Aza-cyclic compounds as aak1 inhibitors

Abstract

Aza-heterocyclic compounds as AAK1 inhibitors. The present invention relates to the use of aza-heterocyclic compounds or pharmaceutically acceptable salts thereof which inhibit the activity of afferin-associated kinase 1 (AAK1 kinase) in a medicament for the treatment or prevention of a disease or disorder mediated by the activity of AAK1. In particular, the present invention provides a compound of formula (I), a stereoisomer or a pharmaceutically acceptable salt thereof, wherein the definitions of the various groups are as described in the specification. The compounds have high AAK1 inhibitory activity.

Description

TECHNICAL FIELD

[0001] The present application belongs to the field of medicine, in particular to nitrogen-containing heterocyclic compounds capable of inhibiting adaptor associated kinase 1 (AAK1) activity, compositions comprising the above-mentioned compounds and the use thereof in the preparation of a medicament for the treatment of diseases or disorders mediated by adaptor associated kinase 1 activity. BACKGROUND

[0002] Adaptor associated kinase 1 (AAK1) is a member of the Ark1 / Prk1 family of serine / threonine kinases. AAK1 mRNA exists in two splice forms known as the short and long forms. The long form predominates and is highly expressed in brain and heart (Henderson and Conner, Mol. Biol. Cell. 2007, 18, 2698-2706). AAK1 is enriched in synaptosomal preparations and co-localizes with endocytic structures in cultured cells. AAK1 regulates clatherin-coated endocytosis, a process that is important in synaptic vesicle recycling and receptor-mediated endocytosis. AAK1 binds to the AP2 complex, a heterotetramer that links receptor cargo to clatherin coats. Binding of clatherin to AAK1 stimulates AAK1 kinase activity (Conner et. al., Traffic 2003, 4, 885-890; Jackson et. al., J. Cell. Biol. 2003, 163, 231-236). AAK1 phosphorylates the mu-2 subunit of AP-2, which promotes the binding of mu-2 to tyrosine-containing sorting motifs on cargo receptors (Ricotta et. al., J. Cell Bio. 2002, 156, 791-795; Conner and Schmid, J. Cell Bio. 2002, 156, 921-929). Mu2 phosphorylation is not essential for receptor uptake, but phosphorylation improves the efficiency of internalization (Motely et. al., Mol. Biol. Cell. 2006, 17, 5298-5308).

[0003] AAK1 has been identified as an inhibitor of neuregulin-1 / ErbB4 signaling in PC12 cells. Loss of AAK1 expression via RNA interference-mediated gene silencing or treatment with the kinase inhibitor K252a, which inhibits AAK1 kinase activity, resulted in an enhancement of neuregulin-1 -induced neurite outgrowth. These treatments resulted in increased ErbB4 expression and increased accumulation of ErbB4 in or near the plasma membrane (Kuai et. al., Chemistry and Biology 2011, 18, 891-906). NRG1 and ErbB4 are putative schizophrenia susceptibility genes (Buonanno, Brain Res. Bull. 2010, 83, 122-131). SNPs in both genes are associated with multiple schizophrenia endophenotypes (Greenwood et. al., Am. J. Psychiatry 2011, 168, 930-946). Neuregulin 1 and ErbB4 KO mouse models have shown schizophrenia-related morphological changes and behavioral phenotypes (Jaaro-Peled et. al., Schizophrenia Bulletin 2010, 36, 301-313; Wen et. al., Proc. Natl. Acad. Sci. USA. 2010, 107, 1211-1216). In addition, a single nucleotide polymorphism in an intron of the AAK1 gene is associated with age of onset of Parkinson’s disease (Latour et. al., BMC Med. Genet. 2009, 10, 98). These results suggest that inhibition of AAK1 activity can be useful in the treatment of schizophrenia, cognitive deficit in schizophrenia, Parkinson’s disease, neuropathic pain, bipolar disorder, and Alzheimers disease.

[0004] AAK1 inhibitor compounds are disclosed in CN106458994A, CN108290843A, WO2023284838A, and others. Compounds that inhibit AAK1 activity, BMS-986176 / LX-9211, are disclosed in J. Med. Chem. 2022, 65, 4457-4480. SUMMARY SUMMARY

[0006] The present inventors have conducted extensive research and found that the compound provided by the present application and stereoisomers or pharmaceutically acceptable salts thereof have strong AAK1 inhibiting function, overcoming the deficiencies of AAK1 inhibiting function in the prior art. The nitrogen-containing heterocyclic compound provided by the present application has good inhibiting effect on AAK1.

[0007] The present application solves the above technical problems through the following technical solutions.

[0008] In one aspect, the present application provides a compound as shown in formula I, a stereoisomer or a pharmaceutically acceptable salt thereof:

[0009]

[0010] wherein,

[0011] X1, X2, X3 are each independently selected from N or CR2;

[0012] Ring A is selected from:

[0013]

[0014] R1, R2, R3 are each independently selected from hydrogen, amino, -CO2H, halogen, fluoromethyl, difluoromethyl, trifluoromethyl, cyano, methylcarbamoyl, C 1-6 alkyl, deuterated C 1-6 alkyl, halogenated C 1-6 alkyl, C 1-6 alkoxy, deuterated C 1-6 alkoxy, halogenated C 1-6 alkoxy, hydroxyl C 1-6 alkyl, C 3-6 cycloalkyl, 4-6 membered heterocycloalkyl containing 1-3 heteroatoms selected from N, S, O;

[0015] R4, R5, R6 are each independently selected from hydrogen, deuterium, halogen, amino, cyano, hydroxyl, alkenyl, fluoromethyl, difluoromethyl, trifluoromethyl, C 1-6 alkyl, halogenated C 1-6 alkyl, C 1-6 alkoxy, halogenated C 1-6 alkoxy, deuterated C 1-6 alkyl, deuterated C 1-6 alkoxy, C 3-6 cycloalkyl, hydroxyl C 1-6 alkyl, 4-6 membered heterocycloalkyl containing 1-3 heteroatoms selected from N, S, O;

[0016] or R4 and R5 together with the carbon atoms to which they are respectively attached form a C 3-6cycloalkyl, 4-6 membered heterocycloalkyl containing 1-3 heteroatoms selected from N, S, O, or a double bond;

[0017] n is selected from 0, 1, 2, 3, 4, and when n is greater than 1, R1may be the same or different.

[0018] As a preferred technical solution, R1is selected from hydrogen, fluorine, chlorine, -CO2H, cyano, methoxy, methyl, difluoromethyl, trifluoromethyl, methionyl.

[0019] As a preferred technical solution, R2is selected from hydrogen, methyl, cyano, difluoromethyl, trifluoromethyl, cyclopropyl.

[0020] As a preferred technical solution, R3is selected from methyl, and R4, R5, R6are each independently selected from hydrogen, methyl, halogen, alkenyl, 4-6 membered heterocycloalkyl containing 1-3 heteroatoms selected from N, S, O.

[0021] As a preferred technical solution, R4and R5together with the carbon atom to which each is attached form a 4-6 membered heterocycloalkyl containing 1-3 heteroatoms selected from N, S, O, or a double bond.

[0022] As a preferred technical solution, R4and R5together with the carbon atom to which each is attached form a 4-6 membered heterocycloalkyl containing 1-3 heteroatoms selected from N, S, O, or a double bond. selected from the following groups:

[0023]

[0024] In a second aspect, the present application provides a compound represented by formula I-1, a stereoisomer or a pharmaceutically acceptable salt thereof:

[0025]

[0026] wherein, R1, R2, ring A, n, R4, R5, R6, X1, X2, X3are as defined above.

[0027] In a third aspect, the present application provides a compound represented by formula I-2, a stereoisomer or a pharmaceutically acceptable salt thereof:

[0028]

[0029] wherein, X1, X2, X3, R1, R2, ring A, n are as defined above.

[0030] In a fourth aspect, the present application provides a compound represented by formula I-3, I-4, I-5, I-6, I-7, I-8, a stereoisomer or a pharmaceutically acceptable salt thereof:

[0031]

[0032] wherein R1, ring A, n are as defined above.

[0033] As a preferred technical solution, the compound provided by the present application includes but is not limited to the following compounds:

[0034]

[0035]

[0036]

[0037] In a fifth aspect, the present application provides a pharmaceutical composition comprising the compound of any one of the preceding technical solutions, a stereoisomer or a pharmaceutically acceptable salt thereof, and a pharmaceutically acceptable carrier.

[0038] The present application also provides the use of the compound of any one of the preceding technical solutions, a stereoisomer or a pharmaceutically acceptable salt thereof, or the pharmaceutical composition in the preparation of a medicament for treating a disease or disorder mediated by the activity of the Connexin-related kinase 1.

[0039] As a preferred technical solution, the disease or disorder is selected from Alzheimer's disease, bipolar disorder, Parkinson's disease, schizophrenia, diabetic peripheral neuropathic pain, post-herpetic neuralgia.

[0040] The amount of the compound, the stereoisomer or the pharmaceutically acceptable salt thereof in the present application is calculated in the form of free base in each case.

[0041] The present inventors have found that the compound is a highly effective AAK1 inhibitor, has a strong AAK1 inhibitory activity, and can be used for preparing a medicament for preventing and / or treating a disease or disorder mediated by the activity of AAK1, including Alzheimer's disease, bipolar disorder, Parkinson's disease, schizophrenia, diabetic peripheral neuropathic pain, post-herpetic neuralgia, fibromyalgia or peripheral neuropathy, etc. The present application is completed based on the above findings. DETAILED DESCRIPTION

[0043] The various aspects and features of the present application are further described below.

[0044] All documents cited in the present application, including during prior art searches, are, in relevant part, incorporated herein by reference; the citation of any document is not to be construed as an admission that it is prior art with respect to the present application. To the extent that any meaning or definition of a term in this document conflicts with the meaning or definition of the same term in a document incorporated herein by reference, the meaning or definition assigned to the term in this document shall control. When values are expressed in a range format, it is intended to include the beginning and ending values, and all the intermediate values of the range. For example, designating a range of "about 0.1% to about 10%," is intended to disclose all individual values and sub-ranges within the specified ranges, e.g., about 1% to about 5%, or about 2% to about 7.5%, within the specified ranges; ranges of "up to 10%"; or "90 to 100%"; and values such as 0.5%, 1.1%, 2.3%, 3.4%, 4.6%, 5.5%, etc. The statement "about X to Y" is intended to include the range of values starting from the lower value "about X" and ending "about Y", inclusive of the values "about X" and "about Y". The use of "about" with respect to a given value should be construed to include not only the value itself but also a range of values around the value, e.g., within 10% of the value. For example, "about 90%" includes the value 90% and a range of values from 81% to 99%.

[0045] The compounds according to the present application can exist in tautomeric forms, and the present application includes all tautomeric forms.

[0046] The compounds of the present application possess asymmetric centers, and the compounds of the present application which contain asymmetrically substituted atoms can be isolated in optically active or racemic forms. Methods for the preparation of optically active forms are known to those skilled in the art, for example, by resolution of a racemic form or by stereoselective synthesis using optically pure starting materials. Unless otherwise specified, specific chemical or isomeric forms of compounds of the present application are not intended to be limiting. The methods for the preparation of the compounds of the present application and intermediates thereof are part of the present application. All tautomers of the compounds of the present application are also part of the present application.

[0047] The singular forms "a," "an," and "the" include plural referents unless the context clearly dictates otherwise.

[0048] As used herein, the terms "optional" or "optionally" mean that the subsequently described event or circumstance can or can not occur, and that the description includes instances where the event or circumstance occurs and instances where it does not. For example, "optionally substituted alkyl" means that the alkyl can be substituted and that the event or circumstance where the alkyl is not substituted is included.

[0049] The term "substituted" means a moiety having a substituent that replaces a hydrogen on one or more carbons of the backbone. It is understood that "substitution" or "substituted by" includes the implicit proviso that such substitution is in accordance with permitted valence of the substituted atom and the substituent, and that the substitution results in a stable compound, e.g., which does not spontaneously undergo transformation such as by rearrangement, cyclization, elimination, etc. The term "substituted" as used herein is contemplated to include all permissible substituents of organic compounds. In a broad aspect, the permissible substituents include noncyclic and cyclic, branched and unbranched, carbocyclic and heterocyclic, aromatic and nonaromatic substituents of organic compounds. The permissible substituents can be one or more and the same or different for appropriate organic compounds. For the purposes of this application, heteroatoms such as nitrogen can have hydrogen substituents and / or any permissible substituents of organic compounds described herein which satisfy the valencies of the heteroatoms. Substituents can include any of the substituents described herein, such as halogen, hydroxyl, alkyl, alkoxy, amino, cyano, heteroaryl, heterocyclyl, and the like. One skilled in the art will appreciate that substituents themselves can be substituted if appropriate.

[0050] As described herein, the terms "halogen," "halo," and the like mean fluorine, chlorine, bromine, or iodine, particularly fluorine, chlorine, bromine, particularly preferred fluorine, chlorine.

[0051] C x-y The expression of a group refers to a group comprising x to y carbon atoms, such as "C 1-6 "Alkyl" refers to an alkyl group containing 1-6 carbon atoms.

[0052] As described herein, the term "alkyl" refers to an alkyl group having the indicated number of carbon atoms, which is straight-chained or branched, and which can include its subgroups, e.g., where reference is made to "C1-C6 alkyl," this also can include groups represented by the subranges C1-C4 alkyl, C1-C3 alkyl, C2-C6 alkyl, C2-C4 alkyl, and the like, as well as specific groups such as methyl, ethyl, n-propyl, i-propyl, n-butyl, sec-butyl, t-butyl, pentyl, hexyl, and the like.

[0053] "HaloC 1-6 "Alkyl" refers to an alkyl group containing 1-6 carbon atoms, wherein one or more hydrogens are replaced by one or more halogen atoms (such as fluorine, chlorine, bromine, iodine), the upper limit of the number of halogen substituents being equal to the sum of the number of hydrogens that can be replaced in the alkyl group. Without particular limitation, the number of halogen substituents is any integer between 1 and this upper limit, preferably 1-5 halogen substituents, 1-3 halogen substituents, 1-2 halogen substituents, or 1 halogen substituent; when the number of halogen substituents is greater than 1, they can be the same or different halogen: including but not limited to -CF3, -CH2Cl, -CH2CF3, -CCl2, CF3, and the like.

[0054] "Heteroalkyl" refers to an alkyl group as defined above, which is substituted with one or more heteroatoms, such as -O-, -S-, or -NR'-, wherein R' is hydrogen, substituted or unsubstituted alkyl, substituted or unsubstituted aryl, or substituted or unsubstituted heteroalkyl. Non-limiting examples of heteroalkyl groups include -CH2OCH3, -CH2OCH2CH3, -CH2NH2, -CH2NHCH3, -CH2N(CH3)2, -CH2NHCH2CH3, -CH2N(CH2CH3)2, -CH2SCH3, -CH2SCH2CH3, -CH2S(O)CH3, -CH2S(O)2CH3, and the like.

[0055] The term "alkoxy" is a term of art and refers to an alkyl group, as defined above, attached to the rest of the molecule through an oxygen atom, and refers to an alkyl group containing an oxygen atom, examples of such groups include methoxy, ethoxy, or propoxy, and the like.

[0056] The term "haloalkyl" refers to an alkyl group, as defined above, attached to the rest of the molecule via a halo linkage. For example, C1-6haloalkyl refers to an alkyl group having from 1 to 6 carbon atoms or 1 to 3 carbon atoms attached to the rest of the molecule via a halo linkage. Preferred haloalkyl groups include, but are not limited to, -CH2Cl, -CHCl2, -CF3, and the like.

[0057] "Haloalkoxy" refers to -O-haloalkyl, which when not otherwise specified, is -O-haloC 1-8 alkyl, preferably -O-haloC 1-6 alkyl, more preferably -O-haloC 1-4 alkyl, further preferably -O-haloC 1-2 alkyl; the upper limit on the number of halo substituents is equal to the total number of hydrogens that can be replaced by a substituent, and no particular consideration is given to this limitation, the number of halo substituents is any integer between 1 and this upper limit, preferably 1-5 halo substitutions, 1-3 halo substitutions, 1-2 halo substitutions, and 1 halo substituent; when the number of halo substituents is greater than 1, the halo substituents can be the same or different; non-limiting examples include monofluoromethoxy, difluoromethoxy, trifluoromethoxy, difluoroethoxy, and the like.

[0058] "Cycloalkyl" refers to a substituted or unsubstituted, saturated or partially unsaturated, non-aromatic hydrocarbon ring, which can be monocyclic, bicyclic, or polycyclic, and the bicyclic or polycyclic can be fused, spiro, or bridged, and when not otherwise specified, typically has from 3 to 20 carbon atoms; when it is a monocyclic cycloalkyl, it preferably has from 3-15 carbon atoms, preferably 3-10 carbon atoms, more preferably 3-8 carbon atoms, more preferably 3-6 carbon atoms, further preferably 3-4 carbon atoms; when it is a bicyclic or polycyclic cycloalkyl, it preferably has from 4-12 carbon atoms, preferably 4-11 carbon atoms, more preferably 5-11 carbon atoms, more preferably 6-11 carbon atoms, further preferably 6-10 carbon atoms; non-limiting examples include cyclopropyl, cyclobutyl, cyclopentyl, cyclohexyl, cycloheptyl, butenyl, cyclopentenyl, cyclohexenyl,

[0059] "Alkenyl" refers to a straight-chain hydrocarbon or branched-chain hydrocarbon group containing at least one carbon-carbon double bond (C=C). Unless otherwise indicated, it contains primarily from 2 to 18 (e.g., 2 to 8, further e.g., 2 to 6, further e.g., 2 to 4) carbon atoms, including, but not limited to, ethenyl, allyl, 1-propenyl, 2-propenyl, 1-butenyl, 2-butenyl, 3-butenyl, 1-pentenyl, 2-pentenyl, 3-pentenyl, 4-pentenyl, 1-methyl-1-butenyl, 2-methyl-1-butenyl, 2-methyl-3-butenyl, 1-hexenyl, 2-hexenyl, 2-hexenyl; the alkenyl group can be optionally further substituted with any group.

[0060] "Heterocycloalkyl" refers to a substituted or unsubstituted saturated or partially unsaturated non-aromatic ring containing at least one heteroatom. Unless otherwise indicated, the heterocycloalkyl is a 3-20 membered ring, when it is a monocyclic heterocycloalkyl, it is preferably 3-15 membered, preferably 3-10 membered, more preferably 3-8 membered, further preferably 3-6 membered; when it is a bicyclic or polycyclic heterocycloalkyl, it is preferably 4-12 membered, preferably 4-11 membered, more preferably 5-11 membered, more preferably 6-11 membered, further preferably 6-10 membered; the heterocycloalkyl can be monocyclic, bicyclic or polycyclic. The ring can be a bridged ring, a parallel ring and a spiro ring, the heteroatom therein is selected from N, S, O, P, Si heteroatom and its oxidation state; when the heterocycloalkyl is bicyclic or polycyclic, at least one ring contains at least one heteroatom, which can be a bicyclic or polycyclic ring formed by a heteroatom-containing ring and a non-heteroatom-containing ring; when connected to other groups, the heteroatom or carbon atom can be connected as a connection point; non-limiting examples include azetidinyl, morpholinyl, piperazinyl, piperidinyl, tetrahydropyranyl, oxetanyl, pyranyl, azacyclopentenyl, azacyclohexenyl, oxacyclopentenyl, oxacyclopentenyl, and the like.

[0061] "Deuterium" refers to the isotope of hydrogen (H).

[0062] As used herein, the term "cyano" refers to a group in which a carbon atom is triple-bonded to a nitrogen atom.

[0063] As used herein, the term "hydroxy" or "hydroxyl", alone or in combination, means -OH.

[0064] "Pharmaceutically acceptable salt" refers to a salt of a compound of the present application that retains the biological effectiveness and properties of the free acids or free bases and is not biologically or otherwise undesirable. The inorganic or organic acids that are used to prepare the pharmaceutically acceptable salts are those that form non-toxic salts.

[0065] A "pharmaceutical composition" refers to a mixture of one or more compounds described herein, or stereoisomers, solvates, pharmaceutically acceptable salts, co-crystals, deuterated compounds, and other components, wherein the other components include physiologically / pharmaceutically acceptable carriers and / or excipients.

[0066] A "stereoisomer" refers to isomers that have the same molecular formula but different physical properties, such as different melting points, different retention times on a chiral column, and / or different optical properties. A stereoisomer can have one or more chiral centers.

[0067] A "solvate" refers to a compound of the present application or a salt thereof, combined with a stoichiometric or non-stoichiometric amount of solvent through intermolecular non-covalent forces. Where the solvent is water, the solvate is a hydrate.

[0068] A "co-crystal" refers to a crystal formed by the combination of a pharmaceutically active ingredient (API) and a co-crystal former (CCF) through hydrogen bonding or other non-covalent bonding, wherein both the API and the CCF are in their pure state and are solids, and there is a fixed stoichiometric ratio between the components. A co-crystal is a multi-component crystal, including binary co-crystals formed between two neutral solids and multi-component co-crystals formed between a neutral solid and a salt or a solvate.

[0069] The term "disease" refers to a physical state of the subject that is associated with the disease described herein. For example, Alzheimer's disease, bipolar disorder, Parkinson's disease, schizophrenia, diabetic peripheral neuropathic pain, postherpetic neuralgia, fibromyalgia, or peripheral neuropathy-related diseases described herein.

[0070] A "carrier" refers to a system that does not cause significant irritation to the organism and does not eliminate the biological activity and characteristics of the administered compound, and can change the way the drug enters the human body and its distribution in the body, control the release rate of the drug and deliver the drug to the target organ, non-limiting examples include liposomes, nanoparticles, etc.

[0071] The compound in the present application or the pharmaceutical composition containing it can be administered in unit dosage form, and the administration route can be enteral or parenteral, such as oral, intravenous injection, intramuscular injection, intravenous infusion, subcutaneous injection, nasal, oral mucosa, eye, lung and respiratory tract, skin, vagina, rectum, etc.

[0072] The administration form can be a liquid form, a solid form or a semi-solid form. The liquid form can be a solution (including true solution and colloidal solution), an emulsion (including o / w type, w / o type and multiple emulsion), a suspension, an injection (including water injection, powder injection and infusion), eye drops, nose drops, lotion and liniment, etc.; the solid form can be a tablet (including ordinary tablet, enteric-coated tablet, buccal tablet, dispersible tablet, chewable tablet, effervescent tablet, oral disintegrating tablet), a capsule (including hard capsule, soft capsule, enteric-coated capsule), a granule, a powder, a pellet, a drop pill, a suppository, a film, a patch, an aerosol (powder) mist, a spray, etc.; the semi-solid form can be an ointment, a gel, a paste, etc.

[0073] To achieve the purpose of administration and enhance the therapeutic effect, the drug or the pharmaceutical composition of the present application can be administered by any known administration method.

[0074] The compound or the composition of the present application can be taken alone or in combination with other therapeutic drugs or symptomatic drugs. When the compound of the present application has a synergistic effect with other therapeutic drugs, the dosage thereof should be adjusted according to the actual situation.

[0075] Advantageous technical effects

[0076] The inventors have unexpectedly found that the compound in the present application has unexpectedly good AKK1 inhibitory activity, and the IC50 or EC50 is equivalent to or less than the positive control drug, i.e. BMS-986176 / LX-9211. The present application provides a class of AKK1 inhibitor compounds with novel structure and strong activity, which has a good application prospect in preventing and / or treating AKK1 inhibition related indications such as Alzheimer's disease, bipolar disorder, Parkinson's disease, schizophrenia, diabetic peripheral neuropathic pain, post-herpetic neuralgia, fibromyalgia or peripheral neuropathy, etc. BRIEF DESCRIPTION OF DRAWINGS

[0077] Figure 1 The effect of compound 26 on the pain threshold of peripheral neuropathic pain of diabetic rats is shown. DETAILED DESCRIPTION

[0078] The following examples are helpful for those skilled in the art to better understand the technical solutions of the present application, but do not limit the present application in any way.

[0079] For all the following examples, standard operations and methods known to those skilled in the art can be used. Unless otherwise specified, all temperatures are in °C (degrees Celsius). The structure of the compound is determined by nuclear magnetic resonance spectrum (NMR) and / or mass spectrum (MS).

[0080] The structure of the compounds of the present application is determined by nuclear magnetic resonance (NMR) or / and liquid-mass spectrometry (LC-MS). The NMR chemical shift (δ) is in units of parts per million (ppm). The nuclear magnetic resonance is determined by using a Bruker avance-400 type nuclear magnetic instrument, the solvents are deuterated dimethyl sulfoxide (DMSO-d6), deuterated methanol (CD3OD) and deuterated chloroform (CDCl3), and the internal standard is tetramethylsilane (TMS).

[0081] The liquid phase mass spectrometry LC-MS measures the liquid phase part using an ACQUITY UPLC ultra-high pressure liquid chromatograph, and the mass spectrometry part uses an Xevo G2-S Qtof mass spectrometer.

[0082] The starting materials in the examples of the present application are known and can be purchased on the market, or can be synthesized using or according to the methods known in the art.

[0083] The compounds of the present application can be prepared by those skilled in the art by combining the documents of WO2017059085, WO2017059080, WO2015153720 and known organic synthesis techniques. The starting materials are commercially available chemicals and / or compounds described in the chemical literature. The "commercially available chemicals" are obtained from regular commercial channels, and the suppliers include: Shanghai Pharmalink, Nanjing Chemical Industry and other companies.

[0084] Example 1: Synthesis of (R)-1-{[2-(difluoromethyl)-6-(pyrazolo[1,5-a]pyrimidin-7-yl)pyridin-3-yl]oxy}-2,4-dimethylpentan-2-amine (Compound 1)

[0085]

[0086] Step 1: 1H-pyrazol-5-amine (6.0 g, 72 mmol, 1.0 eq) and 1,3-dimethylpyrimidine-2,4(1H,3H)-dione (14.2 g, 100.8 mmol, 1.4 eq) were added to sodium ethoxide (60 mL 20 wt%) and reacted at 80°C for 3h. After the reaction was complete, it was cooled to 0°C, filtered, and the filter cake was dissolved in water and washed with dichloromethane three times, then the water phase was adjusted to weakly acidic with acetic acid, cooled to 0°C, and the solid was precipitated, filtered, and dried to obtain white solid pyrazolo[1,5-a]pyrimidin-7(6H)-one (5.8 g, 43.0 mmol, yield 59.7%). LCMS (TOF MS ES + m / z [M+H] + 136.04. 1H NMR (400 MHz, DMSO) δ 12.08 (s, 1H), 8.47 (d, J = 7.9 Hz, 1H), 7.75 (d, J = 1.9 Hz, 1H), 5.93 (d, J = 7.9 Hz, 1H), 5.81 (d, J = 2.0 Hz, 1H).

[0087] Step 2: Pyrazolo[l,5-a]pyrimidin-7(6H)-one (2.0 g, 14.8 mmol, 1.0 eq) and potassium carbonate (6.12 g, 44.4 mmol, 3.0 eq) were added to acetonitrile solution, then phosphorus oxybromide (12.7 g, 44.40 mmol, 3.0 eq) was added, after addition, 80 °C for 5 h. After the reaction was complete, the reaction was poured into ice water, then saturated sodium bicarbonate solution was added to neutralize the phosphorus oxybromide, the aqueous phase was extracted with ethyl acetate three times, the organic phase was combined and concentrated. Purified by normal phase column (ethyl acetate / petroleum ether, ethyl acetate% = 25%), to get yellow solid 7-bromopyrazolo[l,5-a]pyrimidine (800 mg, 4.06 mmol, yield 27.4%). LCMS (TOF MS ES + )m / z [M+H] + : 197.96. 1 H NMR (400 MHz, DMSO) δ 9.08 (d, J = 7.1 Hz, 1H), 8.27 (d, J = 2.3 Hz, 1H), 7.22 (d, J = 7.2 Hz, 1H), 6.73 (d, J = 2.3 Hz, 1H).

[0088] Step 3: (R)-l-{[6-bromo-2-(difluoromethyl)pyridin-3-yl]oxy}-2,4-dimethylpentan-2-amine (200 mg, 0.59 mmol, 1.0 eq) and pinacol diborane (299 mg, 1.18 mmol, 2.0 eq) were dissolved in 1,4-dioxane (12 mL), then potassium acetate (173.7 mg, 1.77 mmol, 3.0 eq) and [l,l'-bis(diphenylphosphino)ferrocene]dichloropalladium (43.2 mg, 0.059 mmol, 0.1 eq) were added successively, under nitrogen protection, 80 °C for 16 hours. After the reaction was complete, the reaction was filtered, the filtrate was concentrated, then purified by reverse phase column (water / acetonitrile, acetonitrile% = 3-5%), to get white oil (R)-{5-[(2-amino-2,4-dimethylpentyl)oxy]-6-(difluoromethyl)pyridin-2-yl}boronic acid (35 mg, 0.11 mmol, yield 17.5%). LCMS (TOF MS ES + )m / z [M+H] + : 303.16. 1H NMR (400 MHz, DMSO-d6) δ 8.28 (d, J = 6.9 Hz, 2H), 7.90 - 7.53 (m, 2H), 7.27 (m, J = 53.7, 10.9 Hz, 1H), 3.95 (s, 2H), 1.79 (m, J = 11.6, 6.1 Hz, 1H), 1.57 - 1.40 (m, 2H), 1.31 (s, 3H), 0.92 (m, J = 14.7, 6.6, 3.4 Hz, 6H).

[0089] Step 4: (R)-{5-[(2-amino-2,4-dimethylpentyl)oxy]-6-(difluoromethyl)pyridin-2-yl}boronic acid (25 mg, 0.082 mmol, 1.0 eq), 7-bromopyrazolo[l,5-a]pyrimidine (16.48 mg, 0.082 mmol, 1.0 eq) were dissolved in 2 mL 1,4-dioxane, then sodium carbonate (35 mg, 0.33 mmol, 4.0 eq), [l,l'-bis(diphenylphosphino)ferrocene]dichloropalladium (6.05 mg, 0.008 mmol, 0.1 eq) were added, the system was replaced with nitrogen twice, and reacted at 120 °C for 6 h. After the reaction was completed, reverse phase preparation was performed to obtain (R)-l-{[2-(difluoromethyl)-6-(pyrazolo[l,5-a]pyrimidin-7-yl)pyridin-3-yl]oxy}-2,4-dimethylpentan-2-amine (1.4 mg, 0.003 mmol, 5.6% yield) as a white solid. LCMS (TOF MS ES + )m / z [M+H] + : 376. 1 H NMR (400 MHz, Methanol-d4) δ 8.99 - 8.91 (m, 1H), 8.67 (d, J = 8.8 Hz, 1H), 8.52 (s, 2H), 8.18 (d, J = 2.4 Hz, 1H), 8.06 (d, J = 7.4 Hz, 1H), 7.80 (d, J = 8.9 Hz, 1H), 7.26 - 6.95 (m, 1H), 6.74 (m, J = 2.3, 0.9 Hz, 1H), 4.24 - 4.09 (m, 2H), 1.85 (m, J = 12.6, 6.3 Hz, 1H), 1.75 (m, J = 14.3, 5.7 Hz, 1H), 1.63 (m, J = 14.2, 5.5 Hz, 1H), 1.41 (s, 3H), 1.02 (m, J = 14.9, 6.6 Hz, 6H).

[0090] Example 2: Synthesis of (R)-l-{[2-(difluoromethyl)-6-(lH-pyrrolo[2,3-b]pyridin-3-yl)pyridin-3-yl]oxy}-2,4-dimethylpentan-2-amine (Compound 2)

[0091]

[0092] Step 1: D-alanine (43 g, 428.90 mmol, 1.0 eq) and sodium carbonate (153.55 g, 1448.70 mmol, 3.8 eq) were dissolved in pure water (540 mL), benzyl chloroformate (93.90 g, 550.50 mmol, 1.3 eq) was slowly added at 0 °C, and the reaction was allowed to proceed at room temperature for 16 hours. After the reaction was completed, water (1 L) and ethyl acetate (1 L) were added to extract the solution, the aqueous phase was adjusted to pH 2 with 4 mol / L dilute hydrochloric acid, and then ethyl acetate (1 L) was added to extract the solution. The organic phase was dried over anhydrous sodium sulfate and concentrated to obtain white solid [(benzyloxy)carbonyl]-D-alanine (55 g, yield 51.40%). LCMS (TOF MS ES + )m / z [M+H] + : 224.08. 1 H NMR (400 MHz, DMSO) δ 12.55 (s, 1H), 7.62 (d, J = 7.6 Hz, 1H), 7.41 - 7.24 (m, 5H), 5.02 (s, 2H), 4.05 - 3.99 (m, 1H), 1.26 (d, J = 7.3 Hz, 3H).

[0093] Step 2: Compound [(benzyloxy)carbonyl]-D-alanine (55 g, 246.40 mmol, 1.0 eq) and benzaldehyde dimethyl acetal (45 g, 295.68 mmol, 1.2 eq) were dissolved in tetrahydrofuran (400 mL), and dichlorosulfoxide (35.20 g, 295.68 mmol, 1.2 eq) was slowly added at 0 °C. After half an hour of reaction at 0 °C, zinc chloride (40.30 g, 295.68 mmol, 1.2 eq) was added, and the reaction was allowed to proceed at 0 °C for 4 hours. After the reaction was completed, the reaction solution was diluted in 1000 ml of water, and ethyl acetate (500 mL) was added to extract the solution three times. The combined ethyl acetate phase was washed twice with water (800 mL) and once with saturated aqueous sodium chloride solution. The organic phase was dried over anhydrous sodium sulfate, filtered, and concentrated. Purification by normal phase column chromatography (petroleum ether: ethyl acetate = 10:1) yielded (4S)-4-methyl-5-oxo-2-phenyloxazolidine-3-carboxylic acid benzyl ester (38 g, 49.54% yield) as a light yellow oil. LCMS (TOF MS ES + )m / z [M+H] + : 312.12. 1H NMR (400 MHz, DMSO) δ 7.53 - 7.41 (m, 5H), 7.41 - 7.18 (m, 5H), 6.58 (s, 1H), 5.06 (d, J = 26.7 Hz, 2H), 4.56 (q, J = 7.0 Hz, 1H), 1.52 (d, J = 7.0 Hz, 3H).

[0094] Step 3: (4S)-4-methyl-5-oxo-2-phenyloxazolidine-3-carboxylic acid benzyl ester (38 g, 122.2 mmol, 1.0 eq) and 3-bromo-2-methyl-1-propene (20 g, 152.6 mmol, 1.25 eq) were dissolved in tetrahydrofuran, and lithium bis(trimethylsilyl)amide (183.07 mL, 183.07 mmol, 1.5 eq) was added dropwise at -78 °C under nitrogen protection. The reaction was stirred for 3 hours. After the reaction was completed, saturated aqueous ammonium chloride solution (15 mL) was added dropwise in an ice bath, and then extracted with ethyl acetate and water. The organic layer was collected and washed with water twice and saturated brine once. Normal phase column separation was performed with 6% ethyl acetate to obtain benzyl (4R)-4-methyl-4-(2-methylallyl)-5-oxo-2-phenyloxazolidine-3-carboxylate (19 g, yield 43.20%) as a transparent oil. LCMS (TOF MS ES + m / z [M+H] + : 366.16. 1 H NMR (400 MHz, DMSO) δ 7.45 (m, J = 11.4, 5.9 Hz, 7H), 7.19 (m, J = 14.6, 7.2 Hz, 2H), 6.79 (d, J = 7.3 Hz, 1H), 6.45 (d, J = 26.5 Hz, 1H), 5.21 - 5.07 (m, 1H), 5.03 - 4.85 (m, 2H), 4.74 - 4.54 (m, 1H), 3.20 - 2.85 (m, 1H), 2.42 (m, J = 13.8, 8.9 Hz, 1H), 1.82 - 1.56 (m, 6H).

[0095] Step 4: Benzyl (4R)-4-methyl-4-(2-methylallyl)-5-oxo-2-phenyloxazolidine-3-carboxylate (19 g, 52 mmol, 1.0 eq) was dissolved in tetrahydrofuran, and lithium hydroxide hydrate (2.4 g, 57.20 mmol, 1, 1 eq) was dissolved in water and added dropwise to the reaction system at 0 °C, and then the reaction was allowed to proceed to room temperature for 16 hours. After the reaction was completed, extraction was performed with ethyl acetate and water, the pH of the aqueous phase was adjusted to 5-6, and then extraction was performed with ethyl acetate and water. The organic phase was concentrated to obtain (R)-2-[(benzyloxy carbonyl)amino]-2,4-dimethylpent-4-enoic acid (9.0 g, yield 42.10%) as a transparent oil. LCMS (TOF MS ES+ m / z [M+H] 278.2 + :278. 1 H NMR (400 MHz, DMSO) δ 12.45 (s, 1H), 7.35 - 7.17 (m, 5H), 4.96 (d, J = 3.7 Hz, 2H), 4.78 (dd, J = 2.7, 1.5 Hz, 1H), 4.63 (d, J = 2.5 Hz, 1H), 2.63 - 2.36 (m, 2H), 1.61 (s, 3H), 1.26 (s, 3H).

[0096] Step 5: (R)-2-[(benzyloxy carbonyl)amino]-2,4-dimethylpent-4-enoic acid (9.0 g, 32.5 mmol, 1.0 eq) and triethylamine (4.9 g, 48.75 mmol, 1.5 eq) were dissolved in THF (30 mL), under nitrogen protection, isobutyl chloroformate (5.3 g, 39 mmol, 1.2 eq) was added dropwise at -15 °C, after 0.5 h at this temperature, filtration was carried out, the filtrate was reserved, sodium borohydride aqueous solution was added to the filtrate at 0 °C, and it was reacted at room temperature for 10 min. After the reaction was completed, water was added to quench the sodium borohydride, and ethyl acetate and water were used for extraction and separation, the organic phase was concentrated, and normal phase column separation was carried out, and the target product was obtained when the ethyl acetate content was 30%, and the concentrated transparent oil was benzyl (R)-(1-hydroxy-2,4-dimethylpent-4-en-2-yl)carbamate (3.8 g, yield 44.46%). LCMS (TOF MS ES + m / z [M+H] 264 / 265 + :264 / 265. 1 H NMR (400 MHz, DMSO) δ 7.42 - 7.25 (m, 5H), 6.73 (s, 1H), 5.07 - 4.91 (m, 2H), 4.79 (m, J = 3.0, 1.6 Hz, 1H), 4.71 (t, J = 5.7 Hz, 1H), 4.64 (d, J = 2.7 Hz, 1H), 3.46 - 3.32 (m, 2H), 2.47 (s, 1H), 2.21 (d, J = 13.2 Hz, 1H), 1.68 (s, 3H), 1.10 (s, 3H).

[0097] Step 6: Compound benzyl (R)-(l-hydroxy-2,4-dimethylpent-4-en-2-yl)carbamate (1.5 g, 5.70 mmol) was dissolved in methanol (20.0 mL), palladium on carbon (150 mg, 10%) was added and the reaction was stirred at room temperature for 16 hours under hydrogen atmosphere. After completion of the reaction, celite was added to the reaction mixture and filtered. The filtrate was concentrated and diluted with water (100 mL) and ethyl acetate (100 mL). The organic layer was separated and washed with water (100 mL) twice and saturated sodium chloride solution (100 mL) once. The organic layer was dried over anhydrous sodium sulfate and concentrated to get the product (R)-2-amino-2,4-dimethylpentan-l-ol (620 mg, 82.90% yield) as a white solid. LCMS (TOF MS ES + m / z [M+H] + : 132.

[0098] Step 7: Compound (R)-2-amino-2,4-dimethylpentan-l-ol (80 mg, 4.42 mmol, 1.0 eq) was taken in tetrahydrofuran (30.0 mL) followed by the addition of potassium tert-butoxide (1.24 g, 11.05 mmol, 2.5 eq) and 6-bromo-2-(difluoromethyl)-3-fluoropyridine (1.49 g, 6.63 mmol, 1.5 eq) and the reaction was stirred at 93 °C for 2 hours. After completion of the reaction, the reaction mixture was filtered and the filtrate was concentrated and purified by normal phase column chromatography to get the product (R)-l-{[6-bromo-2-(difluoromethyl)pyridin-3-yl]oxy}-2,4-dimethylpentan-2-amine (926 mg, 62.11% yield) as a brown oil. LCMS (TOF MS ES + m / z [M+H] + : 337 / 339. 1 H NMR (400 MHz, DMSO) δ 7.79 (d, J = 8.8 Hz, 1H), 7.66 (d, J = 8.9 Hz, 1H), 7.16 (t, J = 53.4 Hz, 1H), 3.81 (s, 2H), 1.79 (m, J = 12.7, 6.3 Hz, 1H), 1.68 (s, 2H), 1.10 (s, 3H), 0.92 (m, J = 8.4, 6.6 Hz, 6H).

[0099] Step 8: (R)-1-{[6-bromo-2-(difluoromethyl)pyridin-3-yl]oxy}-2,4-dimethylpentan-2- amine (40 mg, 0.12 mmol, 1.0 eq), 7-azaindole-3-boronic acid pinacol ester (28.95 mg, 0.12 mmol, 1.0 eq), sodium carbonate (50 mg, 0.48 mmol, 4.0 eq) and [1,1'- bis(diphenylphosphino)ferrocene]dichloropalladium (8.78 mg, 0.12 mmol, 1.0 eq) were added into 1,4-dioxane (3.0 mL) and reacted at 120 °C for 6 h under nitrogen protection. After the reaction was completed, it was filtered and the filtrate was prepared by C18 column reverse phase system to obtain white solid (R)-1-{[2-(difluoromethyl)-6-(1H-pyrrolo[2,3-b]pyridin-3-yl)pyridin-3-yl]oxy}- 2,4-dimethylpentan-2-amine (8.60 mg, yield 19.32%). LCMS (TOF MSES + m / z [M+H] + : 375.2. 1 H NMR (400 MHz, MeOD) δ 8.95 (m, J = 8.0, 1.7 Hz, 1H), 8.55 (s, 1H), 8.29 (d, J = 4.7 Hz, 1H), 8.06 (d, J = 1.7 Hz, 1H), 7.99 (d, J = 8.8 Hz, 1H), 7.69 (d, J = 8.9 Hz, 1H), 7.34 - 7.03 (m, 2H), 4.29 - 4.23 (m, 1H), 4.21 - 4.14 (m, 1H), 1.95 - 1.83 (m, 2H), 1.77 - 1.67 (m, 1H), 1.53 (d, J = 1.7 Hz, 3H), 1.08 (m, J = 15.5, 6.5, 1.8 Hz, 6H).

[0100] Example 16: Synthesis of (S)-1-{[2-(difluoromethyl)-6-(4-methoxy-1H-pyrrolo[2,3-b]pyridin-3- yl)pyridin-3-yl]oxy}-2,4-dimethylpentan-2-amine trifluoroacetate salt (Compound 16)

[0101]

[0102] Step 1: Synthesis of 3-iodo-4-methoxy-1-tosyl-1H-pyrrolo[2,3-b]pyridine

[0103] Compound 4-methoxy-7-azaindole (200 mg, 1.35 mmol, 1.0 eq) and potassium hydroxide (189 mg, 3.37 mmol, 2.5 eq) were added into N,N-dimethylformamide (3.0 mL), and iodine (360 mg, 1.42 mmol, 1.05 eq) was dissolved in N,N-dimethylformamide (2.0 mL) and then slowly added into the above system, and the reaction was carried out at room temperature for 45 minutes, and then additional potassium hydroxide (189 mg, 3.37 mmol, 2.5 eq) was added, and the reaction was carried out at room temperature for 10 minutes, and then p-toluenesulfonyl chloride (540 mg, 2.83 mmol, 2.1 eq) was added, and the reaction was carried out at room temperature for 2.5 hours. After the reaction was completed, it was poured into water (20 mL), and a solid was precipitated, and it was filtered, and the filter cake was washed with water (10 mL) and ethyl acetate (20 mL) respectively, and the filter cake was collected and dried to obtain a yellow solid 3-iodo-4-methoxy-1-tosyl-1H-pyrrolo[2,3-b]pyridine (360 mg, yield 62.28%). LCMS (TOF MS ES+) m / z [M+H]+: 428.9760. 1 HNMR (400 MHz, DMSO) δ 8.29 (d, J = 5.7 Hz, 1H), 8.04 - 7.98 (m, 2H), 7.90 (s, 1H), 7.44 (d, J = 8.1 Hz, 2H), 6.94 (d, J = 5.7 Hz, 1H), 3.94 (s, 3H), 2.36 (s, 3H).

[0104] Step 2: Synthesis of (4-methoxy-1-tosyl-1H-pyrrolo[2,3-b]pyridin-3-yl)boronic acid

[0105] 3-iodo-4-methoxy-1-tosyl-1H-pyrrolo[2,3-b]pyridine (310 mg, 0.72 mmol, 1.0 eq), pinacol borane (463 mg, 3.62 mmol, 5.0 eq), triethylamine (366 mg, 3.62 mmol, 5.0 eq) and [1,1'-bis(diphenylphosphino)ferrocene]palladium dichloride (52 mg, 0.072 mmol, 0.10 eq) were added into 1,4-dioxane (3.0 mL), and the reaction was carried out at 120 °C for 2 hours under nitrogen protection. After the reaction was completed, it was filtered, and the filtrate was concentrated and purified by C18 column reverse phase to obtain a yellow solid (4-methoxy-1-tosyl-1H-pyrrolo[2,3-b]pyridin-3-yl)boronic acid (150 mg, yield 59.86%). LCMS (TOF MS ES+) m / z [M+H]+: 347.1177. 1H NMR (400 MHz, DMSO) δ 8.28 (d, J = 5.7 Hz, 1H), 8.04 - 7.98 (m, 3H), 7.86 (s, 2H), 7.43 (d, J = 8.1 Hz, 2H), 6.99 (d, J = 5.7 Hz, 1H), 4.02 (s, 3H).

[0106] Step 3: Synthesis of (S)-1-{[2-(difluoromethyl)-6-(4-methoxy-1- methylbenzenesulfonyl-1H-pyrrolo[2,3-b]pyridin-3-yl)pyridin-3-yl]oxy}-2,4- dimethylpentan-2-amine

[0107] (4-methoxy-1-methylbenzenesulfonyl-1H-pyrrolo[2,3-b]pyridin-3-yl)boronic acid (100 mg, 0.29 mmol, 1.0 eq), (S)-1-{[6-bromo-2-(difluoromethyl)pyridin-3-yl]oxy}-2,4- dimethylpentan-2-amine (97.41 mg, 0.29 mmol, 1.0 eq), sodium carbonate (122.50 mg, 1.06 mmol, 4.0 eq) and [1,1'-bis(diphenylphosphino)ferrocene]dichloropalladium (21.14 mg, 0.029 mmol, 0.10 eq) were added into 1,4-dioxane (3.0 mL), and reacted at 120 °C for 8 hours under nitrogen protection. After the reaction was completed, it was filtered, and the filtrate was concentrated and subjected to normal phase column chromatography (dichloromethane:methanol = 12:1) to obtain the product (S)-1-{[2-(difluoromethyl)-6-(4-methoxy-1- methylbenzenesulfonyl-1H-pyrrolo[2,3-b]pyridin-3-yl)pyridin-3-yl]oxy}-2,4- dimethylpentan-2-amine (150 mg, yield 92.95%) as brown oil. LCMS (TOFMS ES+) m / z [M+H]+: 559.2274. 1 H NMR (400 MHz, DMSO) δ 8.37 - 8.29 (m, 1H), 8.07 (d, J = 8.0 Hz, 2H), 8.03 - 7.97 (m, 2H), 7.70 (d, J = 8.9 Hz, 1H), 7.45 (d, J = 8.0 Hz, 2H), 7.38 - 7.08 (m, 1H), 7.04 - 6.97 (m, 1H), 4.12 - 3.98 (m, 2H), 3.91 (d, J = 2.2 Hz, 3H), 2.37 (s, 3H), 2.00 (d, J = 2.2 Hz, 2H), 1.81 (d, J = 7.5 Hz, 1H), 1.18 - 1.16 (m, 3H), 0.95 (ddd, J = 9.0, 6.9, 2.1 Hz, 6H).

[0108] Step 4: Synthesis of (S)-1-{[2-(difluoromethyl)-6-(4-methoxy-1H-pyrrolo[2,3- b]pyridin-3-yl)pyridin-3-yl]oxy}-2,4-dimethylpentan-2-amine trifluoroacetate salt

[0109] (S)-1-{[2-(difluoromethyl)-6-(4-methoxy-1-methylsulfonyl-1H-pyrrolo[2,3-b]pyridin-3- yl)pyridin-3-yl]oxy}-2,4-dimethylpentan-2-amine (150 mg, 0.27 mmol, 1.0 eq) was added to hydrogen chloride-dioxane solution (5.0 mL, 4.0 mol / L) and reacted at 60 °C for 16 hours. After the reaction was completed, it was directly filtered, the filter cake was washed with a mixed solvent (petroleum ether: ethyl acetate = 5:1, 20 mL) to remove excess impurities, the filter cake was collected, and reversed phase preparation (formic acid system) was performed on a C18 column to obtain white solid (S)-1-{[2-(difluoromethyl)-6-(4-methoxy-1H-pyrrolo[2,3-b]pyridin-3-yl)pyridin-3-yl]oxy}-2,4-dimethylpentan-2-amine trifluoroacetate salt (6.2 mg, yield 4.26%). LCMS (TOF MS ES+) m / z [M+H]+: 391.2328. 1 H NMR (400 MHz, DMSO) δ 12.12 (s, 1H), 8.23 (d, J = 5.6 Hz, 1H), 8.07 (d, J = 8.9 Hz, 1H), 7.77-7.71 (m, 2H), 7.61-7.30 (m, 1H), 6.85 (d, J = 5.7 Hz, 1H), 4.25-4.11 (m, 2H), 3.98 (s, 3H), 1.89-1.71 (m, 2H), 1.61 (dd, J = 14.1, 5.3 Hz, 1H), 1.40 (s, 3H), 1.04-0.90 (m, 6H).

[0110] Example 17: Synthesis of (R)-1-{[2-(difluoromethyl)-6-(4-methoxy-1H-pyrrolo[2,3- b]pyridin-3-yl)pyridin-3-yl]oxy}-2,4-dimethylpentan-2-amine (Compound 17)

[0111]

[0112] Step 1: Synthesis of [(benzyloxy)carbonyl]-L-alanine

[0113] L-alanine (50 g, 561.15 mmol, 1.0 eq) and sodium carbonate (118.72 g, 1122.96 mmol, 2.0 eq) were dissolved in water (540 mL). Benzyl chloroformate (105.36 g, 550.50 mmol, 1.10 eq) was added to a 1,4-dioxane (83 mL) clock, and benzyl chloroformate (105.36 g, 550.50 mmol, 1.10 eq) was slowly added dropwise at 0 °C. After 0.5 hours of reaction at 0 °C, the reaction was allowed to proceed at room temperature for 16 hours. After the reaction was completed, water (500 mL) and ethyl acetate (750 mL) were added and extracted twice, and the aqueous phase was adjusted to pH 2-4 with 4 mol / L dilute hydrochloric acid, then ethyl acetate (1 L) was added and extracted twice. The organic phase was dried over anhydrous sodium sulfate, and concentrated to obtain a white solid [(benzyloxy)carbonyl]-L-alanine (101.00 g, yield 80.62%). 1 H NMR (400 MHz, DMSO) δ 12.52 (s, 1H), 7.60 (d, J = 7.6 Hz, 1H), 7.44 - 7.16 (m, 65H), 5.02 (s, 2H), 4.07 - 3.94 (m, 1H), 1.28 - 1.13 (m, 3H).

[0114] Step 2: Synthesis of benzyl (4R)-4-methyl-5-oxo-2-phenyloxazolidine-3-carboxylate

[0115] Compound [(benzyloxy)carbonyl]-L-alanine (101 g, 452.71 mmol, 1.0 eq) and benzaldehyde dimethyl acetal (72.29 g, 475.37 mmol, 1.05 eq) were dissolved in tetrahydrofuran (500 mL), and dichlorosulfoxide (64.62 g, 543.24 mmol, 1.2 eq) was slowly added at 0 °C. After half an hour of reaction, zinc chloride (74.04 g, 543.26 mmol, 1.2 eq) was added, and the reaction was continued at 0 °C for 4 hours. After the reaction was completed, the reaction solution was diluted in water (1 L), extracted with ethyl acetate (500 mL) three times, and the combined ethyl acetate phase was washed with water (800 mL) twice and then with saturated sodium chloride aqueous solution once. The organic phase was dried over anhydrous sodium sulfate and concentrated, and (4R)-4-methyl-5-oxo-2-phenyloxazolidine-3-carboxylate benzyl ester was obtained as a light yellow oil (80.7 g, yield 57.29%) by normal phase column chromatography. 1 H NMR (400 MHz, DMSO) δ 7.49 - 7.36 (m, 6H), 7.36 - 7.24 (m, 4H), 6.54 (s, 1H), 5.16 - 4.84 (m, 2H), 4.52 (q, J = 7.0 Hz, 1H), 1.48 (d, J = 7.0 Hz, 3H).

[0116] Step 3: Synthesis of benzyl (4R)-4-methyl-4-(2-methylallyl)-5-oxo-2- phenyloxazolidine-3-carboxylate

[0117] Benzyl (4R)-4-methyl-5-oxo-2-phenyloxazolidine-3-carboxylate (80.7 g, 259.5 mmol, 1.0 eq) and 3-bromo-2-methyl-1-propene (42.5 g, 324.1 mmol, 1.25 eq) were dissolved in tetrahydrofuran (500 mL) under nitrogen protection, and lithium bis(trimethylsilyl)amide (388 mL, 389.1 mmol, 1.5 eq) was added dropwise at -78 °C over 15 min. The reaction was allowed to proceed for 3 h. After the reaction was completed, saturated aqueous ammonium chloride solution (25 mL) was added dropwise in an ice bath, and then extracted with ethyl acetate and water. The organic layer was collected and washed with water twice and saturated brine once. Normal phase column separation was performed, and the product was eluted with ethyl acetate content of 6%, and concentrated to give benzyl (4R)-4-methyl-4-(2-methylallyl)-5-oxo-2-phenyloxazolidine-3-carboxylate (40.76 g, yield 43.03%) as a transparent oil. LCMS (ESI) [M+H]+: 366. 1 H NMR (400 MHz, DMSO) δ 7.47 - 7.32 (m, 7H), 7.16 (m, J = 14.6, 7.2 Hz, 2H), 6.76 (d, J = 7.3 Hz, 1H), 6.49 - 6.33 (m, 1H), 5.16 - 5.04 (m, 1H), 4.98 - 4.83 (m, 2H), 4.70 - 4.51 (m, 1H), 3.16 - 2.78 (m, 1H), 2.39 (dd, J = 13.8, 8.5 Hz, 1H), 1.76 - 1.50 (m, 6H).

[0118] Step 4: Synthesis of (R)-2-[(benzyloxy carbonyl)amino]-2,4-dimethylpent-4-enoic acid

[0119] Benzyl (4R)-4-methyl-4-(2-methylallyl)-5-oxo-2-phenyloxazolidine-3-carboxylate (16.72 g, 45.8 mmol, 1.0 eq) was dissolved in tetrahydrofuran (100 mL), and lithium hydroxide hydrate (2.88 g, 68.67 mmol, 1.15 eq) was dissolved in water (100 mL) and added dropwise to the reaction system at 0 °C, and then the reaction was allowed to proceed at room temperature for 16 h. After the reaction was completed, extraction was performed with ethyl acetate and water, the aqueous phase was adjusted to pH = 5-6, and extraction was performed with ethyl acetate and water three times. The organic phase was concentrated to give a transparent oil (R)-2-[(benzyloxy)carbonyl]amino]-2,4-dimethylpent-4-enoic acid (6.84 g, yield 53.91 %). LCMS (ESI) [M+H]+: 278. 1 H NMR (400 MHz, DMSO) δ 12.59 (s, 1H), 7.35-7.19 (m, 5H), 4.95 (s, 2H), 4.75 (dd, J = 2.7, 1.5 Hz, 1H), 4.63-4.58 (m, 1H), 2.56 (d, J = 13.4 Hz, 1H), 2.44-2.40 (m, 1H), 1.59 (s, 3H), 1.26 (s, 3H).

[0120] Step 5: Synthesis of (R)-(l-hydroxy-2,4-dimethylpent-4-en-2-yl) carbamate

[0121] (R)-2-((benzyloxy)carbonyl)amino)-2,4-dimethylpent-4-enoic acid (6.8 g, 24.5 mmol, 1.0 eq) and triethylamine (3.71 g, 36.7 mmol, 1.5 eq) were dissolved in tetrahydrofuran (25 mL). Under nitrogen protection, isobutyl chloroformate (4.02 g, 24.3 mmol, 1.2 eq) was added dropwise to the system at -15 °C, and the reaction was allowed to proceed at this temperature for 0.5 h, then filtered, and the filtrate was retained. A sodium borohydride (3.7 g, 98.08 mmol, 4.0 eq) solution in water (15 mL) was added to the filtrate at 0 °C, and the reaction was allowed to proceed at room temperature for 10 min. After the reaction was completed, the sodium borohydride was quenched with water, and extraction was performed with ethyl acetate and water. The organic phase was concentrated, and normal phase column separation was performed. When the ethyl acetate content was 23 %, the target product was eluted, and the transparent oil benzyl (R)-(l-hydroxy-2,4-dimethylpent-4-en-2-yl) carbamate (4.26 g, yield 66.01 %) was obtained. LCMS (ESI) [M+H]+: 264. 1H NMR (400 MHz, DMSO) δ 7.37 - 7.22 (m, 5H), 6.66 (s, 1H), 5.01 - 4.88 (m, 2H), 4.76 (m, J = 3.0, 1.5 Hz, 1H), 4.70 - 4.58 (m, 2H), 3.43 - 3.31 (m, 2H), 2.44 (s, 1H), 2.18 (d, J = 13.2 Hz, 1H), 1.65 (s, 3H), 1.08 (s, 3H).

[0122] Step 6: Synthesis of (R)-2-amino-2,4-dimethylpentan-1-ol

[0123] Benzyl (R)-(1-hydroxy-2,4-dimethylpent-4-en-2-yl)carbamate (2.0 g, 7.6 mmol, 1.0 eq) was added to methanol (25 mL) followed by Pd / C (0.2 g, 10% wt) under hydrogen atmosphere. The reaction was stirred at room temperature for 14 h. After completion of the reaction, it was filtered and the filtrate was concentrated to get yellow oil (R)-2-amino-2,4-dimethylpentan-1-ol (1.06 g, crude). No further purification was done and it was used as such in the next step. 1 H NMR (400 MHz, DMSO) δ 3.07 (s, 2H), 1.79 - 1.67 (m, 1H), 1.28 - 1.08 (m, 2H), 0.97 - 0.74 (m, 9H).

[0124] Step 7: Synthesis of (R)-1-{[6-bromo-2-(difluoromethyl)pyridin-3-yl]oxy}-2,4- dimethylpentan-2-amine

[0125] Compound (R)-2-amino-2,4-dimethylpentan-1-ol (200 mg, crude) was taken in tetrahydrofuran (4 mL) followed by potassium tert-butoxide (4.0 g, 11.05 mmol, 2.5 eq) and 6-bromo-2-(difluoromethyl)-3-fluoropyridine (350 mg, 1.55 mmol, 1.0 eq) and the reaction was stirred at 80 °C for 3 h. After completion of the reaction, it was filtered and the filtrate was concentrated to get brown oily liquid (R)-1-{[6-bromo-2-(difluoromethyl)pyridin-3-yl]oxy}-2,4-dimethylpentan-2-amine (255 mg, crude) which was used as such in the next step without further purification. LCMS (TOF MS ES+) m / z [M+H]+: 337. + H] + :337. 1H NMR (400 MHz, DMSO) δ 7.75 (d, J = 8.8 Hz, 1H), 7.63 (d, J = 8.8 Hz, 1H), 7.38 - 6.78 (m, 1H), 3.80 (s, 2H), 2.02 - 1.59 (m, 2H), 1.48 - 1.26 (m, 2H), 1.21 (d, J = 8.7 Hz, 1H), 1.08 (s, 3H), 0.89 (dd, J = 8.7, 6.6 Hz, 6H).

[0126] Step 8: Synthesis of (R)-1-{[2-(difluoromethyl)-6-(4-methoxy-1- methylbenzenesulfonyl-1H-pyrrolo[2,3-b]pyridin-3-yl)pyridin-3-yl]oxy}-2,4- dimethylpentan-2-amine

[0127] (R)-1-{[6-bromo-2-(difluoromethyl)pyridin-3-yl]oxy}-2,4-dimethylpentan-2-amine (255 mg, crude), (4-methoxy-1-methylbenzenesulfonyl-1H-pyrrolo[2,3-b]pyridin-3- yl)boronic acid (261 mg, 0.756 mmol, 1.0 eq), sodium carbonate (320.58 mg, 3.02 mmol, 4.0 eq) and [1,1'-bis(diphenylphosphino)ferrocene]dichloropalladium (55.4 mg, 0.0756 mmol, 0.1 eq) were added into 1,4-dioxane (10 mL) and reacted at 120 °C for 12 h under nitrogen protection. After the reaction was completed, it was filtered and the filtrate was concentrated and purified by normal phase column chromatography (dichloromethane:methanol = 19:1) to give brown solid product (R)-1-{[2-(difluoromethyl)-6-(4-methoxy-1-methylbenzenesulfonyl-1H- pyrrolo[2,3-b]pyridin-3-yl)pyridin-3-yl]oxy}-2,4-dimethylpentan-2-amine (364 mg, yield 86.23%). LCMS (TOF MS ES+) m / z [M+H]+: 559. 1H NMR (400 MHz, DMSO) δ 8.27 (d, J = 5.6 Hz, 1H), 8.13 - 7.96 (m, 3H), 7.93 (d, J = 10.9 Hz, 3H), 7.64 (d, J = 8.9 Hz, 1H), 7.39 (d, J = 8.0 Hz, 2H), 7.29 - 6.98 (m, 1H), 6.95 (d, J = 5.7 Hz, 1H), 3.87 (s, 3H), 3.81 (s, 2H), 2.32 (s, 3H), 1.42 - 1.31 (m, 2H), 1.19 (d, J = 5.5 Hz, 1H), 1.10 (s, 3H), 0.89 (t, J = 7.1 Hz, 6H).

[0128] Step 9: Synthesis of (R)-1-{[2-(difluoromethyl)-6-(4-methoxy-1H-pyrrolo[2,3- b]pyridin-3-yl)pyridin-3-yl]oxy}-2,4-dimethylpentan-2-amine

[0129] (R)-1-{[2-(difluoromethyl)-6-(4-methoxy-1-methylsulfonyl-1H-pyrrolo[2,3-b]pyridin-3- yl)pyridin-3-yloxy]-2,4-dimethylpentan-2-amine (177 mg, 0.44 mmol, 1.0 eq) was added to hydrogen chloride-dioxane solution (3.5 mL, 4.0 mol / L) and reacted at 60 °C for 1 hour, then at room temperature for 16 hours. After the reaction was completed, an insoluble solid was precipitated, and after filtration, the filtrate was concentrated, and the solid and the filtrate were dissolved with methanol (3 mL) to prepare a white solid (R)-1-{[2-(difluoromethyl)-6-(4-methoxy-1H-pyrrolo[2,3-b]pyridin-3-yl)pyridin-3- yl]oxy}-2,4-dimethylpentan-2-amine (16.1 mg, yield 12.56%). LCMS (TOF MS ES+) m / z [M+H]+: 405.2290. + H] + :405.2290. 1 H NMR (400 MHz, DMSO) δ 11.91 (s, 1H), 8.13 (d, J = 5.6 Hz, 1H), 7.99 (d, J = 8.8 Hz, 1H), 7.67 - 7.56 (m, 2H), 7.29 - 6.87 (m, 1H), 6.73 (d, J = 5.7 Hz, 1H), 3.91 (s, 3H), 3.77 (s, 2H), 1.78 (m, J = 6.4 Hz, 1H), 1.41 - 1.30 (m, 2H), 1.09 (s, 3H), 0.90 (t, J = 6.7 Hz, 6H).

[0130] Example 26: Synthesis of (S)-1-((2-(difluoromethyl)-6-(5-fluoro-1H-pyrrolo[2,3-b]pyridin-3- yl)pyridin-3-yl)oxy)-2,4-dimethylpentan-2-amine (Compound 26)

[0131]

[0132] Step 1 : 5-Fluoro-lH-pyrrolo[2,3-b]pyridine (600 mg, 4.4 mmol, 1.0 eq) and potassium hydroxide (616.0 mg, 11.0 mmol, 2.5 eq) were added to N,N-dimethylformamide (5 mL), to the system was slowly added iodine (586.7 mg, 4.62 mmol, 1.05 eq) dissolved in N,N-dimethylformamide (5 mL), and the reaction was carried out for 45 minutes. Potassium hydroxide (616.0 mg, 11.0 mmol, 2.5 eq) was added, and the reaction was continued for 5 minutes, and then p-toluenesulfonyl chloride (1.76 g, 9.24 mmol, 2.1 eq) dissolved in N,N-dimethylformamide (3 mL) was slowly added dropwise to the system, and the reaction was carried out at room temperature for 3 hours. After the reaction was completed, the system was poured into water (50 mL), and after stirring for 10 minutes, the filter cake was directly filtered, washed with water (20 mL) and ethyl acetate (20 mL), and dried to obtain a yellow solid product 5-fluoro-3-iodo-l-p-toluenesulfonyl-lH-pyrrolo[2,3-b]pyridine (957 mg, 69.75%).

[0133] Step 2: 6-Bromo-3-fluoropyridinecarboxaldehyde (20 g, 98.04 mmol, 1.0 eq) was dissolved in super dry dichloromethane (200 mL), and diethylamine sulfide trifluoride (27.2 mL, 196.08 mmol, 2.0 eq) was added dropwise to the system at -20°C, and the reaction was carried out at room temperature for 4 hours. After the reaction was completed, the system was poured into cold saturated aqueous sodium bicarbonate solution (200 mL) to quench, dichloromethane (200 mL) was added to extract the liquid, the organic phase was collected, and washed with water (250 mL) twice, dried and concentrated to obtain a red solid, and then silica gel was added to the filter funnel to decolorize to obtain white crystalline 6-bromo-2-(difluoromethyl)-3-fluoropyridine (19.0 g, yield 85.75%). LCMS (TOF MS ES+) m / z [M+H]+: 225.94. 1 H NMR (400 MHz, DMSO) δ 7.97 (d, J = 6.7 Hz, 2H), 7.33 - 6.78 (m, 1H).

[0134] Step 3: Compound (S)-2-amino-2,4-dimethylpentan-1-ol (17.74 g, 135 mmol, 1 eq) was added to tetrahydrofuran (450 mL), followed by the addition of potassium tert-butoxide (37.87 g, 337.5 mmol, 2.5 eq) and 6-bromo-2-(difluoromethyl)-3-fluoropyridine (36.61 g, 162 mmol, 1.2 eq) at 93 °C for 2 hours. After the reaction was completed, it was filtered, the filtrate was concentrated, and purified by normal phase column chromatography (dichloromethane:methanol = 95:5), followed by reverse phase column again, and the target product was eluted when the ratio of acetonitrile to water (1% formic acid) was 13:87, and concentrated to give the product (S)-1-{[6-bromo-2-(difluoromethyl)pyridin-3-yl]oxy}-2,4-dimethylpentan-2-amine (11.80 g, 25.88%) as a light yellow oil. LCMS (TOF MS ES+) m / z [M+H]+: 337 / 339. + H] + :337 / 339. 1 H NMR (400 MHz, DMSO) δ 7.79 (d, J = 8.8 Hz, 1H), 7.66 (d, J = 8.9 Hz, 1H), 7.16 (t, J = 53.4 Hz, 1H), 3.81 (s, 2H), 1.79 (m, J = 12.7, 6.3 Hz, 1H), 1.38 - 1.34 (m, 2H), 1.10 (s, 3H), 0.92 (m, J = 8.4, 6.6 Hz, 6H).

[0135] Step 4: Compound (S)-1-{[6-bromo-2-(difluoromethyl)pyridin-3-yl]oxy}-2,4- dimethylpentan-2-amine (450 mg, 1.35 mmol, 1.0 eq) and bis(pinacolato)diboron (405 mg, 1.59 mmol, 1.2 eq) were dissolved in 1,4-dioxane (15.0 mL), followed by the addition of potassium acetate (393 mg, 4.02 mmol, 3.0 eq) and [1,1'- bis(diphenylphosphino)ferrocene]dichloropalladium (58.5 mg, 0.08 mmol, 0.06 eq) and reacted at 80 °C for 16 hours under nitrogen protection. After the reaction was completed, it was filtered to obtain a crude product of (S)-{5-[(2-amino-2,4-dimethylpentyl)oxy]-6- (difluoromethyl)pyridin-2-yl}boronic acid (4.8 mL). Then, 5-fluoro-3-iodo-1- p-tolylsulfonyl-1H-pyrrolo[2,3-b]pyridine (588 mg, 1.35 mmol, 1.0 eq), [1,1'- bis(diphenylphosphino)ferrocene]dichloropalladium (99.0 mg, 0.135 mmol, 0.1 eq) and sodium carbonate (426 mg, 4.02 mmol, 3.0 eq) were added to the above solution and reacted at 90 °C for 4 hours under nitrogen protection. After the reaction was completed, it was filtered through diatomite and the filtrate was concentrated and purified by normal phase column chromatography (dichloromethane:methanol = 15:1) to obtain a brown solid product of (S)-1-{[2-(difluoromethyl)-6-(5-fluoro-1- toluenesulfonyl-1H-pyrrolo[2,3-b]pyridin-3-yl)pyridin-3-yl]oxy}-2,4-dimethylpentan-2- amine (260.7 mg, 35.75%). 1 H NMR (400 MHz, DMSO) δ 8.81 (s, 1H), 8.71 (dd, J = 9.3, 2.9 Hz, 1H), 8.48 (dd, J = 2.9, 1.2 Hz, 1H), 8.34 (d, J = 8.8 Hz, 1H), 8.04 (d, J = 8.4 Hz, 2H), 7.78 (d, J = 8.9 Hz, 1H), 7.50 - 7.17 (m, 3H), 3.95 (s, 2H), 2.37 (s, 3H), 1.82 (dt, J = 12.7, 6.3 Hz, 1H), 1.43 - 1.39 (m, 2H), 1.33 (d, J = 9.8 Hz, 1H), 1.22 - 1.17 (m, 1H), 1.14 (s, 3H), 0.94 (dd, J = 8.7, 6.6 Hz, 6H).

[0136] Step 5: (S)-1-{[2-(difluoromethyl)-6-(5-fluoro-1-tosyl-1H-pyrrolo[2,3-b]pyridin-3-yl)pyridin-3-yl]oxy}-2,4-dimethylpentan-2-amine (476 mg, 1.21 mmol, 1.0 eq) was added to a solution of hydrogen chloride-dioxane (3 mL, 4.0 mol / L) and reacted at 60 °C for 16 hours. After the reaction was completed, it was directly filtered, the filter cake was washed with a mixed solvent (petroleum ether: ethyl acetate = 5:1, 20 mL) to remove excess impurities, and the filter cake was collected and dried under reduced pressure to obtain the final product (S)-1-{[2-(difluoromethyl)-6-(5-fluoro-1H-pyrrolo[2,3-b]pyridin-3-yl)pyridin-3-yl]oxy}-2,4-dimethylpentan-2-amine trihydrochloride (246 mg, yield 40.46%). LCMS (TOF MS ES+) m / z [M+H]+: 393.19. + H] + :393.19. 1 HNMR (400 MHz, DMSO) δ 12.22 (d, J = 2.9 Hz, 1H), 8.59 (dd, J = 9.9, 2.9 Hz, 1H), 8.39 (d, J = 2.9 Hz, 4H), 8.29 (dd, J = 2.9, 1.5 Hz, 1H), 8.09 (d, J = 8.9 Hz, 1H), 7.81 - 7.46 (m, 2H), 4.25 - 4.13 (m, 2H), 1.86 - 1.72 (m, 2H), 1.62 (dd, J = 14.0, 5.3 Hz, 1H), 1.39 (s, 3H), 0.96 (d, J = 6.5 Hz, 3H), 0.91 (d, J = 6.4 Hz, 3H).

[0137] Step 6: (S)-1-{[2-(difluoromethyl)-6-(5-fluoro-1H-pyrrolo[2,3-b]pyridin-3-yl)pyridin-3-yl]oxy}-2,4-dimethylpentan-2-amine trihydrochloride (100 mg, 0.2 mmol, 1.0 eq) was dissolved in tetrahydrofuran (1 mL), and then the solution was added to a solution of sodium hydroxide (24 mg, 0.6 mmol, 3 eq) in water (1 mL) and reacted at room temperature for 1 hour. After the reaction was completed, it was directly prepared and separated to obtain a white solid (S)-1-{[2-(difluoromethyl)-6-(5-fluoro-1H-pyrrolo[2,3-b]pyridin-3-yl)pyridin-3-yl]oxy}-2,4-dimethylpentan-2-amine (31.7 mg, yield 40.56%). LCMS (TOF MS ES+) m / z [M+H]+: 393.19.

[0138] Example 28: Synthesis of (S)-1-{[2-(difluoromethyl)-6-[5-(difluoromethyl)-1H- pyrrolo[2,3-b]pyridin-3-yl]pyridin-3-yl]oxy}-2,4-dimethylpentan-2-amine salt (Compound 28)

[0139]

[0140] Step 1: Synthesis of 7-azaindole-5-carbaldehyde

[0141] Dissolve 5-bromo-7-azaindole (5 g, 25 mmol, 1.0 eq) in tetrahydrofuran (50 mL), slowly drop in n-butyllithium (21 mL, 2.5 mol / L, 2.1 eq) at -78 °C under nitrogen protection, react for 40 minutes at -78 °C, and slowly drop in super dry N,N-dimethylformamide (5 mL) at this temperature, react for 2 hours at room temperature. After the reaction is completed, add water (200 mL) and extract with ethyl acetate (200 mL) to separate the liquid, wash the organic phase with water (150 mL) for three times, dry with anhydrous sodium sulfate, and purify by normal phase column chromatography (petroleum ether: ethyl acetate = 2:3) to obtain 7-azaindole-5-carbaldehyde (2.4 g, yield 64.43%) as a yellow solid. LCMS (TOF MS ES+) m / z [M+H]+: 147. 1 H NMR (400 MHz, DMSO) δ 12.20 (s, 1H), 10.11 (s, 1H), 8.78 (d, J = 2.0 Hz, 1H), 8.50 (d, J = 1.9 Hz, 1H), 7.67 (d, J = 3.5 Hz, 1H), 6.69 (d, J = 3.5 Hz, 1H).

[0142] Step 2: Synthesis of 3-iodo-1H-pyrrolo[2,3-b]pyridine-5-carbaldehyde

[0143] Compound 7-azaindole-5-carboxaldehyde (2.4 g, 16.42 mmol, 1.0 eq) was dissolved in N,N-dimethylformamide (25 mL), N-iodosuccinimide (3.69 g, 16.42 mmol, 1.0 eq) was added slowly at 0 °C, and the reaction was allowed to proceed at room temperature for 16 h. After the reaction was completed, the reaction solution was diluted in ethyl acetate (200 mL), washed with water (200 mL) three times, and washed with saturated aqueous sodium chloride solution once. The organic phase was dried over anhydrous sodium sulfate, filtered, and concentrated. Purification by normal phase column chromatography (petroleum ether: ethyl acetate = 3: 1) gave 3-iodo-1H-pyrrolo[2,3-b]pyridine-5-carboxaldehyde (4.0 g, 89.55% yield) as a yellow solid. LCMS (TOF MS ES+) m / z [M+H]+: 272.94.1H NMR (400 MHz, DMSO) δ 12.65 (s, 1H), 10.17 (s, 1H), 8.81 (d, J = 1.9 Hz, 1H), 8.22 (d, J = 1.9 Hz, 1H), 7.94 (d, J = 2.5 Hz, 1H).

[0144] Step 3: Synthesis of tert-butyl 5-formyl-3-iodo-1H-pyrrolo[2,3-b]pyridine-1-carboxylate

[0145] tert-Butyl 3-iodo-1H-pyrrolo[2,3-b]pyridine-5-carboxylate (2 g, 7.35 mmol, 1.0 eq) and triethylamine (3.06 mL, 22.06 mmol, 3.0 eq) were dissolved in dichloromethane (25 mL), 4-dimethylaminopyridine (45 mg, 0.37 mmol, 0.05 eq) and di-tert-butyl dicarbonate (1.76 g, 8.09 mmol, 1.1 eq) were added at 0 °C, and the reaction was allowed to proceed at room temperature for 0.5 h. After the reaction was completed, the reaction solution was diluted in ethyl acetate and water, and the organic layer was collected and washed with water twice and saturated brine once. After drying over anhydrous sodium sulfate, the organic phase was filtered and separated by normal phase column chromatography to give tert-butyl 5-formyl-3-iodo-1H-pyrrolo[2,3-b]pyridine-1-carboxylate (1.7 g, yield 62.16%) as a white solid at 20% ethyl acetate. LCMS (TOF MS ES+) m / z [M+H]+: 373. 1 1H NMR (400 MHz, DMSO) δ 10.23 (s, 1H), 8.96 (d, J = 1.9 Hz, 1H), 8.26 (d, J = 1.9 Hz, 1H), 8.20 (s, 1H), 1.64 (s, 9H).

[0146] Step 4: Synthesis of tert-butyl 5-(difluoromethyl)-3-iodo-1H-pyrrolo[2,3-b]pyridine-1-carboxylate

[0147] To a solution of tert-butyl 5-formyl-3-iodo-lH-pyrrolo[2,3-b]pyridine-l- carboxylate (1.0 g, 2.69 mmol, 1.0 eq) in dichloromethane (15 mL) was added diethylamine sulfide (866 mg, 5.38 mmol, 2.0 eq) dropwise at 0 °C, and then the reaction mixture was stirred at room temperature for 5 h. After completion of the reaction, the reaction mixture was extracted with ethyl acetate and water. The organic layer was washed with water three times and then with saturated sodium chloride solution once. The organic layer was dried over anhydrous sodium sulfate, filtered, and concentrated. The residue was purified by column chromatography on silica gel (petroleum ether: ethyl acetate = 4: 1) to give tert-butyl 5-(difluoromethyl)-3-iodo-lH-pyrrolo[2,3-b]pyridine-l- carboxylate (750 mg, 70.69% yield) as a light yellow solid. LCMS (TOF MS ES+) m / z [M+H]+: 395. 1 H NMR (400 MHz, DMSO) δ 8.69 - 8.64 (m, 1H), 8.17 (s, 1H), 8.00 (dt, J = 2.2, 1.2 Hz, 1H), 7.47 - 7.14 (m, 1H), 1.63 (s, 9H).

[0148] Step 5: Synthesis of tert-butyl 5-(difluoromethyl)-3-(4,4,5,5-tetramethyl-l,3,2- dioxaborolan-2-yl)-lH-pyrrolo[2,3-b]pyridine-l-carboxylate

[0149] To a solution of tert-butyl 5-(difluoromethyl)-3-iodo-lH-pyrrolo[2,3-b]pyridine-l- carboxylate (750 mg, 1.90 mmol, 1.0 eq), 4,4,5,5-tetramethyl-l,3,2-dioxaborolane (414 mg, 3.23 mmol, 1.7 eq) and triethylamine (963 mg, 9.51 mmol, 5.0 eq) in 1,4-dioxane (10.0 mL) was added lastly tetrakis(triphenylphosphine)palladium (220 g, 0.19 mmol, 0.1 eq) under nitrogen atmosphere. The reaction mixture was stirred at 80 °C for 16 h. After completion of the reaction, the reaction mixture was filtered and the filtrate was concentrated. The residue was purified by column chromatography on silica gel to give tert-butyl 5-(difluoromethyl)-3-(4,4,5,5-tetramethyl-l,3,2-dioxaborolan-2-yl)-lH- pyrrolo[2,3-b]pyridine-l-carboxylate (115 mg, 15.33% yield) as a yellow solid. LCMS (TOF MS ES+) m / z [M+H]+: 395. 1 H NMR (400 MHz, DMSO) δ 8.63 (m, J = 1.5 Hz, 1H), 8.33 - 8.26 (m, 1H), 8.10 (s, 1H), 7.46 - 7.11 (m, 1H), 1.62 (s, 9H), 1.33 (s, 12H).

[0150] Step 6: Synthesis of (S)-3-{5-[(2-amino-2,4-dimethylpentyl)oxy]-6- (difluoromethyl)pyridin-2-yl}-5-(difluoromethyl)-1H-pyrrolo[2,3-b]pyridine-1- carboxylate tert-butyl ester

[0151] (S)-1-{[6-bromo-2-(difluoromethyl)pyridin-3-yl]oxy}-2,4-dimethylpentan-2-amine (85 mg, 0.25 mmol, 1.0 eq), 5-(difluoromethyl)-3-(4,4,5,5-tetramethyl-1,3,2-dioxaborolan-2-yl)-1H- pyrrolo[2,3-b]pyridine-1-carboxylate tert-butyl ester (100 mg, 0.25 mmol, 1.0 eq), sodium carbonate (107 mg, 1.00 mmol, 4.0 eq) and [1,1'-bis(diphenylphosphino)ferrocene] palladium dichloride (18 mg, 0.025 mmol, 0.10 eq) were added into 1,4-dioxane (5.0 mL), under nitrogen protection, 120 °C for 8 hours. After the reaction was completed, it was filtered and concentrated, and purified by a reverse phase C18 column to obtain the product (S)-3-{5-[(2-amino-2,4-dimethylpentyl)oxy]-6- (difluoromethyl)pyridin-2-yl}-5-(difluoromethyl)-1H-pyrrolo[2,3-b]pyridine-1-carboxylate tert-butyl ester (45 mg, yield 33.83%) as a yellow solid. LCMS (TOF MS ES+) m / z [M+H]+: 525.24. 1 H NMR (400 MHz, DMSO) δ 9.21 (d, J = 2.2 Hz, 1H), 8.69 (s, 1H), 8.66 (s, 1H), 8.32 (d, J = 8.8 Hz, 1H), 7.77 (d, J = 8.8 Hz, 1H), 7.49 - 7.16 (m, 2H), 3.91 (s, 2H), 1.82 (dd, J = 12.7, 6.3 Hz, 1H), 1.69 (s, 9H), 1.50 - 1.38 (m, 2H), 1.16 (s, 3H), 0.95 (dd, J = 9.1, 6.6 Hz, 6H).

[0152] Step 7: Synthesis of (S)-1-{[2-(difluoromethyl)-6-[5-(difluoromethyl)-1H-pyrrolo[2,3- b]pyridin-3-yl]pyridin-3-yl]oxy}-2,4-dimethylpentan-2-amine hydrochloride

[0153] (S)-3-{5-[(2-amino-2,4-dimethylpentyl)oxy]-6-(difluoromethyl)pyridin-2-yl}-5- (difluoromethyl)-1H-pyrrolo[2,3-b]pyridine-1-carboxylic acid tert-butyl ester (45 mg, 0.086 mmol, 1.0 eq) was added to a solution of hydrogen chloride-dioxane (3.0 mL, 4.0 mol / L) and reacted at room temperature for 2 hours. After the reaction was completed, it was directly concentrated and prepared by C18 reverse phase (formic acid system) to obtain the final product (S)-1-{[2-(difluoromethyl)-6-[5-(difluoromethyl)-1H-pyrrolo[2,3-b]pyridin-3-yl]pyridin-3- yl]oxy}-2,4-dimethylpentan-2-amine formate (32.3 mg, yield 80.03%). LCMS (TOF MS ES+) m / z [M+H]+: 425.2298. 1 H NMR (400 MHz, DMSO) δ 12.34 (s, 1H), 9.08 (s, 1H), 8.50 (s, 1H), 8.40 (s, 1H), 8.30 (s, 1H), 8.11 (d, J = 8.8 Hz, 1H), 7.73 (d, J = 8.9 Hz, 1H), 7.52 - 7.10 (m, 2H), 3.97 (s, 2H), 1.83 (m, J = 12.7, 6.4 Hz, 1H), 1.51 (m, J = 14.1, 5.6 Hz, 2H), 1.23 (s, 3H), 0.95 (dd, J = 12.6, 6.6 Hz, 6H).

[0154] Example 43: Synthesis of (S)-1-{[2-(difluoromethyl)-6-(5-fluoro-1-methyl-1H- pyrrolo[2,3-b]pyridin-3-yl)pyridin-3-yl]oxy}-2,4-dimethylpentan-2-amine (Compound 43)

[0155]

[0156] Step 1: Synthesis of 3-[6-(difluoromethyl)-5-fluoropyridin-2-yl]-5-fluoro-1- toluenesulfonyl-1H-pyrrolo[2,3-b]pyridine

[0157] To a mixture of 5-fluoro-3-(4,4,5,5-tetramethyl-l,3,2-dioxaborolan-2-yl)-l- methyl-lH-pyrrolo[2,3-b]pyridine (200 mg, 0.48 mmol, 1.0 eq), 6-bromo-2- (difluoromethyl)-3-fluoropyridine (108 mg, 0.48 mmol, 1.0 eq) and [l,l'- bis(diphenylphosphino)ferrocene]dichloropalladium (35 mg, 0.048 mmol, 0.1 eq) in 1,4-dioxane (4 mL) and water (1 mL) was heated at 90 °C for 4 h. After the reaction was completed, the mixture was cooled, extracted with ethyl acetate (10 mL) for 3 times, the organic phase was concentrated and purified by normal phase (PE:EA = 3: 1) to give 3-[6-(difluoromethyl)-5-fluoropyridin-2-yl]-5-fluoro-l-methyl-lH- pyrrolo[2,3-b]pyridine (135 mg, yield 64.6%) as a yellow solid. LCMS (TOF MS ES+) m / z [M+H]+: 436.1. 1 HNMR (400 MHz, DMSO) δ 8.95 (s, 1H), 8.72 (dd, J = 9.2, 2.8 Hz, 1H), 8.50 (m, J = 5.9, 4.1 Hz, 2H), 8.05 (d, J = 8.3 Hz, 2H), 7.46 (d, J = 8.2 Hz, 2H), 7.38 - 7.21 (m, 1H), 2.37 (s, 3H).

[0158] Step 2: Synthesis of 3-[6-(difluoromethyl)-5-fluoropyridin-2-yl]-5-fluoro-l- methyl-lH-pyrrolo[2,3-b]pyridine

[0159] To a solution of 3-[6-(difluoromethyl)-5-fluoropyridin-2-yl]-5-fluoro-l- toluenesulfonyl-lH-pyrrolo[2,3-b]pyridine (135 mg, 0.31 mmol, 1.0 eq), sodium hydroxide (49.6 mg, 1.24 mmol, 4.0 eq) and iodomethane (49.6 mg, 0.93 mmol, 3.0 eq) in dichloromethane (5 mL) was heated to 40 °C for 16 h. After the reaction was completed, the mother liquor was concentrated and purified by normal phase (PE:EA = 4: 1) to give 3-[6-(difluoromethyl)-5-fluoropyridin-2-yl]-5-fluoro-l-methyl-lH- pyrrolo[2,3-b]pyridine (65.0 mg, yield 71.0%) as a yellow solid. LCMS (TOF MS ES+) m / z [M+H]+: 296. ¾ NMR (400 MHz, DMSO) d 8.64 (dd, J = 9.7, 2.8 Hz, 1H), 8.53 (s, 1H), 8.44 - 8.34 (m, 1H), 8.12 (dd, J = 8.9, 3.7 Hz, 1H), 7.97 (t, J = 9.5 Hz, 1H), 7.49 - 7.16 (m, 1H), 3.92 (s, 3H).

[0160] Step 3: Synthesis of (S)-l-{[2-(difluoromethyl)-6-(5-fluoro-l-methyl-lH-pyrrolo[2,3- b]pyridin-3-yl)pyridin-3-yl]oxy}-2,4-dimethylpentan-2-amine

[0161] To a solution of 3-[6-(difluoromethyl)-5-fluoropyridin-2-yl]-5-fluoro-l- methyl-lH-pyrrolo[2,3-b]pyridine (88 mg, 0.22 mmol, 1.0 eq) and (S)-2-amino-2,4- dimethylpentan-l-ol (29 mg, 0.22 mmol, 1.0 eq) in tetrahydrofuran (2 mL) was added potassium tert-butoxide (74 mg, 0.66 mmol, 3.0 eq) and the reaction was stirred at 90 °C for 4 h. After completion of the reaction, it was filtered and the filtrate was concentrated and purified by prep to afford (S)-l-{[2-(difluoromethyl)-6-(5-fluoro-l-methyl-lH-pyrrolo[2,3-b]pyridin-3-yl)pyridin-3- yl]oxy}-2,4-dimethylpentan-2-amine (6.9 mg, 7.8% yield) as a white solid. LCMS (TOF MS ES+) m / z [M+H]+: 407.20. ¾ NMR (400 MHz, DMSO) ¾ NMR (400 MHz, DMSO) δ 8.60 (dd, J = 9.8, 2.8 Hz, 1H), 8.38 (s, 1H), 8.33 (dd, J = 2.9, 1.5 Hz, 1H), 7.96 (d, J = 8.8 Hz, 1H), 7.67 (d, J = 8.9 Hz, 1H), 7.40 - 7.09 (m, 1H), 3.88 (s, 3H), 3.81 (s, 2H), 1.80 (m, J = 12.6, 6.2 Hz, 1H), 1.38 (dd, J = 5.5, 2.4 Hz, 2H), 1.11 (s, 3H), 0.92 (t, J = 7.0 Hz, 6H).

[0162] Example 44: Synthesis of (S)-l-{[6-(5-chloro-lH-pyrrolo[2,3-b]pyridin-3-yl)-2- (difluoromethyl)pyridin-3-yl]oxy}-2,4-dimethylpentan-2-amine (Compound 44)

[0163]

[0164] Step 1: Synthesis of 5-chloro-3-(4,4,5,5-tetramethyl-l,3,2-dioxaborolan-2-yl)-lH- pyrrolo[2,3-b]pyridine

[0165] Compound 3-bromo-5-chloro-lH-pyrrolo[2,3-b]pyridine (1.0 g, 4.35 mmol, 1.0 eq), 4,4,5,5-tetramethyl-l,3,2-dioxaborolane (2.2 g, 17.2 mmol, 4.0 eq), [l,l'-bis(diphenylphosphino)ferrocene]dichloropalladium (159 mg, 0.217 mmol, 0.05 eq) and triethylamine (2.19 g, 2.17 mmol, 5 eq) were added into 1,4-dioxane (10 mL) and reacted at 120 °C for 1 h under nitrogen protection. After the reaction was completed, the insoluble matter was removed by filtration, and the filtrate was separated and purified by reverse phase column chromatography to obtain 5-chloro-3-(4,4,5,5-tetramethyl-l,3,2-dioxaborolan-2-yl)-lH-pyrrolo[2,3-b]pyridine (155 mg, yield 12.7%) as a white solid. LCMS (ESI) [M+H]+: 279. 1 H NMR (400 MHz, CDC13) δ 8.37 (dd, J = 4.8, 1.6 Hz, 1H), 8.31 (s, J = 7.8, 1.6 Hz, 1H), 7.68 (s, 1H), 1.39 (s, 12H).

[0166] Step 2: Synthesis of (S)-l-{[6-(5-chloro-lH-pyrrolo[2,3-b]pyridin-3-yl)-2- (difluoromethyl)pyridin-3-yl]oxy}-2,4-dimethylpentan-2-amine

[0167] To a solution of 5-chloro-3-(4,4,5,5-tetramethyl-l,3,2-dioxaborolan-2-yl)-lH- pyrrolo[2,3-b]pyridine (50 mg, 0.180 mmol, 1.0 eq) and (S)-l-{[6-bromo-2- (difluoromethyl)pyridin-3-yl]oxy}-2,4-dimethylpentan-2-amine (60.4 mg, 0.180 mmol, 1.0 eq) in 1,4-dioxane (2 mL) was added [l,l'-bis(diphenylphosphino)ferrocene] palladium dichloride (13.2 mg, 0.018 mmol, 0.05 eq) and sodium carbonate (57.2 mg, 0.54 mmol, 3.0 eq) under nitrogen atmosphere. The reaction mixture was heated at 120 °C for 2 h. After completion of the reaction, the excess solid was filtered off and the filtrate was concentrated to afford (S)-l-{[6-(5-chloro-lH-pyrrolo[2,3-b]pyridin-3-yl)-2- (difluoromethyl)pyridin-3-yl]oxy}-2,4-dimethylpentan-2-amine (4.3 mg, 5.8% yield) as off-white solid. LCMS (ESI) [M+H]+: 409.19. ¾ NMR (400 MHz, DMSO-d6) δ 12.32 (d, J = 2.9 Hz, 1H), 8.89 (d, J = 2.4 Hz, 1H), 8.42 (d, J = 2.8 Hz, 1H), 8.32 (d, J = 2.4 Hz, 1H), 8.14 (s, 1H), 8.11 (s, 2H), 7.79 (d, J = 9.0 Hz, 1H), 7.71 - 7.41 (m, 1H), 4.25 - 4.13 (m, 2H), 1.89 - 1.72 (m, 2H), 1.61 (dd, J = 14.1, 5.3 Hz, 1H), 1.40 (s, 3H), 1.00 (d, J = 6.5 Hz, 3H), 0.94 (d, J = 6.5 Hz, 3H).

[0168] Example 45: Synthesis of (S)-l-{[2-(difluoromethyl)-6-(5-methyl-lH-pyrrolo[2,3- b]pyridin-3-yl)pyridin-3-yl]oxy}-2,4-dimethyl-2-pentanamine bicarbonate (Compound 45)

[0169]

[0170] Step 1: Synthesis of 5-methyl-lH-pyrrolo[2,3-b]pyridine

[0171] Compound 5-bromo-lH-pyrrolo[2,3-b]pyridine (11.0 g, 33.7 mmol, 1.0 eq) and l,l'-bisdiphenylphosphinof errocene palladium dichloride (197.3 mg, 0.27 mmol, 0.008 eq) were added to toluene (100 mL), heated to 100 °C under nitrogen protection, then methyl magnesium chloride (3 M, 28.2 mL, 2.5 eq) was added dropwise to the reaction system, and the reaction was continued for 2 hours. After the reaction was completed, the reaction solution was directly concentrated, water and ethyl acetate were added for liquid-liquid separation, the organic phase was concentrated, and normal phase column separation (PE:EA = 4:1) was performed to obtain the product 5-methyl-lH-pyrrolo[2,3-b]pyridine (4.0 g, yield 53.95%) as colorless oil. LCMS (ESI) [M+H]+: 133.

[0172] Step 2: Synthesis of 3-iodo-5-methyl-lH-pyrrolo[2,3-b]pyridine

[0173] 5-methyl-lH-pyrrolo[2,3-b]pyridine (4.0 g, 30.26 mmol, 1.0 eq) and iodine (3.84 g, 15.13 mmol, 0.5 eq) were dissolved in N,N-dimethylformamide (30 mL), potassium hydroxide (8.56 g, 151.3 mmol, 5 eq) was added portionwise, and the reaction was carried out at room temperature for 16 hours. After the reaction was completed, the reaction solution was poured into ice water, and a yellow solid was generated. Filtration was performed, the filter cake was collected, and oven drying was performed to obtain yellow crude product 3-iodo-5-methyl-lH-pyrrolo[2,3-b]pyridine (3.0 g). LCMS (ESI) [M+H]+: 259.

[0174] Step 3: Synthesis of 5-methyl-3-(4,4,5,5-tetramethyl-l,3,2-dioxaborolan-2-yl)-lH- pyrrolo[2,3-b]pyridine

[0175] 3-iodo-5-methyl-lH-pyrrolo[2,3-b]pyridine (3.0 g, 11.64 mmol, 1.0 eq), 4,4,5,5-tetramethyl-l,3,2-dioxaborolan (7.5 g, 58.2 mmol, 5.0 eq), triethylamine (5.96 g, 58.2 mmol, 5.0 eq), and l,l'-bisdiphenylphosphinof errocene palladium dichloride (852 mg, 1.164 mmol, 0.1 eq) were added to 1,4-dioxane (15 mL), and the reaction was carried out at 120 °C for 2 hours under nitrogen protection. Filtration was performed using diatomite, the solution was concentrated, and normal phase separation purification was performed to obtain yellow solid 5-methyl-3-(4,4,5,5-tetramethyl-l,3,2-dioxaborolan-2-yl)-lH-pyrrolo[2,3-b]pyridine (2.0 g). LCMS (ESI) [M+H]+: 259. 1H NMR (400 MHz, CDC13) δ 8.05 (dd, J = 7.9, 2.0 Hz, 1H), 7.93 (s, 2H), 7.72 (d, J = 2.3 Hz, 1H), 1.30 (s, 3H), 1.07 (s, 12H).

[0176] Step 4: Synthesis of tert-butyl 5-methyl-3-(4,4,5,5-tetramethyl-l,3,2-dioxaborolan-2-yl)- lH-pyrrolo[2,3-b]pyridine-1-carboxylate

[0177] tert-Butyl 5-methyl-3-(4,4,5,5-tetramethyl-l,3,2-dioxaborolan-2-yl)-lH-pyrrolo[2,3- b]pyridine-1-carboxylate (81 mg, yield: 58.4%) was obtained as yellow oil by dissolving 5-methyl-3-(4,4,5,5-tetramethyl-l,3,2-dioxaborolan-2-yl)-lH-pyrrolo[2,3-b]pyridine (100 mg, 0.384 mmol, 1 eq) in dichloromethane (5 mL), adding di-tert-butyl dicarbonate (110 mg, 0.504 mmol, 1.3 eq), triethylamine (78.4 mg, 0.775 mmol, 2.0 eq), DMAP (4.7 mg, 0.038 mmol, 0.1 eq) successively at 0 °C, keeping the reaction at 0 °C for 1 h, adding water (10 mL) to dilute after the reaction was completed, extracting with dichloromethane (20 mL) three times, drying the organic phase over anhydrous sodium sulfate, concentrating the organic phase, and purifying by TLC preparative plate (PE:EA = 2: 1). LCMS (ESI) [M+H]+: 359.

[0178] Step 5: Synthesis of tert-butyl (S)-3-{5-[(2-amino-2,4-dimethylpentyl)oxy]-6- (difluoromethyl)pyridin-2-yl}-5-methyl-lH-pyrrolo[2,3-b]pyridine-1-carboxylate

[0179] To a solution of tert-butyl 5-methyl-3-(4,4,5,5-tetramethyl-l,3,2-dioxaborolan-2-yl)-lH- pyrrolo[2,3-b]pyridine-l-carboxylate (81 mg, 0.226 mmol, 1.0 eq), (S)-l-{[6-bromo-2- (difluoromethyl)pyridin-3-yl]oxy}-2,4-dimethylpentan-2-amine (75.8 mg, 0.226 mmol, 1.0 eq) in 1,4-dioxane (2 mL) and water (0.5 mL) was added l,l'-bis(diphenylphosphino) ferrocene palladium dichloride (16.8 mg, 0.023 mmol, 0.1 eq) and cesium carbonate (222.3 mg, 0.678 mmol, 3.0 eq) and the reaction mixture was heated at 80 °C for 2 h under nitrogen. After the reaction was completed, the reaction mixture was filtered through celite and the filtrate was concentrated and purified by reverse phase column chromatography (50% acetonitrile) to give (S)-tert-butyl 3-{5-[(2-amino-2,4-dimethylpentyl)oxy]-6- (difluoromethyl)pyridin-2-yl}-5-methyl-lH-pyrrolo[2,3-b]pyridine-l-carboxylate (33 mg, crude). The crude was used directly for the next step without further purification. LCMS (ESI) [M+H]+: 489.

[0180] Step 6: Synthesis of (S)-l-{[2-(difluoromethyl)-6-(5-methyl-lH-pyrrolo[2,3-b]pyridin-3- yl)pyridin-3-yl]oxy}-2,4-dimethyl-2-pentanamine bicarbonate

[0181] To a solution of (S)-tert-butyl 3-{5-[(2-amino-2,4-dimethylpentyl)oxy]-6- (difluoromethyl)pyridin-2-yl}-5-methyl-lH-pyrrolo[2,3-b]pyridine-l-carboxylate (33 mg, crude) in dichloromethane (2 mL) was added trifluoroacetic acid (0.5 mL) and the reaction mixture was stirred at room temperature for 1 h. After the reaction was completed, the organic phase was concentrated and purified by preparative purification to give (S)-l-{[2-(difluoromethyl)-6-(5-methyl-lH-pyrrolo[2,3-b]pyridin-3-yl)pyridin-3- yl]oxy}-2,4-dimethyl-2-pentanamine bicarbonate (9.3 mg, 35.4% yield). LCMS (ESI) [M+H]+: 389.21. 1H NMR (400 MHz, DMSO) δ 11.92 (d, J = 2.9 Hz, 1H), 8.65 (d, J = 2.1 Hz, 1H), 8.21 (d, J = 2.8 Hz, 1H), 8.14 (d, J = 2.1 Hz, 1H), 8.13 (s, 2H), 8.06 (d, J = 8.9 Hz, 1H), 7.73 (d, J = 8.9 Hz, 1H), 7.66 - 7.36 (m, 1H), 4.73 (s, 2H), 4.21 - 4.10 (m, 2H), 2.42 (s, 3H), 1.87 - 1.69 (m, 2H), 1.60 (dd, J = 14.1, 5.3 Hz, 1H), 1.38 (s, 3H), 1.00 - 0.89 (m, 6H).

[0182] Example 46: Synthesis of (S)-1-{[3-(difluoromethyl)-5-(1H-pyrrolo[2,3-b]pyridin-3- yl)pyridin-2-yl]oxy}-2,4-dimethylpentan-2-amine carbonate salt (Compound 46)

[0183] Step 1: Synthesis of (S)-1-{[5-bromo-3-(difluoromethyl)pyridin-2-yl]oxy}-2,4- dimethylpentan-2-amine

[0184] Compound (S)-2-amino-2,4-dimethylpentan-1-ol (275 mg, 2.10 mmol, 0.95 eq) was added to tetrahydrofuran (6.0 mL), followed by potassium tert-butoxide (620 mg, 5.53 mmol, 2.5 eq) and 5-bromo-3-(difluoromethyl)-2-fluoropyridine (500 mg, 2.21 mmol, 1.0 eq) and reacted at 93 °C for 2 hours. After the reaction was completed, it was filtered, the filtrate was concentrated and purified by normal phase column chromatography (petroleum ether: ethyl acetate = 3:1) to give the product (S)-1-{[5-bromo-3-(difluoromethyl)pyridin-2-yl]oxy}-2,4-dimethylpentan-2-amine (200 mg, yield 26.85%) as yellow oil. LCMS (TOF MS ES+) m / z [M+H]+: 337 / 339.1H NMR (400 MHz, DMSO) δ 8.49 - 8.43 (m, 1H), 8.14 (d, J = 2.4 Hz, 1H), 7.31 - 7.00 (m, 1H), 4.04 (d, J = 1.7 Hz, 2H), 1.79 (m, J = 12.8, 6.4 Hz, 1H), 1.60 (s, 2H), 1.35 (m, J = 5.8, 3.4 Hz, 2H), 1.09 (s, 3H), 0.90 (d, J = 6.7 Hz, 6H).

[0185] Step 2: Synthesis of (S)-1-{[3-(difluoromethyl)-5-(1H-pyrrolo[2,3-b]pyridin-3- yl)pyridin-2-yl]oxy}-2,4-dimethylpentan-2-amine hydrochloride

[0186] (S)-1-{[5-bromo-3-(difluoromethyl)pyridin-2-yl]oxy}-2,4-dimethylpentan-2-amine (80 mg, 0.24 mmol, 1.0 eq), 7-azaindole-3-boronic acid pinacol ester (57.91 mg, 0.24 mmol, 1.0 eq), sodium carbonate (100 mg, 0.95 mmol, 4.0 eq) and [1,1'-bis(diphenylphosphino)ferrocene]dichloropalladium (17.56 mg, 0.024 mmol, 0.10 eq) were added into 1,4-dioxane (3.0 mL) and reacted at 120 °C for 8 h under nitrogen protection. After the reaction was completed, it was filtered and the filtrate was prepared by C18 column reverse phase (formic acid system) to obtain white solid (S)-1-{[3-(difluoromethyl)-5-(1H-pyrrolo[2,3-b]pyridin-3-yl)pyridin-2-yl]oxy}-2,4-dimethylpentan-2-amine hydrochloride (6.0 mg, yield 6.02%). LCMS (TOF MS ES+) m / z [M+H]+: 375 / 376.1H NMR (400 MHz, DMSO) δ 12.07 (s, 1H), 8.70 (d, J = 2.3 Hz, 1H), 8.44 - 8.36 (m, 1H), 8.31 (dd, J = 4.7, 1.4 Hz, 1H), 8.27 (dd, J = 8.0, 1.6 Hz, 2H), 8.02 (s, 1H), 7.55 - 7.24 (m, 1H), 7.20 (dd, J = 8.0, 4.6 Hz, 1H), 4.28 (s, 2H), 1.81 (m, J = 16.6, 8.2 Hz, 1H), 1.65 - 1.48 (m, 2H), 1.28 (s, 3H), 0.94 (dd, J = 14.3, 6.6 Hz, 6H).

[0187] Example 47: Synthesis of (S)-1-{4-(1H-pyrrolo[2,3-b]pyridin-3-yl)-2- (trifluoromethyl)phenoxy}-2,4-dimethylpentan-2-amine (Compound 47)

[0188]

[0189] Step 1: Synthesis of (S)-1-[4-bromo-2-(trifluoromethyl)phenoxy]-2,4- dimethylpentan-2-amine

[0190] Compound (S)-2-amino-2,4-dimethylpentan-1-ol (200 mg, 1.52 mmol, 0.95 eq) was added to tetrahydrofuran (5.0 mL), followed by potassium tert-butoxide (450 mg, 4.0 mmol, 2.5 eq) and 5-bromo-2-fluorobenzotrifluoride (390 mg, 1.60 mmol, 1.0 eq), and reacted at room temperature for 2 hours. After the reaction was completed, it was filtered, the filtrate was concentrated, and purified by normal phase column chromatography (petroleum ether: ethyl acetate = 3:1) to obtain the product (S)-1-[4-bromo-2-(trifluoromethyl)phenoxy]-2,4-dimethylpentan-2-amine (240 mg, yield 44.50%) as a colorless oil. LCMS (TOF MS ES+) m / z [M+H]+: 354 / 356.1H NMR (400 MHz, CDC13) δ 7.67 (d, J = 2.5 Hz, 1H), 7.57 (dd, J = 8.8, 2.5 Hz, 1H), 6.84 (d, J = 8.8 Hz, 1H), 3.82 - 3.68 (m, 2H), 1.85 - 1.68 (m, J = 6.4 Hz, 1H), 1.47 (dd, J = 5.7, 4.0 Hz, 2H), 1.22 (s, 3H), 0.96 (dd, J = 9.9, 6.7 Hz, 6H).

[0191] Step 2: Synthesis of (S)-1-{4-(1H-pyrrolo[2,3-b]pyridin-3-yl)-2- (trifluoromethyl)phenoxy}-2,4-dimethylpentan-2-amine

[0192] (S)-1-[4-bromo-2-(trifluoromethyl)phenoxy]-2,4-dimethylpentan-2-amine (92 mg, 0.26 mmol, 1.0 eq), 7-azaindole-3-boronic acid pinacol ester (63 mg, 0.26 mmol, 1.0 eq), sodium carbonate (110 mg, 1.04 mmol, 4.0 eq) and [1,1'-bis(diphenylphosphino)ferrocene]dichloropalladium (19 mg, 0.026 mmol, 0.10 eq) were added into 1,4-dioxane (3.0 mL) and reacted at 120 °C for 8 h under nitrogen protection. After the reaction was completed, it was filtered and the filtrate was prepared by C18 column reverse phase (sodium bicarbonate system) to obtain brown solid (S)-1-{4-(1H-pyrrolo[2,3-b]pyridin-3-yl)-2-(trifluoromethyl)phenoxy}-2,4-dimethylpentan-2-amine (3.1 mg, yield 3.05%). LCMS (TOF MS ES+) m / z [M+H]+: 392.2213. 1H NMR (400 MHz, MeOD) δ 8.30 - 8.25 (m, 2H), 7.93 - 7.85 (m, 2H), 7.68 (s, 1H), 7.28 (d, J = 8.5 Hz, 1H), 7.24 (dd, J = 7.8, 5.0 Hz, 1H), 4.01 - 3.90 (m, 2H), 1.90 - 1.80 (m, 1H), 1.58 (m, J = 14.2, 5.5 Hz, 2H), 1.29 (s, 3H), 1.02 (t, J = 7.0 Hz, 6H).

[0193] The compounds of Table 1 were obtained according to the methods of Reference Examples 1 and 2:

[0194] Table 1: Structures and characterization of other compounds

[0195]

[0196]

[0197]

[0198] Experimental Example 1: Test of the binding ability of the compounds of the present application to AAK1 protein

[0199] Purpose of the experiment: to detect the binding ability of the compounds to AAK1 protein by CETSA experimental method

[0200] Background rationale: CETSA experiment is a molecular detection method to measure the affinity of drugs and target proteins. The principle is that after the drug binds to the target protein, the structure becomes more stable. Co-incubate the candidate drug with the target protein, if the candidate drug is an AAK1 inhibitor, then the candidate drug can bind to AAK1, and in the heating process of the sample, AAK1 protein is more stable, and AAK1 protein will be more easily detected by Western blot experiment; on the contrary, the stability of AAK1 protein will be worse after heating, and the amount of protein detected will be lower. Thus, the binding ability of the drug and the target protein is evaluated, which is used for screening AAK1 protein inhibitors.

[0201] Specific experimental procedures:

[0202] AAK1(30-330) protein expression: AAK1 sequence (residues 30-330) was cloned into pET24N vector with n-terminal (His) 6x tag and tobacco etch virus (TEV) protease cleavage site. The constructed vector was transformed into E. coli BL21(DE3), and when the OD was 0.6, 1 mM IPTG was added, and the culture was incubated at 12°C for 16 h. The bacterial cells were collected, ultrasonically broken, and centrifuged to obtain the supernatant. The protein was purified by Ni-NTA, and the protein concentration and purity were determined after dialysis.

[0203] CETSA sample preparation: The sample was incubated with the candidate drug and the control drug and control reagent for 30 min, and the sample was heated at the set temperature points. After recovery to room temperature, the sample was centrifuged at 20,000g, and the supernatant was collected. The protein sample was heated at 100°C for 10 min using sample buffer. After the sample returned to room temperature, the sample was detected by western blot; the protein loading amount was controlled at 20ug. After determining the mutation temperature, the compound concentration gradient was generally set at 9 points, the sample was incubated, and the western blot detection was performed as above. Protein electrophoresis: the concentration gel voltage was set at 60v, and the separation gel voltage was set at 120v; after electrophoresis, electrotransfer was started. The electrotransfer conditions were set at 250mA for 2h; 5% BSA blocking was performed for 1h; specific primary antibody was added, and incubated on a 4°C shaker overnight; TBST was washed 4 times, each for 2.5min; secondary antibody was incubated on a room temperature shaker for 1h; TBST was washed 4 times, each for 2.5min; ECL was used for development, and the AAK1 protein expression of different groups and each temperature point was detected. The western blot band was processed by image J and GraphPad software, and EC50 was calculated.

[0204] EC50 is the concentration for 50% of maximal effect, which refers to the concentration of drug that can cause 50% of individuals effective. The AAK1 inhibitor BMS-986176 / LX-9211 of Bristol-Myers Squibb Company was used as a positive reference compound, and the preparation method is referred to the preparation method of Example 123 in the patent document CN106458994A.

[0205] The structure of BMS-986176 / LX-9211 is as follows:

[0206]

[0207] The test results are shown in Table 2 below, and the EC50 values of each compound are classified according to the following description:

[0208] “+” represents that the EC50 value is greater than 1 μM;

[0209] “++” represents that the EC50 value is less than 1 μM and greater than 100 nM;

[0210] “+++” represents that the EC50 value is less than 100 nM and greater than 10 nM;

[0211] “++++” represents that the EC50 value is less than 10 nM.

[0212] Table 2: EC50 results

[0213] Compound No. EC50 Compound No. EC50 Compound No. EC50 Compound No. EC50 1 +++ 12 +++ 23 ++ 34 +++ 2 +++ 13 +++ 24 ++ 35 +++ 3 +++ 14 +++ 25 +++ 36 ++ 4 +++ 15 +++ 26 +++ 37 +++ 5 +++ 16 +++ 27 +++ 38 +++ 6 +++ 17 +++ 28 +++ 39 +++ 7 ++ 18 +++ 29 ++ 40 +++ 8 +++ 19 +++ 30 +++ 41 ++ 9 +++ 20 +++ 31 +++ 42 +++ 10 +++ 21 ++ 32 +++ 43 ++++ 11 +++ 22 ++ 33 +++ 44 ++++ BMS-986176 ++ 45 ++++

[0214] The experimental results show that the compound of the present application has a strong binding ability to the functional domain of AAK1 protein kinase, and the in vitro binding ability is equivalent to or stronger than that of the positive control compound BMS-986176, which indicates that the compound of the present application has the potential as an AAK1 protein inhibitor.

[0215] Experimental Example 2: Determination of the effect of the compound of the present application on AP2M1 phosphorylation in cells

[0216] Purpose of experiment: The purpose of this test example is to test the determination of the effect of the compound on AP2M1 phosphorylation in cells.

[0217] Background principle: Adapter-related kinase 1 (AAK1), also known as AP2-related kinase 1, is a 104 kDa serine / threonine kinase. AAK1 interacts with the adaptor protein 2 (AP2) complex, phosphorylates the threonine residue in the micro-subunit of AP2 (AP2M1), thereby increasing its binding affinity to certain membrane receptors based on specific tyrosine or di-leucine sorting signals. By detecting the phosphorylation level of AP2M1 in cells, the inhibitory effect of the compound on AAK1 is evaluated, and candidate compounds are screened.

[0218] Specific experimental procedure:

[0219] The 293T cells were seeded in T25, and the confluence reached 60-80% the next day. The next day, 16 ug plasmid (1 ug AAK1 / HA / pIRES and 15 ug Flag / AP2MI / pcDNA) was transfected into each T25 using lip2000 transfection reagent. The third day, the cells were digested, resuspended, centrifuged at 1000 rpm for 5 min, seeded in 24-well plates, 450 ul / well, 2x10^5 cells / well. The test compounds were gradiently diluted, 50 ul was added to each well, mixed, incubated at 37°C for 3 h, DMSO was set as a control, and untransfected plasmid was set as a blank control. After incubation, the cells were collected, washed with PBS twice, and lysed with cell lysis solution containing protease inhibitor PMSF on ice for 30 min; the protein sample was determined for concentration using a BCA kit. Protein electrophoresis: the concentrated gel voltage was set to 60 v, and the separation gel voltage was set to 120 v; after electrophoresis, electrotransfer was started. The electrotransfer conditions were set to 250 mA for 2 h; 5% BSA blocking was performed for 1 h; specific primary antibody anti-p-AP2M1 was added, and incubated on a 4°C shaker overnight; TBST was washed 4 times, each time for 2.5 min; the secondary antibody was incubated on a room temperature shaker for 1 h; TBST was washed 4 times, each time for 2.5 min; ECL was used for development, and the p-AP2M1 expression amount was detected. The western blot band was processed by image J and GraphPad software, and the IC50 was calculated.

[0220] IC50 (half maximal inhibitory concentration) refers to the half-inhibitory concentration of the measured antagonist. It can indicate the half amount of a drug or substance (inhibitor) in inhibiting certain biological programs (or certain substances contained in the program, such as enzymes, cell receptors or microorganisms). Compound BMS-986176 is used as a positive reference compound as described above.

[0221] The test results are shown in Table 3 below, in which the IC50 values of each compound are classified according to the following description:

[0222] “+” indicates that the IC50 value is greater than 1 μM;

[0223] “++” indicates that the IC50 value is less than 1 μM and greater than 100 nM;

[0224] “+++” indicates that the IC50 value is less than 100 nM and greater than 10 nM;

[0225] “++++” indicates that the IC50 value is less than 10 nM.

[0226] The results are shown in Table 3 below:

[0227] Table 3: IC50 experimental data

[0228] Compound No. IC50 Compound No. IC50 Compound No. IC50 Compound No. IC50 1 +++ 12 +++ 23 +++ 34 +++ 2 +++ 13 +++ 24 +++ 35 ++ 3 +++ 14 +++ 25 +++ 36 +++ 4 +++ 15 +++ 26 +++ 37 +++ 5 +++ 16 +++ 27 ++ 38 +++ 6 +++ 17 ++ 28 +++ 39 +++ 7 +++ 18 +++ 29 +++ 40 +++ 8 +++ 19 +++ 30 +++ 41 ++ 9 +++ 20 +++ 31 +++ 42 +++ 10 +++ 21 +++ 32 ++ 43 ++++ 11 +++ 22 +++ 33 +++ 44 ++++ BMS-986176 ++ 45 ++++

[0229] The results show that the compounds of the present application can inhibit the phosphorylation level of AP2M1 in cells, with an IC50 value reaching the nM level, equivalent to or stronger than the positive drug. This strong inhibitory effect has important therapeutic significance for the treatment of diseases or diseases related to AAK1 inhibition.

[0230] Experimental Example 3: Test of the effect of the compounds of the present application on the function of AAK1 kinase

[0231] Purpose of the experiment: The purpose of this test example is to test the effect of the compounds on the function of AAK1 kinase.

[0232] Background principle: AAK1 phosphorylates the threonine residue in the micro-subunit of AP2 (AP2M1), in which ATP will provide phosphate to generate ADP, so by detecting the amount of ADP generated, the ability of AAK1 kinase can be evaluated.

[0233] Specific experimental procedure:

[0234] AAK1 (30-330) protein expression: The AAK1 sequence (residues 30-330) was cloned into the pET24N vector with an n-terminal (His) 6x tag and a tobacco etch virus (TEV) protease cleavage site. The constructed vector was transformed into E. coli BL21 (DE3), and when the OD was 0.6, 1 mM IPTG was added and incubated at 12°C for 16 h. The bacterial cells were collected, ultrasonically broken, centrifuged to obtain the supernatant, and the protein was purified by Ni-NTA. After dialysis, the protein concentration and purity were determined.

[0235] AAK1 kinase experiment: The purified AAK1 protein was added to the test buffer (20 mM Tris, pH 7.80, 10 mM MgCl2, 1 mM DTT, 0.01% Tween20), the substrate (Aha-KEEQSQITSQVTGQIGWR-NH2 and ATP) and the test compound were added. The final concentration of each reagent in the system was: AAK1 was 3.5 nM; Aha-KEEQSQITSQVTGQIGWR-NH2 was 1.5 uM; ATP was 22 uM. The reaction mixture was incubated at 25°C in a metal bath for 3 h, 1% SDS was added to terminate the reaction, and the amount of ADP generated was detected by HPLC. The IC50 was calculated by GraphPad software, and the compounds were screened by comparison with the positive drug BMS-986176.

[0236] The test results are shown in Table 4 below:

[0237] Table 4: IC50 experimental data

[0238] Compound No. IC50 (nM) Compound No. IC50 (nM) Compound No. IC50 (nM) 1 32.1 16 87.2 31 102.1 2 28.3 17 56.9 32 67.8 3 21.2 18 42.5 33 69.5 4 45.9 19 87.3 34 89.2 5 86.1 20 101.6 35 113.4 6 75.2 21 190.2 36 147.6 7 100.5 22 170.3 37 123.7 8 129.3 23 99.6 38 160.2 9 99.2 24 107.4 39 101.3 10 152.1 25 118.5 40 99.5 11 123.4 26 86.3 41 96.7 12 99.2 27 57.8 42 78.4 13 167.8 28 65.3 43 12.1 14 102.3 29 72.1 44 8.2 15 111.2 30 88.9 45 6.5 BMS-986176 260.3

[0239] The results show that the compounds of the present application have very strong inhibitory effect on the function of AAK1 protein kinase, and the IC50 values are in the nM level, equivalent to or less than the positive drug. This strong inhibitory effect has important therapeutic significance for the treatment of diseases or diseases related to AAK1 inhibition.

[0240] Experimental Example 4: In vitro pharmacokinetic test of the compound of the present application in SD rats

[0241] Purpose of the experiment: The purpose of this experimental example is to test the pharmacokinetics of the compound in SD rats

[0242] Background principle: Nonclinical pharmacokinetic studies are conducted through in vivo and in vitro animal and human in vitro research methods to reveal the dynamic changes of drugs in the body, obtain basic pharmacokinetic parameters of drugs, and clarify the processes and characteristics of drug absorption, distribution, metabolism and excretion. Nonclinical pharmacokinetic studies play an important role in the evaluation process of new drug research and development. In pharmacodynamic and toxicological evaluation, drug or active metabolite concentration data and related pharmacokinetic parameters are the basis for generating, determining or clarifying the size of efficacy or toxicity, and can provide the basis for the effect of drugs on target organs (efficacy or toxicity).

[0243] Specific experimental procedures:

[0244] (1) Preparation of test sample: Solvent: 40% PEG300, 15% Tween 80, 10% ethanol, 35% water, prepared on the day of administration;

[0245] (2) Experimental animals: male SD rats, SPF level, 18;

[0246] (3) Experimental design:

[0247] Table 5 Experimental design

[0248]

[0249] Note: *, the animals were fasted overnight (10-14 hours) before administration, and fed 4 hours after administration.

[0250] (4) Administration method: Weigh before administration, calculate the administration amount according to the body weight. Through intravenous, gavage oral administration.

[0251] (5) Blood sampling time points: IV: 0.083h, 0.25h, 0.5h, 1h, 2h, 4h, 8h, 24h, PO: 0.25h, 0.5h, 1h, 2h, 4h, 6h, 8h, 24h after dosing. Blood samples were collected via jugular vein, about 0.2mL for each sample, K2-EDTA anticoagulation, and placed on ice after collection.

[0252] (6) Plasma sample processing: Blood samples were placed on ice after collection and centrifuged to separate plasma (centrifugation condition: 6800g, 6min, 2-8℃) within 1h. Plasma samples were stored at -80℃ when not in use.

[0253] (7) Result analysis: Pharmacokinetic parameters were calculated by Phoenix WinNonlin 7.0 using blood concentration data at different time points, and AUC0-t, AUC0-∞, MRT0-∞, Cmax, Tmax, and T1 / 2, etc. were provided, as well as their mean values and standard deviations.

[0254] The experimental results are shown in Table 6 below:

[0255] Table 6: Pharmacokinetic data of compound 26

[0256] compound Compound 26 BMS-986176 Dose Level mg / kg 3.00 3.00 [TA 1 / 2 h]]> 6.98 6.32 [TA max h]]> 3.10 3.33 C max ng / mL]]> 325.23 85.60 AUC (0-t) h*ng / mL]]> 4012.50 1069.29 AUC (0-∞) ng / mL 4126.32 1159.48 MRT (0-t) h]]> 8.09 7.52 MRT (0-∞) h]]> 10.93 9.50

[0257] Experimental conclusion: The pharmacokinetic data of compound 26 in SD rats showed that, compared with the positive control, the half-life and peak time of compound 26 in SD rats were comparable, but the peak concentration and in vivo exposure were about four times that of the positive control. Overall, the pharmacokinetic data of compound 26 in rats was significantly better than that of BMS-986176.

[0258] Experimental Example 5: Pharmacodynamic test of the compound of the present application on diabetic peripheral neuropathic pain (DPNP) in SD rats

[0259] Experimental purpose: The purpose of this experiment is to study the analgesic effect of the test substance on peripheral neuropathic pain (DPNP) in a streptozotocin (STZ)-induced diabetic rat model and the brain distribution of the compound accompanying the pharmacodynamic effect by using single oral gavage (ig.) administration.

[0260] Experimental method: After the adaptation period of the experimental rats, the baseline pain threshold of the foot was tested by Von Frey (UP-Down method) before modeling. Then, except for the blank control group, the remaining rats were fasted for 16h without water restriction, and then injected intraperitoneally with streptozotocin (65mg / kg; 2mL / kg) for modeling. After one week of modeling, blood was collected from the tail tip to detect 4h fasting blood glucose. Diabetic rats were considered to be those with blood glucose higher than 16.7mmol / L.

[0261] After successful modeling, the rats' baseline pain threshold was tested using Von Frey (UP-Down method), and the rats were evenly grouped according to the pain threshold. The groups were blank control, model, positive control 1 mg / kg, compound 26 (0.5 mg / kg), and compound 26 (1 mg / kg). The next day, the solvent or corresponding drug was administered, and at 2 h, 3 h, 4 h, 5 h, and 6 h after administration, the Von Frey fiber was used to determine the pain threshold of the affected plantar of the rats, to calculate the pain threshold increase rate of the test substance on diabetic rats with neuropathic pain, and to observe the time-effect relationship. After the experiment, the G3-G5 groups of rats were collected, 1-3 rats per group, and the plasma and brain tissue were tested.

[0262] Experimental results: The results are shown in Tables 7, 8, 9, and the accompanying drawings. Figure 1 Before modeling, the baseline pain threshold of each group of rats was 15 g. After modeling, the pain threshold of the rats in each group except the blank control group decreased to about 7.0 g, which showed a statistically significant difference compared with the blank control group (p<0.01).

[0263] Table 7: Effect of single oral gavage ig. of compound 26 on the pain threshold of diabetic rats with peripheral neuropathic pain n=5

[0264]

[0265] ##p<0.01 vs Control *p<0.05, **p<0.01 vs Model

[0266] Table 8: Effect of single oral gavage ig. of compound 26 on the pain threshold increase rate of diabetic rats with peripheral neuropathic pain n=5

[0267]

[0268] Table 9: Blood-brain distribution of compound in DPNP model rats after 6 h of gavage administration

[0269]

[0270] Experimental conclusion: In this test of the analgesic effect of the test substance on diabetic rats with peripheral neuropathic pain, ① After 2 h of administration of the test substances BMS-986176 1 mg / kg, compound 26 (0.5 mg / kg), and compound 26 (1 mg / kg), the pain threshold of the model animals was significantly improved, and the duration of analgesia was maintained for 4 h after administration; ② The analgesic effect and duration of compound 26 were better than those of the positive control BMS-986176. ③ Compound 26 had better brain penetration than the positive control BMS-986176.

Claims

1. A compound represented by the following formula: ###0001### or a pharmaceutically acceptable salt thereof. 。 2. A pharmaceutical composition comprising a compound represented by the formula of claim 1 or a pharmaceutically acceptable salt thereof, and a pharmaceutically acceptable carrier.

3. Use of a compound of claim 1 or a pharmaceutically acceptable salt thereof or a pharmaceutical composition of claim 2 for the manufacture of a medicament for the treatment of a disease or disorder mediated by the activity of Connexin Related Kinase 1.

4. The use according to claim 3, wherein the disease or disorder is selected from the group consisting of Alzheimer's disease, bipolar disorder, Parkinson's disease, schizophrenia, diabetic peripheral neuropathic pain, post-herpetic neuralgia.

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