Pyrazolopyridine compounds, pharmaceutical compositions thereof, methods of preparation and uses thereof

Abstract

The present application relates to pyrazolopyridine compounds, and pharmaceutical compositions, preparation methods and uses thereof, in particular, the present application relates to a compound shown in formula I or its pharmaceutically acceptable form, and pharmaceutical compositions, preparation methods and uses thereof. The compound can be used as a drug for treating cancer or fibrosis disease,

Description

TECHNICAL FIELD

[0001] The present application belongs to the field of pharmaceutical chemistry, and relates to pyrazolopyridine compounds, pharmaceutical compositions thereof, preparation methods and uses thereof. BACKGROUND

[0002] Transforming growth factor-beta (TGF-beta) is a multifunctional cytokine that regulates various cellular responses, such as cell proliferation, differentiation, migration and apoptosis. The TGF-beta superfamily includes TGF-beta 1, TGF-beta 2, TGF-beta 3, activin, inhibin, bone morphogenetic protein, etc. TGF-beta signals through two highly conserved single-pass transmembrane serine / threonine kinases, TGFβR1 and TGFβR2 (ACS Med. Chem. Lett., 2018, 9, 1117).

[0003] Smads are important intracellular TGF-beta signal transduction and regulation molecules that can directly transduce TGF-beta signals from the cell membrane to the nucleus. The TGF-beta / Smads signaling pathway plays an important role in the occurrence and development of tumors. In TGF-beta / Smads signal transduction, activated TGF-beta first binds to TGFβR2 on the surface of the cell membrane to form a heterodimeric complex, which is further recognized and bound by TGFβR1. Activated TGFβR1 further phosphorylates Smad2 / Smad3 protein, which further binds to Smad4 to form a heterotrimeric complex, which enters the nucleus and cooperates with auxiliary activation / inhibitory factors to regulate the transcription of target genes (Nature, 2003, 425, 577). Any change in any link of the TGF-beta / Smads signaling pathway will lead to abnormal signal transduction pathways (PNAS, 2019, 116, 9166).

[0004] Activation of the TGF-beta signaling pathway triggers significant pathological effects in the tumor stroma, including immune suppression, vascular regeneration and connective tissue proliferation. In addition, the TGF-beta signaling pathway can enhance the invasiveness of tumor cells, promote epithelial-mesenchymal transition and increase the tolerance to treatment of tumor epithelial cells (Nat. Neurosci., 2014, 17, 943).

[0005] Currently, the development of inhibitors against the key target TGFβR1 in the TGF-β signaling pathway has gradually been valued in the pharmaceutical industry, and published patent applications include WO 02 / 094833 A1, WO 2009 / 150547 A1, WO 2017 / 035118 A1, WO 2018 / 019106 A1, etc. However, there is still an urgent need in the art for new TGFβR1 inhibitors, especially TGFβR1 inhibitors with high activity and high selectivity. SUMMARY

[0006] The present application, through a large number of studies, unexpectedly found a pyrazolopyridine compound, which can significantly inhibit the activity of TGFβR1 and has good selectivity between TGFβR1 and TGFβR2. As a TGFβR1 inhibitor, it can be used for treating proliferative disorders and apoptosis disorders mediated at least in part by the TGF-β signaling pathway, especially diseases mediated at least in part by TGFβR1, such as cancer or fibrotic diseases.

[0007] In a first aspect, the present application provides a compound having the structure of Formula I or a pharmaceutically acceptable form thereof,

[0008]

[0009] wherein,

[0010] R 1 is selected from C 6-10 aryl and 5-10 membered heteroaryl, said C 6-10 aryl or 5-10 membered heteroaryl is optionally substituted with one or more R 4 ;

[0011] R 2 is selected from C 6-10 aryl and 5-10 membered heteroaryl, said C 6-10 aryl or 5-10 membered heteroaryl is optionally substituted with one or more R 5 ;

[0012] R 3 is selected from hydrogen, halogen, C 1-6 alkyl, C 1-6 haloalkyl, C 3-8 cycloalkyl, hydroxyalkyl, C 1-6 alkoxyalkyl, C 1-6 alkylaminoalkyl, 4-8 membered heterocyclylalkyl, hydroxyl, C 1-6 alkoxy, C 3-6 cycloalkyl-O-, C 1-6 haloalkoxy, amino, C 1-6 alkylamino, C 3-6cycloalkyl, 3-8 membered heterocyclyl, C 6-10 aryl and 5-10 membered heteroaryl, said C 6-10 aryl or 5-10 membered heteroaryl is optionally substituted with one or more R 6 substituents;

[0013] R 4 are each independently selected at each occurrence from hydrogen, deuterium, halogen, cyano, C 1-6 alkyl, C 1-6 haloalkyl, and C 3-8 cycloalkyl;

[0014] R 5 are each independently selected at each occurrence from hydrogen, deuterium, cyano, halogen, C 1-6 alkyl, C 1-6 haloalkyl, hydroxyalkyl, C 1-6 alkoxyalkyl, aminoalkyl, C 1-6 alkylaminoalkyl, C 3-8 cycloalkyl, 3-8 membered heterocyclyl-C 1-6 alkyl-, 3-8 membered heterocyclyl, C 6-10 aryl, 5-10 membered heteroaryl, -OR a , -NR b R c , -NR b C(=O)R a , -C(=O)R a , -C(=O)OR a , -C(=O)NR b R c , -(CR b R c ) m C(=O)NR b R c , -(CR a =CR a ) m C(=O)NR b R c , and -(CºC) m C(=O)NR b R c , said C 6-10 aryl, 5-10 membered heteroaryl, or 3-8 membered heterocyclyl is optionally substituted with one or more R 7 substituents;

[0015] R 6 are each independently selected at each occurrence from hydrogen, deuterium, halogen, C 1-6 alkyl, C 1-6 haloalkyl, C 1-6alkyl, C 1-6 alkyl, C 3-8 cycloalkyl, C 2-6 alkenyl, C 2-6 alkynyl, cyano, carboxyl, ester, 3-8 membered heterocyclyl, 3-8 membered heterocyclyl-C 1-6 alkyl-, C 1-6 alkylsulfonyl, C 3-6 cycloalkylsulfonyl, C 1-6 alkylacyl, C 3-6 cycloalkylacyl, C 4-8 heterocyclylacyl, C 1-6 alkylaminoamido and C 1-6 alkoxyamido;

[0016] R 7 each independently selected from the group consisting of halogen, C 1-6 alkyl, C 1-6 haloalkyl, hydroxyl, hydroxy C 1-6 alkyl, C 1-6 alkoxyalkyl, substituted or unsubstituted carbamoyl, 3-8 membered heterocyclyl, hydrogen, deuterium, -OR a , -NR b R c , -C(=O)R a , -C(=O)OR a , and -C(=O)NR b R c ;

[0017] R a each independently selected from the group consisting of hydrogen, C 1-6 alkyl, C 3-8 cycloalkyl, 3-8 membered heterocyclyl, 3-8 membered heterocyclyl-C 1-6 alkyl, substituted or unsubstituted carbamoyl;

[0018] R b and R c each independently selected from the group consisting of hydrogen, C 1-6 alkyl, halo C 1-6 alkyl, hydroxy C 1-6 alkyl, C 3-8 cycloalkyl and 3-8 membered heterocyclyl, or R b and R c together with the atom to which they are attached form a 3-7 membered ring which is unsubstituted or substituted with a hydroxyl group;

[0019] m is 0, 1, 2 or 3;

[0020] The pharmaceutically acceptable form is selected from the group consisting of pharmaceutically acceptable salts, esters, stereoisomers, tautomers, polymorphs, solvates, nitroxides, isotopically labeled forms, metabolites, and prodrugs.

[0021] In a second aspect, the present application provides a method for preparing the compound, comprising the following steps:

[0022] Step 1-1: Compounds SM1 and SM2 undergo condensation reaction to obtain compound IM1;

[0023]

[0024] Step 1-2: Compound IM1 and sodium azide undergo substitution reaction to obtain compound IM2;

[0025]

[0026] Step 1-3: Compound IM2 undergoes intramolecular cyclization and halogenation reaction to obtain compound IM3;

[0027]

[0028] Step 1-4: Compound IM3 and compound SM3 undergo coupling reaction to obtain compound of formula I;

[0029]

[0030] wherein W is selected from boronic acid group, 4,4,5,5-tetramethyl-1,3,2-dioxaborolan-2-yl, tri-n-butylstannyl group and sodium sulfinic acid group; R 1 , R 2 , R 3 and m are as defined in the first aspect.

[0031] In a third aspect, the present application provides a pharmaceutical composition comprising at least one compound of the first aspect or a pharmaceutically acceptable form thereof, and one or more pharmaceutically acceptable carriers.

[0032] In a fourth aspect, the present application provides a kit product comprising:

[0033] a) at least one compound of the first aspect or a pharmaceutically acceptable form thereof as a first therapeutic agent, or a pharmaceutical composition of the third aspect as a first pharmaceutical composition;

[0034] b) optionally at least one other therapeutic agent as a second therapeutic agent, or a pharmaceutical composition comprising the other therapeutic agent as a second pharmaceutical composition; and

[0035] c) optionally packaging and / or instructions.

[0036] In another aspect, the present application provides the use of a compound of the first aspect, or a pharmaceutically acceptable form thereof, or a pharmaceutical composition of the third aspect, in the manufacture of a medicament for the prevention and / or treatment of a disease or condition mediated at least in part by TGFpR1.

[0037] In another aspect, the present application provides a method of preventing and / or treating a disease or condition mediated at least in part by TGFpR1, comprising the step of administering to a subject in need thereof an effective amount of a compound of the first aspect, or a pharmaceutically acceptable form thereof, or a pharmaceutical composition of the third aspect.

[0038] In another aspect, the present application provides a compound of the first aspect, or a pharmaceutically acceptable form thereof, or a pharmaceutical composition of the third aspect, for use in the prevention and / or treatment of a disease or condition mediated at least in part by TGFpR1.

[0039] In another aspect, the present application provides the use of a compound of the first aspect, or a pharmaceutically acceptable form thereof, or a pharmaceutical composition of the third aspect, in the manufacture of a TGFpR1 inhibitor.

[0040] In another aspect, the present application provides a method of inhibiting the activity of TGFpR1, comprising the step of administering to a subject or a cell in need thereof an effective amount of a compound of the first aspect, or a pharmaceutically acceptable form thereof, or a pharmaceutical composition of the third aspect.

[0041] In another aspect, the present application provides a compound of the first aspect, or a pharmaceutically acceptable form thereof, or a pharmaceutical composition of the third aspect, for use in inhibiting the activity of TGFpR1.

[0042] In another aspect, the present application provides a pharmaceutical combination comprising a compound of the first aspect, or a pharmaceutically acceptable form thereof, or a pharmaceutical composition of the third aspect, and at least one other TGFpR1 inhibitor. DETAILED DESCRIPTION

[0044] In the present application, unless otherwise indicated, the scientific and technical terms used herein have the meanings that would be generally understood by one of ordinary skill in the art. Also, the cell culture, molecular genetics, nucleic acid chemistry, immunological laboratory procedures described herein are in accordance with conventional techniques of the corresponding field. In addition, for better understanding of the present application, the definitions and explanations of relevant terms are provided as follows.

[0045] The terms "comprise", "comprising", "include", "including", "have", "having" or "contain", "containing" or any other variation thereof, are intended to cover a non-exclusive inclusion. For example, a composition, a method, or an apparatus that comprises a list of elements is not necessarily limited to only those elements but can include other elements not expressly listed or inherent to such composition, method, or apparatus.

[0046] The term "pharmaceutically acceptable salt" refers to a salt of a compound of the present application that is substantially non-toxic to the organism. Pharmaceutically acceptable salts generally include, but are not limited to, salts of the compounds of the present application with pharmaceutically acceptable inorganic / organic acids or inorganic / organic bases, which are also known as acid or base addition salts.

[0047] The term "pharmaceutically acceptable ester" refers to an ester of a compound of the present application or its salt, which is substantially non-toxic to the organism and is hydrolyzed in the organism into the compound or its salt. Pharmaceutically acceptable esters generally include, but are not limited to, esters of the compounds of the present application with pharmaceutically acceptable carboxylic or sulfonic acids, which are also known as carboxylic or sulfonic acid esters.

[0048] The term "isomer" refers to a compound having the same molecular formula but different structural arrangement of atoms.

[0049] The term "stereoisomers" (or "optical isomers") refers to stable isomers having a fixed orientation of atoms in space, resulting from the presence of at least one chiral element (including chiral centers, chiral axes, chiral planes, etc.) that are not superimposable with their mirror images. Since the compounds of the present application can have asymmetric centers and other chemical structures that can lead to stereoisomers, the present application also includes these stereoisomers and mixtures thereof. Since the compounds of the present application (or their pharmaceutically acceptable salts) include asymmetric carbon atoms, they can exist in single stereoisomer form, racemates, mixtures of enantiomers, and mixtures of diastereomers. Generally, these compounds can be prepared in racemic form. However, if desired, pure stereoisomers, i.e., single enantiomers or diastereomers, or mixtures enriched (>98%, >95%, >93%, >90%, >88%, >85%, or >80%) in single stereoisomers, can be prepared or isolated from such compounds. As described below, single stereoisomers of the compounds are prepared from optically pure starting materials that contain the desired chiral center, or are prepared by resolution of mixtures of enantiomeric products, for example, by separation of mixtures of diastereomers, by recrystallization, chromatography, using chiral resolving agents, or by direct separation of enantiomers on chiral chromatographic columns. Starting compounds having particular stereochemistry can be obtained either commercially or by methods described herein or by methods known in the art once certain starting materials are available.

[0050] The term "enantiomers" refers to a pair of stereoisomers that are non-superimposable mirror images of one another.

[0051] The term "diastereomers" or "diastereomeric" refers to optical isomers that are not mirror images of one another.

[0052] The term "racemic mixture" or "racemate" refers to a mixture containing equal amounts of single enantiomers (i.e., an equimolar amount of two R and S enantiomers).

[0053] The term "non-racemic mixture" refers to a mixture containing unequal amounts of single enantiomers. Unless otherwise indicated, all stereoisomeric forms of the compounds of the present application are within the scope of the present application.

[0054] The term "tautomer" (or "tautomeric form") refers to structural isomers that exist in equilibrium with one another through low energy barriers. If tautomerism is possible (e.g., in solution), a chemical equilibrium of tautomers can be achieved. For example, prototropic tautomers (or proton-shift tautomers) include, but are not limited to, interconversions by proton migration, such as keto-enol isomerization, imine-enamine isomerization, amide-imidol isomerization, and the like. Unless otherwise specified, all tautomeric forms of the compounds of the application are within the scope of the application.

[0055] The term "polymorph" (or "polymorphic form") refers to a solid crystalline form of a compound or complex. One skilled in the art can obtain polymorphs of a molecule by a number of known methods. These methods include, but are not limited to, melt-recooling, melt- cooling, solvent-recooling, desolvation, rapid evaporation, rapid cooling, slow cooling, vapor diffusion, and sublimation. In addition, polymorphs can be detected, sorted, and characterized using well-known techniques, including, but not limited to, differential scanning calorimetry (DSC), thermogravimetric analysis (TGA), X-ray powder diffraction (XRPD), single crystal X-ray diffraction (SCXRD), solid state nuclear magnetic resonance (NMR), infrared spectroscopy (IR), Raman spectroscopy, and scanning electron microscopy (SEM), and the like.

[0056] The term "solvate" refers to a molecular complex of a compound of the application (or a pharmaceutically acceptable salt thereof) with one or more solvent molecules. Common solvates include, but are not limited to, hydrates (including hemi-hydrates, monohydrates, dihydrates, trihydrates, and the like), ethanolates, acetoneates, and the like.

[0057] The term "nitroxide" refers to a compound formed by oxidation of a nitrogen atom in a tertiary amine or a nitrogen-containing (hetero)aromatic ring. For example, the nitrogen atom at position 1 in the parent nucleus of a compound of Formula I can form a corresponding nitroxide.

[0058] The term "isotopically-labeled" refers to a derivative compound formed by replacing a particular atom in a compound of the application with an isotopic atom thereof. Unless otherwise specified, the compounds of the application include various isotopic atoms of H, C, N, O, F, P, S, Cl, such as 2 H(D), 3 H(T), 13 C, 14 C, 15 N, 17 O, 18 O, 18 F, 31 P, 32 P, 35 S, 36 S, and37 Cl.

[0059] The term "metabolite" refers to a derivative compound formed upon metabolism of a compound of the present application. Further information on metabolism can be found in Goodman and Gilman's: The Pharmacological Basis of Therapeutics (9 th ed.) [M], McGraw-Hill International Editions, 1996.

[0060] The term "prodrug" refers to a derivative compound that, upon administration to a subject, is capable of providing directly or indirectly a compound of the present application. Particularly preferred derivative compounds or prodrugs are compounds that increase the bioavailability of a compound of the present application when administered to a subject (e.g., are more readily absorbed into the blood), or that facilitate delivery of the parent compound to the site of action (e.g., the lymphatic system). Unless otherwise specified, all prodrug forms of a compound of the present application are within the scope of the present application, and various prodrug forms are well known in the art.

[0061] The term "independently" refers to at least two groups (or ring systems) that have the same or similar range of values present in a structure can have the same or different meaning in a particular instance. For example, substituent X and substituent Y are each independently hydrogen, halogen, hydroxyl, cyano, alkyl, or aryl, then when substituent X is hydrogen, substituent Y can be either hydrogen or halogen, hydroxyl, cyano, alkyl, or aryl; similarly, when substituent Y is hydrogen, substituent X can be either hydrogen or halogen, hydroxyl, cyano, alkyl, or aryl.

[0062] The term "halogen" refers to fluorine (F), chlorine (Cl), bromine (Br), and iodine (I).

[0063] The term "ester" refers to a group having the structure "-C(=O)OR", wherein R represents a hydrocarbon group, such as alkyl, aryl, and the like, preferably C 1-6 alkyl. Common ester groups include, but are not limited to, -COOCH3, -COOC2H5, -COOPh, and the like.

[0064] The term "alkyl" refers to a straight-chain or branched-chain saturated aliphatic hydrocarbon group. For example, the term "C 1-6 alkyl" as used in the present application refers to an alkyl group having 1 to 6 carbon atoms. Common alkyl groups include, but are not limited to, methyl, ethyl, n-propyl, i-propyl, n-butyl, sec-butyl, i-butyl, t-butyl, n-pentyl, n-hexyl, and the like. Alkyl groups in the present application are optionally substituted with one or more substituents described in the present application, such as halogen.

[0065] The term "haloalkyl" refers to an alkyl group having from one to six carbon atoms substituted with one or more (such as one to three) same or different halogen atoms. Common haloalkyl groups include, but are not limited to, -CH2F, -CHF2, -CF3, -CH2CF3, -CF2CF3, -CH2CH2CF3, -CH2Cl, and the like. Haloalkyl groups in the present application are optionally substituted with one or more substituents described in the present application. 1-6 The term "haloalkyl" refers to an alkyl group having from one to six carbon atoms substituted with one or more (such as one to three) same or different halogen atoms. Common haloalkyl groups include, but are not limited to, -CH2F, -CHF2, -CF3, -CH2CF3, -CF2CF3, -CH2CH2CF3, -CH2Cl, and the like. Haloalkyl groups in the present application are optionally substituted with one or more substituents described in the present application.

[0066] The term "hydroxyalkyl" refers to an alkyl group substituted with one or more hydroxyl groups, wherein alkyl is defined above. For example, the term "C 1-6 The term "hydroxyalkyl" refers to a hydroxyalkyl group as described above having from one to six carbon atoms. Common hydroxyalkyl groups include, but are not limited to, -CH2OH, -CH2CH2OH, -CH(CH2OH)2, C(CH3)2CH2OH, and the like.

[0067] The term "aminoalkyl" refers to an alkyl group substituted with one or more amino groups (-NH2), wherein alkyl is defined above. For example, the term "C 1-6 The term "aminoalkyl" refers to an aminoalkyl group as described above having from one to six carbon atoms. Common aminoalkyl groups include, but are not limited to, -CH2NH2, -C2H4NH2, -CH(CH2NH2)2, -C(CH3)2CH2NH2, and the like.

[0068] The term "alkoxy" refers to a group having the structure "alkyl-O-," wherein alkyl is defined above. For example, C 1-6 alkoxy, C 1-4 alkoxy, C 1-3 alkoxy, or C 1-2 alkoxy, and the like. Common alkoxy groups include, but are not limited to, methoxy, ethoxy, n-propoxy, i-propoxy, n-butoxy, i-butoxy, t-butoxy, pentoxy, hexoxy, and the like. Alkoxy groups in the present application are optionally substituted with one or more substituents described in the present application, and when further substituted with one or more halogens, form haloalkoxy groups described herein, such as C 1-6 haloalkoxy.

[0069] The term "alkoxyalkyl" refers to an alkyl group substituted with one or more alkoxy groups, wherein alkoxy and alkyl are defined above. For example, the term "C 1-6 alkoxyalkyl" refers to "C 1-6 alkoxy-alkyl," preferably C 1-6 alkoxy-C 1-6 alkyl. For example, C 1-4Alkoxy-C 1-4 Alkyl, C 1-3 Alkoxy-C 1-3 Alkyl or C 1-2 Alkoxy-C 1-2 Alkyl groups, etc. Common alkoxyalkyl groups include (but are not limited to) methoxymethyl, methoxyethyl, ethoxymethyl, ethoxyethyl, methoxypropyl, etc.

[0070] The term "alkoxyamide group" refers to a group having an "alkoxy-C(O)N(R)-" structure, where R represents a hydrogen or hydrocarbon group, and the definition of alkoxy is as described above. For example, the term "C" used in this invention... 1-6 "Alkoxyamide group" refers to "C 1-6 Alkoxy-C(O)N(R)-”, preferably C 1-6 Alkyl group -C(O)NH-. For example, C 1-4 Alkoxy-C(O)NH-, C 1-3 Alkoxy-C(O)NH- or C 1-2 Alkoxy groups include -C(O)NH-, etc. Common alkoxy amide groups include (but are not limited to) CH3OC(O)NH-, C2H5OC(O)NH-, etc.

[0071] The term "alkylamine" refers to an amino group substituted with a monoalkyl or dialkyl group, where the alkyl group is defined as described above. For example, C 1-6 alkylamine group, C 1-4 alkylamine group, C 1-3 alkylamine or C 1-2 Alkylamine groups, etc. Common alkylamine groups include (but are not limited to) methylamino, dimethylamino, ethylamino, diethylamino, methylisopropylamino, etc. The alkylamine group in this invention may optionally be substituted by one or more substituents described in this invention.

[0072] The term "alkylamine alkyl" refers to an alkyl group substituted with one or more alkylamine groups, wherein the definitions of alkylamine and alkyl are as described above. For example, the term "C" as used in this invention... 1-6 "alkylamine alkyl" refers to "C 1-6 "alkylamino-alkyl", preferably C 1-6 Alkylamine-C 1-6 Alkyl group. For example, C6 1-4 Alkylamine-C 1-4 Alkyl, C 1-3 Alkylamine-C 1-3 Alkyl or C 1-2 Alkylamine-C 1-2 Alkyl groups, etc. Common alkylamine alkyl groups include (but are not limited to) methylaminomethyl, methylaminoethyl, ethylaminomethyl, ethylaminoethyl, methylaminopropyl, etc.

[0073] The term "alkylamido" refers to a group having the structure "alkyl-C(=0)N(R)-", wherein R is hydrogen or alkyl, and alkyl is as defined above. For example, the term "C 1-6 alkylamido" as used herein refers to "C 1-6 alkyl-C(=0)N(R)-", preferably C 1-6 alkyl-C(=0)NH-. For example C 1-4 alkyl-C(=0)NH-, C 1-3 alkyl-C(=0)NH-, or C 1-2 alkyl-C(=0)NH-, and the like. Common alkylamido groups include, but are not limited to, CH3NHC(=0)NH-, (CH3)2NC(=0)NH-, and the like.

[0074] The term "alkylacyl" refers to a group having the structure "alkyl-C(=0)-", wherein alkyl is as defined above. For example C 1-6 alkylacyl, C 1-4 alkylacyl, C 1-3 alkylacyl, or C 1-2 alkylacyl, and the like. Common alkylacyl groups include, but are not limited to, formyl, acetyl, and the like.

[0075] The term "cycloalkyl" refers to a saturated, monocyclic or polycyclic (such as bicyclic) non-aromatic cyclic hydrocarbon group. For example, the term "C 3-8 cycloalkyl" as used herein refers to a cycloalkyl group having from 3 to 8 carbon atoms, including C3 cycloalkyl, C4 cycloalkyl, C5 cycloalkyl, C6 cycloalkyl, C7 cycloalkyl, and C8 cycloalkyl, preferably C 3-6 cycloalkyl. Common cycloalkyl groups include, but are not limited to, monocyclic cycloalkyl groups such as cyclopropyl, cyclobutyl, cyclopentyl, cyclohexyl, cycloheptyl, cyclooctyl, cyclononyl, and the like; or bicyclic cycloalkyl groups including fused, bridged, or spirocyclic groups such as bicyclo[l. l. l]pentyl, bicyclo[2.2. l]heptyl, bicyclo[3.2. l]octyl, bicyclo[5.2.0]nonyl, decahydronaphthyl, and the like. Cycloalkyl groups in the present invention are optionally substituted with one or more substituents described in the present invention, such as methyl.

[0076] The term "cycloalkylamido" refers to a group having the structure "cycloalkyl-NH-", wherein cycloalkyl is as defined above. For example, the term "C 3-6 cycloalkylamido" as used herein refers to "C 3-6 cycloalkyl-NH-", including C3 cycloalkylamido, C4 cycloalkylamido, C5 cycloalkylamido, and C6 cycloalkylamido. Common cycloalkyl groups include, but are not limited to, cyclopropylamido, cyclobutylamido, cyclopentylamido, cyclohexylamido, bicyclo[l. l. l]pentylamido, and the like.

[0077] The term "cycloalkylacyl" refers to a group having the structure "cycloalkyl-C(=O)-", wherein cycloalkyl is as defined above. For example, cyclopropylcarbonyl, cyclobutylcarbonyl, cyclopentylcarbonyl, cyclohexylcarbonyl, and the like. 3-6 Common cycloalkylacyls include, but are not limited to, cyclopropylcarbonyl, cyclobutylcarbonyl, cyclopentylcarbonyl, cyclohexylcarbonyl, and the like.

[0078] The term "heterocyclyl" refers to a saturated or partially saturated, monocyclic or polycyclic (such as bicyclic) nonaromatic ring radical whose ring members are comprised of carbon atoms and at least one heteroatom selected from the group consisting of nitrogen, oxygen, and sulfur. The heterocyclyl group can be attached to the remainder of the molecule through any one of the ring atoms, if valence requirements permit. For example, the term "3-8 membered heterocyclyl" as used herein refers to a heterocyclyl group having from 3 to 8 ring atoms, including 3-membered heterocyclyl, 4-membered heterocyclyl, 5-membered heterocyclyl, 6-membered heterocyclyl, 7-membered heterocyclyl, and 8-membered heterocyclyl. Preferred are 4-8 membered heterocyclyl, more preferred are 4-6 membered heterocyclyl. Common heterocyclyls include, but are not limited to, oxiranyl, aziridinyl, azetidinyl, oxetanyl, tetrahydrofuryl, pyrrolidinyl, pyrrolidinonyl, imidazolidinyl, pyrazolidinyl, tetrahydropyranyl, piperidinyl, piperazinyl, morpholinyl, thiomorpholinyl, dithianyl, or trithianyl. The heterocyclyl groups in the present application are optionally substituted with one or more substituents described in the present application.

[0079] The term "heterocyclylalkyl" refers to an alkyl group substituted with one or more heterocyclyl groups, wherein heterocyclyl and alkyl are as defined above. For example, the term "3-8 membered heterocyclylalkyl" as used herein refers to "3-8 membered heterocyclyl-alkyl", preferably 3-8 membered heterocyclyl-C 1-6 alkyl. For example, 4-6 membered heterocyclyl-C 1-6 alkyl, 4-6 membered heterocyclyl-C 1-4 alkyl, 4-6 membered heterocyclyl-C 1-3 alkyl, 4-6 membered heterocyclyl-C 1-2Alkyl groups, etc. Common heterocyclic alkyl groups include (but are not limited to) aziridine methyl, oxadiazine methyl, tetrahydrofuranylmethyl, pyrrolidine methyl, pyrrolidone methyl, imidazolidine methyl, pyrazolidine methyl, tetrahydropyranyl methyl, piperidinyl methyl, piperazinyl methyl, morpholinyl methyl, thiomorpholinyl methyl, etc.

[0080] The term "heterocyclic acyl" refers to a group having a "heterocyclic -C(=O)-" structure, where the heterocyclic group is defined as described above. For example, C 3-8 Heterocyclic acyl group, C 3-6 Heterocyclic acyl group, C 4-6 Heterocyclic acyl groups, etc. Common heterocyclic acyl groups include (but are not limited to) azirrocyclobutylformyl, oxocyclobutylformyl, tetrahydrofuranylformyl, pyrrolylformyl, pyrrolidoneylformyl, imidazoylformyl, pyrazolylalkylformyl, tetrahydropyranylformyl, piperidinylformyl, piperazinylformyl, morpholinylformyl, thiomorpholinylformyl, etc.

[0081] The term "aryl" refers to a monocyclic or fused polycyclic aromatic hydrocarbon group having a conjugated π-electron system. For example, the term "C" as used in this invention... 6-10 "Aryl" refers to an aryl group having 6 to 10 carbon atoms. Common aryl groups include (but are not limited to) phenyl, naphthyl, anthraceneyl, phenanthryl, acenaphthene, azulel, fluorenyl, indene, pyrene, etc. The aryl group in this invention may optionally be replaced by one or more substituents described in this invention (such as halogen, hydroxyl, cyano, nitro, C...). 1-6 Alkyl groups, etc., are substituted.

[0082] The term "heteroaryl" refers to a monocyclic or fused polycyclic (particularly benzo-fused polycyclic) aromatic radical having a conjugated pi-electron system, the ring atoms of which are composed of carbon atoms and at least one heteroatom selected from nitrogen, oxygen, and sulfur. The heteroaryl group can be attached to the remainder of the molecule through any one of the ring atoms, if valence requirements are met. For example, the term "5-10 membered heteroaryl" as used herein refers to a heteroaryl group having 5 to 10 ring atoms. Common heteroaryls include, but are not limited to, thienyl, furyl, pyrrolyl, oxazolyl, thiazolyl, imidazolyl, pyrazolyl, isoxazolyl, isothiazolyl, oxadiazolyl, triazolyl, thiadiazolyl, pyridinyl, pyridazinyl, pyrimidinyl, pyrazinyl, triazinyl, and benzo derivatives thereof, and the like. The heteroaryl groups in the present application are optionally substituted with one or more substituents described in the present application, such as halogen, C 1-6 alkyl, and the like.

[0083] The term "alkenyl" refers to a straight-chain or branched aliphatic hydrocarbon radical having at least one C=C double bond. For example, the term "C 2-6 alkenyl" as used herein refers to an alkenyl group having 2 to 6 carbon atoms. Common alkenyl groups include, but are not limited to, ethenyl, propenyl, n-butenyl, 3-methylbut-2-enyl, n-pentenyl, n-octenyl, n-decenyl, and the like. The alkenyl groups in the present application are optionally substituted with one or more substituents described in the present application.

[0084] The term "alkynyl" refers to a straight-chain or branched aliphatic hydrocarbon radical having at least one C≡C triple bond. For example, the term "C 2-6 alkynyl" as used herein refers to an alkynyl group having 2 to 6 carbon atoms. Common alkynyl groups include, but are not limited to, ethynyl, 2-propynyl, 2-butynyl, 1,3- butadiynyl, and the like. The alkynyl groups in the present application are optionally substituted with one or more substituents described in the present application.

[0085] The term "substituted" means that one or more (e.g., 1, 2, 3, or 4) atoms (e.g., hydrogen atoms) or moieties (e.g., triflate) of the designated group are replaced with other atoms or moieties, provided that the designated group meets the valency requirements in the present context and that a stable compound results after substitution. Combinations of substituents and / or variables are permissible only if such combinations result in stable compounds. If a substituent is described as "optionally substituted" that substituent can be unsubstituted or substituted. If a first substituent is described as optionally being substituted by one or more of a list of second substituents, then one or more hydrogen atoms of the first substituent can be individually or independently replaced with one or more of the list of second substituents, or none are replaced.

[0086] The term "one or more" means 1 or more than 1, e.g., 2, 3, 4, 5, 6, 7, 8, 9, or 10, under reasonable conditions.

[0087] Compounds

[0088] The present application provides a compound having the structure of Formula I, or a pharmaceutically acceptable form thereof,

[0089]

[0090] wherein,

[0091] R 1 is selected from C 6-10 aryl and 5-10 membered heteroaryl, said C 6-10 aryl or 5-10 membered heteroaryl is optionally substituted with one or more R 4 substituents;

[0092] R 2 is selected from C 6-10 aryl and 5-10 membered heteroaryl, said C 6-10 aryl or 5-10 membered heteroaryl is optionally substituted with one or more R 5 substituents;

[0093] R 3 is selected from hydrogen, halogen, C 1-6 alkyl, C 1-6 haloalkyl, C 3-8 cycloalkyl, hydroxyalkyl, C 1-6 alkoxyalkyl, C 1-6 alkylaminoalkyl, 4-8 membered heterocyclylalkyl, hydroxyl, C 1-6 alkoxy, C 3-6 cycloalkyl-O-, C 1-6 haloalkoxy, amino, C 1-6 alkylamino, C3-6 cycloalkylamino, cyano, carboxy, ester, 4-8 membered heterocyclyl, C 6-10 aryl and 5-10 membered heteroaryl, said C 6-10 aryl or 5-10 membered heteroaryl is optionally substituted with one or more R 6 substituents;

[0094] R 4 each occurrence is independently selected from hydrogen, deuterium, halogen, cyano, C 1-6 alkyl, C 1-6 haloalkyl, and C 3-8 cycloalkyl;

[0095] R 5 each occurrence is independently selected from hydrogen, deuterium, cyano, halogen, C 1-6 alkyl, C 1-6 haloalkyl, hydroxyalkyl, C 1-6 alkoxyalkyl, aminoalkyl, C 1-6 alkylaminoalkyl, C 3-8 cycloalkyl, 3-8 membered heterocyclyl-C 1-6 alkyl-, 3-8 membered heterocyclyl, C 6-10 aryl, 5-10 membered heteroaryl, -OR a , -NR b R c , -NR b C(=O)R a , -C(=O)R a , -C(=O)OR a , -C(=O)NR b R c , -(CR b R c ) m C(=O)NR b R c , -(CR a =CR a ) m C(=O)NR b R c , and -(C≡C) m C(=O)NR b R c , said C 6-10 aryl, 5-10 membered heteroaryl, or 3-8 membered heterocyclyl is optionally substituted with one or more R 7 substituents;

[0096] R 6 each occurrence is independently selected from hydrogen, deuterium, halogen, C 1-6 alkyl, C 1-6 haloalkyl, C 1-6alkyl, C 1-6 alkyl, C 3-8 cycloalkyl, C 2-6 alkenyl, C 2-6 alkynyl, cyano, carboxyl, ester, 3-8 membered heterocyclyl, 3-8 membered heterocyclyl-C 1-6 alkyl-, C 1-6 alkylsulfonyl, C 3-6 cycloalkylsulfonyl, C 1-6 alkylacyl, C 3-6 cycloalkylacyl, C 4-8 heterocyclylacyl, C 1-6 alkylaminoamido and C 1-6 alkoxyamido;

[0097] R 7 each independently selected from the group consisting of halogen, C 1-6 alkyl, C 1-6 haloalkyl, hydroxyl, hydroxy C 1-6 alkyl, C 1-6 alkoxyalkyl, substituted or unsubstituted carbamoyl, 3-8 membered heterocyclyl, hydrogen, deuterium, -OR a , -NR b R c , -C(=O)R a , -C(=O)OR a and -C(=O)NR b R c ;

[0098] R a each independently selected from the group consisting of hydrogen, C 1-6 alkyl, C 3-8 cycloalkyl, 3-8 membered heterocyclyl, 3-8 membered heterocyclyl-C 1-6 alkyl, substituted or unsubstituted carbamoyl;

[0099] R b and R c each independently selected from the group consisting of hydrogen, C 1-6 alkyl, halo C 1-6 alkyl, hydroxy C 1-6 alkyl, C 3-8 cycloalkyl and 3-8 membered heterocyclyl, or R b and R c together with the atom to which they are attached form a 3-7 membered ring which is unsubstituted or substituted with a hydroxyl group;

[0100] m is 0, 1, 2 or 3;

[0101] The pharmaceutically acceptable form is selected from the group consisting of pharmaceutically acceptable salts, esters, stereoisomers, tautomers, polymorphs, solvates, nitroxides, isotopically labeled forms, metabolites, and prodrugs.

[0102] In some embodiments, R 1 is selected from the group consisting of phenyl and 5-6 membered heteroaryl, each independently substituted with one or more R 4 substituents;

[0103] R 4 is selected from the group consisting of hydrogen, deuterium, halogen, C 1-6 alkyl, C 1-6 haloalkyl, and C 3-8 cycloalkyl.

[0104] In some embodiments, R 1 is selected from the group consisting of phenyl, pyridyl, and pyrazolyl, each independently substituted with one or more R 4 substituents;

[0105] R 4 is selected from the group consisting of hydrogen, deuterium, halogen, C 1-6 alkyl, C 1-6 haloalkyl, and C 3-8 cycloalkyl.

[0106] In some embodiments, R 1 is selected from the group consisting of phenyl and pyridyl, each independently substituted with one or more R 4 substituents;

[0107] R 4 is selected from the group consisting of halogen, C 1-6 alkyl, and C 1-6 haloalkyl.

[0108] In some embodiments, R 1 is selected from the group consisting of phenyl and pyridyl, each independently substituted with one or more R 4 substituents;

[0109] R 4 is selected from the group consisting of fluorine, chlorine, methyl, and trifluoromethyl.

[0110] In some embodiments, R 1 is selected from the group consisting of the following fragments:

[0111]

[0112] In some embodiments, R 1 is

[0113] In some embodiments, R 2selected from the group consisting of wherein,

[0114] Ring A is a phenyl ring or a 6-membered heteroaromatic ring;

[0115] Ring B is a 5-membered heteroaromatic ring;

[0116] A1to A6and B1to B3are each independently selected from C, CH, N, O, and S;

[0117] R 5 is, at each occurrence, independently selected from hydrogen, deuterium, cyano, halogen, C 1-6 alkyl, C 1-6 haloalkyl, hydroxyalkyl, C 1-6 alkoxyalkyl, aminoalkyl, C 1-6 alkylaminoalkyl, C 3-8 cycloalkyl, 4-8 membered heterocyclyl-C 1-6 alkyl-, 4-8 membered heterocyclyl, phenyl, 5-6 membered heteroaryl, -NR b R c , -C(=O)NR b R c and -(CR a =CR a ) m C(=O)NR b R c , said phenyl, 5-6 membered heteroaryl or 4-8 membered heterocyclyl being optionally substituted with one or more R 7 ;

[0118] R a is, at each occurrence, independently selected from hydrogen, C 1-6 alkyl, C 3-8 cycloalkyl, 3-8 membered heterocyclyl, 3-8 membered heterocyclyl-C 1-6 alkyl and substituted or unsubstituted carbamoyl;

[0119] R b and R c are, at each occurrence, independently selected from hydrogen, C 1-6 alkyl, haloC 1-6 alkyl, hydroxyC 1-6 alkyl, C 3-8 cycloalkyl and 3-8 membered heterocyclyl, or R a and R b together with the atom to which they are attached form a 3-7 membered ring which is unsubstituted or substituted with one or more hydroxyl groups.

[0120] In some embodiments, R 5 is selected from hydrogen, deuterium, hydroxyC 1-6 alkyl, -NR b Rc and -C(=O)NR b R c ;

[0121] R a each independently at each occurrence is selected from hydrogen, C 1-6 alkyl, C 3-8 cycloalkyl, 3-8 membered heterocyclyl, 3-8 membered heterocyclyl-C 1-6 alkyl, and substituted or unsubstituted carbamoyl;

[0122] R b and R c each independently at each occurrence is selected from hydrogen, C 1-6 alkyl, haloC 1-6 alkyl, hydroxyC 1-6 alkyl, C 3-8 cycloalkyl, and 3-8 membered heterocyclyl, or R a and R b together with the atoms to which they are attached form a 3-7 membered ring which is unsubstituted or substituted with one or more hydroxyl groups.

[0123] In some embodiments, R 2 is selected from wherein,

[0124] Ring A is a phenyl ring or a 6-membered heteroaromatic ring;

[0125] Ring B is a 5-membered heteroaromatic ring;

[0126] A1to A6and B1to B3are each independently selected from C, CH, and N;

[0127] R 5 is selected from hydrogen, deuterium, and -C(=O)NR b R c ;

[0128] R b and R c each independently at each occurrence is selected from hydrogen, C 1-6 alkyl, haloC 1-6 alkyl, hydroxyC 1-6 alkyl, C 3-8 cycloalkyl, and 3-8 membered heterocyclyl, or R a and R b together with the atoms to which they are attached form a 3-7 membered ring which is unsubstituted or substituted with one or more hydroxyl groups.

[0129] In some embodiments, R 2 is selected from the following fragments:

[0130]

[0131]

[0132] In some embodiments, R 2 is selected from

[0133] R 5 is selected from hydrogen, deuterium, and -C(=O)NR b R c ;

[0134] R b and R c are each independently at each occurrence selected from hydrogen, C 1-6 alkyl, haloC 1-6 alkyl, and hydroxyC 1-6 alkyl; preferably, R b and R c are each hydrogen.

[0135] In some embodiments, R 2 is selected from

[0136] In some embodiments, R 3 is selected from hydrogen, halo, C 1-6 alkyl, C 1-6 haloalkyl, C 3-8 cycloalkyl, cyano, 4-8 membered heterocyclyl, phenyl, and 5-6 membered heteroaryl, which phenyl or 5-6 membered heteroaryl is optionally substituted with one or more R 6 ;

[0137] R 6 is selected from hydrogen, deuterium, halo, C 1-6 alkyl, C 1-6 haloalkyl, C 1-6 alkoxyalkyl, hydroxyalkyl, C 1-6 alkylaminoalkyl, C 3-8 cycloalkyl, C 2-6 alkenyl, C 2-6 alkynyl, cyano, carboxyl, ester, 3-8 membered heterocyclyl, 3-8 membered heterocyclyl-C 1-6 alkyl, C 1-6 alkylsulfonyl, C 3-6 cycloalkylsulfonyl, C 1-6 alkylacyl, C 3-6 cycloalkylacyl, C 4-8 heterocyclylacyl, C 1-6 alkylamidoamido, and C 1-6 alkoxylamido.

[0138] In some embodiments, R 3 is selected from hydrogen, halo, C1-6 alkyl, C 1-6 haloalkyl, C 3-8 cycloalkyl and 4-8 membered heterocyclyl.

[0139] In some embodiments, R 3 is selected from hydrogen, C 1-6 alkyl and C 1-6 haloalkyl.

[0140] In some embodiments, R 3 is selected from hydrogen, methyl and trifluoromethyl.

[0141] In some embodiments, R 3 is hydrogen.

[0142] In some embodiments, R 1 is selected from phenyl and 5-6 membered heteroaryl, which phenyl or 5-6 membered heteroaryl is optionally substituted with one or more R 4 ;

[0143] R 2 is selected from 5-10 membered heteroaryl, which 5-10 membered heteroaryl is optionally substituted with one or more R 5 ;

[0144] R 3 is selected from hydrogen, halogen, C 1-6 alkyl, C 1-6 haloalkyl, C 3-8 cycloalkyl, cyano, 4-8 membered heterocyclyl, phenyl and 5-6 membered heteroaryl, which phenyl or 5-6 membered heteroaryl is optionally substituted with one or more R 6 ;

[0145] R 4 is at each occurrence independently selected from hydrogen, deuterium, halogen, C 1-6 alkyl, C 1-6 haloalkyl and C 3-8 cycloalkyl;

[0146] R 5 is at each occurrence independently selected from hydrogen, deuterium, cyano, halogen, C 1-6 alkyl, C 1-6 haloalkyl, hydroxyalkyl, C 1-6 alkoxyalkyl, aminoalkyl, C 1-6 alkylaminoalkyl, C 3-8 cycloalkyl, 4-8 membered heterocyclyl-C 1-6 alkyl-, 4-8 membered heterocyclyl, phenyl, 5-6 membered heteroaryl, -NR b R c , -C(=O)NR b R c and -(CRa =CR a ) m C(=O)NR b R c , said phenyl, 5-6 membered heteroaryl or 4-8 membered heterocyclyl is optionally substituted with one or more R 7 substituents;

[0147] R 6 are each independently at each occurrence selected from hydrogen, deuterium, halogen, C 1-6 alkyl, C 1-6 haloalkyl, C 1-6 alkoxyalkyl, hydroxyalkyl, C 1-6 alkylaminoalkyl, C 3-8 cycloalkyl, C 2-6 alkenyl, C 2-6 alkynyl, cyano, carboxyl, ester, 3-8 membered heterocyclyl, 3-8 membered heterocyclyl-C 1-6 alkyl-, C 1-6 alkylsulfonyl, C 3-6 cycloalkylsulfonyl, C 1-6 alkylacyl, C 3-6 cycloalkylacyl, C 4-8 heterocyclylacyl, C 1-6 alkylaminoamido, and C 1-6 alkoxyamido;

[0148] R 7 are each independently at each occurrence selected from halogen, C 1-6 alkyl, C 1-6 haloalkyl, hydroxy, hydroxyC 1-6 alkyl, C 1-6 alkoxyalkyl, substituted or unsubstituted carbamoyl, 4-8 membered heterocyclyl, and -C(=O)NR b R c ;

[0149] R b and R c are each independently at each occurrence selected from hydrogen, C 1-6 alkyl, haloC 1-6 alkyl, hydroxyC 1-6 alkyl, C 3-8 cycloalkyl, and 3-8 membered heterocyclyl, or R b and R c together with the atom to which they are attached form a 4-7 membered ring which is unsubstituted or substituted with one or more hydroxyl groups;

[0150] m is 0, 1, 2, or 3.

[0151] In some embodiments, R 1selected from phenyl and pyridyl, said phenyl or pyridyl being optionally substituted with one or more R 4 substituted;

[0152] R 2 selected from a nitrogen-containing 5-10 membered heteroaryl, said nitrogen-containing 5-10 membered heteroaryl being optionally substituted with one or more R 5 substituted;

[0153] R 3 selected from hydrogen, halogen, C 1-6 alkyl, C 1-6 haloalkyl, C 3-8 cycloalkyl and 4-8 membered heterocyclyl;

[0154] R 4 selected from halogen, C 1-6 alkyl and C 1-6 haloalkyl;

[0155] R 5 selected from hydrogen, cyano, hydroxyalkyl, C 1-6 alkoxyalkyl, C 1-6 alkylaminoalkyl, 4-8 membered heterocyclyl-C 1-6 alkyl-, 4-8 membered heterocyclyl, phenyl, 5-6 membered heteroaryl, -NR b R c and -C(=O)NR b R c , said phenyl, 5-6 membered heteroaryl or 4-8 membered heterocyclyl being optionally substituted with one or more R 7 substituted;

[0156] R 7 selected from halogen, C 1-6 alkyl, C 1-6 haloalkyl, hydroxyC 1-6 alkyl, C 1-6 alkoxyalkyl, substituted or unsubstituted carbamoyl, 4-8 membered heterocyclyl and -C(=O)NR b R c ;

[0157] R b and R c selected from hydrogen, C 1-6 alkyl, haloC 1-6 alkyl, hydroxyC 1-6 alkyl, C 3-8 cycloalkyl and 3-8 membered heterocyclyl, or R b and R c together with the atom to which they are attached form a 4-7 membered ring which is unsubstituted or substituted with one or more hydroxy;

[0158] m is 0, 1, 2 or 3.

[0159] In some embodiments, the compound is selected from:

[0160]

[0161]

[0162]

[0163]

[0164]

[0165]

[0166]

[0167] Preparation process

[0168] In a second aspect, the present application provides a method for preparing the compound of the first aspect, comprising the following steps:

[0169] Step 1-1: condensation reaction of compound SM1 and compound SM2 to obtain compound IM1;

[0170]

[0171] Step 1-2: substitution reaction of compound IM1 and sodium azide to obtain compound IM2;

[0172]

[0173] Step 1-3: intramolecular cyclization and halogenation reaction of compound IM2 to obtain compound IM3;

[0174]

[0175] Step 1-4: coupling reaction of compound IM3 and compound SM3 to obtain the compound of formula I;

[0176]

[0177] wherein W is selected from boronic acid group, 4,4,5,5-tetramethyl-1,3,2-dioxaborolan-2-yl, tri-n-butylstannyl group and sodium sulfinic acid group; R 1 , R 2 , R 3 and m are as defined in the first aspect.

[0178] In some embodiments, step 1-1 is carried out in a suitable organic solvent, which can be selected from halogenated hydrocarbons (such as dichloromethane, chloroform, 1,2-dichloroethane, and the like), nitriles (such as acetonitrile, and the like), N-methylpyrrolidinone, N,N-dimethylformamide, N,N-dimethylacetamide, tetrahydrofuran, 1,4-dioxane, dimethylsulfoxide, and any combination thereof, preferably N,N-dimethylformamide.

[0179] In some embodiments, step 1-1 is carried out in the presence of a suitable condensing agent. The condensing agent can be selected from dicyclohexylcarbodiimide (DCC), 1- ethyl-(3-dimethylaminopropyl)carbodiimide hydrochloride (EDCI), 2-(7-oxadiazolyl)-N,N,N',N'-tetramethyluronium hexafluorophosphate (HATU), O-benzotriazole- tetramethyluronium hexafluorophosphate (HBTU), benzotriazol-1-yl-oxytripyrrolidinophosphonium hexafluorophosphate (PyBOP), preferably 2-(7-oxadiazolyl)-N,N,N',N'- tetramethyluronium hexafluorophosphate (HATU).

[0180] In some embodiments, step 1-1 is carried out in the presence of a suitable organic base, which can be selected from N,N-diisopropylethylamine, triethylamine, 4- dimethylaminopyridine, and pyridine, preferably N,N-diisopropylethylamine.

[0181] In some embodiments, step 1-1 is carried out at a suitable temperature, which is in the range of 0-200 °C, preferably 0-100 °C, for example room temperature.

[0182] In some embodiments, step 1-2 is carried out in a suitable organic solvent, which can be selected from N,N-dimethylformamide, acetonitrile, ethers (such as ethylene glycol dimethyl ether, tetrahydrofuran, 1,4-dioxane), aromatic hydrocarbons (such as toluene), and any combination thereof.

[0183] In some embodiments, step 1-2 is carried out at a suitable temperature, which is in the range of 0-200 °C, preferably 0-100 °C, for example 60-100 °C.

[0184] In some embodiments, step 1-3 is carried out in a suitable organic solvent, which can be selected from halogenated hydrocarbons (such as dichloromethane, chloroform, 1,2- dichloroethane, and the like), N,N-dimethylformamide, acetonitrile, ethers (such as ethylene glycol dimethyl ether, tetrahydrofuran, 1,4-dioxane), aromatic hydrocarbons (such as toluene), and any combination thereof.

[0185] In some embodiments, step 1-3 is carried out at a suitable temperature, which is in the range of 0-200 °C, preferably 0-100 °C, for example 60-100 °C.

[0186] In some embodiments, steps 1-4 are carried out in a suitable organic solvent or a mixed solution of organic solvent and water, which can be selected from halogenated hydrocarbons (e.g., dichloromethane, chloroform, 1,2-dichloroethane, etc.), methanol, ethanol, tert-butanol, N,N-dimethylformamide, acetonitrile, ethers (e.g., ethylene glycol dimethyl ether, tetrahydrofuran, 1,4-dioxane), aromatic hydrocarbons (e.g., toluene), and any combination thereof, preferably toluene or 1,4-dioxane.

[0187] In some embodiments, steps 1-4 are carried out in the presence of a suitable catalyst, which is preferably a palladium catalyst, and can be selected from palladium(0) tetra(triphenylphosphine) (Pd(PPh3)4), palladium(II) acetate (Pd(OAc)2), Pd2(dba)3, Pd(PPh3)2Cl2, Pd(PPh3)2Cl2dichloromethane complex, PdCl2(Amphos)2, and Pd(dppf)Cl2, preferably PdCl2(Amphos)2or Pd(dppf)Cl2.

[0188] In some embodiments, steps 1-4 are carried out in the presence of a suitable ligand, which can be selected from triphenylphosphine (PPh3), BINAP, tri(o-tolyl)phosphine (P(o-tol)3), tricyclohexylphosphine tetrafluoroborate (TCHP), and X-PHOS, preferably PPh3or X-PHOS.

[0189] In some embodiments, steps 1-4 are carried out in the presence of a suitable base, which includes an organic base or an inorganic base, the organic base can be selected from N,N-diisopropylethylamine, triethylamine, 4-dimethylaminopyridine, and pyridine, and the inorganic base can be selected from K3PO4, NaH, K2CO3, Na2CO3, Cs2CO3, and NaOH, preferably K2CO3or Cs2CO3.

[0190] In some embodiments, steps 1-4 are carried out at a suitable temperature, which is 0-200 °C, preferably 50-150 °C.

[0191] Pharmaceutical composition

[0192] In a third aspect, the present application provides a pharmaceutical composition comprising at least one compound of the first aspect or a pharmaceutically acceptable form thereof, and one or more pharmaceutically acceptable carriers.

[0193] The term "pharmaceutical composition" refers to a composition that can be used in the treatment of the human or animal body comprising a pharmaceutically active ingredient (API) (or therapeutic agent) and optionally one or more pharmaceutically acceptable carriers. The term "pharmaceutically acceptable carrier" refers to an excipient with which the therapeutic agent is administered and which is, within the scope of sound medical judgment, suitable for use with humans and / or other animals without undue toxicity, irritation, allergic response, or other problem or complication, commensurate with a reasonable benefit / risk ratio.

[0194] The present application provides a pharmaceutical composition comprising at least one compound of any one of the first aspect or a pharmaceutically acceptable form thereof.

[0195] In some embodiments of the present application, the above-mentioned pharmaceutical composition further comprises one or more pharmaceutically acceptable carriers.

[0196] The above-mentioned pharmaceutical composition can act systemically and / or locally, which can be achieved by a suitable dosage form.

[0197] The above-mentioned pharmaceutical composition can comprise 0.01 mg to 1000 mg of at least one of the compounds or a pharmaceutically acceptable form thereof.

[0198] The present application also provides a method for preparing the above-mentioned pharmaceutical composition or its corresponding preparation form, which comprises combining at least one of the compounds or a pharmaceutically acceptable form thereof with one or more pharmaceutically acceptable carriers.

[0199] Kit product

[0200] In a fourth aspect, the present application provides a kit product comprising:

[0201] a) at least one compound of the first aspect or a pharmaceutically acceptable form thereof as a first therapeutic agent, or a pharmaceutical composition of the third aspect as a first pharmaceutical composition;

[0202] b) optionally at least one other therapeutic agent as a second therapeutic agent, or a pharmaceutical composition comprising the other therapeutic agent as a second pharmaceutical composition; and

[0203] c) optionally packaging and / or instructions.

[0204] The above-mentioned kit product can comprise 0.01 mg to 1000 mg of at least one of the compounds or a pharmaceutically acceptable form thereof.

[0205] The present application also provides a method of preparing the above-mentioned kit, which comprises combining at least one of the above-mentioned compounds or pharmaceutically acceptable forms thereof or the above-mentioned pharmaceutical composition with, optionally, at least one other therapeutic agent or a pharmaceutical composition comprising the other therapeutic agent, packaging and / or instructions.

[0206] Medical use

[0207] The compounds of the present application can exhibit strong inhibitory effect on TGFβR1, with IC50 values mostly reaching below 100 nM, and even below 10 nM in some cases, and weak inhibitory effect on TGFβR2, and can be used as TGFβR1 inhibitors (especially TGFβR1 selective inhibitors).

[0208] Therefore, the present application provides the use of the compounds of any one of the first aspect or pharmaceutically acceptable forms thereof or the above-mentioned pharmaceutical composition as TGFβR1 inhibitors (especially TGFβR1 selective inhibitors).

[0209] In addition, the present application also provides the use of the compounds or pharmaceutically acceptable forms thereof or the pharmaceutical composition in the preparation of a medicament for preventing and / or treating a disease or disorder mediated at least in part by TGFβR1.

[0210] In some embodiments, the disease or disorder mediated at least in part by TGFβR1 refers to a disease in which at least a part of the pathogenesis is related to TGFβR1, and these diseases include (but are not limited to) cancer or fibrotic diseases.

[0211] Therapeutic method

[0212] In another aspect, the present application provides a method of inhibiting the activity of TGFβR1 or preventing and / or treating a disease or disorder mediated at least in part by TGFβR1, which comprises the step of administering to a cell or a subject in need thereof an effective amount of the compounds of the first aspect or pharmaceutically acceptable forms thereof or the pharmaceutical composition of the third aspect.

[0213] In some embodiments, the disease or disorder mediated at least in part by TGFβR1 refers to a disease in which at least a part of the pathogenesis is related to TGFβR1, and these diseases include (but are not limited to) cancer or fibrotic diseases.

[0214] In some embodiments, the term "effective amount" refers to a dosage capable of inducing a biological or medical response in a cell, tissue, organ or organism (e.g. an individual) and sufficient to achieve the desired prophylactic and / or therapeutic effect.

[0215] Dosage regimens can be adjusted to provide the optimum desired response. For example, a single dose can be administered, doses can be administered at intervals, or dosages can be proportionally reduced or increased as actual conditions require, in accordance with the judgment of the practitioner and the professional judgment of the person administering or supervising the administration of the compositions. It will be appreciated that the specific dosage regimen for any particular individual will depend on a variety of factors including the severity and nature of the disease or disorder, the rate of administration, the disposition of the compound, and the judgment of the prescribing physician.

[0216] The amount of a compound of the present application to be administered will depend on the individual case, the severity of the disease or disorder, the rate of administration, the disposition of the compound, and the judgment of the prescribing physician. In general, an effective amount is about 0.001-10000 mg / kg body weight of the subject per day. In appropriate cases, an effective amount is about 0.01-1000 mg / kg body weight of the subject per day. Administration can be about 0.01-1000 mg / kg body weight of the subject per day, typically about 0.1-500 mg / kg body weight of the subject per day, either every day, every two days, or every three days. An exemplary dosage regimen is one or more times per day, or one or more times per week, or one or more times per month. When multiple doses are administered, the intervals between individual doses can be daily, weekly, monthly, or yearly. Alternatively, administration can be in the form of sustained release formulations, in which case less frequent administration is required. The dosage and frequency of administration can vary depending on the half-life of the drug in the subject, and can also vary depending on whether the application is prophylactic or therapeutic. In prophylactic applications, relatively low dosages are administered at relatively infrequent intervals over a long period, until the pathogenesis of the disease is affected, or the subject exhibits only minor or even none of the symptoms of the disease; in therapeutic applications, relatively high dosages can be administered at relatively short intervals to induce a rapid response and / or to counteract a severe attack of the disease.

[0217] The term "treatment" refers to the reduction or elimination of the disease or disorder in question. A subject is successfully "treated" if, after receiving a therapeutic amount of a compound of the present application or a pharmaceutically acceptable form thereof or a pharmaceutical composition of the present application, at least one indicia and symptom of the subject exhibits observable and / or measurable reduction in and / or amelioration of the disease. It will be appreciated that treatment includes not only complete treatment, but also treatment that does not achieve complete treatment, but that achieves some biologically or medically relevant result. In particular, "treatment" means that a compound of the present application or a pharmaceutically acceptable form thereof or a pharmaceutical composition of the present application can achieve at least one of the following effects, for example: (1) preventing the disease from occurring in an animal that can be predisposed to the disease but does not yet experience or display any disease pathology or symptomatology of the disease; (2) inhibiting the disease (i.e., arresting further progression of the pathology and / or symptomatology) in an animal that is experiencing or displaying the pathology or symptomatology of the disease; (3) ameliorating the disease (i.e., reversing the pathology and / or symptomatology) in an animal that is experiencing or displaying the pathology or symptomatology of the disease.

[0218] The term "administrate", "administrating" or "administration" (or "dosing") refers to the process of applying a pharmaceutically active ingredient (such as a compound of the present application) or a pharmaceutical composition comprising a pharmaceutically active ingredient (such as a pharmaceutical composition of the present application) to a site of the body of an individual or a cell, tissue, organ, biological fluid, etc. thereof, so as to bring the pharmaceutically active ingredient or the pharmaceutical composition into contact with the body of the individual or a cell, tissue, organ, biological fluid, etc. thereof. Common modes of administration include, but are not limited to, oral administration, subcutaneous administration, intramuscular administration, subperitoneal administration, ocular administration, nasal administration, sublingual administration, rectal administration, vaginal administration, etc.

[0219] The term "in need thereof" refers to the judgment of a physician or other care provider that an individual needs or will benefit from a prophylactic and / or therapeutic process, which judgment is based on various factors in the area of expertise of the physician or other care provider.

[0220] The term "individual" (or "subject") refers to a human or non-human animal. Individuals of the present application include individuals (patients) suffering from a disease and / or a disorder and normal individuals. Non-human animals of the present application include all vertebrates, for example, non-mammals, such as birds, amphibians, reptiles, etc. and mammals, for example, non-human primates, domestic animals, and / or laboratory animals, e.g., sheep, dogs, cats, cows, pigs, etc.

[0221] Combination

[0222] The compounds of the present application do not significantly inhibit the three major CYP isoforms (CYP1A2, CYP2D6 and CYP3A4), indicating a relatively low potential for drug-drug interactions, and thus the compounds or pharmaceutically acceptable forms thereof or the above pharmaceutical compositions can optionally be administered in combination with other therapeutic agents that are at least somewhat effective in treating various diseases.

[0223] The present application provides pharmaceutical combination or combination preparations of the compounds or pharmaceutically acceptable forms thereof or the above pharmaceutical compositions and at least one other therapeutic agent (especially a TGFβR1 inhibitor) for simultaneous, separate or sequential use in the prevention and / or treatment of a disease or disorder.

[0224] Advantages of the Invention

[0225] The present application provides a novel pyrazolopyridine compound, which can be used as a highly efficient and selective TGFβR1 inhibitor, has anti-tumor activity, and has less toxicity and drug interactions. In addition, the present application also provides a preparation method of the compound, which has mild conditions, simple and easy operation, and is suitable for industrial mass production. DETAILED DESCRIPTION

[0226] To make the objectives and technical solutions of this invention clearer, the embodiments of this invention are described in detail below with reference to examples. However, those skilled in the art will understand that the following examples are for illustrative purposes only and should not be considered as limiting the scope of this invention.

[0227] All reagents and instruments used in the examples are commercially available conventional products. Unless otherwise specified, all conditions were performed under conventional conditions or conditions recommended by the manufacturer. The term "room temperature" as used in this invention refers to 20℃ ± 5℃. When used to modify a numerical value or range, the term "about" as used in this invention refers to the value or range and a range of errors acceptable to those skilled in the art, such as ±10%, ±5%, ±4%, ±3%, ±2%, ±1%, ±0.5%, etc.

[0228] The meanings of the abbreviations in the conventional synthesis methods and in the examples and intermediate synthesis examples are shown in the table below.

[0229]

[0230]

[0231] The structures of the compounds described in the following examples were determined by nuclear magnetic resonance (NMR). 1 Determined by 1H-NMR and / or mass spectrometry (MS).

[0232] Nuclear magnetic resonance (NMR) 1 The H-NMR analysis was performed using a Bruker 400MHz NMR spectrometer. The solvents used were deuterated methanol (CD3OD), deuterated chloroform (CDCl3), and hexadeuterated dimethyl sulfoxide (DMSO-d6). The internal standard was tetramethylsilane (TMS).

[0233] The abbreviations used in the nuclear magnetic resonance (NMR) data in the following examples have the following meanings:

[0234] s: singlet, d: doublet, t: triplet, q: quartet, dd: double doublet, qd: quartet doublet, ddd: double double doublet, dd t: double double triplet, dddd: double double double doublet, m: multiplet, br: broad, J: coupling constant, Hz: hertz, δ: chemical shift.

[0235] All chemical shift (δ) values are given in parts per million (ppm).

[0236] The instrument used for the determination of mass spectrometry (MS) is Agilent 6120B mass spectrometer, and the ion source is electrospray ion source (ESI).

[0237] Embodiments of the present application are prepared by preparative high performance liquid chromatography (Prep-HPLC) purification using the method shown below.

[0238] Method: A

[0239] Column: SunFire Prep C 18 OBD (5 μm * 19 mm * 150 mm)

[0240] Mobile phase A: acetonitrile; mobile phase B: water (containing 0.05% formic acid)

[0241] Time [min] Mobile phase A [%] Mobile phase B [%] Flow rate [mL / min] 0.00 20 80 28 2.00 20 80 28 18.00 90 10 28

[0242] Method: B

[0243] Column: SunFire Prep C 18 OBD (5 μm * 19 mm * 150 mm)

[0244] Mobile phase A: acetonitrile; mobile phase B: water (containing 0.05% formic acid)

[0245] Time [min] Mobile phase A [%] Mobile phase B [%] Flow rate [mL / min] 0.00 10 90 28 2.00 10 90 28 20.00 80 20 28

[0246] Preparation of compounds

[0247] Example 1: Synthesis of 2-(6-methylpyridin-2-yl)-3-(pyrazolo[1,5-a]pyridin-5-yl)-2H-pyrazolo[3,4-b]pyridine (Compound 2)

[0248]

[0249] Step 1: Synthesis of 2-chloro-N-(6-methylpyridin-2-yl)nicotinamide (Compound 2-2)

[0250] To a solution of 2-chloronicotinic acid chloride (Compound 5-1, 800 mg, 4.55 mmol) and 6-methylpyridin-2-ylamine (589.86 mg, 5.45 mmol) in dichloromethane (15 mL) was added triethylamine (1.38 g, 13.64 mmol) at room temperature. The reaction mixture was stirred at room temperature for 1 h. The reaction mixture was diluted with 50 mL of dichloromethane and washed with 20 mL of water and 20 mL of saturated brine. The organic phase was dried over anhydrous sodium sulfate, filtered and concentrated under reduced pressure. The residue was purified by column chromatography (petroleum ether: ethyl acetate = 3:2) to give the title compound (950 mg, 84.07% yield).

[0251] MS (ESI): m / z 248.0 [M+H] +

[0252] Step 2: Synthesis of 2-azido-N-(6-methylpyridin-2-yl)nicotinamide (Compound 2-3)

[0253] To a solution of 2-chloro-N-(6-methylpyridin-2-yl)nicotinamide (Compound 2-2, 950 mg, 3.84 mmol) in N,N-dimethylformamide (15 mL) was added NaN3(748.06 mg, 11.51 mmol) at room temperature. The reaction mixture was stirred at 75 °C for 15 h. The reaction mixture was cooled to room temperature and slowly poured into water (150 mL). The solid was collected by filtration and washed with 50 mL of water, 30 mL of ethanol and 50 mL of petroleum ether. The solid was dried under vacuum to give the title compound (651 mg, 66.75% yield).

[0254] MS (ESI): m / z 255.1 [M+H] +

[0255] Step 3: Synthesis of 3-chloro-2-(6-methylpyridin-2-yl)-2H-pyrazolo[3,4-b]pyridine (Compound 2-4)

[0256] To a solution of 2-azido-N-(6-methylpyridin-2-yl)nicotinamide (compound 2-2, 320 mg, 1.26 mmol) in phosphorus oxychloride (10 mL) was heated to 115 °C for 15 h. The reaction mixture was evaporated under reduced pressure to give an oil, which was quenched with ice water (20 mL), neutralized with saturated aqueous sodium bicarbonate solution to neutral, and extracted with ethyl acetate (30 mL) for three times. The organic phase was combined, washed with 30 mL of saturated brine, dried over anhydrous sodium sulfate, filtered, and the filtrate was evaporated under reduced pressure to give a crude product, which was purified by column chromatography (petroleum ether: ethyl acetate = 1:1) to give the title compound (125 mg, yield 40.58%).

[0257] MS (ESI): m / z 245.1 [M+H] +

[0258] The structure was characterized as follows:

[0259] 1 H NMR (400 MHz, DMSO-d6): δ 8.80 (dd, J = 4.1, 1.7 Hz, 1H), 8.25 (dd, J = 8.4, 1.7 Hz, 1H), 8.07 (t, J = 7.8 Hz, 1H), 7.76 (d, J = 7.8 Hz, 1H), 7.54 (d, J = 7.7 Hz, 1H), 7.29 (dd, J = 8.5, 4.1 Hz, 1H), 2.60 (s, 3H).

[0260] Step four: synthesis of 2-(6-methylpyridin-2-yl)-3-(pyrazolo[1,5-a]pyridin-5-yl)- 2H-pyrazolo[3,4-b]pyridine (compound 2)

[0261] ​​​​​​To a solution of 3-chloro-2-(6-methylpyridin-2-yl)-2H-pyrazolo[3,4-b]pyridine (compound 2-4, 30 mg, 122.61 μmol) and 5-(4,4,5,5-tetramethyl-1,3,2-dioxaborolan-2-yl)pyrazolo[1,5-a]pyridine (44.89 mg, 183.91 μmol) in a mixture of 1,4-dioxane (3 mL) and water (0.8 mL) was added potassium carbonate (33.89 mg, 245.22 μmol) and dichlorobis-(4-dimethylaminophenyl) palladium(II) (8.68 mg, 12.26 μmol). After the addition, the reaction system was purged with nitrogen for 3 times and then heated at 100 °C for 2 hours in a microwave. After the reaction mixture was cooled to room temperature, it was diluted with 20 mL of dichloromethane and washed with 10 mL of water and 10 mL of saturated brine. The organic phase was dried over anhydrous sodium sulfate, filtered and concentrated under reduced pressure to give a crude product. The crude product was purified by preparative high performance liquid chromatography (Method A). The purified fractions were lyophilized to give the title compound (26.32 mg, 65.78% yield).

[0262] MS (ESI): m / z 327.1 [M+H] +

[0263] The structure was characterized as follows:

[0264] 1 H NMR (400 MHz, DMSO-d6): δ 8.79 (dd, J = 4.1, 1.7 Hz, 1H), 8.67 (d, J = 7.2 Hz, 1H), 8.29 (dd, J = 8.5, 1.7 Hz, 1H), 8.07 (d, J = 2.3 Hz, 1H), 8.01 (t, J = 7.8 Hz, 1H), 7.98 (dd, J = 2.0, 0.9 Hz, 1H), 7.80 (d, J = 8.0 Hz, 1H), 7.41 (d, J = 7.6 Hz, 1H), 7.27 (dd, J = 8.4, 4.1 Hz, 1H), 6.74 (dd, J = 2.3, 0.9 Hz, 1H), 6.64 (dd, J = 7.3, 1.9 Hz, 1H), 2.26 (s, 3H).

[0265] Example 2: Synthesis of 5-(2-(6-methylpyridin-2-yl)-2H-pyrazolo[3,4-b]pyridin-3-yl)pyrazolo[1,5-a]pyridine-3-carboxamide (compound 9)

[0266]

[0267] Step one: synthesis of ethyl 5-(4, 4, 5, 5-tetramethyl-l, 3, 2-dioxaborolan-2-yl) pyrazolo[l, 5-a]pyridine-3-carboxylate (compound 9-2)

[0268] Ethyl 5-bromopyrazolo[l, 5-a]pyridine-3-carboxylate (compound 9-1, 950 mg, 3.53 mmol), bis(pinacolato)diboron (1.08 g, 4.24 mmol) were dissolved in 1, 4-dioxane (40 mL), then Pd(dppf)Cl2(129 mg, 0.18 mmol) and methyl acetate (693 mg, 7.06 mmol) were added. After addition, the reaction system was purged with nitrogen for 3 times, and then heated to 95 °C for 15 h under nitrogen atmosphere. The reaction was cooled to room temperature, filtered, and the filtrate was concentrated under reduced pressure to obtain a crude product. The crude product was separated by silica gel column chromatography (eluent: petroleum ether / ethyl acetate = 4 / 1 (v / v)) to obtain the title compound, 945 mg. ESI-MS (m / z): 317.1 [M+H] + .

[0269] Step two: synthesis of ethyl 5-(2-(6-methylpyridin-2-yl)-2H-pyrazolo[3, 4-b]pyridin-3-yl)pyrazolo[l, 5-a]pyridine-3-carboxylate (compound 9-3)

[0270] Ethyl 3-chloro-2-(6-methylpyridin-2-yl)-2H-pyrazolo[3, 4-b]pyridine (compound 2-4, 50 mg, 204.35 μmol) and ethyl 5-(4, 4, 5, 5-tetramethyl-l, 3, 2-dioxaborolan-2-yl)pyrazolo[l, 5-a]pyridine-3-carboxylate (compound 9-2, 96.91 mg, 306.52 μmol) were dissolved in a mixed solvent of 1, 4-dioxane (4 mL) and water (1 mL), then potassium carbonate (56.48 mg, 408.70 μmol) and dichlorobis-(4-dimethylaminophenyl) palladium (II) (14.47 mg, 20.43 μmol) were added. After addition, the reaction system was purged with nitrogen for 3 times, and then heated to 100 °C for 2 h under microwave. The reaction was cooled to room temperature, diluted with 30 mL of dichloromethane, washed with 15 mL of water and 15 mL of saturated brine once, then dried over anhydrous sodium sulfate. The organic phase was filtered and concentrated under reduced pressure to obtain a crude product. The crude product was purified by column chromatography (petroleum ether: ethyl acetate = 3:7) to obtain the title compound (73 mg, yield 89.65%).

[0271] MS (ESI): m / z 399.1 [M+H] +

[0272] Step 3: Synthesis of 5-(2-(6-methylpyridin-2-yl)-2H-pyrazol-3-yl)pyrazolo[1,5- a]pyridine-3-carboxylic acid (Compound 9-4)

[0273] Ethyl 5-(2-(6-methylpyridin-2-yl)-2H-pyrazol-3-yl)pyrazolo[1,5-a]pyridine-3- carboxylate (Compound 9-3, 73 mg, 183.23 μmol) was dissolved in tetrahydrofuran (6 mL), then sodium hydroxide (73.29 mg, 1.83 mmol) and water (2 mL) were added, after addition, the reaction was warmed to 80 °C and stirred for 16 hours. After the reaction solution was cooled to room temperature, the pH was adjusted to about 5 with 3 mol / L aqueous hydrochloric acid solution, then directly evaporated under reduced pressure to obtain a crude product, the crude product was dissolved in dichloromethane / methanol = 10 / 1, inorganic salts were removed by filtration, and the filtrate was evaporated under reduced pressure to obtain the title compound (61 mg, yield 89.89%).

[0274] MS (ESI): m / z 371.1 [M+H] +

[0275] Step 4: Synthesis of 5-(2-(6-methylpyridin-2-yl)-2H-pyrazol-3-yl)pyrazolo[1,5- a]pyridine-3-carboxamide (Compound 9)

[0276] Ethyl 5-(2-(6-methylpyridin-2-yl)-2H-pyrazol-3-yl)pyrazolo[1,5-a]pyridine-3- carboxylate (Compound 9-3, 73 mg, 183.23 μmol) was dissolved in tetrahydrofuran (6 mL), then sodium hydroxide (73.29 mg, 1.83 mmol) and water (2 mL) were added, after addition, the reaction was warmed to 80 °C and stirred for 16 hours. After the reaction solution was cooled to room temperature, the pH was adjusted to about 5 with 3 mol / L aqueous hydrochloric acid solution, then directly evaporated under reduced pressure to obtain a crude product, the crude product was dissolved in dichloromethane / methanol = 10 / 1, inorganic salts were removed by filtration, and the filtrate was evaporated under reduced pressure to obtain the title compound (61 mg, yield 89.89%).

[0277] MS (ESI): m / z 371.1 [M+H] +

[0278] The structure is characterized as follows:

[0279] 1H NMR (400 MHz, DMSO-d6): δ 8.82 (dd, J = 4.1, 1.7 Hz, 1H), 8.79 (dd, J = 7.2, 0.9 Hz, 1H), 8.60 (s, 1H), 8.36 (d, J = 2.2 Hz, 1H), 8.26 (dd, J = 8.5, 1.7 Hz, 1H), 8.01 (t, J = 7.8 Hz, 1H), 7.82 (d, J = 8.0 Hz, 1H), 7.78 (s, 1H), 7.41 (d, J = 7.6 Hz, 1H), 7.31 (dd, J = 8.5, 4.1 Hz, 1H), 7.16 (s, 1H), 6.91 (dd, J = 7.2, 2.1 Hz, 1H), 2.26 (s, 3H).

[0280] Pharmacological activity test

[0281] Test Example 1: In vitro enzymatic activity inhibition test (TGFβR1).

[0282] Experimental method: The inhibitory effect of the compound of the present application on the TGFβR1 enzyme activity was determined according to the instructions of the ADP-Glo™ Kinase Assay Kit (Promega), and the steps were as follows:

[0283] After the TGFβR1 enzyme was pre-incubated with different concentrations of test compounds (1000 nM, 100 nM, 10 nM) at 30°C for 30 min, the TGFβR1 peptide and adenosine triphosphate (ATP) were added to start the reaction. After incubation at 30°C for 3 h, the ADP-Glo™ reagent was added, and after incubation at room temperature for 90 min, the kinase assay reagent was added. After incubation at room temperature for 30 min, the chemiluminescence signal value was detected. The solvent group (DMSO) was used as the negative control, and the buffer group (without TGFβR1 enzyme) was used as the blank control. The percentage inhibition of different concentrations of compounds was calculated according to the following formula:

[0284] Percentage inhibition = (1 - (chemiluminescence signal value of different concentrations of compounds - chemiluminescence signal value of blank control) / (chemiluminescence signal value of negative control - chemiluminescence signal value of blank control)) × 100%;

[0285] When the percentage inhibition is between 30-80%, the half-inhibitory concentration (IC 50 ) or range of the compound is calculated according to the following formula:

[0286] IC 50＝ X × (1 - percentage inhibition (%)) / percentage inhibition (%), where X is the test concentration of the compound with an inhibition rate between 30-80%.

[0287] The experimental results are shown in Table 1 below:

[0288] Table 1. Inhibitory activity of the compounds of the present application on TGFβR1

[0289] Example number IC for TGFβR1 50 (nM) Example 1 10.92±0.74 Example 2 8.81±0.30

[0290] As can be seen from Table 1, the compounds of the present application have significant inhibitory effect on TGFβR1.

[0291] Test Example Two: In vitro enzymatic activity inhibition test (TGFβR2).

[0292] Experimental method: The inhibitory effect of the compounds of the present application on TGFβR2 enzyme activity was determined according to the instructions of the ADP-Glo™ Kinase Assay Kit (Promega), and the steps were as follows:

[0293] After pre-incubation of TGFβR2 enzyme with different concentrations of test compounds (1000 nM, 100 nM, 10 nM) at 30°C for 30 min, myelin basic protein (MBP) and adenosine triphosphate (ATP) were added to start the reaction. After incubation at 30°C for 3 h, ADP-Glo™ reagent was added, and after incubation at room temperature for 90 min, kinase assay reagent was added. After incubation at room temperature for 30 min, the chemiluminescence signal value was detected. The solvent group (DMSO) was used as the negative control, and the buffer group (without TGFβR2 enzyme) was used as the blank control. The percentage inhibition of different concentrations of compounds was calculated according to the following formula:

[0294] Percentage inhibition = (1 - (chemiluminescence signal value of different concentrations of compounds - chemiluminescence signal value of blank control) / (chemiluminescence signal value of negative control - chemiluminescence signal value of blank control)) x 100%;

[0295] When the percentage inhibition is between 30-80%, the half maximal inhibitory concentration (IC 50 ) or range of the compound is calculated according to the following formula:

[0296] IC 50＝ X x (1 - percentage inhibition (%)) / percentage inhibition (%), where X is the test concentration of the compound with an inhibition rate between 30-80%.

[0297] The experimental results are shown in Table 2 below:

[0298] Table 2. Inhibition rate of the compounds of the present application on TGFβR2 enzyme activity

[0299] Example number IC for TGFβR2 50 (nM) <!-- 28 -->]]> Example 1 >1000 Example 2 752.53±10.79

[0300] As can be seen from Table 2, the compounds of the present application have weak inhibitory activity on TGFβR2.

[0301] As can be seen from Table 1 and Table 2, the compounds of the present application have selective inhibitory effect on TGFβR1.

[0302] Test Example 3: In vitro cell activity inhibition test

[0303] Experimental method: The inhibitory effect of the compounds of the present application on the TGFβ / Smads signaling pathway of HEK293-SBE cells was determined according to the instructions of the Bright-Glo Luciferase Assay Kit (Promega), and the steps were as follows: TM The inhibitory effect of the compounds of the present application on the TGFβ / Smads signaling pathway of HEK293-SBE cells was determined according to the instructions of the Bright-Glo Luciferase Assay Kit (Promega), and the steps were as follows:

[0304] HEK293-SBE cells (Bps bioscience) were added to a 96-well plate (10% FBS medium) at 30,000 cells / well, and cultured at 37°C, 5% CO2 overnight. The medium was replaced with 0.5% FBS medium, and test compounds diluted with 0.5% FBS medium were added, with a maximum final concentration of 10 μM, 4-fold dilution of the compounds, and a total of 8 concentration gradients. After 4-5 hours of culture, 10 μl of TGFβ was added. The final concentration of TGFβ was 0.5 ng / ml. 10 μl of medium was added instead of TGFβ as a negative control group. No test compound was added in the blank control group, and TGFβ was added. Bright Glo reagent was added to each well, and the chemiluminescence signal value was read on an enzyme marker.

[0305] The percentage inhibition of different concentrations of compounds was calculated according to the following formula:

[0306] The percentage inhibition of different concentrations of compounds was calculated according to the following formula:

[0307] The percentage inhibition of different concentrations of compounds was calculated according to the following formula: 50

[0308] y = Min + (Max - Min) / (1 + (x / IC 50 )^ (-Hillslope)), where y is the percentage inhibition, Max and Min are the maximum and minimum values of the fitted curve, x is the logarithmic concentration of the compound, and Hillslope is the slope of the curve.

[0309] The experimental results are shown in Table 3 below:

[0310] Table 3. Inhibitory effect of the compounds of the present application on the TGFβ / Smads signaling pathway of HEK293-SBE cells

[0311] Example number IC 50 (nM) Example 1 40.12±3.49 Example 2 40.39±4.19 ​

[0312] As can be seen from Table 3, the compounds of the present application have a significant inhibitory effect on the TGFβ-induced HEK293-SBE cell TGFβ / Smads signaling pathway (luciferase reporter gene method).

[0313] Test Example Four: Biochemical hERG inhibition test

[0314] 1. Test system:

[0315] Kit: Predictor TM hERG fluorescence polarization detection kit (ThermoFisher), which contains the following components: positive control compound hERG potassium channel blocker E4031; hERG cell membrane; affinity tracer Tracer; and hERG buffer.

[0316] 2. Test parameters:

[0317] hERG concentration: 1x; Tracer concentration: 1 nM; incubation time: 2 h; BMG PHERAstar FS FP.

[0318] 3. Test method:

[0319] The test was performed according to the kit instructions, and the steps were as follows:

[0320] Test group: 10 μM and 1 μM of the test compound were added to the microplate containing the hERG cell membrane, and 1 μM of the test compound was added to each well with the high hERG affinity tracer Tracer. After incubation of the microplate at room temperature for 2 hours, the change in fluorescence polarization (excitation wavelength: 540 nm; emission wavelength: 590 nm) value was detected using a multifunctional enzyme label instrument.

[0321] Positive control group: 30 μM of the positive control compound E4031 was used instead of the test compound, and the experimental method was the same as that of the test group.

[0322] Blank control group: hERG buffer was used instead of the test compound, and no hERG cell membrane was added, and the experimental method was the same as that of the test group.

[0323] 4. Data processing:

[0324] According to the data ratio, the percentage inhibition rate (%) of the compounds of the present application on hERG at different concentrations was calculated, and the range of the half-inhibitory concentration (IC 50 ) of the compounds was determined.

[0325] Percentage inhibition (%) = (1 - (fluorescence polarization value of the test compound - fluorescence polarization value of the positive control) / (fluorescence polarization value of the blank control - fluorescence polarization value of the positive control)) x 100%;

[0326] 5. Experimental results:

[0327] The compound was tested for hERG inhibition using the above method, and the results are shown in Table 4 below.

[0328] Table 4. Results of hERG inhibition test

[0329] Example number IC 50 (μM) Example 1 >10 Example 2 >10

[0330] The test results show that the compounds of the present application have low affinity for hERG, and the IC 50 of competition with the affinity tracer Tracer is greater than 10 μM, proving that the compounds of the present application have a low risk of cardiac toxicity related to the hERG ion channel.

[0331] Test Example Five: CYP enzyme (cytochrome P450) inhibition test.

[0332] 1. Test system:

[0333] P450-Glo TM CYP1A2 screening system (Promega);

[0334] P450-Glo TM CYP2D6 screening system (Promega);

[0335] P450-Glo TM CYP3A4 screening system (Promega).

[0336] 2. Test instrument:

[0337] BMG PHERAstar FS Luminescent.

[0338] 3. Test method:

[0339] The test was performed according to the kit instructions, and the steps are as follows:

[0340] 3.1. Inhibition of CYP1A2:

[0341] Test group: Different concentrations of the test compound were added to the microplate, Luciferin-ME (100 μM), K3PO4 (100 mM) and CYP1A2 (0.01 pmol / μL) were added to each well, pre-incubated at room temperature for 10 min, then NADPH regenerating system was added, reacted at room temperature for 30 min, finally equal volume of detection buffer was added, incubated at room temperature for 20 min, then chemiluminescence detection was carried out.

[0342] Negative control group: The experimental method was the same as the test group, but no test compound was added.

[0343] Blank control group: The experimental method was the same as the test group, but no test compound was added, and CYP1A2 Membrance (0.01 pmol / μL) was used instead of CYP1A2.

[0344] 3.2. Inhibition of CYP2D6:

[0345] Test group: Different concentrations of the test compound were added to the microplate, Luciferin-MEEGE (3 μM), K3PO4 (100 mM) and CYP2D6 (5 nM) were added to each well, pre-incubated at room temperature for 10 min, then NADPH regenerating system was added, reacted at 37°C for 30 min, finally equal volume of detection buffer was added, incubated at room temperature for 20 min, then chemiluminescence detection was carried out.

[0346] Negative control group: The experimental method was the same as the test group, but no test compound was added.

[0347] Blank control group: The experimental method was the same as the test group, but no test compound was added, and CYP2D6 Membrance (5 nM) was used instead of CYP2D6.

[0348] 3.3. Inhibition of CYP3A4:

[0349] Test group: Different concentrations of the test compound were added to the microplate, Luciferin-IPA (3 μM), K3PO4 (100 mM) and CYP3A4 (2 nM) were added to each well, pre-incubated at room temperature for 10 min, then NADPH regenerating system was added, reacted at room temperature for 30 min, finally equal volume of detection buffer was added, incubated at room temperature for 20 min, then chemiluminescence detection was carried out.

[0350] Negative control group: The experimental method was the same as the test group, but no test compound was added.

[0351] Blank control group: The experimental method was the same as the test group, but no test compound was added, and CYP3A4 Membrance (2 nM) was used instead of CYP3A4.

[0352] 4. Data processing:

[0353] Percentage inhibition (%) = (1 - (chemiluminescence signal value of the tested compound - chemiluminescence signal value of the blank control) / (chemiluminescence signal value of the negative control - chemiluminescence signal value of the blank control)) x 100%;

[0354] According to the inhibition rates of different concentrations of compounds on CYP enzymes, the half-inhibitory concentration (IC 50 ) or range of the compound is estimated:

[0355] IC 50＝ X x (1 - percentage inhibition (%)) / percentage inhibition (%), wherein: X is the test concentration of the compound.

[0356] 5. Experimental results:

[0357] The inhibition of the three CYPs by the compound of the present application is determined according to the above method, and the results are shown in Table 5 below.

[0358] Table 5. Results of CYPs inhibition test

[0359]

[0360] The test results show that the compound of the present application has weak inhibition on the three CYPs, which proves that the compound of the present application has good drug safety.

[0361] Although the specific embodiments of the present application have been described in detail, those skilled in the art will understand that various modifications and substitutions can be made to those details according to all the teachings disclosed herein, and these changes are within the scope of protection of the present application. The entire scope of the present application is given by the appended claims and any equivalents thereof.

Claims

1. A compound having the structure of Formula I or a pharmaceutically acceptable form thereof, wherein, R 1 selected from pyridyl, said pyridyl being optionally substituted with one or more R 4 substituents; R 2 selected from and ; R 3 is hydrogen; R 4 are each, at each occurrence, independently selected from the group consisting of hydrogen, deuterium, halogen, cyano, C 1-6 alkyl and C 1-6 haloalkyl; m is 0; the pharmaceutically acceptable form is a pharmaceutically acceptable salt.

2. The compound or pharmaceutically acceptable form thereof according to claim 1, wherein, R 1 is selected from the following fragments: 。 3. The compound or pharmaceutically acceptable form thereof according to claim 2, wherein, R 1 For .

4. The compound or pharmaceutically acceptable form thereof according to claim 1, wherein, R 4 selected from halogen, C 1-6 alkyl and C 1-6 haloalkyl.

5. The compound or pharmaceutically acceptable salt thereof according to any one of claims 1-4, wherein the compound is selected from: ， 。 6. A pharmaceutical composition comprising at least one compound or pharmaceutically acceptable form thereof according to any one of claims 1 to 5, and one or more pharmaceutically acceptable carriers.

7. A kit product comprising: a) at least one compound or pharmaceutically acceptable form thereof according to any one of claims 1 to 5 as a first therapeutic agent, or a pharmaceutical composition according to claim 6 as a first pharmaceutical composition; b) optionally at least one other therapeutic agent as a second therapeutic agent, or a pharmaceutical composition comprising the other therapeutic agent as a second pharmaceutical composition; and c) optionally packaging and / or instructions.

8. Use of a compound or pharmaceutically acceptable form thereof according to any one of claims 1 to 5, or a pharmaceutical composition according to claim 6, in the manufacture of a medicament for the prevention and / or treatment of a disease or condition mediated at least in part by TGFpRl.

9. The use according to claim 8, wherein: the disease or condition mediated at least in part by TGFpRl is a cancer or a fibrotic disease.

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