged with nitrogen under sonication. After 20 minutes, Pd / C 10% w / w (200 mg, 0.181 mmol, 0.18 eq.) was added, and the mixture was further sparged with nitrogen under sonication. After 20 minutes, the nitrogen balloon was replaced with a hydrogen balloon, and the reaction mixture was sparged with aeration. After 10 minutes, the reaction mixture was stirred under a hydrogen atmosphere. After 18 hours, complete conversion was demonstrated by LC-MS. The reaction mixture was filtered through Celite, and the Celite was thoroughly washed with MeOH. The filtrate was evaporated under reduced pressure, and the residue was incorporated into siRNA. Heptane was added until a precipitate formed. The suspension was evaporated to dryness to obtain 8 (620 mg, 0.987 mmol, 95% yield) as a pink solid.

[0387] LC-MS method 1: 92.0% purity (at 215 nm), [M+H] + = 578.3. 1H NMR (400 MHz, DMSO-d6) δ ppm 0.81 - 0.87 (m, 1 H), 1.20 - 1.27 (m, 1 H), 1.34 (br s, 9 H), 1.69 - 1.82 (m, 3 H), 1.88 - 1.95 (m, 2 H), 2.07 - 2.23 (m, 1 H), 2.53 - 2.61 (m, 2 H), 3.03 (br d, J = 12.8 Hz, 3 H), 3.37 - 3.51 (m, 1 H), 3.67 (br d, J = 2.1 Hz, 1 H), 4.19 - 4.33 (m, 1 H), 4.45 - 4.87 (m, 1 H), 5.04 (d, J = 46.3 Hz, 1 H), 6.89 (br d, J = 8.1 Hz, 1 H), 6.92 - 7.33 (m, 2 H), 8.29 (s, 1 H), 8.33 (br s, 1 H), 8.82 (d, J = 7.8 Hz, 1 H), 9.32 (br s, 1 H). 19 F NMR (377 MHz, DMSO-d6) δ ppm -187.05 - -180.57 (m, 1 F), -113.62 - -108.43 (m, 2 F).

[0388] Step 6. Preparation of 3-[5-[4-(2,2-dimethoxyethyl)piperazin-1-yl]-1-oxo-isoindorin-2-yl]piperidine-2,6-dione (10) In a round-bottom flask, 3-(1-oxo-5-piperazine-1-ylisoindoline-2-yl)piperidine-2,6-dione hydrochloride C-4 (175 mg, 0.48 mmol, 1 eq.) / CH2Cl2 (6 mL, 0.08 M), followed by 2,2-dimethoxyacetaldehyde 9 (0.12 mL, 0.69 mmol, 1.44 eq.), DIPEA (0.5 mL, 3.25 mmol, 6.8 eq.), and NaBH(OAc)3 (180 mg, 0.85 mmol, 1.8 eq.) were added. The reaction mixture was stirred at room temperature. After 2 hours, LC-MS showed complete conversion to compound 10. The mixture was evaporated to dryness and incorporated into DCM and water. The aqueous phase was extracted with DCM (2x), the organic layer was combined with the aqueous layer, washed with brine, dried over sodium sulfate, filtered, concentrated, and dried to obtain 10 (200 mg, 94% yield) as a yellow solid.

[0389] LC-MS method 2: 93.7% purity (at 215 nm), [M+H] + = 417.2. 1 H NMR (400 MHz, chloroform-d) δ ppm 1.52 - 1.71 (m, 6 H), 2.18 - 2.28 (m, 1 H), 2.29 - 2.43 (m, 1 H), 2.79 - 2.96 (m, 3 H), 3.41 - 3.53 (m, 7 H), 3.61 - 3.82 (m, 3 H), 4.24 - 4.30 (m, 1 H), 4.40 - 4.46 (m, 1 H), 5.20 (dd, J = 13.1, 5.0 Hz, 1 H), 6.90 (s, 1 H), 7.00 (dd, J = 8.6, 2.2 Hz, 1 H), 7.76 (br d, J = 8.1 Hz, 1 H), 7.91 (s, 1 H).

[0390] Step 7. Preparation of 3-[5-[4-(2,2-dihydroxyethyl)piperazin-1-yl]-1-oxo-isoindorin-2-yl]piperidine-2,6-dione (11) 3-[5-[4-(2,2-dimethoxyethyl)piperazin-1-yl]-1-oxo-isoindorin-2-yl]piperidine-2,6-dione 10 (200 mg, 0.45 mmol, 1 eq.) was added to 4 M HCl / dioxane (2 mL, 8 mmol, 18 eq.) and a few drops of water. The reaction mixture was stirred at room temperature. After overnight, LC-MS showed complete conversion to compound 11. The solvent was co-evaporated with MeCN(3x) to obtain 11 (200 mg, quantitative yield) as a brown solid.

[0391] LC-MS method 2: 96.7% purity (at 254 nm), [M+H2O+H] + = 389.1. 1 H NMR (400 MHz, DMSO-d6) δ ppm 1.88 - 2.01 (m, 1 H), 2.32 - 2.47 (m, 1 H), 2.51 - 2.68 (m, 2 H), 2.84 - 2.99 (m, 1 H), 3.03 - 3.43 (m, 5 H), 3.45 - 3.54 (m, 1 H), 3.56 - 3.75 (m, 2 H), 3.92 - 4.06 (m, 1 H), 4.18 - 4.28 (m, 1 H), 4.31 - 4.41 (m, 1 H), 5.06 (dd, J = 13.3, 5.0 Hz, 1 H), 7.07 - 7.22 (m, 2 H), 7.58 (dd, J = 8.4, 1.8 Hz, 1 H), 7.51 - 7.51 (m, 1 H), 10.09 - 10.60 (m, 1 H), 10.95 (s, 1 H).

[0392] Step 8. Preparation of N-[3-(difluoromethyl)-1-[1-[2-[4-[2-(2,6-dioxo-3-piperidyl)-1-oxo-isoindoline-5-yl]piperazine-1-yl]ethyl]-4-piperidyl]pyrazole-4-yl]-5-[(3R,5R)-3-amino-5-fluoro-1-piperidyl]pyrazolo[1,5-a]pyrimidine-3-carboxamide (P-38) 3-[5-[4-(2,2-dihydroxyethyl)piperazin-1-yl]-1-oxo-isoindorin-2-yl]piperidine-2,6-dione 11 (81.48 mg, 0.21 mmol, 1.2 eq.) / MeCN (1.75 mL, 0.1 M) solution to which DIPEA (0.2 mL, 1.3 mmol, 7.4 eq.) and tert-butyl N-[(3R,5R)-1-[3-[[3-(difluoromethyl)-1-(4-piperidyl)pyrazole-4-yl]carbamoyl]pyrazolo[1,5-a]pyrimidine-5-yl]-5-fluoro-3-piperidyl]carbamate 8 (100.97 mg, 0.17 mmol, 1 eq.) are added, followed by NaBH(OAc)3 (74.1 (mg, 0.35 mmol, 2 eq.) was added. The reaction mixture was stirred at room temperature. After 4 hours, LC-MS showed complete conversion to the target product. The solvent was evaporated, and the residue was purified by reverse-phase FC (50 g C18 gold column, holding solution (DMSO), elution: 30 to 60% MeOH / 0.1% HCOOH over 12 CV) to obtain the target product of sufficient purity. The obtained product was dissolved in 4 M hydrochloric acid / dioxane (1 mL, 4 mmol, 23 eq.). After 2 hours, LC-MS showed complete conversion to P-38. The solution was concentrated and purified by reverse-phase FC (50 g C18 column, holding solution (DMSO), 5% MeCN / 0.02 M HCl over 4 CV, then 5 to 20% MeCN / 0.02 M HCl over 15 CV; the target product eluted at approximately 20% MeCN), and P-38 (28.8 mg, 20% yield) was obtained as a white solid dihydrochloride.

[0393] LC-MS method 3: 99.4% purity (at 215 nm), [M+H] + = 832.4. 1H NMR (400 MHz, DMSO-d6) δ ppm 1.90 - 2.10 (m, 2 H), 2.27 - 2.46 (m, 6 H), 2.56 - 2.64 (m, 3 H), 2.86 - 2.98 (m, 2 H), 3.19 - 3.33 (m, 6 H), 3.62 - 3.95 (m, 8 H), 4.05 - 4.17 (m, 2 H), 4.20 - 4.29 (m, 1 H), 4.32 - 4.41 (m, 1 H), 4.53 - 4.70 (m, 2 H), 4.80 - 4.94 (m, 1 H), 5.03 - 5.19 (m, 2 H), 6.90 (br d, J = 8.3 Hz, 1 H), 7.04 - 7.34 (m, 3 H), 7.59 (br d, J = 8.1 Hz, 1 H), 8.25 - 8.38 (m, 4 H), 8.47 (s, 1 H), 8.94 (d, J = 7.8 Hz, 1 H), 9.37 (s, 1 H), 10.62 - 11.07 (m, 2 H). 19 F NMR (377 MHz, DMSO-d6) δ ppm -185.16 - -184.13 (m, 2 F), -111.92 - -111.26 (m, 1 F).

[0394] Table 31

[0395] examples of biology Example B1. BIOCHEM method of biochemistry of IRAK4 PhosphoSens The PhosphoSens® biochemical assay was performed according to the distributor's instructions (AssayQuant Technologies Inc., Marlborough, MA). A 1.25 X stock solution of IRAK4 (PV4002, ThermoFisher Scientific, Waltham, MA), a 5 X stock solution of ATP, and the Sox-conjugated peptide substrate AQT0326 (CSKS-AQT0326B, AssayQuant Technologies) were prepared in 1X kinase reaction buffer [containing 50 mM HEPES (pH 7.5), 0.01% Brij-35, 0.5 mM EGTA, 20 mM MgCl2, and 1 mM DTT]. To obtain final concentrations of 200 μM ATP and 10 μM peptide substrate, a mixture of ATP (10 μL) and substrate solution was added to a Corning 3574 384-well white unbound surface microtiter plate (containing 0.5 μL of serially diluted test compounds prepared in DMSO). The enzyme solution (40 μL) with a final IRAK4 concentration of 1 nM was added to initiate the reaction, and the λ was measured every 71 seconds for 240 minutes at room temperature using a BioTek Synergy H4 plate reader. EX 360 / λ EM Monitoring was performed using 485 (Agilent Technologies, Santa Clara, CA). The initial linear portion of the progression curve was fitted according to a linear equation to determine the slope, and converted to % inhibition relative to the 100% activity value of the control that was not treated with the inhibitor. IC of each compound 50 The values ​​were obtained by fitting a % inhibitor-compound concentration curve using Dotmatics software (Dotmatics, Bishops Stortford, Hertfordshire, England).

[0396] Example B2. Reagent preparation Cell culture media were prepared in a sterile tissue culture hood by adding 10% FBS and 1% penicillin-streptomycin to 500 mL of RPMI1640 medium (without phenol red). The medium was filtered through a Nalgene Bottle Top Filter and stored at 4°C.

[0397] Cell titer Glo (CTG) buffer and substrate (CellTiter-Glo Luminescent Cell Viability Assay, Promega Ref. # G7573) were stored at -20°C. 100 mL of CTG buffer was warmed in a bead bath and added to a bottle containing the CTG substrate in a tissue culture hood. The solution was mixed homogenously using a pipette. The CTG reagent was dispensed into 15 mL Falcon tubes and stored at -20°C.

[0398] For a homogeneous time-resolved fluorescence (HTRF) assay, the Cisbio HTRF kit was used, containing the following: Resolved buffer #1 4X, Blocking reagent #3 100X, 20X antibody 1 (anti-IRAK4 d2), 20X antibody 2 (Anti-IRAK4 k), and detection buffer.

[0399] The 4X decomposition buffer was stored at 4°C. To use as a 1X decomposition buffer, the 4X solution was diluted with deionized water (distilled water, Gibco Cat. # 15230279) and 100X blocking reagent in a volume ratio of 1:3:0.04.

[0400] Aliquots of 20X antibody solution were stored at -80°C, and the detection buffer was stored at 4°C. To use as a 1X antibody solution, aliquots of 20X antibody solution were diluted with detection buffer at a volume ratio of 1:19.

[0401] Example B3. Procedure for homogeneous time-resolved fluorescence (ALE THP1 HTRF) of advanced fatty acid oxidation end product THP1. Cells were lysed with shaking at room temperature for 45 minutes. A BCA protein assay was performed and normalized to the target total protein concentration using 1X lysis buffer. Next, a 1X antibody solution was prepared by adding 380 μL of detection buffer to an aliquot of 20 μL of 20X antibody solution and mixing thoroughly. The 1X antibody solutions were combined in a 1:1 ratio and vortexed briefly. 20 μL of 1X anti-IRAK4-k antibody solution was maintained as the control well. 384-well plates (ProxiPlate-384 Plus, Perkin Elmer Cat.# 6008289) were loaded by adding 4 μL of the mixed antibody solution to empty wells using a single-channel repeater. Using a multi-channel repeater, 16 μL of decomposition solution was added per well, and all formed foam was defoamed using a 20 μL pipette tip and the end of a Kimwipe. Each control was prepared in 3-row configuration in 10 columns. Buffer controls were prepared in wells A10, B10, and C10 by adding 16 μL of degradation buffer and 4 μL of detection buffer. Cryptate controls were prepared in wells D10, E10, and F10 by adding 16 μL of degradation buffer, 2 μL of detection buffer, and 2 μL of 1 X anti-IRAK4-k antibody solution. Negative controls were prepared in wells G10, H10, and I10 by adding 16 μL of degradation buffer and 4 μL of mixed antibody solution. The plate was sealed with a clear seal and covered with an aluminum lid. The plate was centrifuged at 800 g for 5 minutes and incubated overnight in the dark at room temperature. The next day, the plate was centrifuged at 800 g for 5 minutes. Samples were analyzed using a plate reader (Envision, PerkinElmer) with the Desnor 384 HTRF program.

[0402] A summary of the ALE THP1 HTRF data for the tested compounds is shown in Table 8 below. [Table 32] [Table 33]

[0403] Although the present invention is described in some detail by examples and embodiments for the purpose of clarity of understanding, these descriptions and embodiments should not be construed as limiting the scope of the invention. All disclosures of patent and scientific documents cited herein are expressly incorporated herein in their entirety by reference.

Claims

1. Equation (I): 【Chemistry 1】 [In the formula, R 1 C 1 -C 6 Haloalkyl, C 1 -C 6 Alkyl or -CN; R 2a is H or C 1 -C 6 It is alkyl; R 2b is C 3 which may optionally be substituted with 1 to 5 R 5 -C 6 is cycloalkyl; or R 2a and R 2b The dotted line between them represents a ring structure, R 2a and R 2b These combine with the nitrogen atoms bonded to them to form a six-membered heterocycline, which may optionally contain one additional heteroatom selected from N and O, and the heterocycline has 1 to 5 R 3 The base may be replaced as desired; Each R 3 -NH is independent of 2 -OH, halo, C 1 -C 6 Alkyl or C 1 -C 6 It is a haloalkyl; X is either CH or N; L 1 -C(O)N(H)-, -N(H)C(O)-, -C(O)-, -(C 1 -C 6 Alkilen)N(R 4 )-or C 1 -C 6 It is alkylene; R 4 is H or C 1 -C 6 It is alkyl; R 5a and R 5b Each of these is either H, or they together form an oxo group; Ring A is, 【Chemistry 2】 It is either an 8-10 membered spiroheterocyclylene containing 1-3 nitrogen atoms, wherein the heterocyclylene group contains m R 6 It is substituted with the base; Y 1 and Y 2 It is independently CH or N; Each R 6 It is independently, Halo, C 1 -C 6 Alkyl or C 1 -C 6 It is a haloalkyl; m is between 0 and 5; L 2 is a combination or -N(R 7 )-and; and R 7 is H or C 1 -C 6 It is alkyl. Compounds thereof or pharmaceutically acceptable salts thereof.

2. R 1 However, C 1 -C 3 Haloalkyl, C 1 -C 3 A compound according to claim 1, wherein the compound is alkyl or -CN, or a pharmaceutically acceptable salt thereof.

3. R 1 However, -CHF 2 ,-CF 3 ,-CH 3 The compound according to claim 2 or a pharmaceutically acceptable salt thereof, wherein the compound is -CN.

4. R 2a However, H or C 1 -C 3 It is alkyl; and R 2b However, 1 to 3 R 3 A compound according to any one of claims 1 to 3, wherein the compound is a cyclohexyl group which may be optionally substituted.

5. R 2a and R 2b These, together with the nitrogen atoms bonded to them, form a six-membered heterocycline which may optionally contain one additional heteroatom selected from N and O, and the heterocycline comprises 1 to 2 R 3 A compound according to any one of claims 1 to 3 or a pharmaceutically acceptable salt thereof, which may be optionally substituted with a base.

6. Each R 3 However, independently, -NH 2 -OH, halo, C 1 -C 3 Alkyl or C 1 -C 3 A compound according to any one of claims 1 to 5, which is a haloalkyl compound, or a pharmaceutically acceptable salt thereof.

7. Each R 3 However, independently, -NH 2 -OH, F, or -CH 3 The compound according to claim 6 or a pharmaceutically acceptable salt thereof. 【Request Item 8】 【Chemistry 3】 but, 【Chemistry 4】 A compound according to any one of claims 1 to 7 or a pharmaceutically acceptable salt thereof.

9. L 1 is -C(O)N(H)-, -N(H)C(O)-, -C(O)-, -(C 1 -C 3 Alkilen)N(R 4 )-or C 1 -C 3 It is alkylene; and R 4 However, H or C 1 -C 3 A compound according to any one of claims 1 to 8, which is alkyl, or a pharmaceutically acceptable salt thereof.

10. L 1 is -C(O)N(H)-, -N(H)C(O)-, -C(O)-, -CH 2 N(CH 3 )-,-CH 2 - or -CH 2 CH 2 -The compound according to claim 9 or a pharmaceutically acceptable salt thereof.

11. Ring A is 【Transformation 5】 It is either a 10-membered spiroheterocyclylene containing two nitrogen atoms, and the spiroheterocyclylene contains m R 6 A compound according to any one of claims 1 to 10, which is substituted with a group, or a pharmaceutically acceptable salt thereof.

12. Each R 6 is independently halo, C 1 -C 3 alkyl or C 1 -C 3 haloalkyl, a compound according to any one of claims 1 to 11 or a pharmaceutically acceptable salt thereof.

13. A compound according to any one of claims 1 to 12 or a pharmaceutically acceptable salt thereof, wherein m is 0, 1, or 2.

14. Ring A is 【Transformation 6】 A compound according to any one of claims 1 to 13 or a pharmaceutically acceptable salt thereof.

15. (i) L 2 is a combination; or (ii) L 2 is -N(R 7 )-; and R 7 However, H or C 1 -C 3 A compound according to any one of claims 1 to 14, which is alkyl, or a pharmaceutically acceptable salt thereof. 【Request Item 16】 【Chemistry 7】 but 【Transformation 8】 A compound according to any one of claims 1 to 15 or a pharmaceutically acceptable salt thereof.

17. The compound is of formula (IIa), (IIb), (IIIa), or (IIIb): 【Chemistry 9】 A compound according to any one of claims 1 to 16 or a pharmaceutically acceptable salt thereof.

18. A compound selected from the compounds in Table 1 or their pharmaceutically acceptable salts.

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

20. (i) A method for modulating interleukin-1 (IL1) receptor-related kinase 4 (IRAK4), characterized by contacting IRAK4 with an effective amount of a compound or a pharmaceutically acceptable salt thereof according to any one of claims 1 to 18 or the pharmaceutical composition according to claim 19; or, (ii) A method for treating an inflammatory disease or autoimmune disease in a subject requiring treatment, characterized by administering an effective amount of a compound according to any one of claims 1 to 18 or a pharmaceutically acceptable salt thereof, or a pharmaceutical composition according to claim 19, to the subject, wherein the inflammatory disease or autoimmune disease may optionally be atopic dermatitis, asthma, lupus, rheumatoid arthritis, familial Mediterranean fever, psoriasis, generalized pustular psoriasis, cryopyrin-associated periodic syndromes, hidradenitis suppurativa, Beckett syndrome, or familial cold autoinflammatory syndrome.

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