CN115557945A - Triazolopyridine compounds as AXL inhibitors - Google Patents

Abstract

The present invention relates to triazolopyridine compounds which are AXL inhibitors. These compounds are useful for the treatment or prevention of AXL receptor tyrosine kinase induced disorders.

Description

Triazolopyridine compounds as AXL inhibitors

Technical Field

The present invention is in the field of medicinal chemistry, and in particular relates to triazolopyridine compounds as AXL inhibitors, which are useful for the treatment of AXL-mediated diseases and conditions.

Background

AXL (also known as UFO, ARK and Tyro7 or JTK 11), is one of the members of the TAM family of Receptor Tyrosine Kinases (RTKs). AXL consists of an extracellular domain, a transmembrane domain and an intracellular domain. The extracellular ligand binding region includes two immunoglobulin (Ig) -like repeats and two fibronectin type III (froiii) -like repeats. The intracellular segment is essential for autophosphorylation and subsequent kinase activity (Zhu C, wei Y, wei X. Mol cancer.2019 Nov 4 (1): 153.). AXL, like other TMA family members, is partially activated by interaction with the vitamin K-dependent protein ligand growth arrest-specific protein 6 ((Gas 6)). The AXL protein is expressed in normal tissues, particularly in bone marrow stroma and bone marrow cells, as well as in tumor cells and tumor vasculature (Gay CM, balaji K, byers LA. Br J cancer.2017 Feb14;116 (4): 415-423.).

AXL signaling stimulates cellular responses including activation of phosphoinositide 3-kinase-Akt, the extracellular signal-regulated kinase (ERK) and p38 mitogen-activated protein kinase cascades, the NF- κ B pathway, and Signal Transducer and Activator of Transcription (STAT) signaling (3) AXL signaling is, in addition, an important pathway to drive invasion, migration, survival signaling, angiogenesis, resistance to chemotherapy and targeted drugs, cellular transformation and proliferation. High AXL expression is observed in many human tumors and is associated with tumor progression in cancer patients. AXL may therefore be a potential target for cancer therapy. The first selective inhibitor against AXL kinase is compound R428 (BGB-324). The action mechanism is to inhibit the phosphorylation of AXL and break the steady regulation of AXL kinase on the membrane, thereby reducing the expression quantity on the AXL membrane. (Apatira A, chua J, et al. Cancer Res.2010 Feb 15 (4): 1544-54.). BGB-324 is a similar pioneering, high-selectivity and bioavailable oral AXL small-molecule inhibitor developed by BerGenBio, and has the following structure:

furthermore, patent document WO2009047514A1 also discloses compounds as AXL inhibitors.

Disclosure of Invention

In one aspect, the invention provides compound I, or a pharmaceutically acceptable salt thereof:

wherein ring A is selected from phenyl or 5-to 10-membered heteroaryl, optionally substituted with one or more R ¹ Substituent group, R ¹ Selected from the following groups: -SO ₂ NR ² R ³ Halogen, C1-C6 alkyl, (C1-C6 alkyl) -O-, (5-to 10-membered heterocyclyl) -O-, phenyl-O-, (5-to 6-membered heteroaryl) -O-, cyano or nitro, wherein (5-to 10-membered heterocyclyl) -O-, phenyl-O-, and (5-to 6-membered heteroaryl) -O-are optionally substituted with one or more R ⁴ Substituent(s) substituted, R ⁴ Selected from C1-C3 alkyl, halogen, cyano or nitro, R ² And R ³ Each independently selected from hydrogen or C1-C6 alkyl.

In some embodiments, ring a is selected from phenyl or 5-to 6-membered heteroaryl, optionally substituted with one or more R ¹ Substituent(s) substituted, R ¹ As defined above.

In some embodiments, ring A is selected from phenyl, imidazolyl, oxazolyl, isoxazolyl, thiazolyl, isothiazolyl, triazolyl, oxadiazolyl, thiadiazolyl, tetrazolyl, oxatriazolyl, thiatriazolyl, pyridyl, pyrimidinyl, pyrazinyl, pyridazinyl, 1,2, 3-triazinyl, 1,2, 4-triazinyl, or 1,3, 5-triazinyl, optionally substituted with one or more R ¹ Substituent(s) substituted, R ¹ As defined above; preferably, ring A is selected from phenyl, imidazolyl, oxazolyl, thiazolyl, pyridyl or pyrimidinyl, optionally substituted with one or more R ¹ Substituent group, R ¹ As defined above; more preferably, ring a is selected from phenyl or pyridyl, optionally substituted with one or more R ¹ The substituent group is used for substitution,R ¹ as defined above.

In some embodiments, ring a is selected from

Optionally substituted by one or more R ¹ Substituent(s) substituted, R ¹ Selected from the following groups: -SO ₂ NR ² R ³ Halogen, C1-C6 alkyl, (C1-C6 alkyl) -O-, (5-to 10-membered heterocyclyl) -O-, or phenyl-O-, wherein (5-to 10-membered heterocyclyl) -O-and phenyl-O-are optionally substituted with one or more R ⁴ Substituent group, R ² 、R ³ And R ⁴ As defined above.

In some embodiments, ring a is selected from

Optionally substituted by one or more R ¹ Substituent(s) substituted, R ¹ Selected from the following groups: -SO ₂ N(CH ₃ ) ₂ 、-SO ₂ N(CH ₂ CH ₃ ) ₂ Halogen, methyl, ethyl, ⁱ Pr-O-、CH ₃ O-、CH ₃ CH ₂ O-, piperidinyl, pyrrolidinyl, piperazinyl, morpholinyl thiomorpholinyl, 1, 3-dioxolanyl, and tetrahydrofuranyl, tetrahydrothienyl, tetrahydrothiopyranyl, tetrahydropyridinyl, dihydropyridinyl,

Wherein piperidinyl, pyrrolidinyl, piperazinyl, morpholinyl, thiomorpholinyl, 1, 3-dioxolanyl, tetrahydrofuryl, tetrahydrothienyl, tetrahydrothiopyranyl, tetrahydropyridinyl, dihydropyridinyl, or mixtures thereof,

Optionally substituted by one or more R ⁴ Substituent group, R ⁴ As defined above; preferably, in some embodiments, ring a is selected from

Optionally substituted by one or more R ¹ Substituent(s) substituted, R ¹ Selected from the following groups: -SO ₂ N(CH ₃ ) ₂ 、-SO ₂ N(CH ₂ CH ₃ ) ₂ Halogen, methyl, ethyl, ⁱ Pr-O-、CH ₃ O-、CH ₃ CH ₂ O-, piperidyl, pyrrolidinyl, piperazinyl, morpholinyl, thiomorpholinyl, 1, 3-dioxolanyl, or a pharmaceutically acceptable salt thereof tetrahydrofuranyl, tetrahydrothienyl, tetrahydrothiopyranyl, tetrahydropyridinyl, dihydropyridinyl,

Wherein piperidinyl, pyrrolidinyl, piperazinyl, morpholinyl, thiomorpholinyl, 1, 3-dioxolanyl, tetrahydrofuryl, tetrahydrothienyl, tetrahydrothiopyranyl, tetrahydropyridinyl, dihydropyridinyl, or mixtures thereof,

Optionally substituted by one or more R ⁴ Substituent(s) substituted, R ⁴ As defined above.

In some embodiments, ring a is selected from

Optionally substituted by one or more R ¹ Substituent(s) substituted, R ¹ Selected from the following groups: -Cl,

ⁱ Pr-O-、-SO ₂ N(CH ₃ ) ₂ Or

Wherein, the first and the second end of the pipe are connected with each other,

optionally is covered withA plurality of R ⁴ Substituent(s) substituted, R ⁴ Is methyl; preferably, R ¹ Selected from the following groups: ⁱ Pr-O-、-SO ₂ N(CH ₃ ) ₂ or

In some embodiments, ring a is

It is covered ⁱ Pr-O-substitution; preferably, ring A is

In another aspect, the present invention provides the following compounds or pharmaceutically acceptable salts thereof:

in some embodiments, the present invention provides a pharmaceutical composition comprising a therapeutically effective amount of compound I, or a pharmaceutically acceptable salt thereof, and a pharmaceutically acceptable carrier.

In some embodiments, the present invention provides the use of compound I or a pharmaceutically acceptable salt thereof for the manufacture of a medicament for the treatment and/or prevention of AXL receptor tyrosine kinase induced disorders.

In some embodiments, the AXL receptor tyrosine kinase induced disorder is a disorder caused by, associated with, and/or accompanied by AXL kinase hyperactivity.

In some embodiments, the AXL receptor tyrosine kinase-induced disorder is selected from breast cancer, colon cancer, prostate cancer, gastric cancer, ovarian cancer, endometrial cancer, renal cancer, hepatocellular carcinoma, thyroid cancer, uterine cancer, esophageal cancer, squamous cell carcinoma, osteosarcoma, melanoma, glioblastoma, and neuroblastoma.

In another aspect, the invention provides a process for preparing compound I, comprising reacting compound I-1 with compound I-2 in the presence of a base, a catalyst and a solvent to obtain compound I,

correlation definition

Unless specifically stated otherwise, the following terms used in the specification and claims have the following meanings:

the "compounds" of the invention may be asymmetric, e.g., having one or more chiral centers. Unless otherwise indicated, "compound" of the present invention refers to any one stereoisomer or a mixture of two or more stereoisomers. Stereoisomers include, but are not limited to, enantiomers and diastereomers. The compounds of the present invention containing asymmetric carbon atoms can be isolated in optically active pure form or as a mixture of two or more stereoisomers. The optically active pure form can be resolved from a mixture of two or more stereoisomers or synthesized by using chiral starting materials or chiral reagents.

The term "cyano" refers to the group — CN.

The term "nitro" means-NO ₂ A group.

The term "halogen" refers to fluorine, chlorine, bromine and iodine.

The term "alkyl" refers to a saturated aliphatic hydrocarbon group, including straight or branched chain saturated hydrocarbon groups having the indicated number of carbon atoms. As the term "C1-C6 alkyl" includes C1 alkyl, C2 alkyl, C3 alkyl, C4 alkyl, C5 alkyl, C6 alkyl, examples include, but are not limited to, methyl, ethyl, n-propyl, isopropyl, n-butyl, isobutyl, t-butyl, n-pentyl, 2-pentyl, 3-pentyl, n-hexyl, 2-hexyl, 3-hexyl and the like.

The term "(C1-C3 alkyl) -O-" refers to a group having the structure alkyl-O-, which alkyl group is a saturated monovalent hydrocarbon group including straight or branched chains. Including but not limited to methoxy, ethoxy, n-propoxy, or isopropoxy.

The term "member" refers to the number of backbone atoms or groups of atoms that make up the ring. For example, "5-6 membered" means that the number of skeleton atoms or atom groups constituting the ring is 5 or 6. Thus, for example, pyridine, piperidine, piperazine and benzene are six-membered rings, while thiophene, pyrrole are five-membered rings.

The term "heteroaryl" refers to an aromatic carbocyclic group in which at least one ring atom is a heteroatom independently selected from nitrogen, oxygen and sulfur, and the other ring atoms are carbon. Heteroaryl groups may be optionally substituted independently with one or more substituents described herein. In one example, the heteroaryl is a 5-to 10-membered heteroaryl; in another example, the heteroaryl is a 5-to 6-membered heteroaryl. Examples of heteroaryl groups include, but are not limited to, phenyl, imidazolyl, oxazolyl, isoxazolyl, thiazolyl, isothiazolyl, triazolyl, oxadiazolyl, thiadiazolyl, tetrazolyl, oxatriazolyl, thiatriazolyl, pyridyl, pyrimidinyl, pyrazinyl, pyridazinyl, 1,2, 3-triazinyl, 1,2, 4-triazinyl, or 1,3, 5-triazinyl.

The term "heterocyclyl" refers to a saturated or partially unsaturated cyclic group in which at least one ring atom is a heteroatom independently selected from nitrogen, oxygen, phosphorus and sulfur, the remaining ring atoms being C. In one embodiment, heterocyclyl includes monocyclic or bicyclic rings having 5-10 members. Examples of heterocyclic groups include, but are not limited to

Piperidinyl, pyrrolidinyl, piperazinyl, morpholinyl, thiomorpholinyl, 1, 3-dioxolanyl, tetrahydrofuranyl, tetrahydrothienyl, tetrahydrothiopyranyl, tetrahydropyridinyl, dihydropyridinyl. The heterocyclic group may be optionally substituted unless specifically prohibited.

The terms "optionally" or "optionally" mean that the subsequently described event or circumstance may or may not occur, and that the description includes instances where said event or circumstance occurs and instances where it does not.

Numerical ranges herein refer to each integer in the given range. For example, "C1-C3" means that the group can have 1 carbon atom, 2 carbon atoms, or 3 carbon atoms; by "C1-C6" is meant that the group can have 1 carbon atom, 2 carbon atoms, 3 carbon atoms, 4 carbon atoms, 5 carbon atoms, or 6 carbon atoms.

The term "substituted" means that any one or more hydrogen atoms on a specified group is replaced with a substituent, so long as the valence of the specified group is normal and the substituted compound is stable. For example, "substituted with halogen" means that any one or more hydrogen atoms on a specified group is substituted with halogen, as long as the valence of the specified group is normal and the substituted compound is stable.

In

Refers to the chemical bond connection.

The term "pharmaceutically acceptable salt" refers to salts that retain the biological effectiveness of the free acids and bases of the particular compound without biologically adverse effects. Such as acid (including organic and inorganic) addition salts or base addition salts (including organic and inorganic bases).

The pharmaceutically acceptable salts of the present invention can be synthesized from the parent compound, which contains an acid or base, by conventional chemical methods. In general, such salts are prepared by the following method: prepared by reacting these compounds in free acid or base form with a stoichiometric amount of the appropriate base or acid, in water or an organic solvent or a mixture of the two.

The term "effective amount" or "therapeutically effective amount" refers to a sufficient amount of a drug or pharmaceutical agent that is non-toxic but achieves the desired effect.

The term "pharmaceutically acceptable carrier" refers to those carriers which do not significantly irritate the body and which do not impair the biological activity and performance of the active compound. Including, but not limited to, any diluents, disintegrants, binders, glidants, and wetting agents approved by the national food and drug administration for use in humans or animals.

Abbreviations used in the claims and specification have the following meanings:

"v/v" means weight ratio;

M:mol/L；

mM：mmol/L；

μM：μmol/L；

nM：nmol/L；

DMSO, DMSO: dimethyl sulfoxide (DMSO).

Detailed Description

The following more specifically describes the preparation of the compounds of the present invention, but these specific preparation methods do not set any limit to the scope of the present invention. Further, the reaction conditions such as reactants, solvent, base, amount of the compound used, reaction temperature, reaction time and the like are not limited to the following examples.

The compounds of the present invention may also be conveniently prepared by optionally combining various synthetic methods described in the present specification or known in the art, and such combinations may be readily carried out by those skilled in the art.

Example 1: (S) -5- (4-chloro-2-isopropoxyphenyl) -N- (7- (pyrrolidin-1-yl) -6,7,8, 9-tetrahydro-5H-benzo [7] rotanen-2-yl) - [1,2,4] triazolo [1,5-a ] pyridin-2-amine

a) 2- (4-chloro-2-isopropoxyphenyl) -4, 5-tetramethyl-1, 3, 2-dioxaborane

1-bromo-4-chloro-2-isopropoxybenzene (32.1 mmol), bis-pinacolborate (38.5 mmol), 1' -bisdiphenylphosphinoferrocene palladium dichloride (3.21 mmol), potassium acetate (128.4 mmol) and 1, 4-dioxane (40 mL) were placed in a 100mL three-necked flask and protected with nitrogen. Followed by reaction for 15h at 85 ℃ in an oil bath. The reaction was stopped. Water (50 mL) was added to the reaction solution, and extraction was performed with ethyl acetate (50 mLx 2). The organic layers were combined, washed with water and saturated sodium chloride in this order, the organic layer was concentrated to dryness under reduced pressure, and the resulting residue was subjected to silica gel column chromatography (dichloromethane: methanol =10: 1v/v) to give the title product (12.6 mmol).

b) 5- (4-chloro-2-isopropoxyphenyl) - [1,2,4] triazolo [1,5-a ] pyridin-2-amine

2- (4-chloro-2-isopropoxyphenyl) -4, 5-tetramethyl-1, 3, 2-dioxaborane (1.13 mmol), 5-bromo- [1,2,4] triazolo [1,5-a ] pyridin-2-amine (1.47 mmol), 1' -bisdiphenylphosphinoferrocene palladium dichloride (0.12 mmol), potassium carbonate (2.26 mmol), 1, 4-dioxane (10 mL) and water (2 mL) were placed in a 50mL three-necked flask and protected with nitrogen. Followed by reaction for 5h at 100 ℃ in an oil bath. After that, the reaction was stopped, the reaction liquid was concentrated to dryness under reduced pressure, and the obtained residue was subjected to silica gel column chromatography (dichloromethane: methanol =10: 1v/v) to obtain the title product (0.66 mmol). MS (ESI +): 303.1 (M + H).

c) 5- (4-chloro-2-isopropoxyphenyl) -2-iodo- [1,2,4] triazolo [1,5-a ] pyridine

5- (4-chloro-2-isopropoxyphenyl) - [1,2,4] triazolo [1,5-a ] pyridin-2-amine (0.67 mmol,; p-toluenesulfonic acid monohydrate (3.4 mmol) and acetonitrile (20 mL) were added to a 100mL single-neck flask, stirred and cooled to-20 ℃. Subsequently, sodium nitrite (1.68 mmol) and potassium iodide (1.34 mmol) were dissolved in water (1 mL) and added dropwise to the reaction mixture. Stirring for 30min under heat preservation, and then moving to room temperature to stir for 2h. When the reaction of the raw materials is completed, the reaction is stopped. Ethyl acetate (30 mL) and water (30 mL) were added to the reaction mixture, followed by addition of a saturated solution of sodium persulfate (30 mL), stirring, and extraction. The aqueous layer was re-extracted with ethyl acetate (30 mL). The organic phases were combined, washed successively with water, brine and dried over anhydrous sodium sulfate. Filtration, the filtrate was concentrated to dryness under reduced pressure, and the resulting residue was subjected to silica gel column chromatography (dichloromethane: methanol = 10. MS (ESI +): 414.2 (M + H).

d) (S) -5- (4-chloro-2-isopropoxyphenyl) -N- (7- (pyrrolidin-1-yl) -6,7,8, 9-tetrahydro-5H-benzo [7] annulen-2-yl) - [1,2,4] triazolo [1,5-a ] pyridin-2-amine

5- (4-chloro-2-isopropoxyphenyl) -2-iodo- [1,2,4] triazolo [1,5-a ] pyridine (0.68 mmol), (S) -7- (pyrrolidin-1-yl) -6,7,8, 9-tetrahydro-5H-benzo [7] annulen-2-amine (0.57 mmol), tris (dibenzylideneacetone) dipalladium (0.07 mmol), sodium tert-butoxide (1.36 mmol), 4, 5-bis (diphenylphosphino) -9, 9-dimethylxanthene (0.17 mmol) and 1, 4-dioxane were placed in a 100mL single-necked flask and protected with nitrogen. The reaction solution was stirred in an oil bath at 100 ℃ for 5 hours. When the reaction was completed, the reaction solution was concentrated to dryness under reduced pressure, and the obtained residue was subjected to silica gel column chromatography (dichloromethane: methanol =10: 1v/v) to obtain the title product (0.06 mmol).

¹ H NMR(400MHz,DMSO-d ₆ ,ppm)δ9.44(s,1H),7.63-7.53(m,3H),7.42(d,J＝2.1Hz,1H),7.38(dd,J＝8.1,2.2Hz,1H),7.31(d,J＝1.9Hz,1H),7.15(dd,J＝8.2,1.9Hz,1H),7.02–6.97(m,2H),4.67(dt,J＝12.1,6.0Hz,1H),3.18-3.04(m,4H),2.82–2.54(m,5H),2.28-2.14(m,2H),1.85(br,4H),1.49-1.34(m,2H),1.10(s,3H),1.08(s,3H).MS(ESI+):516.3(M+H).

Example 2 (S) -5- (3-Isopropoxyphenyl) -N- (7- (pyrrolidin-1-yl) -6,7,8, 9-tetrahydro-5H-benzo [7] annulen-2-yl) - [1,2,4] triazolo [1,5-a ] pyridin-2-amine

Reference example 1 steps b) to d) 2- (4-chloro-2-isopropoxyphenyl) -4, 5-tetramethyl-1, 3, 2-dioxaborane from step b) in example 1 was replaced with (3-isopropoxyphenyl) boronic acid.

¹ H NMR(400MHz,DMSO-d ₆ ,ppm)δ9.46(s,1H),8.03–7.99(m,1H),7.89–7.78(m,2H),7.67–7.59(m,3H),7.39–7.31(m,2H),7.05–7.00(m,1H),6.95(d,J＝8.1Hz,1H),3.6-3.23(m,6H),3.15-2.92(m,6H),2.81–2.57(m,4H),2.20-2.05(m,2H),1.82(s,4H),1.48-1.32(m,2H).MS(ESI+):482.3(M+H).

Example 3 (S) -N, N-dimethyl-3- (2- ((7- (pyrrolidin-1-yl) -6,7,8, 9-tetrahydro-5H-benzo [7] annulen-2-yl ] amino) - [1,2,4] triazolo [1,5-a ] pyridin-5-yl) benzenesulfonamide

Reference example 1 steps b) to d) 2- (4-chloro-2-isopropoxyphenyl) -4, 5-tetramethyl-1, 3, 2-dioxaborane from step b) in example 1 was replaced with (3- (N, N-dimethylsulfamoyl) phenyl) boronic acid.

¹ H NMR(400MHz,DMSO-d ₆ ,ppm)δ9.49(s,1H),8.51(s,1H),8.35(d,J＝7.7Hz,1H),7.96-7.85(m,2H),7.71–7.59(m,2H),7.46(dd,J＝8.0,2.1Hz,1H),7.38(d,J＝1.9Hz,1H),7.30(dd,J＝7.1,1.1Hz,1H),7.02(d,J＝8.2Hz,1H),3.12-2.94(br,5H),2.86-2.74(m,2H),2.68(s,6H),2.66-2.54(m,2H),2.22-2.06(m,2H),1.82(br,4H),1.52-1.36(m,2H).MS(ESI+):531.3(M+H).

Example 4: (S) -N, N-dimethyl-2- (2- ((7- (pyrrolidin-1-yl) -6,7,8, 9-tetrahydro-5H-benzo [7] annulen-2-yl) amino) - [1,2,4] triazolo [1,5-a ] pyridin-5-yl) benzenesulfonamide

Reference example 1 steps b) to d) 2- (4-chloro-2-isopropoxyphenyl) -4, 5-tetramethyl-1, 3, 2-dioxaborane from step b) in example 1 was replaced with (2- (N, N-dimethylsulfamoyl) phenyl) boronic acid.

¹ H NMR(400MHz,DMSO-d ₆ ,ppm)δ9.46(s,1H),8.03–7.99(m,1H),7.89–7.78(m,2H),7.67–7.59(m,3H),7.39–7.31(m,2H),7.05–7.00(m,1H),6.95(d,J＝8.1Hz,1H),3.6-3.23(m,5H),3.15-2.92(m,6H),2.81–2.57(m,4H),2.20-2.05(m,2H),1.82(s,4H),1.48-1.32(m,2H).MS(ESI+):531.3(M+H).

Example 5 (S) -5- (4-Isopropoxyphenyl) -N- (7- (pyrrolidinyl-1-yl) -6,7,8, 9-tetrahydro-5H-benzo [7] annulen-2-yl) - [1,2,4] triazolo [1,5-a ] pyridin-2-amine

Reference example 1 steps b) to d) 2- (4-chloro-2-isopropoxyphenyl) -4, 5-tetramethyl-1, 3, 2-dioxaborane from step b) in example 1 was replaced with (4-isopropoxyphenyl) boronic acid.

¹ H NMR(400MHz,DMSO-d ₆ ,ppm)δ9.46(s,1H),8.05(d,J＝8.8Hz,2H),7.68–7.57(m,1H),7.58–7.45(m,2H),7.39(dd,J＝8.1,2.1Hz,1H),7.22–7.07(m,3H),7.03(d,J＝8.2Hz,1H),4.77(dt,J＝12.0,6.0Hz,1H),3.01(br,5H),2.88-2.74(m,2H),2.75–2.59(m,2H),2.48-2.06(m,2H),1.82(s,4H),1.57–1.40(m,2H),1.34(s,3H),1.33(s,3H).MS(ESI+):482.3(M+H).

Example 6 (S) -N, N-dimethyl-4- (2- (((7- (pyrrolidin-1-yl) -6,7,8, 9-tetrahydro-5H-benzo [7] annulen-2-yl ] amino) - [1,2,4] triazolo [1,5-a ] pyridin-5-yl) benzenesulfonamide

Reference example 1 steps b) to d) 2- (4-chloro-2-isopropoxyphenyl) -4, 5-tetramethyl-1, 3, 2-dioxaborane from step b) in example 1 was replaced with (4- (N, N-dimethylsulfamoyl) phenyl) boronic acid.

¹ H NMR(400MHz,DMSO-d ₆ ,ppm)δ9.54(s,1H),8.35(d,J＝8.5Hz,2H),7.96(d,J＝8.5Hz,2H),7.72-7.63(m,2H),7.56(d,J＝2.2Hz,1H),7.38–7.28(m,2H),7.02(d,J＝8.2Hz,1H),3.13-2.93(m,5H),2.86–2.74(m,2H),2.72(s,6H),2.68-2.55(m,2H),2.12(br,2H),1.82(s,4H),1.45(br,2H).MS(ESI+):531.3(M+H).

Example 7 (S) -5- (4-Phenoxyphenyl) -N- (7- (pyrrolidin-1-yl) -6,7,8, 9-tetrahydro-5H-benzo [7] annulen-2-yl) - [1,2,4] triazolo [1,5-a ] pyridin-2-amine

Reference example 1 steps b) to d) 2- (4-chloro-2-isopropoxyphenyl) -4, 5-tetramethyl-1, 3, 2-dioxaborane from step b) in example 1 was replaced with (4-phenoxyphenyl) boronic acid.

¹ H NMR(400MHz,DMSO-d ₆ ,ppm)δ9.48(s,1H),8.13(d,J＝8.8Hz,2H),7.64(dd,J＝8.7,7.4Hz,1H),7.58-7.52(m,2H),7.51–7.44(m,2H),7.35(dd,J＝8.1,2.1Hz,1H),7.24(t,J＝7.4Hz,1H),7.20-7.13(m,5H),7.01(d,J＝8.2Hz,1H),2.93(br,5H),2.90-2.72(m,2H),2.67–2.57(m,2H),2.07(br,2H),1.80(s,4H),1.45(br,2H).MS(ESI+):516.3(M+H).

Example 8 (S) -5- (2-Phenoxyphenyl) -N- (7- (pyrrolidin-1-yl) -6,7,8, 9-tetrahydro-5H-benzo [7] annulen-2-yl) - [1,2,4] triazolo [1,5-a ] pyridin-2-amine

Reference example 1 steps b) to d) the 2- (4-chloro-2-isopropoxyphenyl) -4, 5-tetramethyl-1, 3, 2-dioxaborane from step b) in example 1 was replaced with (2-phenoxyphenyl) boronic acid.

¹ H NMR(400MHz,DMSO-d ₆ ,ppm)δ9.44(s,1H),7.68(dd,J＝7.6,1.6Hz,1H),7.59–7.51(m,3H),7.43(s,1H),7.38(d,J＝8.0Hz,1H),7.32(t,J＝7.5Hz,1H),7.28–7.22(m,2H),7.08–7.03(m,2H),7.02–6.94(m,4H),2.86(br,7H),2.58(br,2H),2.05(br,2H),1.78(s,4H),1.45(br,2H).MS(ESI+):516.3(M+H).

Example 9 (S) -N- (7- (pyrrolidin-1-yl) -6,7,8, 9-tetrahydro-5H-benzo [7] annulen-2-yl) -5- (3- (o-tolyloxy) phenyl) - [1,2,4] triazolo [1,5-a ] pyridin-2-amine

Referring to example 1, 1-bromo-4-chloro-2-isopropoxybenzene of step a) in example 1 was replaced with 1- (3-bromophenoxy) -2-toluene.

¹ H NMR(400MHz,DMSO-d ₆ ,ppm)(s,1H),7.81(s,1H),7.76(d,J＝7.8Hz,1H),7.66–7.60(m,1H),7.59–7.54(m,2H),7.51(s,1H),7.35(t,J＝6.3Hz,2H),7.21(d,J＝7.2Hz,2H),7.12(t,J＝7.2Hz,1H),7.03–6.95(m,3H),2.80(s,7H),2.65-2.55(m,2H),2.51(s,3H),2.05–1.97(m,2H),1.75(br,4H),1.46(br,2H).MS(ESI+):530.3(M+H).

Example 10 (S) -N- (7- (pyrrolidin-1-yl) -6,7,8, 9-tetrahydro-5H-benzo [7] annulen-2-yl) -5- (6- ((tetrahydro-2H-pyran-4-yl) oxy) pyridin-3-yl) - [1,2,4] triazolo [1,5-a ] pyridin-2-amine

Reference example 1 steps b) to d) 2- (4-chloro-2-isopropoxyphenyl) -4, 5-tetramethyl-1, 3, 2-dioxaborane from step b) in example 1 was replaced with 2- ((tetrahydro-2H-pyran-4-yl) oxy) -5- (4, 5-tetramethyl-1, 3, 2-dioxaborane-2-yl) pyridine.

¹ H NMR(400MHz,DMSO-d ₆ ,ppm)δ9.54(s,1H),8.85(d,J＝2.4Hz,1H),8.46(dd,J＝8.7,2.5Hz,1H),7.65(dd,J＝8.8,7.3Hz,1H),7.61–7.50(m,2H),7.40(dd,J＝8.2,2.3Hz,1H),7.24(dd,J＝7.2,1.3Hz,1H),7.04(t,J＝8.2Hz,2H),5.31(tt,J＝8.8,4.1Hz,1H),3.91(dt,J＝11.6,4.3Hz,2H),3.54(ddd,J＝12.0,9.7,2.7Hz,2H),3.22(s,5H),2.87–2.60(m,4H),2.26(s,2H),2.14–2.01(m,2H),1.89(s,4H),1.71(dtd,J＝13.3,9.4,4.2Hz,2H),1.41(s,2H).MS(ESI+):525.3(M+H).

Example 11 (S) -5- (6-Isopropoxypyridin-3-yl) -N- (7- (pyrrolidin-1-yl) -6,7,8, 9-tetrahydro-5H-benzo [7] annulen-2-yl) - [1,2,4] triazolo [1,5-a ] pyridin-2-amine

Reference example 1 steps b) to d) the 2- (4-chloro-2-isopropoxyphenyl) -4, 5-tetramethyl-1, 3, 2-dioxaborane from step b) in example 1 was replaced with (6-isopropoxypyridin-3-yl) boronic acid.

¹ H NMR(400MHz,DMSO-d ₆ ,ppm)δ9.53(s,1H),8.86(d,J＝2.5Hz,1H),8.43(dd,J＝8.7,2.6Hz,1H),7.64(dd,J＝8.8,7.2Hz,1H),7.59–7.49(m,2H),7.39(dd,J＝8.1,2.3Hz,1H),7.24(dd,J＝7.2,1.3Hz,1H),7.05(d,J＝8.2Hz,1H),6.97(d,J＝8.7Hz,1H),5.37(hept,J＝6.1Hz,1H),3.16(s,5H),2.88–2.59(m,4H),2.23(s,2H),1.88(d,J＝6.2Hz,4H),1.54–1.40(m,2H),1.37(d,J＝6.2Hz,6H).MS(ESI+):483.3(M+H).

Biological activity assay

The specific structure of the positive drug (BGB-324) used in the activity test is as follows:

this compound was purchased from Shanghai Shenghong Biotech Co., ltd.

1. Compound AXL kinase inhibitory Activity

1. Experimental procedure

a) AXL enzyme (Carna, 08-107) configuration and addition: using 1 Xenzyme buffer (200. Mu.L of enzymic buffer kinase 5X, 10. Mu.L of 500mM MgCl ₂ 10 μ L of 100mM DTT,6.26 μ L of 2500nM SEB, 773.75 μ L of H was added ₂ O, 1mL of 1 Xenzyme buffer was prepared. ) AXL enzyme 33.33ng/uL was diluted to 0.027 ng/uL (1.67 x, final conc. =0.016 ng/uL), and 6 uL of 1.67-fold final concentration enzyme solution was added to each of the compound well and the positive control well using a BioTek (MultiFlo FX) autosterometer; mu.L of 1 × enzyme buffer was added to the negative control wells.

b) Compound preparation and addition: the compounds prepared in the examples and the positive drugs were diluted from 10mM to 100. Mu.M using DMSO and titrated using a compound titrator (Tecan, D300 e) which automatically sprays the desired concentration into each well at 1. Mu.M, 1/2log gradient dilution, for a total of 8 concentrations. Centrifuge at 2500rpm for 30s and incubate at room temperature for 15min.

c) ATP, substrate preparation and addition: ATP (Sigma, A7699) was diluted in 1 Xenzyme buffer from 10mM to 75. Mu.M (5X) to a final concentration of 15. Mu.M; substrate TK Substrate 3-biotin (Cisbio, 61TK0 BLC) was diluted from 500. Mu.M to 5. Mu.Ma 5X in 1 Xenzyme buffer to a final concentration of 1. Mu.M; ATP was mixed with the substrate in equal volumes and added to each well using a BioTek autostergrator 4 μ L; the reaction mixture was centrifuged at 2500rpm for 30s and reacted at 25 ℃ for 45min.

d) Preparing and adding a detection reagent: streptavidin-XL665 (Cisbio, 610 SAXLG) was diluted from 16.67. Mu.M to 250nM (4X) with HTRF KinEASE detection buffer (Cisbio) to a final concentration of 62.5nM; TK Antibody-Cryptate (Cisbio) was diluted from 100X to 5X with HTRF KinEASE detection buffer (Cisbio) to a final concentration of 1X; XL665 was mixed with equal volumes of Antibody, added to each well using a BioTek autosterfer 10. Mu.L, centrifuged 30s at 2500rpm, and reacted for 1 hour at 25 ℃. After the reaction is finished, detection is carried out by using a multifunctional plate reading instrument HTRF.

2. Data analysis

The IC of the compound for inhibiting AXL kinase is obtained by fitting a dose-effect curve with GraphPad Prism 5 software log (inhibitor) vs. stress-Variable slope ₅₀ The value is obtained.

The inhibition rate calculation formula is as follows:

conversion% _ sample: is the conversion reading for the sample;

conversion% _ min: a conversion reading representing no enzyme live wells;

conversion% _ max: a conversion reading representing no compound inhibited the well.

3. Results of the experiment

The results are shown in Table 1, wherein the IC of AXL of the compound ₅₀ Has three grades of A, B and C, wherein A represents<10nM; b represents 10 to 50nM; c represents>50nM。

AXL IC ₅₀ Data of

2. Compound-based cell proliferation inhibition assay

1. Experimental procedure

MV-4-11 (human myelomonocytic leukemia cell line, culture medium: IMDM +10% fetal bovine serum) was purchased from Bai Biotech Ltd, kyoto, and was incubated at 37 ℃ for 5% CO ₂ The incubator of (2). Cells in the logarithmic growth phase were plated in 96-well plates at cell densities of 8000, 6000, 2000 and 3000 cells per well, respectively, while blank control groups were set.

The test compound and the positive drug were dissolved in dimethyl sulfoxide to prepare a 10mM stock solution, and stored in a refrigerator at-80 ℃ for a long period. After 24h of cell plating, 10mM stock solution of the compound was diluted with DMSO to give 200-fold concentration working solution (maximum concentration 200 or 2000. Mu.M, 3-fold gradient, 10 concentrations total), 3. Mu.L of each concentration was added to 197. Mu.L of complete medium, diluted to give 3-fold concentration working solution, and 50. Mu.L of each concentration was added to 100. Mu.L of cell culture solution (final concentration of DMSO is 0.5%, v/v), with two duplicate wells per concentration. After 72h of dosing, 50. Mu.l of each well was added

(purchased from Promega) fluorescence signals were measured on Envision (PerkinElmer) according to the protocol of the instructions and dose-response curves were fitted using GraphPad Prism 5 software log (inhibitor) vs. response-Variable slope to obtain IC of the compounds for inhibition of cell proliferation ₅₀ The value is obtained. Inhibition rate calculation formula:

wherein:

test substance signal value: mean value of fluorescence signals of cell + culture medium + compound group;

blank set signal value: mean fluorescence signal of medium group (containing 0.5% DMSO);

signal values of negative control group: mean fluorescence signal of cell + medium group (containing 0.5% DMSO).

2. Results of the experiment

The results of the experiment are shown in Table 2, in which the IC of the compound for cell proliferation inhibition ₅₀ The value is divided into three grades A, B and C, wherein A represents<100nM; b represents 100 to 200nM; c represents>200nM。

TABLE 2 antiproliferative Activity of Compounds on MV4-11 cells

EXAMPLES title Compounds	IC 50 (MV4-11,nM)
		Example 3	A
Example 5	A
		Positive drug (BGB-324)	208.1

Claims (10)

1. Compound I or a pharmaceutically acceptable salt thereof:

wherein ring A is selected from phenyl or 5-to 10-membered heteroaryl, optionally substituted with one or more R ¹ Substituent(s) substituted, R ¹ Selected from the following groups: -SO ₂ NR ² R ³ Halogen, C1-C6 alkyl, (C1-C6 alkyl) -O-, (5-to 10-membered heterocyclyl) -O-, phenyl-O-, (5-to 6-membered heteroaryl) -O-, cyano or nitro, wherein (5-to 10-membered heterocyclyl) -O-, phenyl-O-, and (5-to 6-membered heteroaryl) -O-are optionally substituted with one or more R ⁴ Substituent(s) substituted, R ⁴ Selected from C1-C3 alkyl, halogen, cyano or nitro, R ² And R ³ Each independently selected from hydrogen or C1-C6 alkyl.

2. Compound I or a pharmaceutically acceptable salt thereof according to claim 1, wherein Ring A is selected from phenyl or 5-to 6-membered heteroaryl, optionally substituted by one or more R ¹ Substituent groupSubstituted, R ¹ Selected from the following groups: -SO ₂ NR ² R ³ Halogen, C1-C6 alkyl, (C1-C6 alkyl) -O-, (5-to 10-membered heterocyclyl) -O-, phenyl-O-, (5-to 6-membered heteroaryl) -O-, cyano or nitro, wherein (5-to 10-membered heterocyclyl) -O-, phenyl-O-, and (5-to 6-membered heteroaryl) -O-are optionally substituted with one or more R ⁴ Substituent(s) substituted, R ⁴ Selected from C1-C3 alkyl, halogen, cyano or nitro, R ² And R ³ Each independently selected from hydrogen or C1-C6 alkyl.

3. Compound I according to claim 1, or a pharmaceutically acceptable salt thereof, ring A being selected from phenyl, imidazolyl, oxazolyl, isoxazolyl, thiazolyl, isothiazolyl, triazolyl, oxadiazolyl, thiadiazolyl, tetrazolyl, oxatriazolyl, thiatriazolyl, pyridyl, pyrimidinyl, pyrazinyl, pyridazinyl, 1,2, 3-triazinyl, 1,2, 4-triazinyl or 1,3, 5-triazinyl, optionally substituted with one or more R ¹ Substituent(s) substituted, R ¹ Selected from the following groups: -SO ₂ NR ² R ³ Halogen, C1-C6 alkyl, (C1-C6 alkyl) -O-, (5-to 10-membered heterocyclyl) -O-, phenyl-O-, (5-to 6-membered heteroaryl) -O-, cyano or nitro, wherein (5-to 10-membered heterocyclyl) -O-, phenyl-O-, and (5-to 6-membered heteroaryl) -O-are optionally substituted with one or more R ⁴ Substituent(s) substituted, R ⁴ Selected from C1-C3 alkyl, halogen, cyano or nitro, R ² And R ³ Each independently selected from hydrogen or C1-C6 alkyl; preferably, ring A is selected from phenyl, imidazolyl, oxazolyl, thiazolyl, pyridyl or pyrimidinyl, optionally substituted with one or more R ¹ Substituent(s) substituted, R ¹ Selected from the following groups: -SO ₂ NR ² R ³ Halogen, C1-C6 alkyl, (C1-C6 alkyl) -O-, (5-to 10-membered heterocyclyl) -O-, phenyl-O-, (5-to 6-membered heteroaryl) -O-, cyano or nitro, wherein (5-to 10-membered heterocyclyl) -O-, phenyl-O-, and (5-to 6-membered heteroaryl) -O-are optionally substituted with one or more R ⁴ Substituent(s) substituted, R ⁴ Selected from C1-C3 alkyl, halogen, cyano or nitro, R ² And R ³ Each independently selected from hydrogen or C1-C6 alkyl; more preferably, ring A is selected from phenylOr pyridyl, optionally substituted by one or more R ¹ Substituent group, R ¹ Selected from the following groups: -SO ₂ NR ² R ³ Halogen, C1-C6 alkyl, (C1-C6 alkyl) -O-, (5-to 10-membered heterocyclyl) -O-, phenyl-O-, (5-to 6-membered heteroaryl) -O-, cyano or nitro, wherein (5-to 10-membered heterocyclyl) -O-, phenyl-O-, and (5-to 6-membered heteroaryl) -O-are optionally substituted with one or more R ⁴ Substituent group, R ⁴ Selected from C1-C3 alkyl, halogen, cyano or nitro, R ² And R ³ Each independently selected from hydrogen or C1-C6 alkyl.

4. Compound I according to claim 1, or a pharmaceutically acceptable salt thereof, wherein Ring A is selected from

Optionally substituted by one or more R ¹ Substituent(s) substituted, R ¹ Selected from the following groups: -SO ₂ NR ² R ³ Halogen, C1-C6 alkyl, (C1-C6 alkyl) -O-, (5-to 10-membered heterocyclyl) -O-, or phenyl-O-, wherein (5-to 10-membered heterocyclyl) -O-and phenyl-O-are optionally substituted with one or more R ⁴ Substituent group, R ⁴ Selected from C1-C3 alkyl, halogen, cyano or nitro, R ² And R ³ Each independently selected from hydrogen or C1-C6 alkyl.

5. Compound I according to claim 1, or a pharmaceutically acceptable salt thereof, wherein Ring A is selected from

Optionally substituted by one or more R ¹ Substituent(s) substituted, R ¹ Selected from the following groups: -SO ₂ N(CH ₃ ) ₂ 、-SO ₂ N(CH ₂ CH ₃ ) ₂ Halogen, methyl, ethyl, ⁱ Pr-O-、CH ₃ O-、CH ₃ CH ₂ O-, piperidyl, pyrrolidinyl, piperazinyl, morpholinyl, thiomorpholinyl, 1, 3-dioxolanyl, tetrahydrofuryl, tetrahydrothienyl, tetrahydrothiopyranyl, tetrakisHydropyridyl, dihydropyridyl, pyridinyl, and the like,

<xnotran> , , , , ,1,3- , , , , , , </xnotran>

Optionally substituted by one or more R ⁴ Substituent group, R ⁴ Selected from C1-C3 alkyl, halogen, cyano or nitro; preferably, ring A is selected from

Optionally substituted by one or more R ¹ Substituent group, R ¹ Selected from the following groups: -SO ₂ N(CH ₃ ) ₂ 、-SO ₂ N(CH ₂ CH ₃ ) ₂ Halogen, methyl, ethyl, ⁱ Pr-O-、CH ₃ O-、CH ₃ CH ₂ O-, piperidyl, pyrrolidinyl, piperazinyl, morpholinyl, thiomorpholinyl, 1, 3-dioxolanyl, or a pharmaceutically acceptable salt thereof tetrahydrofuranyl, tetrahydrothienyl, tetrahydrothiopyranyl, tetrahydropyridinyl, dihydropyridinyl,

Wherein piperidinyl, pyrrolidinyl, piperazinyl, morpholinyl, thiomorpholinyl, 1, 3-dioxolanyl, tetrahydrofuryl, tetrahydrothienyl, tetrahydrothiopyranyl, tetrahydropyridinyl, dihydropyridinyl, or mixtures thereof,

Optionally substituted by one or more R ⁴ Substituent(s) substituted, R ⁴ Is a halogen.

6. Compound I according to claim 1, or a pharmaceutically acceptable salt thereof, wherein Ring A is selected from

Optionally substituted by one or more R ¹ Substituent group, R ¹ Selected from the following groups: -Cl,

ⁱ Pr-O-、-SO ₂ N(CH ₃ ) ₂ Or

Wherein, the first and the second end of the pipe are connected with each other,

optionally substituted by one or more R ⁴ Substituent(s) substituted, R ⁴ Is methyl; preferably, R ¹ Selected from the following groups: ⁱ Pr-O-、-SO ₂ N(CH ₃ ) ₂ or

7. Compound I according to claim 1, or a pharmaceutically acceptable salt thereof, wherein Ring A is

It is covered ⁱ Pr-O-; preferably, ring A is

8. A compound or a pharmaceutically acceptable salt thereof:

9. a pharmaceutical composition comprising a therapeutically effective amount of compound I or a pharmaceutically acceptable salt thereof and a pharmaceutically acceptable carrier.

10. Compound I or a pharmaceutically acceptable salt thereof for the manufacture of a medicament for the treatment and/or prevention of AXL receptor tyrosine kinase induced disorders.