WO2019056990A1

WO2019056990A1 - 1,2-dihydro-1,6-naphthyridine derivative, preparation method therefor, and application thereof in medicine

Info

Publication number: WO2019056990A1
Application number: PCT/CN2018/105718
Authority: WO
Inventors: 别平彦; 陈磊; 白骅
Original assignee: 浙江海正药业股份有限公司
Priority date: 2017-09-20
Filing date: 2018-09-14
Publication date: 2019-03-28
Also published as: TWI707853B; CN111094286B; TW201914999A; CN111094286A

Abstract

A 1,2-dihydro-1,6-naphthyridine derivative, a preparation method therefor, and an application thereof in medicine. Specifically provided are a 1,2-dihydro-1,6-naphthyridine derivative as represented by formula (I), a preparation method therefor, pharmaceutically acceptable salts thereof, and application thereof as therapeutic agents, particularly c-KIT inhibitors, wherein the definitions of the substituents in the formula (I) are the same as the definitions in the description.

Description

1,2-dihydro-1,6-naphthyridine derivative, preparation method thereof and use thereof in medicine

Technical field

The present invention relates to a novel 1,2-dihydro-1,6-naphthyridine derivative, a process for the preparation thereof, and a pharmaceutical composition containing the same, and as a therapeutic agent, particularly as a c-KIT inhibitor use.

Background technique

c-KIT (also known as KIT, CD117 and stem cell factor receptor) is a 145 kDa transmembrane tyrosine kinase protein that acts as a type III receptor. The c-KIT proto-oncogene located on chromosome 4q11-21 encodes c-KIT and its ligand is a stem cell factor. The receptor has tyrosine protein kinase activity and binding to the ligand SCF results in autophosphorylation of c-KIT and its association with a substrate such as phosphatidylinositol 3-kinase (PI3K). Phosphorylation of tyrosine by protein tyrosine kinases is particularly important in cell signaling and can mediate signals of major cellular processes such as proliferation, survival, differentiation, apoptosis, ligation, invasion and migration. c-KIT mutations are commonly found in DNA encoding the membrane proximal domain (exon 11). They also appear in exons 7, 8, 9, 11, 13, 14, 17, and 18 at a lower frequency. Mutations make c-KIT function independent of activation by SCF, resulting in high cell division rates and possible genomic instability. Functionally increased mutations in the c-KIT gene and expression of constitutive phosphorylation c-KIT can be found in most gastrointestinal stromal tumors (GIST), mastocytosis, and acute myeloid leukemia. It has mutations at different positions in different exons. The first generation of c-KIT mutations and related drugs are mainly imatinib, sunitinib, dasatinib and PKC412.

c-KIT has germ cell tumors in gastrointestinal stromal tumors, acute myeloid leukemia, melanoma, breast adenoma, cervical tumor, glioblastoma multiforme, ovarian tumor, seminoma or dysplasia It has been found in teratoma, mast cell leukemia and other tissues, and its protein expression level is closely related to the occurrence and development of tumors. Gastrointestinal stromal tumor (GIST) is the most common mesenchymal origin of the gastrointestinal tract. According to the current GIST diagnostic criteria, epidemiological studies show an incidence rate of 0.66 to 2.20/100,000. GIST is extremely insensitive to traditional chemotherapy, with less than 5% effective chemotherapy drugs and a median survival rate of only about 18 months. Even with complete tumor resection, the 5-year survival rate of GIST is only 35%-65%, and the recurrence and metastasis rate is 40%-50% within 2 years, and as many as 15%-50% of patients in the first diagnosis have metastasis. It was found that the transmembrane tyrosine kinase receptor c-KIT on the surface of stem cell factor and the functional activating mutation of the platelet-derived growth factor receptor PDGFRα gene are the key to the development of GIST. The platelet-derived growth factor receptor (PDGF-R) is a cell surface tyrosine kinase receptor that is a member of the platelet-derived growth factor (PDGF) family. The PDGF subunits PDGFα and PDGFβ are important regulators of cell proliferation, cell differentiation, cell growth, development, and many diseases including cancer.

With the clinical application of the first-generation inhibitor, Imatinib, the acquired resistance of imatinib has become a serious challenge in the clinical use of such inhibitors. Therefore, there is an urgent need to research and develop new c-KIT inhibitors to meet market demand. A series of c-KIT inhibitor patents have been published, including WO2014039714, WO2014100620, WO2015134536A1 and WO2013184119. The research and application of c-KIT inhibitors have made some progress, but the space for improvement is still huge, and it is still necessary. Continue to research and develop new c-KIT inhibitors.

Summary of the invention

In order to overcome the deficiencies of the prior art, it is an object of the present invention to provide a novel class of 1,2-dihydro-1,6-naphthyridine derivatives of the formula (I), or a stereo thereof Isomers, tautomers or pharmaceutically acceptable salts thereof, the compounds of the invention have large structural differences from the compounds specifically disclosed in the prior art, and can be treated or prevented by modulating c-KIT and PDGFα activity Diseases such as gastrointestinal stromal tumors, acute myeloid leukemia, and systemic mast cell hyperplasia.

A compound of the formula (I) of the present invention, or a stereoisomer, tautomer or pharmaceutically acceptable salt thereof:

among them:

R ^{1 is} selected from a hydrogen atom, an alkyl group or -C(O)R ⁵ , wherein the alkyl group is optionally further further selected from one or more selected from the group consisting of halogen, hydroxy, alkoxy, cycloalkyl, heterocyclic, -C(O)R ⁵ , -OC(O)R ⁵ , -C(O)OR ⁵ , -NR ⁶ R ⁷ , -C(O)NR ⁶ R ⁷ , -S(O) _n NR ⁶ R ⁷ Or substituted with a substituent of -NR ⁶ C(O)R ⁷ ;

R ^{2 is} selected from alkyl, wherein said alkyl group is further substituted with one or more halogens;

R ^{3 is the} same or different and is each independently selected from the group consisting of a hydrogen atom, an alkyl group, an alkoxy group, a hydroxyl group, a cyano group, a nitro group, a halogen group, a cycloalkyl group, a heterocyclic group, -C(O)R ⁵ , -OC ( O) R ⁵ , -C(O)OR ⁵ , -NR ⁶ R ⁷ , -C(O)NR ⁶ R ⁷ , -S(O) _n NR ⁶ R ⁷ or -NR ⁶ C(O)R ⁷ , Wherein the alkyl, alkoxy, cycloalkyl or heterocyclic group is optionally further selected from one or more selected from the group consisting of hydroxyl, halogen, nitro, cyano, alkyl, alkoxy, cycloalkyl, hetero a cyclic group, an aryl group, a heteroaryl group, -C(O)R ⁸ , -C(O)OR ⁸ , -OC(O)R ⁸ , -NR ⁹ R ¹⁰ , -C(O)NR ⁹ R ¹⁰ , Substituted by a substituent of -S(O) _n NR ⁹ R ¹⁰ or -NR ⁹ C(O)R ¹⁰ ;

R ^{4 is} selected from aryl or heteroaryl, wherein said aryl or heteroaryl is optionally further selected from one or more selected from the group consisting of alkyl, alkoxy, hydroxy, cyano, nitro, halogen, cycloalkane. Base, heterocyclic group, -C(O)R ⁵ , -OC(O)R ⁵ , -C(O)OR ⁵ , -NR ⁶ R ⁷ , -C(O)NR ⁶ R ⁷ , -S(O Substituting a substituent of _n NR ⁶ R ⁷ or -NR ⁶ C(O)R ⁷ ; wherein said alkyl or alkoxy group is optionally further substituted with one or more halogens;

R ⁵ , R ⁶ and R ⁷ are each independently selected from a hydrogen atom, a hydroxyl group, a halogen, a nitro group, a cyano group, an alkyl group, an alkoxy group, a cycloalkyl group, a heterocyclic group, an aryl group or a heteroaryl group. The alkyl, alkoxy, cycloalkyl, heterocyclyl, aryl or heteroaryl group optionally further selected from one or more selected from the group consisting of hydroxyl, halogen, nitro, cyano, alkyl, alkoxy, Cycloalkyl, heterocyclyl, aryl, heteroaryl, -C(O)R ⁸ , -C(O)OR ⁸ , -OC(O)R ⁸ , -NR ⁹ R ¹⁰ , -C(O) Substituted by a substituent of NR ⁹ R ¹⁰ , -S(O) _n NR ⁹ R ¹⁰ or -NR ⁹ C(O)R ¹⁰ ;

Alternatively, R ⁶ and R ⁷ together with the N atom to which they are bonded form a 4 to 8 membered heterocyclic group, wherein the 4 to 8 membered heterocyclic ring contains one or more N, O or S(O) _n and 4 to The 8-membered heterocyclic ring is optionally further selected from one or more selected from the group consisting of hydroxyl, halogen, nitro, cyano, alkyl, alkoxy, cycloalkyl, heterocyclyl, aryl, heteroaryl, =0, -C(O)R ⁸ , -C(O)OR ⁸ , -OC(O)R ⁸ , -NR ⁹ R ¹⁰ , -C(O)NR ⁹ R ¹⁰ , -S(O) _n NR ⁹ R ¹⁰ Or substituted with a substituent of -NR ⁹ C(O)R ¹⁰ ;

R ⁸ , R ⁹ and R ¹⁰ are each independently selected from a hydrogen atom, an alkyl group, a cycloalkyl group, a heterocyclic group, an aryl group or a heteroaryl group, wherein the alkyl group, the cycloalkyl group, the heterocyclic group, and the aromatic group Or a heteroaryl group optionally further selected from one or more selected from the group consisting of hydroxyl, halogen, nitro, cyano, alkyl, alkoxy, cycloalkyl, heterocyclyl, aryl, heteroaryl, carboxy or carboxy Substituted by a substituent of the acid ester group;

m is selected from 1, 2, 3 or 4;

n is selected from 0, 1 or 2.

A preferred embodiment of the invention, a compound of the formula (I), or a stereoisomer, tautomer thereof or a pharmaceutically acceptable salt thereof, which is a compound of the formula (II) or a stereoisomer, a tautomer or a pharmaceutically acceptable salt thereof,

among them:

R ^{3 is} selected from the group consisting of a hydrogen atom, an alkyl group, an alkoxy group, a hydroxyl group, a cyano group, a nitro group, a halogen, a cycloalkyl group, a heterocyclic group, -C(O)R ⁵ , -OC(O)R ⁵ , -C (O)OR ⁵ , -NR ⁶ R ⁷ , -C(O)NR ⁶ R ⁷ , -S(O) _n NR ⁶ R ⁷ or -NR ⁶ C(O)R ⁷ , wherein the alkyl group, The alkoxy, cycloalkyl or heterocyclic group is optionally further selected from one or more selected from the group consisting of hydroxyl, halogen, nitro, cyano, alkyl, alkoxy, cycloalkyl, heterocyclyl, aryl, hetero Aryl, -C(O)R ⁸ , -C(O)OR ⁸ , -OC(O)R ⁸ , -NR ⁹ R ¹⁰ , -C(O)NR ⁹ R ¹⁰ , -S(O) _n NR Substituted by a substituent of ⁹ R ¹⁰ or -NR ⁹ C(O)R ¹⁰ ;

R ¹ , R ² , R ⁴ to R ¹⁰ and n are as defined in the formula (I).

A preferred embodiment of the invention, a compound of the formula (I) or (II), or a stereoisomer, tautomer thereof or a pharmaceutically acceptable salt thereof, wherein:

R ^{1 is} selected from a hydrogen atom or a C _1-4 alkyl group, wherein said C _1-4 alkyl group is optionally further substituted with one or more C _1-6 alkoxy groups; wherein said C _1-6 The alkoxy group is preferably a methoxy group.

R ^{2 is} selected from C _1-4 alkyl, wherein said C _1-4 alkyl group is further substituted by one or more halogens; wherein said halogen is preferably fluorine, chlorine or bromine; more preferably fluorine.

A preferred embodiment of the invention is a compound of the formula (I) or (II), or a stereoisomer, tautomer thereof or a pharmaceutically acceptable salt thereof, wherein R ² is a trifluoroethyl group.

R ^{3 is} selected from a hydrogen atom, a C _1-4 alkyl group or a halogen;

Wherein the C _1-4 alkyl group is preferably a methyl group or an ethyl group;

The halogen described therein is preferably fluorine, chlorine or bromine.

R ^{4 is} selected from phenyl or benzothienyl, wherein said phenyl or benzothienyl is optionally further further selected from one or more selected from the group consisting of halogen, C _1-4 alkyl or C _1-6 alkoxy. Substituted by a substituent;

Wherein the C _1-4 alkyl or C _1-6 alkoxy group is optionally further substituted with one or more halogens;

Wherein the C _1-4 alkyl group is preferably a methyl group or an ethyl group;

The halogen described therein is preferably fluorine, chlorine or bromine; more preferably fluorine.

R ^{1 is} selected from a hydrogen atom or an alkyl group, preferably a C _1-4 alkyl group, wherein the alkyl group is optionally further substituted with one or more C _1-6 alkoxy groups; wherein the C _1-6 The alkoxy group is preferably a methoxy group.

R ^{2 is} selected from alkyl, wherein said alkyl group is further substituted by one or more halogens; preferably said alkyl group is a C _1-4 alkyl group, more preferably an ethyl group; said halogen is preferably fluorine, chlorine or Bromine is more preferably fluorine.

R ^{3 is} selected from a hydrogen atom, an alkyl group or a halogen, wherein the alkyl group is optionally further substituted by one or more halogens; the alkyl group is preferably a C _1-4 alkyl group, more preferably a methyl group or an ethyl group. And the halogen is preferably fluorine, chlorine or bromine.

R ^{4 is} selected from phenyl or benzothienyl, wherein said phenyl or benzothienyl is optionally further substituted with one or more substituents selected from halogen, alkyl or alkoxy; The alkyl or alkoxy group is optionally further substituted with one or more halogens; the alkyl group is preferably a C _1-4 alkyl group; the alkoxy group is preferably a C _1-6 alkoxy group; The halogen is preferably fluorine, chlorine or bromine; more preferably fluorine.

Typical compounds of the invention include, but are not limited to:

Or a stereoisomer, tautomer thereof or a pharmaceutically acceptable salt thereof.

The present invention provides a process for the preparation of a compound of the formula (I) or a stereoisomer, tautomer or a pharmaceutically acceptable salt thereof, the method comprising:

The compound of the formula (IA) is reacted with a compound of the formula (IB) under basic conditions to give a compound of the formula (I);

Wherein: R ¹ to R ⁴ and m are as defined in the formula (I).

In the above preparation method, the basic condition is provided by an organic base or an inorganic base selected from the group consisting of diisopropylethylamine, pyridine, triethylamine, piperidine, N-methylpiperazine, 4-dimethylaminopyridine, Preference is given to diisopropylethylamine or triethylamine; the inorganic base is selected from the group consisting of sodium carbonate, potassium carbonate, cesium carbonate, sodium hydride, potassium hydride, preferably cesium carbonate or potassium carbonate.

The present invention provides a method for producing a compound of the formula (II), or a stereoisomer, tautomer thereof or a pharmaceutically acceptable salt thereof, the method comprising:

The compound of the formula (IIA) is reacted with a compound of the formula (IB) under basic conditions to give a compound of the formula (II);

Wherein: R ¹ to R ⁴ are as defined in the formula (II).

In the above preparation method, the basic condition is provided by an organic base or an inorganic base selected from the group consisting of diisopropylethylamine, pyridine, triethylamine, piperidine, N-methylpiperazine, 4-dimethylaminopyridine, Preference is given to diisopropylethylamine and/or triethylamine; the inorganic base is selected from the group consisting of sodium carbonate, potassium carbonate, cesium carbonate, sodium hydride, potassium hydride, preferably cesium carbonate and/or potassium carbonate.

Further, the present invention provides an intermediate compound of the formula (IA), or a stereoisomer, tautomer thereof or a pharmaceutically acceptable salt thereof,

Wherein: R ¹ to R ³ and m are as defined in the formula (I).

The present invention provides an intermediate compound of the formula (IA), or a stereoisomer, tautomer thereof or a pharmaceutically acceptable salt thereof, which is an intermediate compound of the formula (IIA) or a stereoisomer, tautomer or pharmaceutically acceptable salt thereof,

Wherein: R ¹ to R ³ are as defined in the formula (II).

Typical intermediate compounds of the formula (IA) of the present invention include, but are not limited to:

The present invention provides a process for the preparation of an intermediate compound of the formula (IA), or a stereoisomer, tautomer or a pharmaceutically acceptable salt thereof, which comprises:

The compound of the formula (Ia) is subjected to a condensation reaction with a compound of the formula (Ib) under heating to obtain a compound of the formula (Ic); the compound of the formula (Ic) is reacted with a compound of the formula (Id) under heating, Obtaining an intermediate compound of the formula (IA);

among them:

X is a halogen;

R ^a is an alkyl group;

R ¹ to R ³ and m are as defined in the formula (IA), and R ^{1 is} not a hydrogen atom.

The present invention provides a method for producing an intermediate compound of the formula (IA), or a stereoisomer thereof, a tautomer or a pharmaceutically acceptable salt thereof, the method comprising:

The compound of the formula (Ia) is subjected to a condensation reaction with a compound of the formula (Ib) under heating to obtain a compound of the formula (Ic); and the compound of the formula (Ic) and the compound of the formula (Ie) are subjected to heating under heating a condensation reaction to obtain a compound of the formula (If); a compound of the formula (If) is reacted in the presence of trifluoroacetic acid to give an intermediate compound of the formula (IA);

among them:

X is a halogen;

R ¹ is a hydrogen atom;

R ^a is an alkyl group; R ^b is an alkyl group;

R ² , R ³ and m are as defined in the general formula (IA).

The present invention provides a method for producing an intermediate compound of the formula (IIA), or a stereoisomer thereof, a tautomer or a pharmaceutically acceptable salt thereof, the method comprising:

The compound of the formula (Ia) is subjected to a condensation reaction with a compound of the formula (IIa) under heating to obtain a compound of the formula (IIb); the compound of the formula (IIb) and the compound of the formula (Id) are reacted under heating. , obtaining an intermediate compound of the formula (IIA);

among them:

X is a halogen;

R ^a is an alkyl group;

R ¹ to R ³ are as defined in the formula (IIA), and R ^{1 is} not a hydrogen atom.

The compound of the formula (Ia) is subjected to a condensation reaction with a compound of the formula (IIa) under heating to obtain a compound of the formula (IIb); the compound of the formula (IIb) and the compound of the formula (Ie) are subjected to a condensation reaction under heating. To obtain a compound of the formula (IIc); a compound of the formula (IIc) is reacted in the presence of trifluoroacetic acid to give an intermediate compound of the formula (IIA);

among them:

X is a halogen;

R ¹ is a hydrogen atom;

R ^a is an alkyl group; R ^b is an alkyl group;

R ² and R ³ are as defined in the formula (IIA).

Furthermore, the present invention provides a pharmaceutical composition comprising an effective amount of a compound of the formula (I) or (II) or a stereoisomer, tautomer thereof or Pharmaceutically acceptable salts, and pharmaceutically acceptable carriers, excipients or combinations thereof.

The present invention provides a method for inhibiting c-KIT, which comprises reacting a c-KIT receptor with a compound of the formula (I) or (II) or a stereoisomer, tautomer thereof or a drug thereof The salt used, or a pharmaceutical composition thereof, is contacted.

The present invention provides a compound of the formula (I) or (II), or a stereoisomer, tautomer thereof or a pharmaceutically acceptable salt thereof, or a pharmaceutical composition thereof, in the preparation of a therapeutic by c-KIT Or the use of a medicament for mutating c-KIT mediated diseases, wherein the c-KIT or mutated c-KIT mediated disease is preferably gastrointestinal stromal tumor, systemic mastocytosis, Acute myeloid leukemia, ovarian cancer, melanoma, cervical cancer, seminoma, dysgerminoma, glioblastoma multiforme, teratoma, mast cell leukemia; more preferably gastrointestinal stromal tumor, Systemic mastocytosis, glioblastoma multiforme, and acute myeloid leukemia, most preferably gastrointestinal stromal tumors, glioblastoma multiforme, and systemic mastocytosis; preferably said The mutated c-KIT mutation is located at 9, 9, 13, 14, 17, and/or 18 of the exon, or at the 816th, 670th, 560th, and/or 654th amino acid residues, wherein The mutation at the amino acid residue at position 816 is preferably D816V or D816H; wherein the mutation at the amino acid residue at position 670 is preferred T670I; wherein the mutation at the amino acid residue at position 560 is preferably V560G; wherein the mutation at the amino acid residue at position 654 is preferably V654A.

The present invention provides a compound of the formula (I) or (II) or a stereoisomer, tautomer thereof or a pharmaceutically acceptable salt thereof, or a pharmaceutical composition thereof for preparing a c-KIT inhibitor Use in.

The present invention provides a compound of the formula (I) or (II), or a stereoisomer, tautomer thereof or a pharmaceutically acceptable salt thereof, or a pharmaceutical composition thereof, in the preparation of a PDFGRα inhibitor use.

The present invention provides a compound of the formula (I) or (II) or a stereoisomer, tautomer thereof or a pharmaceutically acceptable salt thereof, or a pharmaceutical composition thereof, in the preparation of a therapeutic mutation or Use of a wild-type PDFGRα-mediated disease, wherein the disease mediated by mutated or wild-type PDFGRα is preferably gastrointestinal stromal tumor, systemic mastocytosis, acute myeloid leukemia, ovary Cancer, melanoma, cervical cancer, seminoma, dysgerminoma, glioblastoma multiforme, teratoma, mast cell leukemia; more preferably gastrointestinal stromal tumor, systemic mastocytosis , glioblastoma multiforme and acute myeloid leukemia, most preferably gastrointestinal stromal tumors, glioblastoma multiforme and systemic mastocytosis; preferably the mutant PDFGRα mutation is located At position 18 and/or at position 842 of the amino acid residue, wherein the mutation at amino acid residue position 842 is preferably a D842V mutation.

The present invention provides a method of treating a disease mediated by c-KIT or a mutant c-KIT comprising administering to a patient a therapeutically effective amount of a compound of the formula (I) or (II) or a stereoisomer thereof. a tautomer or a pharmaceutically acceptable salt thereof, or a pharmaceutical composition thereof, wherein the c-KIT or mutant c-KIT mediated disease is preferably a gastrointestinal stromal tumor or a systemic mast cell Hyperplasia, acute myeloid leukemia, ovarian cancer, melanoma, cervical cancer, seminoma, dysgerminoma, glioblastoma multiforme, teratoma, mast cell leukemia; more preferably between the gastrointestinal tract A tumor, systemic mastocytosis, glioblastoma multiforme, and acute myeloid leukemia, most preferably gastrointestinal stromal tumors, glioblastoma multiforme, and systemic mastocytosis; The mutant of the mutant c-KIT is located at 9, 9, 13, 14, 17, and/or 18 of the exon, or amino acid residues 816, 670, 560, and/or 654. Wherein the mutation at the amino acid residue at position 816 is preferably D816V or D816H; wherein the amino acid at position 670 The mutation at the residue is preferably T670I; wherein the mutation at the amino acid residue at position 560 is preferably V560G; wherein the mutation at the amino acid residue at position 654 is preferably V654A.

The present invention provides a method of treating a disease mediated by PDFGRα or a mutant PDFGRα, which comprises administering to a patient a therapeutically effective amount of a compound of the formula (I) or (II) or a stereoisomer thereof, a tautomer thereof. A constitutive or a pharmaceutically acceptable salt thereof, or a pharmaceutical composition thereof, wherein the PDFGRα or mutated PDFGRα mediated disease is preferably gastrointestinal stromal tumor, systemic mastocytosis, acute myeloid leukemia, Ovarian cancer, melanoma, cervical cancer, seminoma, dysgerminoma, glioblastoma multiforme, teratoma, mast cell leukemia; more preferably gastrointestinal stromal tumor, systemic mast cell hyperplasia Symptoms, glioblastoma multiforme and acute myeloid leukemia, most preferably gastrointestinal stromal tumors, glioblastoma multiforme, and systemic mastocytosis; wherein the mutation of PDFGRα is mutated Located at exon 18 and/or amino acid residue position 842, wherein the mutation at amino acid residue position 842 is preferably a D842V mutation.

Detailed description of the invention

Unless otherwise stated, some of the terms used in the specification and claims of the invention are defined as follows:

When "alkyl" as a group or part of a group is meant to include C ₁ -C ₂₀ linear or branched aliphatic hydrocarbon group with a chain. It is preferably a C ₁ -C ₁₀ alkyl group, more preferably a C ₁ -C ₆ alkyl group. Examples of alkyl groups include, but are not limited to, methyl, ethyl, n-propyl, isopropyl, n-butyl, isobutyl, t-butyl, sec-butyl, n-pentyl, 1, 1-di Methylpropyl, 1,2-dimethylpropyl, 2,2-dimethylpropyl, 1-ethylpropyl, 2-methylbutyl, 3-methylbutyl, n-hexyl, 1 -ethyl-2-methylpropyl, 1,1,2-trimethylpropyl, 1,1-dimethylbutyl, 1,2-dimethylbutyl, 2,2-dimethyl Butyl, 1,3-dimethylbutyl, 2-ethylbutyl, 2-methylpentyl, 3-methylpentyl, 4-methylpentyl, 2,3-dimethylbutyl Wait. The alkyl group can be substituted or unsubstituted.

"Alkenyl" means an aliphatic hydrocarbon group containing a carbon-carbon double bond, which may be straight or branched, and representative examples include, but are not limited to, vinyl, 1-propenyl, 2-propenyl, 1- , 2- or 3-butenyl, and the like. The alkenyl group can be optionally substituted or unsubstituted.

"Alkynyl" means an aliphatic hydrocarbon group containing a carbon-carbon triple bond, either straight or branched. Preference is given to C ₂ -C ₁₀ alkynyl groups, more preferably C ₂ -C ₆ alkynyl groups, most preferably C ₂ -C ₄ alkynyl groups. Examples of alkynyl groups include, but are not limited to, ethynyl, 1-propynyl, 2-propynyl, 1-, 2- or 3-butynyl, and the like. An alkynyl group can be substituted or unsubstituted.

"Cycloalkyl" refers to a saturated or partially saturated monocyclic, fused, bridged, and spiro carbon ring, but none of the rings have a fully conjugated π-electron aromatic system. It is preferably a C ₃ -C ₁₂ cycloalkyl group, more preferably a C ₃ -C ₈ cycloalkyl group, and most preferably a C ₃ -C ₆ cycloalkyl group. Examples of monocyclic cycloalkyl groups include, but are not limited to, cyclopropyl, cyclobutyl, cyclopentyl, cyclopentenyl, cyclohexyl, cyclohexenyl, cyclohexadienyl, cycloheptyl, cycloheptatriene The alkenyl group, the cyclooctyl group and the like are preferably a cyclopropyl group or a cyclohexenyl group.

"Spirocyclyl" refers to a polycyclic group of 5 to 18 members, two or more cyclic structures, and a single ring sharing a carbon atom (referred to as a spiro atom), one or more of which may contain One or more double bonds, but none of the rings have a fully conjugated π-electron aromatic system. It is preferably 6 to 14 members, more preferably 7 to 10 members. The spirocycloalkyl group is classified into a monospiro, a spiro- or a spirocycloalkyl group, preferably a mono- and bi-spirocycloalkyl group, preferably 4 yuan/5 yuan, 4, depending on the number of common spiro atoms between the rings. Yuan / 6 yuan, 5 yuan / 5 yuan or 5 yuan / 6 yuan. Non-limiting examples of "spirocycloalkyl" include, but are not limited to, spiro[4.5]decyl, spiro[4.4]decyl, spiro[3.5]decyl, spiro[2.4]heptyl.

"Fused ring group" means a 5- to 18-membered, all-carbon polycyclic group containing two or more ring structures that share a carbon atom with each other, and one or more rings may contain one or more double bonds, but None of the rings have a fully conjugated π-electron aromatic system, preferably 6 to 12 members, more preferably 7 to 10 members. Depending on the number of constituent rings, it may be classified into a bicyclic ring, a tricyclic ring, a pyridone or a polycyclic fused ring alkyl group, preferably a bicyclic ring or a tricyclic ring, more preferably a 5-membered/5-membered or 5-membered/6-membered bicycloalkyl group. Non-limiting examples of "fused cycloalkyl" include, but are not limited to, bicyclo[3.1.0]hexyl, bicyclo[3.2.0]hept-1-enyl, bicyclo[3.2.0]heptyl, Decalinyl or tetradecafluorophenanyl.

"Bridge ring group" means 5 to 18 members, containing two or more cyclic structures, sharing two carbon-polycyclic groups which are not directly bonded to each other, and one or more rings may contain one or more A double bond, but none of the rings have a fully conjugated π-electron aromatic system, preferably 6 to 12 members, more preferably 7 to 10 members. It is preferably 6 to 14 members, more preferably 7 to 10 members. Depending on the number of constituent rings, it may be classified into a bicyclic ring, a tricyclic ring, a pyridone or a polycyclic bridged cycloalkyl group, preferably a bicyclic ring, a tricyclic ring or a pyridone, and more preferably a bicyclic ring or a tricyclic ring. Non-limiting examples of "bridged cycloalkyl" include, but are not limited to: (1s, 4s)-bicyclo[2.2.1]heptyl, bicyclo[3.2.1]octyl, (1s,5s)-di Ring o [3.3.1] fluorenyl, bicyclo [2.2.2] octyl, (1r, 5r)-bicyclo[3.3.2] fluorenyl.

The cycloalkyl ring may be fused to an aryl, heteroaryl or heterocyclyl ring, wherein the ring to which the parent structure is attached is a cycloalkyl group, non-limiting examples include indanyl, tetrahydronaphthalene Base, benzocycloheptyl and the like. The cycloalkyl group can be optionally substituted or unsubstituted.

"Heterocyclyl", "heterocyclic" or "heterocyclic" are used interchangeably herein to refer to a non-aromatic heterocyclic group wherein one or more of the ring-forming atoms are heteroatoms such as oxygen, Nitrogen, sulfur atoms, etc., including monocyclic, fused, bridged, and spiro rings. It preferably has a 5- to 7-membered monocyclic ring or a 7- to 10-membered double- or tricyclic ring which may contain 1, 2 or 3 atoms selected from nitrogen, oxygen and/or sulfur. Examples of "heterocyclyl" include, but are not limited to, morpholinyl, oxetane, thiomorpholinyl, tetrahydropyranyl, 1,1-dioxo-thiomorpholinyl, piperidine , 2-oxo-piperidinyl, pyrrolidinyl, 2-oxo-pyrrolidinyl, piperazin-2-one, 8-oxa-3-aza-bicyclo[3.2.1]octyl and Piperazinyl. The heterocyclic group may be substituted or unsubstituted.

"Spiroheterocyclyl" means a polycyclic group of 5 to 18 members, two or more cyclic structures, and a single ring sharing one atom with each other, and the ring contains one or more double bonds, but no a ring having a fully conjugated π-electron aromatic system wherein one or more ring atoms are selected from the group consisting of nitrogen, oxygen or S(O) _n (where n is selected from 0, 1 or 2) heteroatoms, the remaining ring atoms are carbon. It is preferably 6 to 14 members, more preferably 7 to 10 members. The spirocycloalkyl group is classified into a monospiroheterocyclic group, a dispiroheterocyclic group or a polyspirocyclic group according to the number of common spiro atoms between the ring and the ring, and is preferably a monospiroheterocyclic group and a dispiroheterocyclic group. More preferably, it is 4 yuan / 4 yuan, 4 yuan / 5 yuan, 4 yuan / 6 yuan, 5 yuan / 5 yuan or 5 yuan / 6-membered monospiroheterocyclic group. Non-limiting examples of "spiroheterocyclyl" include, but are not limited to, 1,7-dioxaspiro[4.5]fluorenyl, 2-oxa-7-azaspiro[4.4]decyl, 7-oxo Heterospiro[3.5]decyl and 5-oxaspiro[2.4]heptyl.

"Fused heterocyclic group" means an all-carbon polycyclic group containing two or more cyclic structures that share a pair of atoms with each other, and one or more rings may contain one or more double bonds, but none of the rings have complete A conjugated π-electron aromatic system in which one or more ring atoms are selected from the group consisting of nitrogen, oxygen or a hetero atom of S(O) _n (where n is selected from 0, 1 or 2) and the remaining ring atoms are carbon. It is preferably 6 to 14 members, more preferably 7 to 10 members. Depending on the number of constituent rings, it may be classified into a bicyclic ring, a tricyclic ring, a pyridone or a polycyclic fused heterocyclic group, preferably a bicyclic ring or a tricyclic ring, and more preferably a 5-membered/5-membered or 5-membered/6-membered bicyclic fused heterocyclic group. Non-limiting examples of "fused heterocyclic groups" include, but are not limited to, octahydropyrrolo[3,4-c]pyrrolyl, octahydro-1H-isoindenyl, 3-azabicyclo[3.1. 0] hexyl, octahydrobenzo[b][1,4]dioxine.

"Bridge heterocyclyl" means 5 to 14 members, 5 to 18 members, containing two or more cyclic structures, sharing two polycyclic groups which are not directly connected to each other, and one or more rings may be used. An aromatic system containing one or more double bonds, but none of which has a fully conjugated π-electron, wherein one or more ring atoms are selected from nitrogen, oxygen or S(O) _n (where n is selected from 0, 1 or 2) Heteroatoms, the remaining ring atoms are carbon. It is preferably 6 to 14 members, more preferably 7 to 10 members. Depending on the number of constituent rings, it may be classified into a bicyclic ring, a tricyclic ring, a pyridone or a polycyclic bridged heterocyclic group, preferably a bicyclic ring, a tricyclic ring or a pyridone, and more preferably a bicyclic ring or a tricyclic ring. Non-limiting examples of "fused heterocyclic groups" include, but are not limited to, 2-azabicyclo[2.2.1]heptyl, 2-azabicyclo[2.2.2]octyl and 2-aza-di Ring [3.3.2] sulfhydryl. The heterocyclyl ring may be fused to an aryl, heteroaryl or cycloalkyl ring wherein the ring to which the parent structure is attached is a heterocyclic group. The heterocyclic group may be optionally substituted or unsubstituted.

"Aryl" means a carbocyclic aromatic system containing one or two rings wherein the rings may be joined together in a fused manner. The term "aryl" includes aryl groups such as phenyl, naphthyl, tetrahydronaphthyl. Preferably, the aryl group is a C ₆ -C ₁₀ aryl group, more preferably the aryl group is a phenyl group and a naphthyl group, and most preferably a phenyl group. The aryl group can be substituted or unsubstituted. The "aryl" may be fused to a heteroaryl, heterocyclyl or cycloalkyl group, wherein the parent structure is attached to an aryl ring, non-limiting examples include, but are not limited to:

"Heteroaryl" means an aromatic 5 to 6 membered monocyclic or 9 to 10 membered bicyclic ring containing from 1 to 4 atoms selected from nitrogen, oxygen and/or sulfur. Examples of "heteroaryl" include, but are not limited to, furyl, pyridyl, 2-oxo-1,2-dihydropyridyl, pyridazinyl, pyrimidinyl, pyrazinyl, thienyl, isoxazolyl , oxazolyl, oxadiazolyl, imidazolyl, pyrrolyl, pyrazolyl, triazolyl, tetrazolyl, thiazolyl, isothiazolyl, 1,2,3-thiadiazolyl, benzo Dioxolyl, benzothienyl, benzimidazolyl, fluorenyl, isodecyl, 1,3-dioxo-isoindenyl, quinolinyl, oxazolyl, benzo Isothiazolyl, benzoxazolyl and benzisoxazole. Heteroaryl groups can be substituted or unsubstituted. The heteroaryl ring can be fused to an aryl, heterocyclic or cycloalkyl ring wherein the ring to which the parent structure is attached is a heteroaryl ring, non-limiting examples include, but are not limited to:

"Alkoxy" means a group of (alkyl-O-). Among them, the alkyl group is defined in the relevant definition herein. Alkoxy groups of C ₁ -C ₆ are preferred. Examples thereof include, but are not limited to, methoxy, ethoxy, n-propoxy, isopropoxy, n-butoxy, isobutoxy, tert-butoxy and the like.

"Hydroxy" refers to an -OH group.

"Halogen" means fluoro, chloro, bromo and iodo.

"Amino" means -NH ₂ .

"Cyano" means -CN.

"Nitro" means -NO ₂ .

"Benzyl" refers to -CH ₂ - phenyl.

"Carboxy" refers to -C(O)OH.

"Carboxylic acid ester group" means -C(O)O-alkyl or -C(O)O-cycloalkyl, wherein alkyl, cycloalkyl are as defined above.

"DMSO" refers to dimethyl sulfoxide.

"Substituted" refers to one or more hydrogen atoms in the group, preferably up to 5, more preferably 1 to 3, hydrogen atoms, independently of each other, substituted by a corresponding number of substituents. It goes without saying that the substituents are only in their possible chemical positions, and those skilled in the art will be able to determine (by experiment or theory) substitutions that may or may not be possible without undue effort. For example, an amino group or a hydroxyl group having a free hydrogen may be unstable when combined with a carbon atom having an unsaturated (e.g., olefinic) bond.

As used herein, "substituted" or "substituted", unless otherwise indicated, means that the group may be substituted by one or more groups selected from the group consisting of alkyl, alkenyl, alkynyl, alkoxy. , alkylthio, alkylamino, halogen, sulfhydryl, hydroxy, nitro, cyano, cycloalkyl, heterocyclyl, aryl, heteroaryl, cycloalkoxy, heterocycloalkoxy, cycloalkane Thio, heterocycloalkylthio, amino, haloalkyl, hydroxyalkyl, carboxyl, carboxylate, =O, -C(O)R ⁵ , -OC(O)R ⁵ , -C(O)OR ⁵ , -NR ⁶ R ⁷ , -C(O)NR ⁶ R ⁷ , -S(O) _n NR ⁶ R ⁷ or -NR ⁶ C(O)R ⁷ .

Alternatively, R ⁶ and R ⁷ together with the N atom to which they are attached form a 4 to 8 membered heterocyclic group, wherein the 4 to 8 membered heterocyclic ring contains one or more N, O, S(O) _n atoms, and 4 Optionally, one or more selected from the group consisting of hydroxyl, halogen, nitro, cyano, alkyl, alkoxy, cycloalkyl, heterocyclyl, aryl, heteroaryl, =0 , -C(O)R ⁸ , -C(O)OR ⁸ , -OC(O)R ⁸ , -NR ⁹ R ¹⁰ , -C(O)NR ⁹ R ¹⁰ , -S(O) _n NR ⁹ R ¹⁰ or -NR ⁹ C (O) R ¹⁰ is substituted by a substituent.

n is selected from 0, 1 or 2.

"Pharmaceutically acceptable salt" refers to certain salts of the above compounds which retain their original biological activity and are suitable for pharmaceutical use. The pharmaceutically acceptable salt of the compound represented by the formula (I) may be a metal salt or an amine salt formed with a suitable acid.

"Pharmaceutical composition" means a mixture comprising one or more of the compounds described herein, or a physiologically pharmaceutically acceptable salt or prodrug thereof, and other chemical components, as well as other components such as physiologically pharmaceutically acceptable carriers and Shape agent. The purpose of the pharmaceutical composition is to promote the administration of the organism, which facilitates the absorption of the active ingredient and thereby exerts biological activity.

Method for synthesizing the compound of the present invention

In order to accomplish the object of the present invention, the present invention adopts the following technical solutions:

The preparation method of the compound of the formula (I) or a stereoisomer, tautomer or a pharmaceutically acceptable salt thereof of the present invention comprises the following steps:

The compound of the formula (Ia) is subjected to a condensation reaction with a compound of the formula (Ib) under heating to obtain a compound of the formula (Ic); and the compound of the formula (Ic) is reacted with a compound of the formula (Id) under heating to obtain a compound of the formula (IA); reacting a compound of the formula (IA) with a compound of the formula (IB) under basic conditions to give a compound of the formula (I);

among them:

X is a halogen;

R ^a is an alkyl group;

R ¹ to R ⁴ and m are as defined in the formula (I), and R ^{1 is} not a hydrogen atom.

The compound of the formula (Ic) is subjected to a condensation reaction with a compound of the formula (Ie) under heating to obtain a compound of the formula (If); the compound of the formula (If) is reacted in the presence of trifluoroacetic acid to give a formula (IA). An intermediate compound; reacting a compound of the formula (IA) with a compound of the formula (IB) under basic conditions to give a compound of the formula (I);

among them:

X is a halogen;

R ¹ is a hydrogen atom;

R ^b is an alkyl group;

R ² to R ⁴ are as defined in the formula (I).

The preparation method of the compound of the formula (II) or a stereoisomer, tautomer or a pharmaceutically acceptable salt thereof of the present invention comprises the following steps:

The compound of the formula (Ia) is subjected to a condensation reaction with a compound of the formula (IIa) under heating to obtain a compound of the formula (IIb); the compound of the formula (IIb) is reacted with a compound of the formula (Id) under heating, Obtaining a compound of the formula (IIA); reacting a compound of the formula (IIA) with a compound of the formula (IB) under basic conditions to obtain a compound of the formula (II);

among them:

X is a halogen;

R ^a is an alkyl group;

R ¹ to R ⁴ are as defined in the formula (II), and R ^{1 is} not a hydrogen atom.

The compound of the formula (IIb) is reacted with a compound of the formula (Ie) under heating to obtain a compound of the formula (IIc); and the compound of the formula (IIc) is reacted in the presence of trifluoroacetic acid to give an intermediate of the formula (IIA) a compound of the formula (IIA) and a compound of the formula (IB) are reacted under basic conditions to give a compound of the formula (II);

among them:

X is a halogen;

R ¹ is a hydrogen atom;

R ^b is an alkyl group;

R ² to R ⁴ are as defined in the formula (II).

Detailed ways

The invention is further described in the following examples, but these examples are not intended to limit the scope of the invention.

Example

The examples give the preparation of representative compounds represented by formula (I) and related structural identification data. It is to be understood that the following examples are intended to illustrate the invention and not to limit the invention. ^{The 1} H NMR spectrum was determined using a Bruker instrument (400 MHz) and the chemical shift was expressed in ppm. The internal standard of tetramethylsilane (0.00 ppm) was used. ¹ H NMR representation: s = singlet, d = doublet, t = triplet, m = multiplet, br = broadened, dd = doublet of doublet, dt = doublet of triplet. If a coupling constant is provided, its unit is Hz.

Mass spectrometry was measured by LC/MS, and the ionization method was ESI or APCI.

Thin layer chromatography silica gel plate using Yantai Yellow Sea HSGF254 or Qingdao GF254 silica gel plate, thin layer chromatography (TLC)

The specifications of the silica gel plate used are 0.15 mm to 0.2 mm, and the specifications for separation and purification of thin layer chromatography are 0.4 mm to 0.5 mm.

Column chromatography generally uses Yantai Huanghai silica gel 200-300 mesh silica gel as a carrier.

In the following examples, all temperatures are in degrees Celsius unless otherwise indicated, and unless otherwise indicated, the various starting materials and reagents are either commercially available or synthesized according to known methods, and the commercially available materials and reagents are not further processed. Purification is used directly, unless otherwise indicated, and commercially available, including but not limited to Aldrich Chemical Company, ABCR GmbH & Co. KG, Acros Organics, Guangzan Chemical Technology Co., Ltd. and Jingyan Chemical Technology Co., Ltd., etc.

CD ₃ OD: Deuterated methanol.

CDCl ₃ : deuterated chloroform.

DMSO-d ₆ : deuterated dimethyl sulfoxide.

The argon atmosphere means that the reaction flask is connected to an argon balloon having a volume of about 1 L.

There is no particular description in the examples, and the solution in the reaction means an aqueous solution.

The compound is purified by silica gel column chromatography eluent system and thin layer chromatography, wherein the eluent system is selected from the group consisting of: A: petroleum ether and ethyl acetate system; B: dichloromethane and methanol system; C: two Methyl chloride: ethyl acetate; wherein the volume ratio of the solvent varies depending on the polarity of the compound, and it may be adjusted by adding a small amount of an acidic or alkaline agent such as acetic acid or triethylamine.

Example 1

1-(4-Chloro-2-fluoro-5-(7-(methylamino)-2-oxo-1-(2,2,2-trifluoroethyl)-1,2-dihydro-1 ,6-naphthyridin-3-yl)phenyl)-3-phenylurea

first step

Ethyl 6-chloro-4-((2,2,2-trifluoroethyl)amino)nicotinic acid

Ethyl 4,6-dichloronicotinate 1a (10.1 g, 45.9 mmol), 2,2,2-trifluoroethylamine (15.5 g, 157 mmol) and N,N-diisopropylethylamine (17.8 g) , 138 mmol) was dissolved in 200 mL of dimethyl adipate and reacted at 100 ° C for 8 hours. The residue was purified by silica gel column chromatography (eluent: A) to give ethyl 6-chloro-4-((2,2,2-trifluoroethyl) 1b (4 g, white solid), yield: 31%.

MS m/z (ESI): 282.9 [M+1]

Second step

(6-chloro-4-((2,2,2-trifluoroethyl)amino)pyridin-3-yl)methanol

Ethyl 6-chloro-4-((2,2,2-trifluoroethyl)amino)nicotinic acid 1b (4 g, 14.18 mmol) was dissolved in 50 mL of tetrahydrofuran, and lithium tetrahydroaluminum was added portionwise at -50 °C. (1.89 g, 51 mmol), slowly warmed to 0 ° C, and reacted at 0 ° C for 1.5 hours. The reaction was quenched by slowly adding 20 mL of 10% sodium hydroxide solution at 0 ° C, filtered, and the filtrate was extracted with ethyl acetate (30 mL), the aqueous layer was separated, and the organic phase was washed with saturated sodium chloride solution (30 mL×2) Drying over anhydrous sodium sulfate, EtOAc (EtOAc m.) , Yield: 100%.

MS m/z (ESI): 240.9 [M+1]

third step

6-chloro-4-((2,2,2-trifluoroethyl)amino)nicotin

(6-Chloro-4-((2,2,2-trifluoroethyl)amino)pyridin-3-yl)methanol 1c (3.4 g, 14.18 mmol) and active manganese dioxide (18.5 g, 212.7 mmol) Dissolved in 52 mL of dichloromethane and stirred at room temperature overnight. The reaction mixture was filtered, and the filtrate was evaporated tolulujjjjjjjjjjjjjjjjjjjjj Amino) Nicotinaldehyde 1d (2.4 g, white solid), Yield: 71%.

MS m/z (ESI): 238.9 [M+1]

the fourth step

3-(5-Amino-2-chloro-4-fluorophenyl)-7-chloro-1-(2,2,2-trifluoroethyl)-1,6-naphthyridin-2(1H)-one

6-Chloro-4-((2,2,2-trifluoroethyl)amino)nicotin 1d (1 g, 4.2 mmol), 2-(5-amino-2-chloro-4-fluorophenyl)acetic acid Ethyl ester 1e (970 mg, 4.2 mmol, prepared according to published patent application WO2013184119), potassium carbonate (1.74 g, 12.6 mmol) dissolved in 70 mL of N,N-dimethylformamide and toluene (V:V=2:5) The reaction was carried out at 150 ° C for 12 hours in a mixed solvent. After adding 100 mL of ethyl acetate, the mixture was washed with water (100 mL×2) and brine (100 mL). Purification: A system) was purified to give 3-(5-amino-2-chloro-4-fluorophenyl)-7-chloro-1-(2,2,2-trifluoroethyl)-1,6 -naphthyridin-2(1H)-one 1f (870 mg, yellow solid), yield: 48%.

MS m/z (ESI): 405.8 [M+1]

the fifth step

3-(5-Amino-2-chloro-4-fluorophenyl)-7-(methylamino)-1-(2,2,2-trifluoroethyl)-1,6-naphthyridine-2 ( 1H)-ketone

3-(5-Amino-2-chloro-4-fluorophenyl)-7-chloro-1-(2,2,2-trifluoroethyl)-1,6-naphthyridin-2(1H)- The ketone 1f (300 mg, 0.74 mmol) and a solution of methylamine in tetrahydrofuran (3 mL, 6 mmol) were dissolved in 20 mL of 1,4-dioxane and reacted in a sealed tube at 100 ° C for 16 hours. The organic layer was concentrated under reduced pressure and purified to purified crystals crystals crystals -(2,2,2-Trifluoroethyl)-1,6-naphthyridin-2(1H)-one 1 g (150 mg, yellow solid), yield: 50%.

MS m/z (ESI): 400.8 [M+1]

Step 6

3-(5-Amino-2-chloro-4-fluorophenyl)-7-(methylamino)-1-(2,2,2-trifluoroethyl)-1,6-naphthyridine-2 (1H)-ketone 1g (100mg, 0.25mmol), phenyl isocyanate 1h (33mg, 0.28mmol, prepared according to published patent application WO2005113626) and triethylamine (126mg, 1.25mmol) dissolved in 2mL of dichloromethane The reaction was allowed to proceed overnight at room temperature. The organic layer was concentrated under reduced pressure. 1-(2,2,2-Trifluoroethyl)-1,2-dihydro-1,6-naphthyridin-3-yl)phenyl)-3-phenylurea 1 (20 mg, white solid) Yield: 18%.

MS m/z (ESI): 520.1 [M+1]

^{1 H NMR (400MHz, DMSO-} d 6) δ9.17 (s, 1H), 8.76 (s, 1H), 8.46 (s, 1H), 8.21 (d, J = 8.8Hz, 1H), 7.87 (s, 1H), 7.56 (d, J = 10.8 Hz, 1H), 7.43 (d, J = 8.4 Hz, 2H), 7.28 (t, J = 7.2 Hz, 2H), 7.21-7.20 (m, 1H), 6.99 ( t, J = 7.6 Hz, 1H), 6.37 (s, 1H), 5.13-5.11 (m, 2H), 2.87 (d, J = 4.8 Hz, 3H).

Example 2

1-(4-Chloro-2-fluoro-5-(7-(methylamino)-2-oxo-1-(2,2,2-trifluoroethyl)-1,2-dihydro-1 ,6-naphthyridin-3-yl)phenyl)-3-(3-fluorophenyl)urea

first step

3-(5-Amino-2-chloro-4-fluorophenyl)-7-(methylamino)-1-(2,2,2-trifluoroethyl)-1,6-naphthyridine-2 (1H)-ketone 1g (100mg, 0.25mmol), 1-fluoro-3-isocyanatophenyl 2a (51mg, 0.375mmol) and triethylamine (75mg, 0.75mmol) in 2mL dichloromethane, room temperature The reaction was overnight. The organic layer was concentrated under reduced pressure. 1-(2,2,2-trifluoroethyl)-1,2-dihydro-1,6-naphthyridin-3-yl)phenyl)-3-(3-fluorophenyl)urea 2 (13 mg , white solid), yield: 10%.

MS m/z (ESI): 537.9 [M+1]

^{1 H NMR (400MHz, DMSO-} d 6) δ9.35 (s, 1H), 8.80 (s, 1H), 8.45 (s, 1H), 8.19 (d, J = 6.8Hz, 1H), 7.87 (s, 1H), 7.58 (d, J = 11.2 Hz, 1H), 7.48 (d, J = 10.8 Hz, 1H), 7.32-7.29 (m, 1H), 7.21 (s, 1H), 7.10-7.08 (m, 1H) ), 6.81-6.79 (m, 1H), 6.37 (s, 1H), 5.76 (s, 1H), 5.13-5.09 (m, 1H), 2.87 (d, J = 4.8 Hz, 3H).

Example 3

1-(2-Fluoro-4-methyl-5-(7-(methylamino)-2-oxo-1-(2,2,2-trifluoroethyl)-1,2-dihydro- 1,6-naphthyridin-3-yl)phenyl)-3-(3-fluorophenyl)urea

first step

3-(5-Amino-4-fluoro-2-methylphenyl)-7-chloro-1-(2,2,2-trifluoroethyl)-1,6-naphthyridin-2(1H)- ketone

6-Chloro-4-((2,2,2-trifluoroethyl)amino)nicotin 1d (1 g, 4.2 mmol), 2-(5-amino-4-fluoro-2-methylphenyl) Ethyl acetate 3a (890 mg, 4.2 mmol, prepared according to published patent application WO2013184119), potassium carbonate (1.74 g, 12.6 mmol) dissolved in 70 mL of N,N-dimethylformamide and toluene (V:V=2) The reaction mixture of 5:5 was reacted at 150 ° C for 8 hours. After adding 50 ml of ethyl acetate, the mixture was washed with water (50 mL×2) and brine (50 mL). (eluent: A system) was purified to give 3-(5-amino-4-fluoro-2-methylphenyl)-7-chloro-1-(2,2,2-trifluoroethyl)-1 , 6-naphthyridin-2(1H)-one 3b (620 mg, yellow solid), yield: 39%.

MS m/z (ESI): 385.8 [M+1]

Second step

3-(5-Amino-4-fluoro-2-methylphenyl)-7-(methylamino)-1-(2,2,2-trifluoroethyl)-1,6-naphthyridine-2 (1H)-ketone

3-(5-Amino-4-fluoro-2-methylphenyl)-7-chloro-1-(2,2,2-trifluoroethyl)-1,6-naphthyridine-2 (1H) A solution of the ketone 3b (620 mg, 1.61 mmol) and methylamine in tetrahydrofuran (6 mL, 12 mmol) was dissolved in 36 mL of 1,4-dioxane and reacted in a sealed tube at 100 ° C for 48 hours. The residue was purified by silica gel column chromatography (eluent: A) to give 3-(5-amino-4-fluoro-2-methylphenyl)-7-(methylamino) 1-(2,2,2-Trifluoroethyl)-1,6-naphthyridin-2(1H)-one 3c (550 mg, yellow solid), yield: 89%.

MS m/z (ESI): 380.9 [M+1]

third step

3-(5-Amino-4-fluoro-2-methylphenyl)-7-(methylamino)-1-(2,2,2-trifluoroethyl)-1,6-naphthyridine- 2(1H)-one 3c (183mg, 0.48mmol), 1-fluoro-3-isocyanatophenyl 2a (198mg, 1.44mmol) and triethylamine (243mg, 2.41mmol) were dissolved in 20mL of tetrahydrofuran at room temperature overnight. The residue was purified by silica gel column chromatography (eluent: A) to give 1-(2-fluoro-4-methyl-5-(7-(methylamino)-2- Oxo-1-(2,2,2-trifluoroethyl)-1,2-dihydro-1,6-naphthyridin-3-yl)phenyl)-3-(3-fluorophenyl)urea 3 (50 mg, white solid), yield: 20%.

MS m/z (ESI): 517.8 [M+1]

^{1 H NMR (400MHz, DMSO-} d 6) δ9.25 (s, 1H), 8.57 (s, 1H), 8.45 (s, 1H), 7.95 (d, J = 8.4Hz, 1H), 7.79 (s, 1H), 7.51-7.47 (m, 1H), 7.33-7.27 (m, 1H), 7.18-7.13 (m, 2H), 7.07 (d, J = 7.6 Hz, 1H), 6.79 (dt, J = 8.8, 2.4 Hz, 1H), 6.37 (s, 1H), 5.14 - 5.12 (m, 2H), 2.86 (d, J = 4.8 Hz, 3H), 2.08 (s, 3H).

Example 4

1-(2-Fluoro-4-methyl-5-(7-(methylamino)-2-oxo-1-(2,2,2-trifluoroethyl)-1,2-dihydro- 1,6-naphthyridin-3-yl)phenyl)-3-(4-fluorophenyl)urea

first step

3-(5-Amino-4-fluoro-2-methylphenyl)-7-(methylamino)-1-(2,2,2-trifluoroethyl)-1,6-naphthyridine- 2(1H)-one 3c (183mg, 0.48mmol), phenyl 1-fluoro-4-isocyanate 4a (198mg, 1.44mmol) and triethylamine (243mg, 2.41mmol) were dissolved in 20mL of tetrahydrofuran at room temperature overnight. The residue was purified by silica gel column chromatography (eluent: A) to give 1-(2-fluoro-4-methyl-5-(7-(methylamino)-2- Oxo-1-(2,2,2-trifluoroethyl)-1,2-dihydro-1,6-naphthyridin-3-yl)phenyl)-3-(4-fluorophenyl)urea 4 (50 mg, white solid), yield: 20%.

MS m/z (ESI): 517.8 [M+1]

^{1 H NMR (400MHz, DMSO-} d 6) δ9.07 (s, 1H), 8.50 (s, 1H), 8.44 (s, 1H), 7.95 (d, J = 8.4Hz, 1H), 7.78 (s, 1H), 7.46-7.42 (m, 2H), 7.17-7.09 (m, 4H), 6.37 (s, 1H), 5.14-5.12 (m, 2H), 2.86 (d, J = 4.8 Hz, 3H), 2.07 (s, 3H).

Example 5

1-(2,4-difluoro-5-(7-(methylamino)-2-oxo-1-(2,2,2-trifluoroethyl)-1,2-dihydro-1, 6-naphthyridin-3-yl)phenyl)-3-(3-fluorophenyl)urea

first step

3-(5-Amino-2,4-difluorophenyl)-7-chloro-1-(2,2,2-trifluoroethyl)-1,6-naphthyridin-2(1H)-one

6-Chloro-4-((2,2,2-trifluoroethyl)amino)nicotin 1d (300 mg, 1.26 mmol), 2-(5-amino-2,4-difluorophenyl)acetic acid Ester 5a (271.7 mg, 1.26 mmol, prepared according to published patent application WO2013184119) and potassium carbonate (522 mg, 3.78 mmol) dissolved in 28 mL of N,N-dimethylformamide and toluene (V:V=2:5) The reaction was carried out at 150 ° C for 8 hours in a mixed solvent. After adding 50 ml of ethyl acetate, the mixture was washed with water (50 mL×2) and brine (50 mL). (eluent: A system) was purified to give 3-(5-amino-2,4-difluorophenyl)-7-chloro-1-(2,2,2-trifluoroethyl)-1,6 - Naphthyridine-2(1H)-one 5b (237 mg, pale yellow solid), yield: 48%.

MS m/z (ESI): 389.8 [M+1]

Second step

3-(5-Amino-2,4-difluorophenyl)-7-(methylamino)-1-(2,2,2-trifluoroethyl)-1,6-naphthyridine-2 (1H )-ketone

3-(5-Amino-2,4-difluorophenyl)-7-chloro-1-(2,2,2-trifluoroethyl)-1,6-naphthyridin-2(1H)-one 5b (237 mg, 0.61 mmol) and a solution of methylamine in tetrahydrofuran (3 mL, 6 mmol) were dissolved in 20 mL of 1,4-dioxane and reacted in a sealed tube at 100 ° C for 12 hours. The residue was purified by silica gel column chromatography (eluent: A) to give 3-(5-amino-2,4-difluorophenyl)-7-(methylamino)- 1-(2,2,2-Trifluoroethyl)-1,6-naphthyridin-2(1H)-one 5c (168 mg, pale yellow solid).

MS m/z (ESI): 384.9 [M+1]

^{1 H NMR (400MHz, DMSO-} d 6) δ8.43 (s, 1H), 7.86 (s, 1H), 7.17 (d, J = 4.0Hz, 1H), 7.07 (t, J = 8.0Hz, 1H) , 6.82 (t, J = 8.0 Hz, 1H), 6.34 (s, 1H), 5.11-5.09 (m, 2H), 5.04 (s, 2H), 2.85 (d, J = 4.0 Hz, 3H).

third step

3-(5-Amino-2,4-difluorophenyl)-7-(methylamino)-1-(2,2,2-trifluoroethyl)-1,6-naphthyridine-2 ( 1H)-ketone 5c (168 mg, 0.44 mmol), 1-fluoro-3-isocyanatophenyl 2a (90 mg, 0.66 mmol) and triethylamine (727 mg, 7.2 mmol) were dissolved in 10 mL dichloromethane. overnight. The residue was purified by silica gel column chromatography (eluent: A) to give 1-(2,4-difluoro-5-(7-(methylamino)-2-oxo 1-(2,2,2-trifluoroethyl)-1,2-dihydro-1,6-naphthyridin-3-yl)phenyl)-3-(3-fluorophenyl)urea 5 ( 4.5 mg, pale yellow solid), yield: 2%.

MS m/z (ESI): 521.8 [M+1]

^{1 H NMR (400MHz, DMSO-} d 6) δ9.32 (s, 1H), 8.66 (s, 1H), 8.46 (s, 1H), 8.13 (t, J = 8.5Hz, 1H), 7.93 (s, 1H), 7.49 (d, J = 11.8 Hz, 1H), 7.41 (t, J = 10.3 Hz, 1H), 7.31 (q, J = 7.9 Hz, 1H), 7.23 (d, J = 5.1 Hz, 1H) , 7.10 (d, J = 8.2 Hz, 1H), 6.80 (t, J = 8.1 Hz, 1H), 6.37 (s, 1H), 5.14 - 5.11 (m, 2H), 2.87 (d, J = 4.7 Hz, 3H).

Example 6

1-(2,4-difluoro-5-(7-(methylamino)-2-oxo-1-(2,2,2-trifluoroethyl)-1,2-dihydro-1, 6-naphthyridin-3-yl)phenyl)-3-(4-fluorophenyl)urea

first step

3-(5-Amino-2,4-difluorophenyl)-7-(methylamino)-1-(2,2,2-trifluoroethyl)-1,6-naphthyridine-2 ( 1H)-ketone 5c (300 mg, 0.78 mmol), phenyl 1-fluoro-4-isocyanate 4a (321 mg, 2.34 mmol) and triethylamine (394 mg, 3.9 mmol) were dissolved in 20 mL of THF. The residue was purified by silica gel column chromatography (eluent: A) to give 1-(2,4-difluoro-5-(7-(methylamino)-2-oxo 1-(2,2,2-trifluoroethyl)-1,2-dihydro-1,6-naphthyridin-3-yl)phenyl)-3-(4-fluorophenyl)urea 6 ( 160 mg, white solid), yield: 39%.

MS m/z (ESI): 521.8 [M+1]

^{1 H NMR (400MHz, DMSO-} d 6) δ9.58 (s, 1H), 8.83 (s, 1H), 8.65 (s, 1H), 8.40 (s, 1H), 8.18 (t, J = 8.4Hz, 1H), 8.10 (s, 1H), 7.86-7.45 (m, 3H), 7.13 (t, J = 8.4 Hz, 2H), 6.76 (s, 1H), 5.21 (s, 2H), 3.00 (s, 3H) ).

Example 7

1-(4-bromo-2-fluoro-5-(7-(methylamino)-2-oxo-1-(2,2,2-trifluoroethyl)-1,2-dihydro-1 ,6-naphthyridin-3-yl)phenyl)-3-phenylurea

first step

3-(5-Amino-2-bromo-4-fluorophenyl)-7-chloro-1-(2,2,2-trifluoroethyl)-1,6-naphthyridin-2(1H)-one

6-Chloro-4-((2,2,2-trifluoroethyl)amino)nicotin 1d (600 mg, 2.52 mmol), 2-(5-amino-2-bromo-4-fluorophenyl)acetic acid Ethyl ester 7a (698 mg, 2.52 mmol, prepared according to published patent application WO2013184119) and potassium carbonate (1.04 g, 7.56 mmol) dissolved in 70 mL of N,N-dimethylformamide and toluene (V:V=2:5) The reaction was carried out at 150 ° C for 12 hours in a mixed solvent. After adding 100 mL of ethyl acetate, the mixture was washed with water (100 mL×2), EtOAc (EtOAc) Purification: A system) was purified to give 3-(5-amino-2-bromo-4-fluorophenyl)-7-chloro-1-(2,2,2-trifluoroethyl)-1,6 - Naphthyridine-2(1H)-one 7b (660 mg, pale yellow solid), yield: 58%.

MS m/z (ESI): 449.7 [M+1]

Second step

3-(5-Amino-2-bromo-4-fluorophenyl)-7-(methylamino)-1-(2,2,2-trifluoroethyl)-1,6-naphthyridine-2 ( 1H)-ketone

3-(5-Amino-2-bromo-4-fluorophenyl)-7-chloro-1-(2,2,2-trifluoroethyl)-1,6-naphthyridin-2(1H)- A solution of the ketone 7b (160 mg, 0.35 mmol) and methylamine in tetrahydrofuran (1.5 mL, 3 mmol) was dissolved in 5 mL of 1,4-dioxane and reacted in a sealed tube at 100 ° C for 8 hours. The residue was purified by silica gel column chromatography (eluent: A) to give 3-(5-amino-2-bromo-4-fluorophenyl)-7-(methylamino) 1-(2,2,2-Trifluoroethyl)-1,6-naphthyridin-2(1H)-one 7c (123 mg, yellow solid), yield: 78%.

MS m/z (ESI): 444.8 [M+1]

third step

3-(5-Amino-2-bromo-4-fluorophenyl)-7-(methylamino)-1-(2,2,2-trifluoroethyl)-1,6-naphthyridine-2 (1H)-ketone 7c (114 mg, 0.26 mmol), phenyl isocyanate 1 h (34 mg, 0.28 mmol) and triethylamine (130 mg, 1.28 mmol) dissolved in 20 mL of toluene and dichloromethane (V:V=1: The mixed solution of 1) was allowed to react at room temperature overnight. The residue was purified by silica gel column chromatography (eluent: A) to give 1-(4-bromo-2-fluoro-5-(7-(methylamino)-2- ox 1-(2,2,2-Trifluoroethyl)-1,2-dihydro-1,6-naphthyridin-3-yl)phenyl)-3-phenylurea 7 (20 mg, white solid ), yield: 12%.

MS m/z (ESI): 563.8 [M+1]

^{1 H NMR (400MHz, DMSO-} d 6) δ9.16 (s, 1H), 8.76 (s, 1H), 8.45 (s, 1H), 8.22 (d, J = 8.8Hz, 1H), 7.84 (s, 1H), 7.69 (d, J = 10.8 Hz, 1H), 7.43 (d, J = 7.6 Hz, 2H), 7.28 (t, J = 7.6 Hz, 2H), 7.22-7.20 (m, 1H), 6.99 ( t, J = 7.2 Hz, 1H), 6.37 (s, 1H), 5.14 - 5.11 (m, 2H), 2.86 (d, J = 4.8 Hz, 3H).

Example 8

1-(Benzo[b]thiophen-3-yl)-3-(2-fluoro-4-methyl-5-(7-(methylamino)-2-oxo-1-(2,2, 2-trifluoroethyl)-1,2-dihydro-1,6-naphthyridin-3-yl)phenyl)urea

first step

3-isocyanatobenzo[b]thiophene ester

Benzo[b]thiophene-3-carboxylic acid 8a (500 mg, 2.8 mmol) and triethylamine (319 mg, 3.2 mmol) were dissolved in 10 mL of toluene and stirred at room temperature until 8a to dissolve. Diphenylphosphoryl azide (847 mg, 3.1 mmol) was added, and the reaction was carried out at 50 ° C for 1 hour, and the reaction was continued at 100 ° C for 0.5 hour. The organic layer was concentrated under reduced pressure to give EtOAc (EtOAc m.

Second step

3-(5-Amino-4-fluoro-2-methylphenyl)-7-(methylamino)-1-(2,2,2-trifluoroethyl)-1,6-naphthyridine- 2(1H)-one 3c (183mg, 0.48mmol), 3-isocyanatobenzo[b]thiophene ester 8b (252mg, 1.44mmol) and triethylamine (243mg, 2.41mmol) in 20mL of tetrahydrofuran, room temperature The reaction was overnight. The residue was purified by silica gel column chromatography (eluent: A) to give 1-(benzo[b]thiophen-3-yl)-3-(2-fluoro-4-methyl) 5-(7-(methylamino)-2-oxo-1-(2,2,2-trifluoroethyl)-1,2-dihydro-1,6-naphthyridin-3-yl Phenyl)urea 8 (60 mg, yellow solid), yield: 22%.

MS m/z (ESI): 555.8 [M+1]

^{1 H NMR (400MHz, DMSO-} d 6) δ9.42 (s, 1H), 8.86 (s, 1H), 8.46 (s, 1H), 8.10 (d, J = 8.8Hz, 1H), 7.97 (d, J = 8.0 Hz, 1H), 7.89 (d, J = 8.4 Hz, 1H), 7.81 (s, 1H), 7.70 (s, 1H), 7.51 (t, J = 6.4 Hz, 1H), 7.43 (t, J=8.0 Hz, 1H), 7.20 (d, J=12 Hz, 1H), 7.15-7.13 (m, 1H), 6.38 (s, 1H), 5.15-5.10 (m, 2H), 2.87 (d, J = 4.8 Hz, 3H), 2.09 (s, 3H).

Example 9

1-(Benzo[b]thiophen-3-yl)-3-(2,4-difluoro-5-(7-(methylamino)-2-oxo-1-(2,2,2-) Trifluoroethyl)-1,2-dihydro-1,6-naphthyridin-3-yl)phenyl)urea

first step

3-(5-Amino-2,4-difluorophenyl)-7-(methylamino)-1-(2,2,2-trifluoroethyl)-1,6-naphthyridine-2 ( 1H)-ketone 5c (200 mg, 0.52 mmol), 3-isocyanatobenzo[b]thiophene ester 8b (91 mg, 0.52 mmol) and triethylamine (157 mg, 1.56 mmol) were dissolved in 5 mL of THF. . The residue was purified by silica gel column chromatography (eluent: A) to give 1-(benzo[b]thiophen-3-yl)-3-(2,4-difluoro- 5-(7-(methylamino)-2-oxo-1-(2,2,2-trifluoroethyl)-1,2-dihydro-1,6-naphthyridin-3-yl)benzene Urea 9 (50 mg, white solid), yield: 17%.

MS m/z (ESI): 559.8 [M+1]

¹ H NMR (400 MHz, DMSO-d ₆ ) δ 9.41 (s, 1H), 8.89 (s, 1H), 8.48 (s, 1H), 8.28 (t, J = 8.4 Hz, 1H), 7.99-7.96 ( m, 2H), 7.89 (d, J = 8.0 Hz, 1H), 7.71 (s, 1H), 7.51 (t, J = 7.6 Hz, 1H), 7.46-7.41 (m, 2H), 7.23-7.20 (m , 1H), 6.37 (s, 1H), 5.14 - 5.09 (m, 2H), 2.87 (d, J = 4.8 Hz, 3H).

Example 10

1-(4-bromo-2-fluoro-5-(7-((2-methoxyethyl)amino)-2-oxo-1-(2,2,2-trifluoroethyl)-1 ,2-dihydro-1,6-naphthyridin-3-yl)phenyl)-3-(3-fluorophenyl)urea

first step

3-(5-Amino-2-bromo-4-fluorophenyl)-7-((2-methoxyethyl)amino)-1-(2,2,2-trifluoroethyl)-1, 6-naphthyridine-2(1H)-one

3-(5-Amino-2-bromo-4-fluorophenyl)-7-chloro-1-(2,2,2-trifluoroethyl)-1,6-naphthyridin-2(1H)- Ketone 7b (340 mg, 0.76 mmol), 2-methoxyethylamine (568 mg, 7.57 mmol) and 1,8-diazabicycloundec-7-ene (DBU) (230 mg, 1.51 mmol) were dissolved. In 7 mL of N-methylpyrrolidone, microwave reaction was carried out at 180 ° C for 1.5 hours. After adding 70 mL of water, and extracting with dichloromethane (100 mL × 2), the organic phase was combined, washed with a saturated sodium chloride solution (100 mL), dried over anhydrous sodium sulfate, filtered and evaporated. Purification by chromatography (eluent: A system) to give 3-(5-amino-2-bromo-4-fluorophenyl)-7-((2-methoxyethyl)amino)-1-( 2,2,2-Trifluoroethyl)-1,6-naphthyridin-2(1H)-one 10a (370 mg, yellow oil), yield: 100%.

MS m/z (ESI): 488.8 [M+1]

Second step

3-(5-Amino-2-bromo-4-fluorophenyl)-7-((2-methoxyethyl)amino)-1-(2,2,2-trifluoroethyl)-1 6-naphthyridine-2(1H)-one 10a (100 mg, 0.21 mmol), 1-fluoro-3-isocyanatophenyl 2a (42 mg, 0.31 mmol) and triethylamine (62 mg, 0.62 mmol) The reaction was carried out overnight at room temperature in 3 mL of dichloromethane. The organic layer was concentrated under reduced pressure and purified to purified crystals crystals crystals )-2-oxo-1-(2,2,2-trifluoroethyl)-1,2-dihydro-1,6-naphthyridin-3-yl)phenyl)-3-(3-fluoro Phenyl)urea 10 (30 mg, white solid), yield: 23%.

MS m/z (ESI): 625.7 [M+1]

^{1 H NMR (400MHz, DMSO-} d 6) δ9.36 (s, 1H), 8.81 (s, 1H), 8.45 (s, 1H), 8.18 (d, J = 8.8Hz, 1H), 7.83 (s, 1H), 7.71 (d, J = 10.4 Hz, 1H), 7.48 (d, J = 12.4 Hz, 1H), 7.31 (q, J = 8.4 Hz, 1H), 7.27-7.23 (m, 1H), 7.08 ( d, J = 8.0 Hz, 1H), 6.81 (t, J = 8.8 Hz, 1H), 6.51 (s, 1H), 5.06-5.04 (m, 2H), 3.53 - 3.50 (m, 4H), 3.29 (s) , 3H).

Example 11

1-(4-bromo-2-fluoro-5-(7-((2-methoxyethyl)amino)-2-oxo-1-(2,2,2-trifluoroethyl)-1 ,2-dihydro-1,6-naphthyridin-3-yl)phenyl)-3-(4-fluorophenyl)urea

first step

3-(5-Amino-2-bromo-4-fluorophenyl)-7-((2-methoxyethyl)amino)-1-(2,2,2-trifluoroethyl)-1 6-naphthyridine-2(1H)-one 10a (100 mg, 0.21 mmol), phenyl 1-fluoro-4-isocyanatoate 4a (42 mg, 0.31 mmol) and triethylamine (62 mg, 0.62 mmol) The reaction was carried out overnight at room temperature in 3 mL of dichloromethane. The organic layer was concentrated under reduced pressure and purified to purified crystals crystals crystals )-2-oxo-1-(2,2,2-trifluoroethyl)-1,2-dihydro-1,6-naphthyridin-3-yl)phenyl)-3-(4-fluoro Phenyl)urea 11 (10 mg, white solid), yield: 8%.

MS m/z (ESI): 625.7 [M+1]

¹ H NMR (400 MHz, DMSO-d ₆ ) δ 9.21 (s, 1H), 8.76 (s, 1H), 8.44 (s, 1H), 8.19 (d, J = 8.4 Hz, 1H), 7.83 (s, 1H), 7.69 (d, J = 10.8 Hz, 1H), 7.46-7.43 (m, 2H), 7.25-7.24 (m, 1H), 7.13 (t, J = 8.8 Hz, 2H), 6.50 (s, 1H) ), 5.07-5.02 (m, 2H), 3.53-3.50 (m, 4H), 3.29 (s, 3H).

Example 12

1-(4-Chloro-2-fluoro-5-(7-((2-methoxyethyl)amino)-2-oxo-1-(2,2,2-trifluoroethyl)-1 ,2-dihydro-1,6-naphthyridin-3-yl)phenyl)-3-(3-fluorophenyl)urea

first step

3-(5-Amino-2-chloro-4-fluorophenyl)-7-((2-methoxyethyl)amino)-1-(2,2,2-trifluoroethyl)-1, 6-naphthyridine-2(1H)-one

3-(5-Amino-2-chloro-4-fluorophenyl)-7-chloro-1-(2,2,2-trifluoroethyl)-1,6-naphthyridin-2(1H)- Ketone 1f (400 mg, 1 mmol), 2-methoxyethylamine (750 mg, 10 mmol) and 1,8-diazabicycloundec-7-ene (DBU) (304 mg, 2 mmol) dissolved in 8 mL N In the methylpyrrolidone, microwave reaction was carried out at 180 ° C for 1 hour. After adding 100 mL of water, and extracting with dichloromethane (50 mL × 3), the organic phase was combined, dried over anhydrous sodium sulfate, filtered, and concentrated under reduced pressure. Purification to give 3-(5-amino-2-chloro-4-fluorophenyl)-7-((2-methoxyethyl)amino)-1-(2,2,2-trifluoroethyl) -1,6-naphthyridin-2(1H)-one 12a (400 mg, yellow oil), yield: 90%.

MS m/z (ESI): 444.9 [M+1]

Second step

3-(5-Amino-2-chloro-4-fluorophenyl)-7-((2-methoxyethyl)amino)-1-(2,2,2-trifluoroethyl)-1 6-naphthyridine-2(1H)-one 12a (70 mg, 0.16 mmol), 1-fluoro-3-isocyanatophenyl 2a (26 mg, 0.19 mmol) and triethylamine (48 mg, 0.47 mmol) The reaction was carried out overnight at room temperature in 3 mL of dichloromethane. The organic layer was concentrated under reduced pressure and purified to purified crystals crystals eluted )-2-oxo-1-(2,2,2-trifluoroethyl)-1,2-dihydro-1,6-naphthyridin-3-yl)phenyl)-3-(3-fluoro Phenyl)urea 12 (9 mg, white solid), yield: 10%.

MS m/z (ESI): 581.8 [M+1]

^{1 H NMR (400MHz, DMSO-} d 6) δ9.36 (s, 1H), 8.80 (s, 1H), 8.45 (s, 1H), 8.18 (d, J = 8.8Hz, 1H), 7.86 (s, 1H), 7.58 (d, J = 10.8 Hz, 1H), 7.50-7.43 (m, 2H), 7.31 (q, J = 6.8 Hz, 1H), 7.08 (d, J = 8.4 Hz, 1H), 6.83 6.79 (m, 1H), 6.51 (s, 1H), 5.06-5.03 (m, 2H), 3.51-3.50 (m, 4H), 3.29 (s, 3H).

Example 13

1-(4-Chloro-2-fluoro-5-(7-((2-methoxyethyl)amino)-2-oxo-1-(2,2,2-trifluoroethyl)-1 ,2-dihydro-1,6-naphthyridin-3-yl)phenyl)-3-(4-fluorophenyl)urea

first step

3-(5-Amino-2-chloro-4-fluorophenyl)-7-((2-methoxyethyl)amino)-1-(2,2,2-trifluoroethyl)-1 6-naphthyridine-2(1H)-one 12a (100 mg, 0.225 mmol), phenyl 1-fluoro-4-isocyanate 4a (37 mg, 0.27 mmol) and triethylamine (68 mg, 0.675 mmol) The reaction was carried out in 3 mL of dichloromethane at room temperature overnight. The organic layer was concentrated under reduced pressure and purified to purified crystals crystals eluted )-2-oxo-1-(2,2,2-trifluoroethyl)-1,2-dihydro-1,6-naphthyridin-3-yl)phenyl)-3-(4-fluoro Phenyl)urea 13 (9 mg, white solid), yield: 10%.

MS m/z (ESI): 581.8 [M+1]

^{1 H NMR (400MHz, DMSO-} d 6) δ9.15 (s, 1H), 8.71 (s, 1H), 8.45 (s, 1H), 8.19 (d, J = 8.8Hz, 1H), 7.86 (s, 1H), 7.57 (d, J = 11.2 Hz, 1H), 7.46-7.43 (m, 2H), 7.25-7.23 (m, 1H), 7.13 (d, J = 8.8 Hz, 2H), 6.51 (s, 1H) ), 5.06-5.04 (m, 2H), 3.53-3.48 (m, 4H), 3.29 (s, 3H).

Example 14

1-(4-Chloro-2-fluoro-5-(7-((2-methoxyethyl)amino)-2-oxo-1-(2,2,2-trifluoroethyl)-1 ,2-dihydro-1,6-naphthyridin-3-yl)phenyl)-3-(4-(trifluoromethyl)phenyl)urea

first step

3-(5-Amino-2-chloro-4-fluorophenyl)-7-((2-methoxyethyl)amino)-1-(2,2,2-trifluoroethyl)-1 6-naphthyridine-2(1H)-one 12a (100 mg, 0.225 mmol), 1-isocyanato-4-(trifluoromethyl)phenyl ester 14a (50 mg, 0.27 mmol) and triethylamine (68 mg, 0.675 mmol) was dissolved in 3 mL of dichloromethane and allowed to react at room temperature overnight. The organic layer was concentrated under reduced pressure and purified to purified crystals crystals eluted )-2-oxo-1-(2,2,2-trifluoroethyl)-1,2-dihydro-1,6-naphthyridin-3-yl)phenyl)-3-(4-( Trifluoromethyl)phenyl)urea 14 (16 mg, white solid), yield: 11%.

MS m/z (ESI): 631.8 [M+1]

^{1 H NMR (400MHz, DMSO-} d 6) δ9.67 (s, 1H), 8.91 (s, 1H), 8.45 (s, 1H), 8.18 (d, J = 8.4Hz, 1H), 7.87 (s, 1H), 7.64 (s, 4H), 7.59 (d, J = 11.2 Hz, 1H), 7.25-7.24 (m, 1H), 6.51 (s, 1H), 5.06-5.04 (m, 2H), 3.53-3.50 (m, 4H), 3.29 (s, 3H).

Example 15

(R)-1-(4-chloro-2-fluoro-5-(7-((1-methoxypropan-2-yl)amino)-2-oxo-1-(2,2,2- Trifluoroethyl)-1,2-dihydro-1,6-naphthyridin-3-yl)phenyl)-3-(3-fluorophenyl)urea

first step

(R)-3-(5-Amino-2-chloro-4-fluorophenyl)-7-((1-methoxypropan-2-yl)amino)-1-(2,2,2-tri Fluoroethyl)-1,6-naphthyridin-2(1H)-one

3-(5-Amino-2-chloro-4-fluorophenyl)-7-chloro-1-(2,2,2-trifluoroethyl)-1,6-naphthyridin-2(1H)- Ketone 1f (202 mg, 0.4 mmol), (R)-1-methoxypropan-2-amine (223 mg, 2.5 mmol) and 1,8-diazabicycloundec-7-ene (DBU) ( 152 mg, 1 mmol) was dissolved in 4 mL of N-methylpyrrolidone and subjected to microwave reaction at 180 ° C for 1.5 hours. After adding 40 mL of water, and extracting with dichloromethane (30 mL × 2), the organic phase is combined, dried over anhydrous sodium sulfate, filtered, and concentrated under reduced pressure. Purification afforded (R)-3-(5-amino-2-chloro-4-fluorophenyl)-7-((1-methoxypropan-2-yl)amino)-1-(2,2, 2-Trifluoroethyl)-1,6-naphthyridin-2(1H)-one 15a (229 mg, yellow solid), yield: 100%.

MS m/z (ESI): 458.9 [M+1]

Second step

(R)-3-(5-Amino-2-chloro-4-fluorophenyl)-7-((1-methoxypropan-2-yl)amino)-1-(2,2,2- Trifluoroethyl)-1,6-naphthyridin-2(1H)-one 15a (100 mg, 0.22 mmol), 1-fluoro-3-isocyanatophenyl 2a (45 mg, 0.33 mmol) and triethylamine ( 66 mg, 0.65 mmol) was dissolved in 2 mL of dichloromethane and allowed to react at room temperature overnight. The organic layer was concentrated under reduced pressure, and the obtained residue was purified (jjjjjjjj Prop-2-yl)amino)-2-oxo-1-(2,2,2-trifluoroethyl)-1,2-dihydro-1,6-naphthyridin-3-yl)phenyl) -3-(3-Fluorophenyl)urea 15 (12 mg, white solid), yield: 10%.

MS m/z (ESI): 595.8 [M+1]

^{1 H NMR (400MHz, DMSO-} d 6) δ9.38 (s, 1H), 8.81 (s, 1H), 8.44 (s, 1H), 8.18 (d, J = 8.4Hz, 1H), 7.85 (s, 1H), 7.58 (d, J = 10.8 Hz, 1H), 7.48 (d, J = 11.6 Hz, 1H), 7.32 - 7.29 (m, 1H), 7.11 - 7.08 (m, 2H), 6.83 - 6.79 (m , 1H), 6.48 (m, 1H), 5.06-5.034 (m, 2H), 4.25-4.22 (m, 1H), 3.53-3.50 (m, 2H), 3.30 (s, 3H), 1.16 (d, J) =7.2Hz, 3H).

Example 16

1-(5-(7-Amino-2-oxo-1-(2,2,2-trifluoroethyl)-1,2-dihydro-1,6-diazaphthalidin-3-yl) -2,4-difluorophenyl)-3-(4-fluorophenyl)urea

first step

3-(5-Amino-2,4-difluorophenyl)-7-((4-methoxybenzyl)amino)-1-(2,2,2-trifluoroethyl)-1,6 -naphthyridine-2(1H)-one

3-(5-Amino-2,4-difluorophenyl)-7-chloro-1-(2,2,2-trifluoroethyl)-1,6-naphthyridin-2(1H)-one 5b (500 mg, 1.28 mmol) was dissolved in 5 mL of 4-methoxybenzylamine 16a and allowed to react at 130 ° C overnight. After completion of the reaction, the mixture was cooled to room temperature, and a mixed solvent of 10 mL of ethyl acetate and water (V: V = 1:1) was added, and the mixture was stirred at room temperature for 30 minutes. The mixture was extracted with EtOAc (EtOAc (EtOAc) (EtOAc. Purification: A system) purified to give 3-(5-amino-2,4-difluorophenyl)-7-((4-methoxybenzyl)amino)-1-(2,2,2 -Trifluoroethyl)-1,6-naphthyridin-2(1H)-one 16b (375 mg, pale yellow solid), yield: 60%.

MS m/z (ESI): 490.9 [M+1]

Second step

7-Amino-3-(5-amino-2,4-difluorophenyl)-1-(2,2,2-trifluoroethyl)-1,6-naphthyridin-2(1H)-one

3-(5-Amino-2,4-difluorophenyl)-7-((4-methoxybenzyl)amino)-1-(2,2,2-trifluoroethyl)-1, 6-Naphthyridine-2(1H)-one 16b (375 mg, 0.76 mmol) was dissolved in 4 mL of trifluoroacetic acid and allowed to react at 60 ° C overnight. After completion of the reaction, the mixture was cooled to room temperature, EtOAc (EtOAc) (EtOAc) Drying with anhydrous sodium sulfate, filtration, and concentration under pressure, the obtained residue was purified by silica gel column chromatography (eluent: A system) to give 7-amino-3-(5-amino-2,4- Difluorophenyl)-1-(2,2,2-trifluoroethyl)-1,6-naphthyridin-2(1H)-one 16c (200 mg, pale yellow solid), yield: 71%.

MS m/z (ESI): 370.9 [M+1]

third step

Triethylamine (40.4 mg, 0.4 mmol) was dissolved in 1 mL of tetrahydrofuran, and 7-amino-3-(5-amino-2,4-difluorophenyl)-1-(2,2,2) was added at 0 °C. -trifluoroethyl)-1,6-naphthyridin-2(1H)-one 16c (50 mg, 0.1 mmol), then 1-fluoro-4-isocyanatophenyl ester 4a (13.7 mg, 0.1 mmol) The reaction was allowed to proceed overnight at room temperature. After completion of the reaction, the mixture was concentrated under reduced vacuolulululululululululu ,2-dihydro-1,6-diazaphthalidin-3-yl)-2,4-difluorophenyl)-3-(4-fluorophenyl)urea 16 (9 mg, pale yellow solid) Rate: 15%.

MS m/z (ESI): 507.8 [M+1]

¹ H NMR (400 MHz, DMSO-d ₆ ) δ 9.10 (s, 1H), 8.58 (s, 1H), 8.42 (s, 1H), 8.10 (t, J = 12.0 Hz, 1H), 7.94 (s, 1H), 7.47-7.37 (m, 3H), 7.12 (t, J = 8.0 Hz, 2H), 6.78 (s, 2H), 6.42 (s, 1H), 5.01 (d, J = 8.0 Hz, 2H).

Example 17

1-(5-(7-Amino-2-oxo-1-(2,2,2-trifluoroethyl)-1,2-dihydro-1,6-diazaphthalidin-3-yl) -2,4-difluorobenzene

first step

1-(5-(7-Amino-2-oxo-1-(2,2,2-trifluoroethyl)-1,2-dihydro-1,6-diazaphthalidin-3-yl) -2,4-difluorophenyl)-3-(3-fluorophenyl)urea

Triethylamine (54.6 mg, 0.54 mmol) was dissolved in 3 mL of tetrahydrofuran, and 7-amino-3-(5-amino-2,4-difluorophenyl)-1-(2,2,2) was added at 0 °C. -trifluoroethyl)-1,6-naphthyridin-2(1H)-one 16c (100 mg, 0.27 mmol), then 1-fluoro-3-isocyanatophenyl ester 2a (40.76 mg, 0.29 mmol) The reaction was allowed to proceed overnight at room temperature. After completion of the reaction, the mixture was concentrated under reduced vacuolulululululululululu ,2-dihydro-1,6-diazaphthalidin-3-yl)-2,4-difluorophenyl)-3-(3-fluorophenyl)urea 17 (54 mg, white solid) : 41%.

MS m/z (ESI): 507.9 [M+1]

^{1 H NMR (400MHz, DMSO-} d 6) δ9.31 (s, 1H), 8.68 (s, 1H), 8.48 (s, 1H), 8.10 (t, J = 8.0Hz, 1H), 7.99 (s, 1H), 7.51-7.40 (m, 2H), 7.34-7.28 (q, J = 8.0 Hz, 1H), 7.10 (d, J = 8.0 Hz, 3H), 6.82-6.78 (m, 1H), 6.53 (s) , 1H), 5.04 (d, J = 8.0 Hz, 2H).

Biological evaluation

Test Example 1. Determination of recombinant human c-KIT [D816V] kinase activity by the compound of the present invention

The following method was used to determine the degree of inhibition of the kinase activity of the recombinant human c-KIT [D816V] (D816V mutation) by the compounds of the invention under in vitro conditions.

This method uses the company of Cisbio

Tyrosine Kinase Kit (Cat. No. 62TK0PEB), based on time-resolved fluorescence energy resonance transfer (TF-FRET), reflects compound-to-protein by measuring the degree of phosphorylation of protein-mediated biotinylated peptide substrates The inhibition of kinase activity is strong or weak. For detailed experimental procedures, refer to the kit instructions. Recombinant human c-KIT [D816V] protein kinase was purchased from Carna bioscience (Japan, article number c-KIT [D816V] #08-505).

The experimental procedure is briefly described as follows: Test compounds (the compounds listed in Table 1) are first dissolved in DMSO to prepare a stock solution, followed by gradient dilution with the buffer provided in the kit, and the test compound is finally in the reaction system. The concentration range is from 10 μM to 0.1 nM. The concentration of ATP solution (Bio Bioengineering (Shanghai) Co., Ltd., #A600311) used in the test was a pre-determined ATP Km concentration corresponding to each kinase, wherein the concentration of ATP Km of c-KIT [D816V] was 30 μM. . The reaction was carried out in a 384-well microplate. First, the test compound and 0.66 ng of the test protein were added to the well, and incubated at room temperature for 5 minutes, and then the ATP solution and the biotinylated polypeptide substrate were added to the reaction solution. After incubating for 50 minutes at room temperature with shaking at room temperature, an anti-phosphotyrosine antibody conjugated with a lanthanide compound and streptavidin coupled with modified allophycocyanin XL665 were added to the reaction. Incubate for 1 hour at room temperature with continued shaking. After the end of the incubation, the fluorescence intensity values of the respective wells at an excitation wavelength of 304 nm and emission wavelengths of 620 nM and 665 nM were measured in a TF-FRET mode by a microplate reader. The percentage inhibition of the compound at each concentration was calculated by comparison with the fluorescence intensity ratio of the control group (0.1% DMSO), and the compound IC was obtained by nonlinear regression analysis of the compound concentration-inhibition rate by GraphPad Prism 5 software. ₅₀ values, see Table 1.

Table 1 Compound of the invention ₅₀ values for c-KIT [D816V] inhibition IC

化合物编号Compound number	IC ₅₀(nM)/c-KIT D816V IC ₅₀ (nM)/c-KIT D816V
伊马替尼Imatinib	>5,000>5,000
参考化合物1Reference compound 1	6565
参考化合物2Reference compound 2	5151
参考化合物3Reference compound 3	2020
参考化合物4Reference compound 4	9393
33	33
44	99
55	66

66	1313
77	1414
88	77
99	88

Conclusion: The compound of the present invention has a good inhibitory effect on c-KIT D816V.

Remarks: Imatinib was purchased from Jingyan Chemical (CAS: 152459-95-5, purity: 98.87%, batch number 1604105, off-white solid);

Reference compounds 1, 2 and 3 are the compounds prepared in Example 31, Example 59 and Example 15 disclosed in WO2013184119, respectively; the specific structure is as follows:

The preparation method and structure identification thereof are respectively referred to Example 31, Example 59 and Example 15 of WO2013184119;

The preparation method of Reference Compound 4 was prepared by referring to Example 15 of WO2013184119, and the structure was identified as follows:

MS m/z (ESI): 468.8 [M + 1];

1H NMR (400MHz, DMSO-d6) δ 9.07 (s, 1H), 8.55 (s, 1H), 8.45 (s, 1H), 8.12 (t, J = 12.0 Hz, 1H), 7.84 (s, 1H) , 7.45-7.35 (m, 3H), 7.15-7.08 (m, 3H), 6.25 (s, 1H), 4.16 (q, J = 4.0 Hz, 2H), 2.88 (d, J = 8.0 Hz, 3H), 1.23 (t, J = 4.0 Hz, 3H).

Test Example 2: Determination of Recombinant Human C-KIT [T670I] and c-KIT [V560G/D816V] Kinase Activity by Compounds of the Invention

The following method was used to determine the degree of inhibition of the kinase activity of recombinant human c-KIT [T670I] and c-KIT [V560G/D816V] protein kinases in vitro by the compounds of the invention.

This method uses the company of Cisbio

Tyrosine Kinase Kit (Cat. No. 62TK0PEB), which is based on time-resolved fluorescence energy resonance transfer (TF-FRET), which reacts to protein by measuring the degree of phosphorylation of protein-mediated biotinylated peptide substrates. The inhibition of kinase activity is strong or weak. For detailed experimental procedures, refer to the kit instructions. Recombinant human c-KIT [T670I] protein kinase and c-KIT [V560G/D816V] were purchased from Carna bioscience (Japan, article number c-KIT[T670I]#08-195 and c-KIT[V560G/D816V#08- 535).

The experimental procedure is briefly described as follows: The test compound (the compound described in Table 2) is first dissolved in DMSO to prepare a stock solution, followed by gradient dilution with the buffer provided in the kit, and the final concentration of the test compound in the reaction system. The range is from 10 μM to 0.1 nM. The concentration of the ATP solution (Bio Bioengineering (Shanghai) Co., Ltd., #A600311) used in the test was a pre-determined ATP Km concentration corresponding to each kinase, wherein the concentration of ATP Km of c-KIT [T670I] was 30 μM. The concentration of ATP Km of c-KIT [V560G/D816V] was 10 μM. The reaction was carried out in a 384-well microplate. First, the test compound and the test protein (c-KIT[T670I]0.66 ng or c-KIT[V560G/D816V]0.03 ng) were added to the well and incubated at room temperature. After 5 minutes, an ATP solution and a biotinylated polypeptide substrate solution were added to the reaction solution, and after incubation for 50 minutes at room temperature with shaking, an anti-phosphotyrosine antibody conjugated with a lanthanide compound was added to the reaction. The streptavidin coupled with the modified allophycocyanin XL665 was incubated for 1 hour at room temperature with continued shaking. After the end of the incubation, the fluorescence intensity values of the respective wells at an excitation wavelength of 304 nm and emission wavelengths of 620 nM and 665 nM were measured in a TF-FRET mode by a microplate reader. The percentage inhibition of the compound at each concentration was calculated by comparison with the fluorescence intensity ratio of the control group (0.1% DMSO), and the compound IC was obtained by nonlinear regression analysis of the compound concentration-inhibition rate by GraphPad Prism 5 software. ₅₀ values, see Table 2.

Table Compound 2 of the present invention c-KIT [T670I] IC ₅₀ values, and c-KIT [V560G / D816V] inhibition

Conclusion: The compounds of the present invention have a good inhibitory effect on c-KIT T760I and c-KIT V560G/D816V.

Test Example 3: Determination of Recombinant Human PDFGRα[D842V] Kinase Activity by Compounds of the Invention

The following method was used to determine the inhibitory effect of the representative compounds of the present application on the kinase activity of recombinant human PDGFRα [D842V] (D842V mutation) under in vitro conditions.

This method uses the company of Cisbio

Tyrosine Kinase Kit (Cat. No. 62TK0PEB), based on time-resolved fluorescence energy resonance transfer (TF-FRET), reflects compound-to-protein by measuring the degree of phosphorylation of protein-mediated biotinylated peptide substrates The inhibition of kinase activity is strong or weak. For detailed experimental procedures, refer to the kit instructions. Recombinant human PDGFRα [D842V] protein kinase was purchased from Carna bioscience (Japan, article number PDFGRα [D842V] #08-506).

The experimental procedure is briefly described as follows: The test compound is first dissolved in DMSO to prepare a stock solution, followed by serial dilution with a buffer provided in the kit, and the final concentration of the test compound in the reaction system ranges from 10 μM to 0.1 nM. The concentration of the ATP solution (Bio Bioengineering (Shanghai) Co., Ltd., #A600311) used for the test was a pre-measured ATP Km concentration of 30 μM. The reaction was carried out in a 384-well microplate. First, the test compound and 0.66 ng of the test protein were added to the well, and incubated at room temperature for 5 minutes, and then the ATP solution and the biotinylated polypeptide substrate were added to the reaction solution. After incubating for 50 minutes at room temperature with shaking at room temperature, an anti-phosphotyrosine antibody conjugated with a lanthanide compound and streptavidin coupled with modified allophycocyanin XL665 were added to the reaction. Incubate for 1 hour at room temperature with continued shaking. After the end of the incubation, the fluorescence intensity values of the respective wells at an excitation wavelength of 304 nm and emission wavelengths of 620 nM and 665 nM were measured in a TF-FRET mode by a microplate reader. The percentage inhibition of the compound at each concentration was calculated by comparison with the fluorescence intensity ratio of the control group (0.1% DMSO), and the compound IC was obtained by nonlinear regression analysis of the compound concentration-inhibition rate by GraphPad Prism 5 software. ₅₀ values, see Table 3.

Table ₅₀ value of the compound represented by IC 3 of the present application PDGFRα [D842V] inhibition

实施例编号Example number	IC ₅₀(nM)/PDGFRα[D842V] IC ₅₀ (nM)/PDGFRα[D842V]
1616	6161
1717	1818

Conclusion: Representative compounds 16 and 17 of the present application have a good inhibitory effect on PDGFRα [D842V].

Test Example 4: Determination of P815 activity in mouse mastocytoma by the compound of the present invention

The following method was used to determine the effect of the compound of the present invention on tumor cell proliferation by using a Cell Counting Kit-8 kit (Dojindo, Toray Chemical Technology), and the specific procedure was carried out in accordance with the instructions. For c-KIT [D816V], mouse mastocytoma P815 (purchased from the Cell Resource Center of Shanghai Institutes for Biological Sciences, Chinese Academy of Sciences) was used for culture.

The experimental methods are briefly described as follows: The test compound (the compound described in Table 3) is first dissolved in DMSO to prepare a stock solution, and then serially diluted with the corresponding medium of the cells to prepare a test sample, and the final concentration of the compound is in the range of 30 μM. 0.01 nM. The tumor cells in the logarithmic growth phase were seeded at a density of 1000 cells/well into a 96-well cell culture plate, and after overnight at 37 ° C in a 5% CO ₂ incubator, the test compound samples were added and the cells were further cultured for 48 hours. After the completion of the culture, 10 μL of CCK-8 test solution was added to each well, and incubated at 37 ° C for 1 to 2 hours, and then the absorbance values of the respective wells at 450 nM were read on a microplate reader. The percentage inhibition of the compound at each concentration point was calculated by comparison with the absorbance values of the control group (0.3% DMSO), followed by nonlinear regression analysis of the compound concentration log-inhibition rate in GraphPad Prism 5 software to obtain compound inhibitory cells. The IC ₅₀ values for proliferation are shown in Table 4.

Table IC ₅₀ values of active compounds of the invention on mouse mast cell tumor inhibition 4

化合物编号Compound number	IC ₅₀(nM)/P815 IC ₅₀ (nM)/P815
伊马替尼Imatinib	32523252
参考化合物1Reference compound 1	4444
55	2525
66	1919
99	2020

Conclusion: The compounds of the present invention have a significant inhibitory effect on the proliferation of mouse mastocytoma P815.

Test Example 5, Pharmacokinetic Testing of Compounds of the Invention

1. Summary

SD rats were used as test animals, and the concentration of the drug in plasma was measured by LC/MS/MS method in rats after intragastric administration of reference compound 1 and compound 6 of the present invention. Pharmacokinetic characteristics.

2, the experimental program

2.1 Experimental drugs and animals

Reference compound 1 and compound 6 of the invention;

Six healthy adult SD male rats were divided into three groups and purchased from Xipuer-Beikai Experimental Animal Co., Ltd.

2.2 drug configuration and drug delivery

Weigh an appropriate amount of the experimental drug, add 0.5% sodium carboxymethylcellulose (CMC-Na), sonicate to dissolve, pipet 100 μL for concentration determination, and set the solution concentration to 0.3 mg/mL.

Six healthy adult SD male rats were divided into three groups. After fasting overnight, the rats were intragastrically administered at a dose of 3 mg/kg and the administration volume was 10 mL/kg.

2.3 operation

0.25 mL of blood was collected from the neck before 0.083 hours, 0.25 hours, 0.5 hours, 1 hour, 2 hours, 4 hours, 8 hours and 24 hours after administration, and placed in heparinized tubes at 2-8 °C. At 8000 rpm, centrifuge for 6 minutes, store at -70 ° C, and take it 2 hours after administration. The content of the test compound in the plasma of SD male rats after intragastric administration of different compounds was determined by LC-MS/MS. The pharmacokinetic parameters were calculated using WinNonlin through blood concentration data at different time points.

3, pharmacokinetic parameters results

The pharmacokinetic parameters of the compounds of the invention are shown in Table 5.

Table 5 Pharmacokinetic data sheets for the compounds of the invention

Conclusion: Compared with Reference Compound 1, Compound 6 of the present invention has a higher blood concentration and an area under the curve of the drug, and has a longer half-life and better pharmacokinetic properties.

Test Example 6, Pharmacokinetic Test of Preferred Compound ICR Mouse of the Present Invention

1. Summary

Using ICR mice as test animals, the concentration of the drug in plasma at different times after the mice were administered with the compounds of Reference Compound 1, Example 5 and Example 6 by tail vein injection and intragastric administration by LC/MS/MS method. The pharmacokinetic profile of the compounds of the invention in mice was investigated.

2, the experimental program

2.1 Experimental drugs and animals

Reference compound 1, compound 5 and compound 6 of the invention;

54 healthy adult ICR male mice, 31.3～35.6g, were divided into 6 groups, A～F group, purchased from Beijing Weitong Lihua Experimental Animal Technology Co., Ltd. Animal quality certificate number: 11400700310546.

2.2 drug configuration and drug delivery

2.2.1 Preparation of intravenous (IV-A) preparations

Weigh the appropriate amount of experimental drug, add DMSO: Solutol HS-15: Saline = 5:10:85 (v / v / v), vortex and mix, filter with Nylon filter membrane (0.45 μm), pipette 100 μL × 2 was used for concentration measurement, and the solution concentration was 0.2 mg/mL, and stored at 2 to 8 °C.

2.2.2 Preparation of intragastric (IG-B) preparation

Weigh the appropriate amount of experimental drug, add 0.5% sodium carboxymethylcellulose (CMC-Na), sonicate to dissolve, pipet 100μL × 2 for concentration determination, the concentration of the solution is 0.3mg / mL, 2 ~ Store at 8 ° C.

2.2.3 Administration

54 healthy adult ICR male mice, 31.3～35.6g, were divided into 6 groups, group A～F. After fasting overnight, group A, group C and group E were administered via tail vein respectively. The dose was 1mg. /kg, the administration volume was 5 mL/kg; Group B, Group D and Group F were intragastrically administered at a dose of 3 mg/kg and a dose of 10 mL/kg.

2.3 operation

Before intravenous administration and 0.083 hours, 0.25 hours, 0.5 hours, 1 hour, 2 hours, 4 hours, 8 hours, 12 hours, and 24 hours after administration or before and after administration and 0.25 hours after administration, 80 μL of blood was collected through the orbital vein at 0.5 hour, 1 hour, 2 hours, 4 hours, 8 hours, 12 hours, and 24 hours, placed in an EDTA-K2 anticoagulant tube, and centrifuged at 1500 g for 10 minutes at -40 to -20 ° C. Store and eat 4 hours after administration. The content of the test compound in the plasma of ICR male mice after tail vein injection and intragastric administration of different compounds was determined by LC-MS/MS. The pharmacokinetic parameters were calculated using WinNonlin through blood concentration data at different time points.

3, pharmacokinetic parameters results

The pharmacokinetic parameters of the compounds of the invention are shown in Tables 6 and 7.

Table 6 Pharmacokinetic data sheets of the compounds of the present invention administered by intragastric administration

Conclusion: Compared with the reference compound 1, the mice were orally administered with the highest blood concentration and the area under the curve of the compound of the present invention, and had good pharmacokinetic properties.

Table 7 Pharmacokinetic data sheets for intravenous administration of the compounds of the present invention

Conclusion: Compared with the reference compound 1, the mice administered the compound 5 and the compound 6 of the present invention have a higher drug-time curve area and have better pharmacokinetic properties.

Claims

A compound of the formula (I): or a stereoisomer, tautomer or pharmaceutically acceptable salt thereof:

among them:

R 1 is selected from a hydrogen atom, an alkyl group or -C(O)R 5 , wherein the alkyl group is optionally further further selected from one or more selected from the group consisting of halogen, hydroxy, alkoxy, cycloalkyl, heterocyclic, -C(O)R 5 , -OC(O)R 5 , -C(O)OR 5 , -NR 6 R 7 , -C(O)NR 6 R 7 , -S(O) n NR 6 R 7 Or substituted with a substituent of -NR 6 C(O)R 7 ;

R 2 is selected from alkyl, wherein said alkyl group is further substituted with one or more halogens;

R 3 is the same or different and is each independently selected from the group consisting of a hydrogen atom, an alkyl group, an alkoxy group, a hydroxyl group, a cyano group, a nitro group, a halogen group, a cycloalkyl group, a heterocyclic group, -C(O)R 5 , -OC ( O) R 5 , -C(O)OR 5 , -NR 6 R 7 , -C(O)NR 6 R 7 , -S(O) n NR 6 R 7 or -NR 6 C(O)R 7 , Wherein the alkyl, alkoxy, cycloalkyl or heterocyclic group is optionally further selected from one or more selected from the group consisting of hydroxyl, halogen, nitro, cyano, alkyl, alkoxy, cycloalkyl, hetero a cyclic group, an aryl group, a heteroaryl group, -C(O)R 8 , -C(O)OR 8 , -OC(O)R 8 , -NR 9 R 10 , -C(O)NR 9 R 10 , Substituted by a substituent of -S(O) n NR 9 R 10 or -NR 9 C(O)R 10 ;

R 4 is selected from aryl or heteroaryl, wherein said aryl or heteroaryl is optionally further selected from one or more selected from the group consisting of alkyl, alkoxy, hydroxy, cyano, nitro, halogen, cycloalkane. Base, heterocyclic group, -C(O)R 5 , -OC(O)R 5 , -C(O)OR 5 , -NR 6 R 7 , -C(O)NR 6 R 7 , -S(O Substituting a substituent of n NR 6 R 7 or -NR 6 C(O)R 7 ; wherein said alkyl or alkoxy group is optionally further substituted with one or more halogens;

R 5 , R 6 and R 7 are each independently selected from a hydrogen atom, a hydroxyl group, a halogen, a nitro group, a cyano group, an alkyl group, an alkoxy group, a cycloalkyl group, a heterocyclic group, an aryl group or a heteroaryl group. The alkyl, alkoxy, cycloalkyl, heterocyclyl, aryl or heteroaryl group optionally further selected from one or more selected from the group consisting of hydroxyl, halogen, nitro, cyano, alkyl, alkoxy, Cycloalkyl, heterocyclyl, aryl, heteroaryl, -C(O)R 8 , -C(O)OR 8 , -OC(O)R 8 , -NR 9 R 10 , -C(O) Substituted by a substituent of NR 9 R 10 , -S(O) n NR 9 R 10 or -NR 9 C(O)R 10 ;

Alternatively, R 6 and R 7 together with the N atom to which they are bonded form a 4 to 8 membered heterocyclic group, wherein the 4 to 8 membered heterocyclic ring contains one or more N, O or S(O) n and 4 to The 8-membered heterocyclic ring is optionally further selected from one or more selected from the group consisting of hydroxyl, halogen, nitro, cyano, alkyl, alkoxy, cycloalkyl, heterocyclyl, aryl, heteroaryl, =0, -C(O)R 8 , -C(O)OR 8 , -OC(O)R 8 , -NR 9 R 10 , -C(O)NR 9 R 10 , -S(O) n NR 9 R 10 Or substituted with a substituent of -NR 9 C(O)R 10 ;

R 8 , R 9 and R 10 are each independently selected from a hydrogen atom, an alkyl group, a cycloalkyl group, a heterocyclic group, an aryl group or a heteroaryl group, wherein the alkyl group, the cycloalkyl group, the heterocyclic group, and the aromatic group Or a heteroaryl group optionally further selected from one or more selected from the group consisting of hydroxyl, halogen, nitro, cyano, alkyl, alkoxy, cycloalkyl, heterocyclyl, aryl, heteroaryl, carboxy or carboxy Substituted by a substituent of the acid ester group;

m is selected from 1, 2, 3 or 4;

n is selected from 0, 1 or 2.
The compound according to claim 1 or a stereoisomer, tautomer or pharmaceutically acceptable salt thereof, wherein said R 1 is selected from alkyl or -C(O)R 5 , wherein said The alkyl group is optionally further selected from one or more selected from the group consisting of halogen, hydroxy, alkoxy, cycloalkyl, heterocyclyl, -C(O)R 5 , -OC(O)R 5 , -C(O)OR 5, -NR 6 R 7, -C (O) NR 6 R 7, -S (O) n NR 6 R 7 or -NR 6 C (O) R 7 is substituted with a substituent;

Preferably, said R 1 is selected from C 1-4 alkyl, wherein said C 1-4 alkyl group is optionally further substituted with one or more C 1-6 alkoxy groups, wherein said C 1 - The 6 alkoxy group is preferably a methoxy group.
The compound according to claim 1 or 2, or a stereoisomer, tautomer thereof or a pharmaceutically acceptable salt thereof, which is a compound of the formula (II) or a stereoisomer thereof or a tautomer thereof. Isomer or a pharmaceutically acceptable salt thereof,

among them:

R 3 is selected from the group consisting of a hydrogen atom, an alkyl group, an alkoxy group, a hydroxyl group, a cyano group, a nitro group, a halogen, a cycloalkyl group, a heterocyclic group, -C(O)R 5 , -OC(O)R 5 , -C (O)OR 5 , -NR 6 R 7 , -C(O)NR 6 R 7 , -S(O) n NR 6 R 7 or -NR 6 C(O)R 7 , wherein the alkyl group, The alkoxy, cycloalkyl or heterocyclic group is optionally further selected from one or more selected from the group consisting of hydroxyl, halogen, nitro, cyano, alkyl, alkoxy, cycloalkyl, heterocyclyl, aryl, hetero Aryl, -C(O)R 8 , -C(O)OR 8 , -OC(O)R 8 , -NR 9 R 10 , -C(O)NR 9 R 10 , -S(O) n NR Substituted by a substituent of 9 R 10 or -NR 9 C(O)R 10 ;

R 1 , R 2 , R 4 to R 10 and n are as defined in claim 1 or 2.
A compound according to any one of claims 1 to 3, or a stereoisomer, tautomer or pharmaceutically acceptable salt thereof, wherein:

R 2 is selected from C 1-4 alkyl, wherein said C 1-4 alkyl group is further substituted by one or more halogens; wherein said halogen is preferably fluorine, chlorine or bromine; more preferably fluorine; R 2 is preferably a trifluoroethyl group.
A compound according to any one of claims 1 to 3, or a stereoisomer, tautomer or pharmaceutically acceptable salt thereof, wherein:

R 3 is selected from a hydrogen atom, a C 1-4 alkyl group or a halogen;

Wherein the C 1-4 alkyl group is preferably a methyl group or an ethyl group;

The halogen described therein is preferably fluorine, chlorine or bromine.
A compound according to any one of claims 1 to 3, or a stereoisomer, tautomer or pharmaceutically acceptable salt thereof, wherein:

R 4 is selected from phenyl or benzothienyl, wherein said phenyl or benzothienyl is optionally further further selected from one or more selected from the group consisting of halogen, C 1-4 alkyl or C 1-6 alkoxy. Substituted by a substituent;

Wherein the C 1-4 alkyl or C 1-6 alkoxy group is optionally further substituted with one or more halogens;

Wherein the C 1-4 alkyl group is preferably a methyl group or an ethyl group;

The halogen described therein is preferably fluorine, chlorine or bromine; more preferably fluorine.
A compound according to any one of claims 1 to 3, or a stereoisomer, tautomer or pharmaceutically acceptable salt thereof, wherein:

R 1 is selected from a hydrogen atom or a C 1-4 alkyl group, wherein said C 1-4 alkyl group is optionally further substituted with one or more C 1-6 alkoxy groups; said C 1-6 alkoxy group The group is preferably a methoxy group;

R 2 is selected from C 1-4 alkyl, wherein said C 1-4 alkyl group is further substituted with one or more halogens selected from fluorine, chlorine or bromine;

R 3 is selected from a hydrogen atom, a C 1-4 alkyl group or a halogen, wherein said C 1-4 alkyl group is optionally further substituted with one or more halogens, preferably halogen, chlorine or bromine;

R 4 is selected from phenyl or benzothienyl, wherein said phenyl or benzothienyl is optionally further further selected from one or more selected from the group consisting of halogen, C 1-4 alkyl or C 1-6 alkoxy. Substituted with a substituent; wherein the C 1-4 alkyl or C 1-6 alkoxy group is optionally further substituted with one or more halogens; and the halogen is preferably fluorine, chlorine or bromine; more preferably fluorine.
The compound according to claim 1 or 2, or a stereoisomer, tautomer or pharmaceutically acceptable salt thereof, wherein the compound is:
A process for the preparation of a compound of the formula (I), or a stereoisomer, tautomer or a pharmaceutically acceptable salt thereof, according to claim 1 or 2, which comprises:

A compound of the formula (IA) is reacted with a compound of the formula (IB) to give a compound of the formula (I);

Wherein: R 1 to R 4 and m are as defined in claim 1 or 2.
A process for the preparation of a compound of the formula (II), or a stereoisomer, tautomer or a pharmaceutically acceptable salt thereof, according to claim 3, which comprises:

A compound of the formula (IIA) is reacted with a compound of the formula (IB) to give a compound of the formula (II);

Wherein: R 1 to R 4 are as defined in claim 3.
An intermediate compound of the formula (IA): or a stereoisomer, tautomer or pharmaceutically acceptable salt thereof,

Wherein: R 1 to R 3 and m are as defined in claim 1 or 2.
The compound according to claim 11 or a stereoisomer, tautomer or pharmaceutically acceptable salt thereof, which is a compound of the formula (IIA) or a stereoisomer thereof, tautomerizable Or a pharmaceutically acceptable salt thereof,

Wherein: R 1 to R 3 are as defined in claim 11.
The intermediate compound according to claim 11 or a stereoisomer, tautomer or pharmaceutically acceptable salt thereof, wherein the compound is:
A process for the preparation of an intermediate compound of the formula (IA), or a stereoisomer, tautomer or a pharmaceutically acceptable salt thereof, according to claim 11, the method comprising:

1) When R1 is not a hydrogen atom:

The compound of the formula (Ia) is subjected to a condensation reaction with a compound of the formula (Ib) under heating to obtain a compound of the formula (Ic); a compound of the formula (Ic) is reacted with a compound of the formula (Id) to give a formula (IA). Intermediate compound;

among them:

X is a halogen;

R a is an alkyl group;

R 1 to R 3 and m are as defined in claim 11, and R 1 is not H;

2) When R1 is a hydrogen atom:

The compound of the formula (Ia) is subjected to a condensation reaction with a compound of the formula (Ib) under heating to obtain a compound of the formula (Ic); and a compound of the formula (Ic) is reacted with a compound of the formula (Ie) to give a formula ( If) a compound of the formula (If) is reacted in the presence of trifluoroacetic acid to give an intermediate compound of the formula (IA);

among them:

X is a halogen;

R 1 is a hydrogen atom;

R a is an alkyl group;

R b is an alkyl group;

The definitions of R 2 , R 3 and m are as set forth in claim 11.
A process for the preparation of an intermediate compound of the formula (IIA), or a stereoisomer, tautomer or a pharmaceutically acceptable salt thereof, according to claim 12, which comprises:

1) When R1 is not a hydrogen atom:

A compound of the formula (Ia) is subjected to a condensation reaction with a compound of the formula (IIa) under heating to obtain a compound of the formula (IIb); a compound of the formula (IIb) is reacted with a compound of the formula (Id) to give a formula (IIA) Intermediate compound;

among them:

X is a halogen;

R a is an alkyl group;

R 1 to R 3 are as defined in claim 12, and R 1 is not H;

2) When R 1 is a hydrogen atom:

The compound of the formula (Ia) is subjected to a condensation reaction with a compound of the formula (IIa) under heating to obtain a compound of the formula (IIb); a compound of the formula (IIb) is reacted with a compound of the formula (Ie) to give a formula (IIc) a compound of the formula (IIc) is reacted in the presence of trifluoroacetic acid to give an intermediate compound of the formula (IIA);

among them:

X is a halogen;

R 1 is a hydrogen atom;

R a is an alkyl group;

R b is an alkyl group;

The definitions of R 2 and R 3 are as set forth in claim 12.
A pharmaceutical composition comprising an effective amount of a compound according to any one of claims 1 to 8, or a stereoisomer, tautomer thereof or a pharmaceutically acceptable salt thereof, And pharmaceutically acceptable carriers, excipients or combinations thereof.
The compound according to any one of claims 1 to 8, or a stereoisomer, tautomer thereof or a pharmaceutically acceptable salt thereof, or the pharmaceutical composition according to claim 16 in the preparation of a treatment by c Use of a KIT or a mutant c-KIT mediated disease, wherein the c-KIT or mutant c-KIT mediated disease is preferably gastrointestinal stromal tumor, systemic mast cell hyperplasia Symptoms, acute myeloid leukemia, ovarian cancer, melanoma, cervical cancer, seminoma, dysgerminoma, glioblastoma multiforme, teratoma, mast cell leukemia; more preferably gastrointestinal interstitial Tumor, systemic mastocytosis, glioblastoma multiforme, and acute myeloid leukemia, most preferably gastrointestinal stromal tumors, glioblastoma multiforme, and systemic mastocytosis; The mutant c-KIT mutation is located at 9, 9, 13, 14, 17, and/or 18 of the exon, or at the 816th, 670th, 560th, and/or 654th amino acid residues Wherein the mutation at the amino acid residue at position 816 is preferably D816V or D816H; wherein the 670th ammonia is The mutation at the residue of the acid group is preferably T670I; wherein the mutation at the amino acid residue at position 560 is preferably V560G; wherein the mutation at the amino acid residue at position 654 is preferably V654A.
The compound according to any one of claims 1 to 8, or a stereoisomer, tautomer thereof or a pharmaceutically acceptable salt thereof, or the pharmaceutical composition according to claim 16 in the preparation of c-KIT Use in inhibitors.
The compound according to any one of claims 1 to 8, or a stereoisomer, tautomer thereof or a pharmaceutically acceptable salt thereof, or a pharmaceutical composition according to claim 16 in the preparation of a therapeutic Use of a medicament for mutating or wild-type PDFGRα-mediated diseases, wherein the mediated or wild-type PDFGRα-mediated disease is preferably gastrointestinal stromal tumor, systemic mastocytosis, acute myeloid Leukemia, ovarian cancer, melanoma, cervical cancer, seminoma, dysgerminoma, glioblastoma multiforme, teratoma, mast cell leukemia; more preferably gastrointestinal stromal tumor, systemic hypertrophy Cell hyperplasia, glioblastoma multiforme, and acute myeloid leukemia, most preferably gastrointestinal stromal tumors, glioblastoma multiforme, and systemic mastocytosis; preferably the mutant PDFGRα The mutation is located at exon 18 and/or amino acid residue 842, wherein the mutation at amino acid residue position 842 is preferably a D842V mutation.
A compound according to any one of claims 1 to 8, or a stereoisomer, tautomer or pharmaceutically acceptable salt thereof, or a pharmaceutical composition according to claim 16 for preparing a PDFGRα inhibitor Use in.