CN112538038A - 2-indolone ERR alpha inverse agonist and pharmaceutical composition and application thereof - Google Patents

Abstract

The invention provides an indolone compound with a structure shown in a formula (I) or pharmaceutically acceptable salt thereof or a stereoisomer thereof or a prodrug molecule thereof and application thereof. The compounds can effectively inhibit the combination of estrogen related receptor alpha (ERR alpha) and coactivator PGC-1 alpha, are brand-new ERR alpha inverse agonists, have certain proliferation inhibition activity on tumor cells, and obviously improve the in-vivo sugar tolerance of db/db mice.

Description

2-indolone ERRα inverse agonist and pharmaceutical composition and application thereof

technical field

The invention relates to the field of medicinal chemistry, in particular to an indolinone compound and an application thereof.

Background technique

Estrogen-related receptors (ERRs) belong to the nuclear receptor superfamily of transcription factors, including three subtypes of ERRα, ERRβ and ERRγ, because of their high sequence and structural homology with estrogen receptor ERs sex, hence the name. Unlike ERs, ERRs do not bind to hormonal ligands such as estrogen, and no other endogenous ligands have been found, so they are a class of "orphan nuclear receptors". Among them, ERRα (NR3B1) was first discovered by Giguère V et al. in 1988. It is widely distributed in metabolically active tissues and organs such as skeletal muscle, heart, kidney, liver and adipose tissue, and is a transcriptional regulator that maintains the balance of cellular energy metabolism.

Studies have shown that ERRα can be expressed to varying degrees in normal cells and tumor cells. In normal cells, receptor tyrosine kinases (EGFR or/and ERBB2) can up-regulate the post-translational modification or phosphorylation process of ERRα, thereby increasing its transcriptional activity and promoting ERRα protein and co-activators (PGC-1/SRC, etc.) Or co-repressor (RIP-140, etc.), and then identify the estrogen-related receptor response elements (ERR response elements, ERRE) on the target gene, activate the transcription level of the target gene, participate in the regulation of estrogen receptor-related signaling pathways, affect Differentiation of chondrocytes, formation of bone and joints, regulation of fatty acid oxidation, mitochondrial biosynthesis and oxidative capacity, glucose metabolism, etc.

ERRα protein is highly expressed in ovarian cancer, endometrial cancer, prostate cancer, colorectal cancer, especially breast cancer. In breast cancer cells, the over-activation of EGFR or/and ErbB2 signaling pathway leads to the overexpression of ERRα protein in cells. After ERRα binds to coactivator proteins (such as PGC-1), it upregulates a variety of downstream factors that are closely related to the occurrence and development of breast cancer. Target genes, including genes PDK4 and ACADM related to oxidative metabolism, tricarboxylic acid cycle, SOD2 and UCP2 related to mitochondrial dysfunction, ENO1 and ALDOA related to sugar degradation, CCNE1 and NEK2 related to cell cycle, and cell invasion CXCR4 associated with metastasis, and TFF1, S100P, etc. associated with poor prognosis, thereby regulating intracellular oxidative metabolic pathways, activating VEGF, aromatase, and WNT signaling pathways, causing metabolic reprogramming, cell proliferation, and inflammatory response in breast cancer cells. Angiogenesis, and promote the epithelial-mesenchymal transition of tumor cells, tumor invasion and metastasis, leading to tumor resistance, deterioration or recurrence, resulting in poor prognosis.

Experiments found that ERRα is overexpressed in ERα-deficient triple-negative breast cancer cells, and its expression is positively correlated with the malignancy, recurrence rate, and poor clinical prognosis of triple-negative breast cancer. It is a new type of biomarker for breast cancer; The introduction of ERRα small interfering RNA (siRNA) can effectively inhibit the invasion and metastasis of triple-negative breast cancer cells MDA-MB-231 in vitro, and show good proliferation inhibition activity in vivo. In addition, Genevieve et al. found that Lapatinib can cause the degradation of ERRα protein in sensitive breast cancer cells, but under the pressure of continuous chemotherapy, breast cancer cells significantly upregulate the expression of ERRα protein by activating the mTOR signaling pathway and restore intracellular glutamine Metabolic level, promote mitochondrial energy metabolism, and improve ROS self-detoxification ability (ROS detoxification), thereby maintaining the energy homeostasis of tumor cells and ensuring cell survival; in a mouse model of breast cancer with high expression of HER2, the intervention of ERRα inverse agonist is effective Overcoming lapatinib resistance by disrupting the energy homeostasis established by tumor cells.

With the gradual elucidation of the key roles of ERRα in fatty acid oxidation, mitochondrial biosynthesis and oxidative capacity, glucose metabolism, and metabolic reprogramming in tumor cells, ERRα inverse agonists play a role in metabolic diseases, tumors and other diseases. The application in is paid attention to. However, the reported structures of ERRα inverse agonists are sparse, and no candidate molecule has entered clinical research, which limits the further development of ERRα inverse agonists.

Therefore, there is an urgent need in the art for new types of compounds, especially compounds with novel skeletons, to solve the current problem of low drugability of ERRα inverse agonists, and to fill the gap in the clinical application of ERRα inverse agonists.

SUMMARY OF THE INVENTION

In view of the above problems, the present invention provides a new class of indolinone compounds, which can effectively inhibit the binding of estrogen-related receptor α (ERRα) and co-activator PGC-1α, and are a new type of ERRα inverse Agonist, has a certain proliferation inhibitory activity on tumor cells.

The specific technical solutions are as follows:

Indolinone compounds having the structure represented by formula (I) or pharmaceutically acceptable salts or stereoisomers or prodrug molecules thereof:

in,

m is selected from: 0, 1 and 2;

选自：s个R₀取代或者未取代的C₆-C₁₀芳基、s个R₀取代或者未取代的C₅-C₁₀杂芳基；ring

Selected from: s R ₀ substituted or unsubstituted C ₆ -C ₁₀ aryl, s R ₀ substituted or unsubstituted C ₅ -C ₁₀ heteroaryl;

R ₁ , R ₂ , R ₃ , R ₄ , R ₅ , R ₆ , R ₇ , R ₈ , R ₀ are each independently selected from: hydrogen, halogen, cyano, nitro, amino, hydroxy, _Rf substituted or Unsubstituted C ₁ -C ₆ alkyl, R _f substituted or unsubstituted C ₃ -C ₆ cycloalkyl, R _f substituted or unsubstituted C ₁ -C ₆ alkoxy, R _f substituted or unsubstituted C ₃ -C ₆ cycloalkoxy, R _f substituted or unsubstituted C ₂ -C ₆ alkenyl, R _f substituted or unsubstituted C ₂ -C ₆ alkynyl, R _f substituted or unsubstituted phenyl, -(CH ₂ ) _p NR _a R _b , -C(=O)OR, -CONHR, or R ₃ and R ₅ are linked together by chemical bonds to form -O-, -NH-, or -(CH ₂ ) _n ;

_R9 is selected from: hydrogen, -( _{CH2 ) qNRdRe} , -( _{CH2 ) qORd} , - _{( CH2 ) qOCRcRdRe , - ( CH2 ) qCRcRd Re} ;

s is selected from: an integer between 0 and 5;

n is selected from: 1 or 2;

p is selected from; an integer between 0 and 6;

q is selected from: an integer between 1 and 6;

R _a and R _b are each independently selected from: hydrogen, R _f substituted or unsubstituted C ₁ -C ₆ alkyl, R _f substituted or unsubstituted C ₁ -C ₆ heteroalkyl, or R _a , R _b together with the N atom to which it is attached to form R _substituted or unsubstituted heteroatom-containing monocyclic, fused, spiro or bridged rings;

R is selected from: hydrogen, C ₁ -C ₆ alkyl;

R _c is selected from: hydrogen, C ₁ -C ₃ alkyl;

R _d and R _e are each independently selected from: hydrogen, R _f substituted or unsubstituted C ₁ -C ₆ alkyl, R _f substituted or unsubstituted C ₁ -C ₆ heteroalkyl, or R _d , R _e together with the N atom or C atom to which it is attached, to form an R _substituted or unsubstituted heteroatom-containing monocyclic, fused, spiro or bridged ring;

Each R _f is independently selected from: halogen, hydroxy, amino, C ₁ -C ₃ alkyl, hydroxy substituted C ₁ -C ₃ alkyl, C ₁ -C ₃ alkoxy, -NH(C ₁ -C ₃ alkyl), -N(C ₁ -C ₃ alkyl) (C ₁ -C ₃ alkyl), -C(=O)(C ₁ -C ₃ alkyl), R substituted or unsubstituted 5- 7-membered heterocyclyl.

选自：苯环、吡啶环、萘环、呋喃环、噻吩环、吡咯环、吡唑环、噻唑环、异噻唑环、噁唑环、异噁唑环、吡嗪环、哒嗪环、咪唑环或嘧啶环。In some of these embodiments, the ring

Selected from: benzene ring, pyridine ring, naphthalene ring, furan ring, thiophene ring, pyrrole ring, pyrazole ring, thiazole ring, isothiazole ring, oxazole ring, isoxazole ring, pyrazine ring, pyridazine ring, imidazole ring or pyrimidine ring.

In some of these embodiments, R ₁ , R ₂ , R ₃ , R ₄ , R ₅ , R ₆ , R ₇ , R ₈ , R ₀ are each independently selected from: hydrogen, halogen, cyano, nitro, amino , hydroxyl, C ₁ -C ₆ alkyl, halogenated C ₁ -C ₆ alkyl, C ₃ -C ₆ cycloalkyl, C ₁ -C ₆ alkoxy, halogenated C ₁ -C ₆ alkoxy, C ₃ -C ₆ cycloalkoxy, C ₂ -C ₆ alkenyl, C ₂ -C ₆ alkynyl, phenyl, -(CH ₂ ) _p NR _a R _b , -C(=O)OR, -CONHR , or R ₃ and R ₅ are linked together by chemical bonds to form -O-;

R _a and R _b are each independently selected from: hydrogen, R _f substituted or unsubstituted C ₁ -C ₃ alkyl, or R _a , R _b together with the N atom to which they are attached form R _f substituted or unsubstituted 3-8 membered monocyclic ring containing 1, 2 or 3 heteroatoms, or R _a , R _b together with the N atom to which it is attached to form R _f substituted or unsubstituted 8 containing 1, 2 or 3 heteroatoms -12-membered fused ring, spiro ring or bridged ring; heteroatom is selected from: O, N, S.

In some of these embodiments, R ₁ , R ₂ , R ₃ , R ₄ , R ₅ , R ₆ , R ₇ , R ₈ , R ₀ are each independently selected from: hydrogen, halogen, cyano, nitro, hydroxyl , C ₁ -C ₃ alkyl, halogenated C ₁ -C ₃ alkyl, C ₃ -C ₆ cycloalkyl, C ₁ -C ₃ alkoxy, phenyl, -C(=O)OR, -CONHR , or R ₃ and R ₅ are linked together by chemical bonds to form -O-; wherein, R is selected from: hydrogen, C ₁ -C ₃ alkyl.

-C(＝O)OH、-C(＝O)OCH₃、-C(＝O)OCH₂CH₃、-C(＝O)OCH₂(CH₃)CH₃、-CONH₂、-CONHCH₃，或者R₃和R₅通过化学键相连一起形成-O-；其中，p选自0～3之间的整数。In some of these embodiments, R ₁ , R ₂ , R ₃ , R ₄ , R ₅ , R ₆ , R ₇ , R ₈ , R ₀ are each independently selected from: hydrogen, fluorine, chlorine, bromine, iodine, cyanide group, nitro, amino, hydroxyl, methyl, ethyl, isopropyl, propyl, cyclopropyl, butyl, tert-butyl, cyclobutyl, cyclohexyl, trifluoromethyl, difluoromethyl, Methoxy, ethoxy, propoxy, trifluoromethoxy, cyclopropoxy, vinyl, ethynyl, phenyl, -(CH ₂ ) _p -NH ₂ ,

-C(=O)OH, -C(=O)OCH ₃ , -C(=O)OCH ₂ CH ₃ , -C(=O)OCH ₂ (CH ₃ )CH ₃ , -CONH ₂ , -CONHCH ₃ , or R ₃ and R ₅ are linked together by chemical bonds to form -O-; wherein, p is selected from an integer between 0 and 3.

In some of these embodiments, R ₉ is selected from: hydrogen, -(CH ₂ ) _q NR _{d Re} ; wherein,

q is selected from: 1, 2, 3 or 4;

R _d , R _e are each independently selected from: hydrogen, R _f substituted or unsubstituted C ₁ -C ₆ alkyl, or R _a , R _b together with the N atom to which they are attached form R _f substituted or unsubstituted C 1 -C 6 alkyl 3-8 membered monocyclic ring containing 1, 2 or 3 heteroatoms, or R _a , R _b together with the N atom to which it is attached to form R _f substituted or unsubstituted 8 containing 1, 2 or 3 heteroatoms -12-membered fused ring, spiro ring or bridged ring; heteroatom is selected from: O, N, S.

In some of these embodiments, R _d , R _e are each independently selected from: hydrogen, C ₁ -C ₃ alkyl, or R _a , R _b together with the N atom to which they are attached form R _f substituted or unsubstituted A 5-7 membered monocyclic ring containing 1, 2 or 3 heteroatoms; heteroatoms are selected from: O, N; R _{f is} selected from: H, C ₁ -C ₃ alkyl, -NH(C ₁ -C ₃ alkane group), -N(C ₁ -C ₃ alkyl) (C ₁ -C ₃ alkyl), 5-7 membered heterocyclic group, C ₁ -C ₃ alkyl substituted 5-7 membered heterocyclic group.

-(CH₂)_q-OH、-(CH₂)_q-OCH₃、-(CH₂)_q-OCH₂CH₃；其中，q选自:1、2、3或4。In some of these embodiments, R ₉ is selected from the group consisting of: hydrogen, -(CH ₂ ) _q -NH ₂ ,

-( _CH2 ) _q -OH, -( _CH2 ) _q - _OCH3 , -( _CH2 ) _q - _OCH2CH3 ; wherein q is selected from: 1, 2, ₃ or 4.

选自：苯环、萘环；In some of these embodiments, the ring

Selected from: benzene ring, naphthalene ring;

R ₁ , R ₂ , R ₃ , R ₄ , R ₅ , R ₆ , R ₇ , R ₈ are each independently selected from: hydrogen, methoxy, hydroxy, fluorine, chlorine, bromine, methyl, cyclopropyl, Phenyl, or R ₃ and R ₅ are linked together by chemical bonds to form -O-;

R ₀ is selected from: hydrogen, cyano, trifluoromethyl, nitro, fluorine, chlorine, bromine, carboxamido, formate, methyl formate, difluoromethyl, methoxy;

其中，q选自:1、2、3或4。R ₉ is selected from: hydrogen, -(CH ₂ ) _q -NH ₂ ,

wherein q is selected from: 1, 2, 3 or 4.

选自：苯环、萘环；In some of these embodiments, the ring

Selected from: benzene ring, naphthalene ring;

R ₁ , R ₂ , R ₃ , R ₄ , R ₅ , R ₆ , R ₇ , R ₈ are each independently selected from: hydrogen, methoxy, hydroxy, fluorine, chlorine, bromine, methyl, cyclopropyl, Phenyl, or R ₃ and R ₅ are linked together by chemical bonds to form -O-;

R ₀ is selected from: hydrogen, cyano, trifluoromethyl, nitro, fluorine, chlorine, bromine, methyl formate, difluoromethyl, methoxy;

其中，q选自:1、2、3或4。其中，R₄进一步优选为甲氧基。R ₉ is selected from: hydrogen,

wherein q is selected from: 1, 2, 3 or 4. Among them, R ₄ is more preferably a methoxy group.

In some of these embodiments, the indolinone compound has the structure represented by formula (II) or formula (III):

Wherein, s is selected from: an integer between 0 and 4.

In some of these embodiments, R ₁ , R ₂ , R ₃ , R ₄ , R ₆ , R ₇ , R ₈ are each independently selected from the group consisting of: hydrogen, methoxy, hydroxy, fluoro, chloro, bromo, methyl, Cyclopropyl, phenyl;

R ₅ is selected from: hydrogen, phenyl; and, when R ₅ is hydrogen, neither R ₂ nor R ₃ is hydroxyl;

R ₀ is selected from: hydrogen, cyano, trifluoromethyl, nitro, fluorine, chlorine, bromine, methyl formate, difluoromethyl, methoxy;

R ₉ is selected from: hydrogen,

q is selected from: 1, 2, 3 or 4.

In some of these embodiments, the indolinone compound is selected from the following compounds:

The present invention also provides applications of the above compounds.

The specific technical solutions are as follows:

Use of the above indolinone compounds or pharmaceutically acceptable salts or stereoisomers or prodrug molecules thereof in the preparation of ERRα inverse agonists.

Use of the above indolinone compounds or pharmaceutically acceptable salts or stereoisomers or prodrug molecules thereof in the preparation of medicines for preventing and/or treating diseases related to abnormal expression of ERRα protein activity.

In some of the embodiments, the diseases associated with abnormal expression of ERRα protein activity are: tumors, metabolic diseases.

In some of these embodiments, the tumor is: non-small cell lung cancer, small cell lung cancer, lung adenocarcinoma, lung squamous cell carcinoma, pancreatic cancer, breast cancer, prostate cancer, liver cancer, skin cancer, epithelial cell carcinoma, gastrointestinal stromal tumor, endometrial cancer, histiocytic lymphoma, nasopharyngeal cancer, head and neck tumor, colon cancer, rectal cancer, glioma, malignant melanoma, kidney cancer, bladder cancer, ovarian cancer, cervical cancer, throat cancer Cancer or multiple myeloma, B lymphoma, leukemia;

The metabolic diseases are: hyperglycemia, diabetes, obesity, atherosclerosis, metabolic syndrome, liver fibrosis, non-alcoholic fatty liver, gallstones, hyperlipidemia, hypercholesterolemia, hyperlipoproteinemia Symptoms, hypertriglyceridemia, hypertension, hyperinsulinemia, hyperuricemia, Parkinson's disease, Alzheimer's disease.

The present invention also provides a pharmaceutical composition for preventing or treating diseases related to abnormal expression of ERRα protein activity.

The specific technical solutions are as follows:

A pharmaceutical composition for preventing and/or treating diseases related to abnormal expression of ERRα protein activity, prepared from active ingredients and pharmaceutically acceptable excipients, the active ingredients comprising the above-mentioned indolinone compounds or their pharmaceutically acceptable ingredients. Acceptable salts or stereoisomers or prodrug molecules thereof.

Based on the above technical solutions, the present invention has the following beneficial effects:

After extensive and in-depth research, the inventors of the present application have unexpectedly developed a novel indolinone compound for the first time, which can exhibit nanomolar ERRα inhibitory activity in time-resolved-fluorescence resonance energy transfer experiments. The indolinone compounds of the present invention can effectively inhibit the combination of estrogen-related receptor alpha (ERRα) and co-activator PGC-1α, are a new class of ERRα inverse agonists, and can be used for preventing or treating the activity of ERRα protein Diseases related to abnormal expression, such as tumors or metabolic diseases.

Description of drawings

Figure 1 is a graph showing the blood glucose curve of DB/DB mice after continuous administration for 28 days in the OGTT experiment.

Detailed ways

In the compounds of the present invention, when any variable (eg, R, etc.) occurs more than once in any component, its definition at each occurrence is independent of the definition at each other occurrence. Likewise, combinations of substituents and variables are permissible so long as such combinations stabilize the compound. A line drawn into a ring system from a substituent indicates that the indicated bond may be attached to any substitutable ring atom. If the ring system is polycyclic, it means that such bonds are only attached to any suitable carbon atoms adjacent to the ring. It will be appreciated that one of ordinary skill in the art can select substituents and substitution patterns for the compounds of the present invention to provide compounds that are chemically stable and readily synthesized from readily available starting materials by the skill of the art and the methods set forth below. If a substituent is itself substituted with more than one group, it is understood that these groups may be on the same carbon atom or on different carbon atoms, so long as the structure is stabilized.

As used herein, the phrase " _Rf substituted", "R substituted" is considered equivalent to the phrase "substituted with at least one substituent" and in this case preferred embodiments will have 1-4 substituents.

The term "alkyl" as used herein is meant to include branched and straight chain saturated aliphatic hydrocarbon groups having the specified number of carbon atoms. For example, the definition of " _C1 - _C6 " in " _C1 - _C6 alkyl" includes groups having 1, 2, 3, 4, 5 or 6 carbon atoms in a straight or branched chain arrangement. For example, " _C1 - _C6 alkyl" specifically includes methyl, ethyl, n-propyl, isopropyl, n-butyl, tert-butyl, isobutyl, pentyl, hexyl. The term "cycloalkyl" refers to a monocyclic saturated aliphatic hydrocarbon group having the specified number of carbon atoms. For example, "cycloalkyl" includes cyclopropyl, cyclobutyl, cyclopentyl or cyclohexyl, and the like. The term "alkoxy" refers to groups having an -O-alkyl structure, such as -OCH ₃ , -OCH ₂ CH ₃ , -OCH ₂ CH ₂ CH ₃ , -O-CH ₂ CH(CH ₃ ) ₂ , - OCH ₂ CH ₂ CH ₂ CH ₃ , -O-CH(CH ₃ ) ₂ and the like. The term "heteroaryl" refers to an aromatic ring containing 1, 2 or 3 heteroatoms selected from O, N or S, heteroaryl groups within the scope of the present invention include, but are not limited to: imidazolyl, triazolyl, pyrazole radical, furyl, thienyl, oxazolyl, isoxazolyl, pyrazinyl, pyridazinyl, pyridyl, pyrimidinyl, pyrrolyl.

As understood by those skilled in the art, "halo" ("halo") or "halo" as used herein means chlorine, fluorine, bromine and iodine.

是指方向不固定的单键，表示所述化合物包括顺式结构和反式结构两种异构体。例如，本发明的式(I)化合物

包括如下两种顺反异构体：

The chemical bond in the chemical structural formula of the present invention

It refers to a single bond whose direction is not fixed, indicating that the compound includes two isomers of cis-structure and trans-structure. For example, compounds of formula (I) of the present invention

Including the following two cis-trans isomers:

包括如下两种顺反异构体：For another example,

Including the following two cis-trans isomers:

That is, the 2-indolone compounds of the present invention include (Z)-2-indolone compounds and (E)-2-indolone compounds, and a single (Z)-2-indolone can be obtained by separation Ketone compounds or (E)-2-indolone compounds, but these two isomers will be transformed into each other in chemical solvents, that is, the 2-indolone compounds of the present invention are in chemical solvents. It exists in the mixed form of cis-trans isomer interconversion, that is, in the mixed form of (Z)-2-indolone compounds and (E)-2-indolone compounds. The "chemical solvent" includes, but is not limited to, dimethyl sulfoxide, methanol, ethanol, isopropanol, ethyl acetate, dichloromethane, chloroform, acetone, acetonitrile, and water.

The present invention provides a compound of formula I, its pharmaceutically acceptable salt or stereoisomer or its prodrug molecule,

in,

m is selected from: 0, 1 and 2;

选自：s个R₀取代或者未取代的C₆-C₁₀芳基、s个R₀取代或者未取代的C₅-C₁₀杂芳基；ring

Selected from: s R ₀ substituted or unsubstituted C ₆ -C ₁₀ aryl, s R ₀ substituted or unsubstituted C ₅ -C ₁₀ heteroaryl;

R ₁ , R ₂ , R ₃ , R ₄ , R ₅ , R ₆ , R ₇ , R ₈ , R ₀ are each independently selected from: hydrogen, halogen, cyano, nitro, amino, hydroxy, _Rf substituted or Unsubstituted C ₁ -C ₆ alkyl, R _f substituted or unsubstituted C ₃ -C ₆ cycloalkyl, R _f substituted or unsubstituted C ₁ -C ₆ alkoxy, R _f substituted or unsubstituted C ₃ -C ₆ cycloalkoxy, R _f substituted or unsubstituted C ₂ -C ₆ alkenyl, R _f substituted or unsubstituted C ₂ -C ₆ alkynyl, R _f substituted or unsubstituted phenyl, -(CH ₂ ) _p NR _a R _b , -C(=O)OR, -CONHR, or R ₃ and R ₅ are linked together by chemical bonds to form -O-, -NH-, or -(CH ₂ ) _n ;

_R9 is selected from: hydrogen, -( _{CH2 ) qNRdRe} , -( _{CH2 ) qORd} , - _{( CH2 ) qOCRcRdRe , - ( CH2 ) qCRcRd Re} ;

s is selected from: an integer between 0 and 5;

n is selected from: 1 or 2;

p is selected from; an integer between 0 and 6;

q is selected from: an integer between 1 and 6;

R _a and R _b are each independently selected from: hydrogen, R _f substituted or unsubstituted C ₁ -C ₆ alkyl, R _f substituted or unsubstituted C ₁ -C ₆ heteroalkyl, or R _a , R _b together with the N atom to which it is attached to form R _substituted or unsubstituted heteroatom-containing monocyclic, fused, spiro or bridged rings;

R is selected from: hydrogen, C ₁ -C ₆ alkyl;

R _c is selected from: hydrogen, C ₁ -C ₃ alkyl;

R _d and R _e are each independently selected from: hydrogen, R _f substituted or unsubstituted C ₁ -C ₆ alkyl, R _f substituted or unsubstituted C ₁ -C ₆ heteroalkyl, or R _d , R _e together with the N atom or C atom to which it is attached, to form an R _substituted or unsubstituted heteroatom-containing monocyclic, fused, spiro or bridged ring;

Each R _f is independently selected from: halogen, hydroxy, amino, C ₁ -C ₃ alkyl, hydroxy substituted C ₁ -C ₃ alkyl, C ₁ -C ₃ alkoxy, -NH(C ₁ -C ₃ alkyl), -N(C ₁ -C ₃ alkyl) (C ₁ -C ₃ alkyl), -C(=O)(C ₁ -C ₃ alkyl), R substituted or unsubstituted 5- 7-membered heterocyclyl.

The present invention includes free forms of compounds of formula (I), as well as pharmaceutically acceptable salts and stereoisomers thereof. Some of the specific exemplary compounds herein are protonated salts of amine compounds. The term "free form" refers to the amine compound in non-salt form. Included pharmaceutically acceptable salts include not only exemplary salts of the particular compounds described herein, but also typical pharmaceutically acceptable salts of all compounds of formula (I) in free form. The free forms of specific salts of the compounds can be isolated using techniques known in the art. For example, the free form can be regenerated by treating the salt with a suitable dilute aqueous base, such as dilute aqueous NaOH, dilute aqueous potassium carbonate, dilute aqueous ammonia, and dilute aqueous sodium bicarbonate. The free forms differ somewhat from their respective salt forms in certain physical properties such as solubility in polar solvents, but for the purposes of the invention such acid and base salts are otherwise pharmaceutically equivalent to their respective free forms.

The pharmaceutically acceptable salts of the present invention can be synthesized from compounds of the present invention containing a basic or acidic moiety by conventional chemical methods. Generally, salts of basic compounds are prepared by ion exchange chromatography or by reacting the free base and a stoichiometric amount or excess of an inorganic or organic acid in the desired salt form in a suitable solvent or combination of solvents. Similarly, salts of acidic compounds are formed by reaction with a suitable inorganic or organic base.

Accordingly, pharmaceutically acceptable salts of the compounds of the present invention include conventional non-toxic salts of the compounds of the present invention formed by reacting a basic compound of the present invention with an inorganic or organic acid. For example, conventional non-toxic salts include those derived from inorganic acids such as hydrochloric acid, hydrobromic acid, sulfuric acid, sulfamic acid, phosphoric acid, nitric acid, and the like, as well as those derived from organic acids such as acetic acid, propionic acid, succinic acid, glycolic acid, hard Fatty acid, lactic acid, malic acid, tartaric acid, citric acid, ascorbic acid, pamoic acid, maleic acid, hydroxymaleic acid, phenylacetic acid, glutamic acid, benzoic acid, salicylic acid, p-aminobenzenesulfonic acid, 2-acetyl Salts prepared from oxymonobenzoic acid, fumaric acid, toluenesulfonic acid, methanesulfonic acid, ethanedisulfonic acid, oxalic acid, isethionic acid, trifluoroacetic acid, etc.

If the compound of the present invention is acidic, appropriate "pharmaceutically acceptable salts" refer to salts prepared from pharmaceutically acceptable non-toxic bases including inorganic and organic bases. Salts derived from inorganic bases include aluminum salts, ammonium salts Salts, calcium salts, copper salts, iron salts, ferrous salts, lithium salts, magnesium salts, manganese salts, manganous salts, potassium salts, sodium salts, zinc salts, etc. Ammonium, calcium, magnesium, potassium and sodium salts are particularly preferred. Salts derived from pharmaceutically acceptable organic non-toxic bases including salts of primary, secondary and tertiary amines, substituted amines including naturally occurring substituted amines, cyclic amines and basic ion exchange resins such as Amino acid, betaine, caffeine, choline, N,N'-dibenzylethylenediamine, diethylamine, 2-diethylaminoethanol, 2-dimethylaminoethanol, aminoethanol, ethanolamine, ethyl Diamine, N-Ethylmorpholine, N-Ethylpiperidine, Glucosamine, Glucosamine, Histidine, Hydroxocobalamin, Isopropylamine, Lysine, Methylglucamine, Morpholine, Piperazine , piperidine, quack, polyamine resin, procaine, purine, theobromine, triethylamine, trimethylamine, tripropylamine, tromethamine, etc.

Berg et al., "Pharmaceutical Salts" J. Pharm. Sci. '1977:66:1-19 describes the preparation of the pharmaceutically acceptable salts described above and other typical pharmaceutically acceptable salts in more detail.

Since deprotonated acidic moieties such as carboxyl groups in compounds can be anionic under physiological conditions, such charged charges can then be replaced by internally cationic protonated or alkylated basic moieties such as tetravalent The nitrogen atoms balance out, so it should be noted that the compounds of the present invention are potential inner salts or zwitterions.

In addition to standard methods known in the literature or exemplified in experimental procedures, the compounds of this invention can be prepared using reactions as shown in the following schemes. Accordingly, the following illustrative schemes are for illustrative purposes and are not limited to the listed compounds or any particular substituents. The numbers of substituents shown in the schemes do not necessarily correspond to the numbers used in the claims, and for clarity, mono-substituents are shown attached to compounds that allow for multi-substituents under the definition of formula (I) above.

The compound of formula (I) as described in the present invention can be prepared by the following reaction scheme:

的定义同前述发明内容所述；A选自：氯、溴、碘。wherein R ₁ , R ₂ , R ₃ , R ₄ , R ₅ , R ₆ , R ₇ , R ₈ , R ₉ , m and

The definition of A is the same as described in the foregoing content of the invention; A is selected from the group consisting of chlorine, bromine and iodine.

Wherein, the compound of formula (VI) and the compound of formula (VII) undergo a nucleophilic substitution reaction under basic conditions to obtain an ether compound of formula (VIII). The base used in the reaction conditions can be potassium carbonate, cesium carbonate, sodium carbonate, triethylamine, etc., and the solvent used can be acetonitrile, N,N-dimethylformamide, N,N-dimethylacetamide, dimethylformamide, etc. One or more of the sulfoxides, the reaction temperature is preferably 80-120° C., and the reaction time is 5-12 hours.

The compound of the formula (VIII) and the indolinone compound of the formula (VIII) undergo an aldol condensation reaction under acid or basic catalysis conditions to obtain the α-β unsaturated indolinone compound of the formula (X). The basic catalysts used in the reaction conditions can be oxides, hydroxides, bicarbonates, carbonates and carboxylates of alkali metals or alkaline earth metals, organic amine compounds and anion exchange resins, such as sodium carbonate, hydrogen carbonate, etc. Sodium, sodium acetate, sodium hydroxide, calcium hydroxide, sodium hydride, sodium alkoxide, triethylamine, diethylamine, tri-n-butylamine, tripropylamine, diisopropylamine, diisopropylethylamine ( DIPEA), trimethylamine, piperidine, pyrrolidine, etc.; the acid catalyst used in the reaction conditions can be sulfuric acid, hydrochloric acid, p-toluenesulfonic acid, cation exchange resin, boron trifluoride, etc., and the solvent used can be methanol, ethanol, etc., The reaction temperature is preferably 60 to 80°C, and the reaction time is 2 to 5 hours.

The compound of formula (X) and the compound of formula (XI) undergo a nucleophilic substitution reaction under basic conditions to obtain the target product of formula (I). The base used in the reaction conditions can be potassium carbonate, cesium carbonate, sodium carbonate, triethylamine, etc., and the solvent used can be acetonitrile, N,N-dimethylformamide, N,N-dimethylacetamide, dimethylformamide, etc. One or more of the sulfoxides, the reaction temperature is preferably 80-120° C., and the reaction time is 5-12 hours.

The salt of the compound of formula XI and the salt of the compound of formula I are selected from inorganic acid salts or organic acid salts, such as hydrochloride, sulfate, hydrobromide, phosphate, nitrate, acetate, maleate , p-toluenesulfonate, mesylate or trifluoroacetic acid.

In one embodiment, the present application provides a compound with the structure represented by formula (I) and pharmaceutically acceptable salts thereof for the treatment of human or other mammalian tumors and other transitional proliferative diseases or symptoms, metabolic diseases or symptoms Methods.

In one embodiment, the compounds designed in the present application and their pharmaceutically acceptable salts can be used for the treatment or control of non-small cell lung cancer, small cell lung cancer, lung adenocarcinoma, lung squamous cell carcinoma, pancreatic cancer, breast cancer, prostate cancer , liver cancer, skin cancer, epithelial cell cancer, gastrointestinal stromal tumor, endometrial cancer, histiocytic lymphoma, nasopharyngeal cancer, head and neck tumor, colon cancer, rectal cancer, glioma, malignant melanoma, Kidney cancer, bladder cancer, ovarian cancer, cervical cancer, throat cancer or multiple myeloma, B lymphoma, leukemia, hyperglycemia, diabetes, obesity, atherosclerosis, metabolic syndrome, liver fibrosis, non-alcoholic Fatty liver, gallstones, hyperlipidemia, hypercholesterolemia, hyperlipoproteinemia, hypertriglyceridemia, hypertension, hyperinsulinemia, hyperuricemia, Parkinson's disease, Alzheimer's disease Wait.

Drug Metabolites and Prodrugs

The metabolites of the compounds involved in the present application and their pharmaceutically acceptable salts, as well as prodrugs that can be converted into the structures of the compounds involved in the present application and their pharmaceutically acceptable salts in vivo, are also included in the claims of the present application middle.

pharmaceutical composition

The present invention also provides a pharmaceutical composition comprising the active ingredient in a safe and effective amount, and a pharmaceutically acceptable carrier.

The "active ingredient" of the present invention includes the compounds of formula I of the present invention.

The "active ingredients" and pharmaceutical compositions of the present invention can be used as ERRα inverse agonists. In another preferred example, it is used to prepare a medicament for preventing and/or treating tumors/metabolic diseases.

A "safe and effective amount" refers to an amount of the active ingredient sufficient to significantly improve the condition without causing serious side effects. Typically, the pharmaceutical composition contains 1-2000 mg of active ingredient/dose, more preferably 10-200 mg of active ingredient/dose. Preferably, the "one dose" is one tablet.

"Pharmaceutically acceptable carrier" refers to one or more compatible solid or liquid filler or gel substances which are suitable for human use and which must be of sufficient purity and sufficiently low toxicity.

"Compatibility" as used herein means that the components of the composition can be blended with the active ingredients of the present invention and with each other without significantly reducing the efficacy of the active ingredients.

)、润湿剂(如十二烷基硫酸钠)、着色剂、调味剂、稳定剂、抗氧化剂、防腐剂、无热原水等。Examples of pharmaceutically acceptable carrier moieties include cellulose and its derivatives (such as sodium carboxymethyl cellulose, sodium ethyl cellulose, cellulose acetate, etc.), gelatin, talc, solid lubricants (such as stearic acid) , magnesium stearate), calcium sulfate, vegetable oils (such as soybean oil, sesame oil, peanut oil, olive oil, etc.), polyols (such as propylene glycol, glycerol, mannitol, sorbitol, etc.), emulsifiers (such as

), wetting agents (such as sodium lauryl sulfate), colorants, flavors, stabilizers, antioxidants, preservatives, pyrogen-free water, etc.

In another preferred embodiment, the compound of formula I of the present invention can form a complex with a macromolecular compound or macromolecule through non-bonding interaction. In another preferred embodiment, the compound of formula I of the present invention, as a small molecule, can also be linked with a macromolecular compound or a macromolecule through chemical bonds. The macromolecular compounds can be biological macromolecules such as polysaccharides, proteins, nucleic acids, polypeptides, and the like.

The mode of administration of the active ingredient or pharmaceutical composition of the present invention is not particularly limited, and representative modes of administration include (but are not limited to): oral, intratumoral, rectal, parenteral (intravenous, intramuscular or subcutaneous) and the like.

Solid dosage forms for oral administration include capsules, tablets, pills, powders and granules.

In these solid dosage forms, the active ingredient is mixed with at least one conventional inert excipient (or carrier), such as sodium citrate or dicalcium phosphate, or with the following:

(a) fillers or extenders such as starch, lactose, sucrose, glucose, mannitol and silicic acid;

(b) binders such as hydroxymethylcellulose, alginate, gelatin, polyvinylpyrrolidone, sucrose and acacia;

(c) humectants, for example, glycerin;

(d) disintegrants, for example, agar, calcium carbonate, potato or tapioca starch, alginic acid, certain complex silicates, and sodium carbonate;

(e) slow solvents, for example, paraffin;

(f) absorption accelerators, for example, quaternary amine compounds;

(g) humectants such as cetyl alcohol and glyceryl monostearate;

(h) adsorbents, for example, kaolin; and

(i) Lubricants such as talc, calcium stearate, magnesium stearate, solid polyethylene glycol, sodium lauryl sulfate, or mixtures thereof. In capsules, tablets and pills, the dosage form may also contain buffering agents.

The solid dosage forms can also be prepared using coatings and shell materials, such as enteric coatings and other materials known in the art. They may contain opacifying agents, and the release of the active ingredient in such compositions may be in a certain part of the digestive tract in a delayed manner. Examples of embedding components that can be employed are polymeric substances and waxes.

Liquid dosage forms for oral administration include pharmaceutically acceptable emulsions, solutions, suspensions, syrups or tinctures. In addition to the active ingredient, liquid dosage forms may contain inert diluents conventionally employed in the art, such as water or other solvents, solubilizers and emulsifiers, for example, ethanol, isopropanol, ethyl carbonate, ethyl acetate, propylene glycol, 1 , 3-butanediol, dimethylformamide and oils, especially cottonseed oil, peanut oil, corn germ oil, olive oil, castor oil and sesame oil or mixtures of these substances, and the like. Besides these inert diluents, the compositions can also contain adjuvants such as wetting agents, emulsifying and suspending agents, sweetening, flavoring and perfuming agents.

In addition to the active ingredient, suspensions may contain suspending agents such as ethoxylated isostearyl alcohols, polyoxyethylene sorbitol and sorbitan esters, microcrystalline cellulose, aluminum methoxide and agar, or mixtures of these substances and the like.

Compositions for parenteral injection may comprise physiologically acceptable sterile aqueous or anhydrous solutions, dispersions, suspensions or emulsions, and sterile powders for reconstitution into sterile injectable solutions or dispersions. Suitable aqueous and non-aqueous carriers, diluents, solvents or excipients include water, ethanol, polyols and suitable mixtures thereof.

The compounds of the present invention may be administered alone or in combination with other therapeutic agents such as hypoglycemic agents.

When the pharmaceutical composition is used, a safe and effective amount of the compound of the present invention is suitable for mammals (such as people) in need of treatment, and the dose is a pharmaceutically considered effective dose when administered, and for a person with a body weight of 60kg, the daily dose is The administration dose is usually 1 to 2000 mg, preferably 20 to 500 mg. Of course, the specific dosage should also take into account the route of administration, the patient's health and other factors, which are all within the skill of the skilled physician.

Combination medication

The compounds of formula (I) may be used in combination with other drugs known to treat or ameliorate similar conditions. In the combined administration, the mode of administration and dosage of the original drug remains unchanged, while the compound of formula (I) is administered concurrently or subsequently. When the compound of formula (I) is administered concomitantly with one or more other drugs, it is preferred to use a pharmaceutical composition containing one or more known drugs and the compound of formula I at the same time. Drug combinations also include administration of a compound of formula (I) with one or more other known drugs at overlapping time periods. When the compound of formula (I) is administered in combination with one or more other drugs, the dose of the compound of formula (I) or known drugs may be lower than when they are administered alone.

Drugs or active ingredients that can be used in combination with the compounds of formula (I) include, but are not limited to:

Estrogen receptor modulators, androgen receptor modulators, retinal-like receptor modulators, cytotoxic/cytostatics, antiproliferative agents, protein transferase inhibitors, HMG-CoA reductase inhibitors, HIV protein kinase inhibitors agents, reverse transcriptase inhibitors, angiogenesis inhibitors, cell proliferation and survival signal inhibitors, drugs that interfere with cell cycle checkpoints and apoptosis inducers, cytotoxic drugs, tyrosine protein inhibitors, EGFR inhibitors, VEGFR inhibitor, serine/threonine protein inhibitor, Bcr-Abl inhibitor, c-Kit inhibitor, Met inhibitor, Raf inhibitor, MEK inhibitor, MMP inhibitor, topoisomerase inhibitor, histidine Acid deacetylase inhibitors, proteasome inhibitors, CDK inhibitors, Bcl-2 family protein inhibitors, MDM2 family protein inhibitors, IAP family protein inhibitors, STAT family protein inhibitors, PI3K inhibitors, AKT inhibitors , Integrin blocker, interferon-α, interleukin-12, COX-2 inhibitor, p53, p53 activator, VEGF antibody, EGF antibody, etc.

In one embodiment, drugs or active ingredients that can be used in combination with the compound of formula I include, but are not limited to: aldesleukin, alendronic acid, interferon, atranoin, allopurinol, allopurinol sodium, palonosetron hydrochloride, hexamethylmelamine, aminoglumitide, amifostine, amrubicin, amidine, anastrozole, dolasetron, aranesp, arglabin, arsenic trioxide, Arnoxin, 5-azacytidine, azathioprine, BCG or tic BCG, betadine, betamethasone acetate, betamethasone sodium phosphate preparation, bexarotene, bleomycin sulfate, bromuridine, bortezomib, busulfan, calcitonin, alezolizumab injection, capecitabine, carboplatin, casodex, cefesone, simoleukin, daunorubicin, chlorambucil, cisplatin, cladrix Bine, cladribine, clodronate, cyclophosphamide, cytarabine, dacarbazine, actinomycin D, daunorubicin liposome, dexamethasone, dexamethasone phosphate, estradiol valerate Alcohol, Denisole-2, Dipomet, Deslorelin, Delazoxan, Diethylstilbestrol, Diflucan, Docetaxel, Deoxyfluridine, Doxorubicin, Dronabinol, Chin-166-Shell Glycan complex, eligard, rasburicase, epirubicin hydrochloride, aprepitant, epirubicin, epoetin alfa, erythropoietin, eplatin, levamisole tablets, estradiol preparations, 17-β-estradiol, estramustine sodium phosphate, ethinyl estradiol, amifostine, hydroxyphosphate, fenbifur, etoposide, fadrozole, tamoxifen preparations, filgrastim, Nalasti, Fellastide, Flouridine, Fluconazole, Fludarabine, 5-Fluorodeoxyuridine Monophosphate, 5-Fluorouracil, Fluoxymesterone, Flutamide, Formestan, 1-β-D-arabinofuranocythidine-5'-stearoyl phosphate, fomustine, fulvestrant, gamma globulin, gemcitabine, gemtuzumab, imatinib mesylate Nitrile, Carazepine Wax Paper Capsules, Goserelin, Granisirone Hydrochloride, Histrelin, Hemethicone, Hydrocortisone, Erythro-Hydroxynonyl Adenine, Hydroxyurea, Tetanisibib Momumab, idarubicin, ifosfamide, interferon alpha, interferon-alpha2, interferon alpha-2A, interferon alpha-2B, interferon alpha-nl, interferon alpha-n3, interferon beta , Interferon γ-la, Interleukin-2, Intron A, Iressa, Irinotecan, Kaiteri, Lentinan Sulfate, Letrozole, Leucovorin, Leuprolide, Leuprolide Relin Acetate, Levotetramisole, Calcium Levofolinate, Levothyroxine Sodium, Levothyroxine Sodium Preparation, Lomustine, Lonidamine, Dronabinol, Nitrogen Mustard, Methylcobalamin, Mehydroxyl Progesterone acetate, megestrol acetate, melphalan, esterified estrogen, 6-mercaptopurine, mesna, methotrexate, methyl aminolevulinate, miltefosine, minocycline , mitomycin C, mitotane, mitosodium quinone, trilostane, doxorubicin citrate liposome, nedaplatin, pegylated filgrastim, opreleukin, neupogen, Nilutamide, Tamoxifen, NSC-63157 0. Recombinant human interleukin-1-beta, octreotide, ondansetron hydrochloride, dehydrocortisone oral solution, oxaliplatin, paclitaxel, prednisone sodium phosphate preparation, pegaspargase, perol Xin, pentostatin, streptolytic preparations, pilocarpine hydrochloride, pirorubicin, prukamycin, porfim sodium, prednimustine, steprednisolone, prednisone, Premarin, Procarba, Recombinant Human Erythropoietin, Raltitrexed, Ribbi, Etidronate, Rhenium-186, Rituxan, Rituxan-A, Romotide, Pilocarpine Hydrochloride Tablets, Octreotide, Shamo Stim, Semustine, Sizoran, Sobuzoxan, Methylprednisolone, Paphos Acid, Stem Cell Therapy, Streptozocin, Strontium Chloride-89, Levothyroxine Sodium, Tamoxifen, Tamsulosin, Tasonamin, Tastolactone, Taxotere, Tecethiazine, Temozolomide, Teniposide, Testosterone Propionate, Methyltestosterone, Thioguanine, Thiatepa, Thyroid Stimulating Hormone, Tiludronate acid, topotecan, toremifene, tosilimumab, trastuzumab, triosulfan, tretinoin, methotrexate tablets, trimethylmelamine, trimetrexate, acetic acid Triptorelin, Triptorelin pamoate, Eufodine, Uridine, Valrubicin, Vesrinone, Vinblastine, Vincristine, Vincamide, Vinorelbine, Velulizine, Dextropropimide, net statin, zofenin, paclitaxel protein stabilizer, acolbifene, interferon r-lb, affinitak, aminopterin, azoxifene, asoprisnil, atamestane, atraxen Tan, BAY43-9006, Avastin, CCI-779, CDC-501, Celebrex, Cetuximab, Crinator, Cyproterone Acetate, Decitabine, DN-101, Doxorubicin -MTC, dSLIM, dutasteride, edotecarin, eflunithine, ixitecan, fenretinide, histamine dihydrochloride, histrelin hydrogel implant, holmium-166DOTMP, ibandronic acid, interferon gamma, intron-PEG, ixabepilone, keyhole limpet hemocyanin, L-651582, lanreotide, lasoxifene, libra, lonafamib, milpredoxifene, minoxifene Ester, MS-209, liposomal MTP-PE, MX-6, nafarelin, naimrubicin, novarestat, nolatrexide, olimerson, onco-TCS, osidem, paclitaxel Polyglutamic acid ester, sodium bromide, PN-401, QS-21, quasiyang, R-1549, raloxifene, leopardinase, 13-cis-retinoic acid, saplatin, theocalcidol , T-138067, tarceva, docosahexaenoate paclitaxel, thymosin alfa, gazofurin, tipifarnib, tirapazamine, TLK-286, toremifene, trans MID-lo7R, valspor Dalta, vaprotide, vatalanib, verteporfin, vinflunine, Z-100 and zolelinic acid or combinations thereof.

The above features mentioned in the present invention or the features mentioned in the embodiments can be combined arbitrarily. All features disclosed in this specification may be used in combination with any composition, and each feature disclosed in the specification may be replaced by any alternative feature serving the same, equivalent or similar purpose. Therefore, unless otherwise stated, the disclosed features are only general examples of equivalent or similar features.

The benefits of the present invention are:

(1) A novel 2-indolone compound is provided.

(2) The compounds can effectively inhibit the combination of estrogen-related receptor α and co-activator PGC-1α, and can be used for preparing anti-tumor drugs and/or drugs for treating metabolic diseases.

The present invention will be further described below in conjunction with specific embodiments. It should be understood that these examples are only used to illustrate the present invention and not to limit the scope of the present invention. The experimental method of unreceipted specific conditions in the following examples, usually according to conventional conditions such as people such as Sambrook, molecular cloning: the conditions described in the laboratory manual (New York:Cold Spring Harbor LaboratoryPress, 1989), or according to the manufacturer's recommended conditions. Percentages and parts are by weight unless otherwise indicated.

Unless otherwise defined, all professional and scientific terms used herein have the same meanings as those familiar to those skilled in the art. In addition, any methods and materials similar or equivalent to those described can be used in the methods of the present invention. Methods and materials for preferred embodiments described herein are provided for illustrative purposes only.

Example 1 4-(4-((5-Fluoro-1-(2-ethylmorpholine)-2-indolone-3-ylide)methyl)-2-methoxyphenoxy)-3 -Trifluoromethylbenzonitrile (LS 1-86)

4-(4-((5-fluoro-1-(2-morpholinoethyl)-2-oxoindolin-3-ylidene)methyl)-2-methoxyphenoxy)-3-(trifluoromethyl)benzonitrile

Step 1 4-(4-Aldol-2-methoxyphenoxy)-3-(trifluoromethyl)benzonitrile (3)

4-Fluoro-3-(trifluoromethyl)benzonitrile (1,5g, 26.5mmol), vanillin (2,4g, 26mmol), potassium carbonate (6.1g, 44mmol) were dissolved in anhydrous N,N- In dimethylformamide (50 mL), the mixture was stirred at room temperature overnight. When the reaction was complete, ice water (200 mL) was added under stirring, and a large amount of solid was precipitated. Filtration under reduced pressure and drying in vacuo gave a white solid (7.6 g, yield 92%).

¹ H NMR (400MHz, DMSO-d ₆ ): δ 10.02 (s, 1H), 8.36 (d, J=2.0Hz, 1H), 8.04 (dd, J=8.4, 2.0Hz, 1H), 7.72 (d ,J=2.0Hz,1H),7.66(dd,J=8.0,2.0Hz,1H),7.45(d,J=8.4Hz,1H),6.95(d,J=8.8Hz,1H),3.83(s , 3H).

MS(ESI): m/z 322[M+H] ⁺ .

Step 2 4-(4-((5-Fluoro-1-H-2-indolone-3-ylide)methyl)-2-methoxyphenoxy)-3-trifluoromethylbenzonitrile (LS 1-63)

4-(4-((5-fluoro-2-oxoindolin-3-ylidene)methyl)-2-methoxyphenoxy)-3-(trifluoromethyl)benzonitrile

Combine 4-(4-aldehyde-2-methoxyphenoxy)-3-(trifluoromethyl)benzonitrile (3, 1.08 g, 3.3 mmol) and 5-fluoroindolinone (4, 0.48 g, 3.2 mmol) was dissolved in 20 mL of absolute ethanol, and stirred until the reaction system was clear. After adding a catalytic amount of piperidine dropwise to the reaction system, the reaction temperature rose to 60 degrees Celsius. After the reaction was completed, it was cooled, and a large amount of yellow solid was precipitated. Filtration under reduced pressure and vacuum drying gave a yellow solid (1.28 g, yield 90%).

Isomer A ¹ H NMR (400 MHz, DMSO-d ₆ ): δ 10.69 (s, 1H), 8.77 (s, 1H), 8.34 (s, 1H), 8.03 (d, J=8.8 Hz, 1H) ,7.98-7.90(m,2H),7.67(dd,J=8.8Hz,2.0Hz,1H),7.33(d,J=8.4Hz,1H),7.10-7.02(m,1H),6.94(d, J=8.8Hz, 1H), 6.83(dd, J=8.4, 4.4Hz, 1H), 3.81(s, 3H).

Isomer B ¹ H NMR (400 MHz, DMSO-d ₆ ): δ 10.68 (d, J=7.2 Hz, 1H), 8.35 (d, J=2.0 Hz, 1H), 8.06 (dd, J=8.8, 2.0Hz, 1H), 7.73(s, 1H), 7.61(d, J=2.0Hz, 1H), 7.48-7.37(m, 3H), 7.18-7.08(m, 1H), 7.02(d, J=8.8 Hz,1H),6.89(dd,J=8.8,4.8Hz,1H),3.78(s,3H).

MS(ESI): m/z 455[M+H] ⁺ .

Step 3 4-(4-((5-Fluoro-1-(2-ethylmorpholine)-2-indolone-3-ylide)methyl)-2-methoxyphenoxy)-3- Trifluoromethylbenzonitrile (LS 1-86)

4-(4-((5-fluoro-1-(2-morpholinoethyl)-2-oxoindolin-3-ylidene)methyl)-2-methoxyphenoxy)-3-(trifluoromethyl)benzonitrile

4-(4-((5-Fluoro-1-H-2-indolone-3-ylide)methyl)-2-methoxyphenoxy)-3-trifluoromethylbenzonitrile ( LS 1-63, 0.50 g, 1.1 mmol), 4-(2-chloroethyl)morpholine hydrochloride (5, 0.41 g, 2.2 mmol) and potassium carbonate (0.46 g, 3.3 mmol) were dissolved in 20 mL of water in acetonitrile, heated to 90 degrees Celsius and reacted overnight. Cool to room temperature, spin dry most of the acetonitrile solvent, extract three times with dichloromethane, combine the organic phases, wash with saturated brine, spin dry, and separate by column chromatography to obtain a yellow solid (0.47 g, 76%).

¹ H NMR (400MHz, DMSO-d ₆ ) δ 8.75(s, 1H), 8.34(s, 2H), 8.13-7.93(m, 4H), 7.81(s, 1H), 7.73(d, J=8.8 Hz,1H),7.62(s,1H),7.46(d,J＝8.8Hz,2H),7.39(d,J＝8.0Hz,1H),7.34(d,J＝8.4Hz,1H),7.26- 7.06(m, 4H), 7.02(d, J=8.4Hz, 1H), 6.94(d, J=8.8Hz, 1H), 3.95-3.83(m, 4H), 3.81(s, 3H), 3.77(s ,3H),3.62-3.46(m,8H),2.54(s,4H),2.45(s,8H).

Further column chromatography separation was obtained:

Isomer A ¹ H NMR (400 MHz, DMSO-d ₆ ): δ 8.75 (d, J=1.6 Hz, 1H), 8.34 (d, J=2.0 Hz, 1H), 8.06-7.99 (m, 2H) ,7.97(dd,J＝8.8,1.7Hz,1H),7.73(dd,J＝8.8,2.4Hz,1H),7.35(d,J＝8.4Hz,1H),7.16(td,J＝8.8,2.4 Hz, 1H), 7.09(dd, J＝8.4, 4.4Hz, 1H), 6.95(d, J＝8.4Hz, 1H), 3.89(t, J＝6.8Hz, 2H), 3.81(s, 3H), 3.51(t, J＝4.0Hz, 4H), 2.54(t, J＝6.8Hz, 2H), 2.48-2.37(m, 4H).

Isomer B ¹ H NMR (400 MHz, DMSO-d ₆ ): δ 8.34 (d, J=1.6 Hz, 1H), 8.05 (dd, J=8.8, 2.0 Hz, 1H), 7.81 (s, 1H) ,7.62(d,J＝2.0Hz,1H),7.49-7.43(m,2H),7.39(d,J＝8.4Hz,1H),7.21(td,J＝8.8,2.8Hz,1H),7.15( dd, J=8.8, 4.4Hz, 1H), 7.02 (d, J=8.8Hz, 1H), 3.89 (t, J=6.4Hz, 2H), 3.77 (s, 3H), 3.53 (t, J=4.4 Hz, 4H), 2.55(t, J＝6.4Hz, 2H), 2.47-2.39(m, 4H).

HRMS(ESI): m/z 568.1838[M+H] ⁺ .

Example 2 4-(2-methoxy-4-((2-indolone-3-ylide)methyl)phenoxy)-3-trifluoromethylbenzonitrile (LS 1-61)

4-(2-methoxy-4-((2-oxoindolin-3-ylidene)methyl)phenoxy)-3-(trifluoromethyl)benzonitrile

The synthetic method is as in Example 1, step 2.

¹ H NMR (400MHz, DMSO-d ₆ ): δ 10.67(s, 1H), 10.63(s, 0.9H), 8.75(d, J=2.0Hz, 1H), 8.33(d, J=1.6Hz, 1.8H), 8.03(td, J＝8.8, 2.0Hz, 2H), 7.95(dd, J＝8.4, 1.6Hz, 1H), 7.87(s, 1H), 7.73(d, J＝7.6Hz, 1H) ,7.69-7.62(m,1.8H),7.59(d,J=1.6Hz,0.9H),7.43(dd,J=8.8,1.6Hz,0.9H),7.36(d,J=8.4Hz,0.9H) ),7.31(d,J＝8.4Hz,1H),7.27-7.20(m,1.8H),7.05-6.96(m,2H),6.96-6.81(m,3.9H),3.80(s,3H), 3.77(s, 2.7H).

Further separation gave the isomers A ¹ H NMR (400 MHz, DMSO-d ₆ ): δ 10.68 (s, 1H), 8.76 (d, J=2.0 Hz, 1H), 8.34 (d, J=1.6 Hz, 1H) ),8.03(dd,J＝8.4,2.0Hz,1H),7.96(dd,J＝8.8,2.0Hz,1H),7.88(s,1H),7.74(d,J＝7.2Hz,1H),7.32 (d, J=8.4Hz, 1H), 7.24 (td, J=7.6Hz, 1.2Hz, 1H), 7.02 (td, J=7.6Hz, 0.8Hz, 1H), 6.94 (d, J=8.8Hz, 1H), 6.86(d, J=7.6Hz, 1H), 3.81(s, 3H).

MS(ESI): m/z 437[M+H] ⁺ .

Example 3 4-(4-((5-Bromo-1-H-2-indolone-3-ylide)methyl)-2-methoxyphenoxy)-3-trifluoromethylbenzyl Nitrile (LS 1-59)

4-(4-((5-bromo-2-oxoindolin-3-ylidene)methyl)-2-methoxyphenoxy)-3-(trifluoromethyl)benzonitrile

The synthetic method is as in Example 1, step 2.

¹ H NMR (400MHz, DMSO-d ₆ ): δ 10.81(s, 1H), 10.80(s, 0.4H), 8.75(d, J=2.0Hz, 1H), 8.39-8.30(m, 1.4H) ,8.11-7.93(m,3.8H),7.79-7.69(m,1H),7.63(s,0.6H),7.49-7.37(m,2.4H),7.33(d,J=8.4Hz,1H), 6.96(dd,J=17.2,8.8Hz,1.4H),6.90-6.77(m,1.4H),3.80(s,3H),3.79(s,1.2H).

MS(ESI): m/z 515[M+H] ⁺ .

Example 4 4-(4-((5-Fluoro-1-H-2-indolone-3-ylide)methyl)-phenoxy)-3-trifluoromethylbenzonitrile (LS1-108 )

4-(4-((5-fluoro-2-oxoindolin-3-ylidene)methyl)phenoxy)-3-(trifluoromethyl)benzonitrile

The synthetic method is as in Example 1, step 2.

¹ H NMR (400MHz, DMSO-d ₆ ): δ 10.67 (s, 1H), 10.66 (s, 1H), 8.58-8.49 (m, 2H), 8.41 (d, J=2.0Hz, 2H), 8.19 -8.09(m, 2H), 7.93(s, 1H), 7.87-7.79(m, 2H), 7.71(s, 1H), 7.66(dd, J=9.2, 2.8Hz, 1H), 7.40-7.20(m ,7H),7.11(td,J＝8.8,2.4Hz,1H),7.08-6.99(m,1H),6.88(dd,J＝8.8,4.8Hz,1H),6.80(dd,J＝8.8,4.8 Hz, 1H).

MS(ESI): m/z 425[M+H] ⁺ .

Example 5 4-(4-((5-Fluoro-1-H-2-indolone-3-ylide)methyl)-3-methoxyphenoxy)-3-trifluoromethylbenzyl Nitrile (LS 1-104)

4-(4-((5-fluoro-2-oxoindolin-3-ylidene)methyl)-3-methoxyphenoxy)-3-(trifluoromethyl)benzonitrile

The synthetic method is as in Example 1, step 2.

Column chromatography separation yields:

Isomer A ¹ H NMR (400 MHz, DMSO-d ₆ ): δ 10.58 (s, 1H), 8.61 (d, J=8.4 Hz, 1H), 8.40 (d, J=1.6 Hz, 1H), 8.13 (dd, J＝8.8, 2.4Hz, 1H), 7.93(s, 1H), 7.58(dd, J＝8.8, 2.8Hz, 1H), 7.25(d, J＝8.8Hz, 1H), 7.08-6.99( m,1H),6.98(d,J＝2.4Hz,1H),6.79(dd,J＝8.4Hz,4.4Hz,1H),6.74(dd,J＝8.8Hz,2.4Hz,1H),3.90(s , 3H).

Isomer B ¹ H NMR (400 MHz, DMSO-d ₆ ): δ 10.63 (s, 1H), 8.41 (d, J=1.6 Hz, 1H), 8.15 (dd, J=8.8, 2.0 Hz, 1H) ,7.78(d,J＝8.4Hz,1H),7.67(s,1H),7.33(d,J＝8.8Hz,1H),7.19(dd,J＝9.2,2.4Hz,1H),7.15-7.04( m,2H),6.93-6.81(m,2H),3.87(s,3H).

MS(ESI): m/z 455[M+H] ⁺ .

Example 6 4-(4-((5-Fluoro-1-H-2-indolone-3-ylide)methyl)-3-hydroxyphenoxy)-3-trifluoromethylbenzonitrile ( LS 1-109)

4-(4-((5-fluoro-2-oxoindolin-3-ylidene)methyl)-3-hydroxyphenoxy)-3-(trifluoromethyl)benzonitrile

4-(4-((5-Fluoro-1-H-2-indolone-3-ylide)methyl)-3-methoxyphenoxy)-3-trifluoromethylbenzonitrile ( LS1-104, 30 mg, 0.066 mmol) was dissolved in anhydrous dichloromethane, and 0.5 ml of 1.0 MBBr ₃ dichloromethane solution was slowly added dropwise at 0 degrees Celsius. The reaction was continued for about 3 hours and quenched by dropwise addition of ice water. Dichloromethane was extracted three times, the organic phases were combined, washed with saturated brine, spun dry, and separated by column chromatography to obtain a yellow solid (16 mg, 55%).

¹ H NMR (400MHz, DMSO-d ₆ ): δ 10.78(s, 0.5H), 10.77(s, 1H), 10.65-10.53(m, 1.5H), 8.76(d, J=8.8Hz, 0.5H ),8.41(d,J＝2.0Hz,1H),8.20-8.10(m,1.5H),7.95(s,0.5H),7.74(d,J＝8.4Hz,1H),7.70(s,1H) ,7.52(dd,J＝8.8,2.4Hz,0.5H),7.37(d,J＝8.8Hz,1H),7.30(d,J＝8.8Hz,0.5H),7.25(dd,J＝9.6,2.8 Hz,1H),7.08(td,J＝9.2,2.8Hz,1H),7.05-6.99(m,0.5H),6.86(dd,J＝8.4,4.4Hz,1H),6.79(dd,J＝8.4 ,4.4Hz,0.5H),6.76(dd,J＝8.0,2.0Hz,1H),6.69(d,J＝2.4Hz,1H),6.65(dd,J＝8.8,2.4Hz,0.5H),6.61 (d, J=2.4Hz, 0.5H), 6.63(s, 0.5H).

MS(ESI): m/z 439[MH] ^- .

Example 7 4-(3-methoxy-4-((4-methyl-2-indolone-3-ylide)methyl)phenoxy)-3-trifluoromethylbenzonitrile (LS 1-128)

4-(3-methoxy-4-((4-methyl-2-oxoindolin-3-ylidene)methyl)phenoxy)-3-(trifluoromethyl)benzonitrile

The synthetic method is as in Example 1, step 2.

Column chromatography separation yields:

Isomer A ¹ H NMR (400 MHz, CDCl ₃ ): δ 8.38 (d, J=9.6 Hz, 1H), 7.99 (d, J=1.6 Hz, 1H), 7.95 (s, 1H), 7.73 (dd , J=8.8, 2.0Hz, 1H), 7.54(s, 1H), 7.17-7.08(m, 2H), 6.84(d, J=8.0Hz, 1H), 6.73-6.64(m, 3H), 3.88( s,3H),2.61(s,3H).

Isomer B ¹ H NMR (400 MHz, CDCl ₃ ): δ 8.00 (d, J=2.0 Hz, 1H), 7.93 (s, 1H), 7.76 (dd, J=8.4, 2.0 Hz, 1H), 7.56 (s,1H),7.21-7.09(m,2H),6.99(d,J=8.4Hz,1H),6.77(d,J=7.6Hz,1H),6.72(d,J=7.6Hz,1H) ,6.68(d,J＝2.0Hz,1H),6.63(dd,J＝8.0,2.0Hz,1H),3.87(s,3H),1.82(s,3H).

MS(ESI): m/z 451[M+H] ⁺ .

Example 8 4-(4-((4-Bromo-2-indolone-3-ylide)methyl)-3-methoxyphenoxy)-3-trifluoromethylbenzonitrile (LS1- 131)

4-(4-((4-bromo-2-oxoindolin-3-ylidene)methyl)-3-methoxyphenoxy)-3-(trifluoromethyl)benzonitrile

The synthetic method is as in Example 1, step 2.

Further column chromatography separation was obtained:

Isomer A ¹ HNMR for (400 MHz, DMSO-d ₆ ): 10.81 (s, 1H), 8.79 (s, 1H), 8.42 (d, J=8.4 Hz, 1H), 8.40 (d, J=2.0 Hz ,1H),8.13(dd,J＝8.8,2.0Hz,1H),7.25(d,J＝8.8Hz,1H),7.20(dd,J＝8.0,0.8Hz,1H),7.13(t,J＝ 8.0Hz,1H),6.97(d,J=2.0Hz,1H),6.85(dd,J=7.6,1.2Hz,1H),6.73(dd,J=8.4,2.4Hz,1H),3.89(s, 3H).

MS(ESI): m/z 515[M+H] ⁺ .

Example 9 4-(4-((4-Fluoro-2-indolone-3-ylide)methyl)-3-methoxyphenoxy)-3-trifluoromethylbenzonitrile (LS1- 140)

4-(4-((4-fluoro-2-oxoindolin-3-ylidene)methyl)-3-methoxyphenoxy)-3-(trifluoromethyl)benzonitrile

The synthetic method is as in Example 1, step 2.

¹ H NMR (400MHz, DMSO-d ₆ ): δ 10.94-10.80 (m, 1.85H), 8.65 (d, J=8.8Hz, 1H), 8.45-8.36 (m, 1.85H), 8.19-8.10 ( m,1.85H),8.03(d,J=3.2Hz,1H),7.74(s,0.85H),7.39(dd,J=8.4,5.6Hz,0.85H),7.33-7.20(m,3H), 7.18(d,J=8.8Hz,0.85H),7.02-6.95(m,1.85H),6.82(dd,J=10.4,8.4Hz,1H),6.77-6.67(m,4.4H),3.89(s ,3H),3.84(s,2.55H).

MS(ESI): m/z 455[M+H] ⁺ .

Example 10 4-(4-((5-Fluoro-1-(2-ethylmorpholine)-2-indolone-3-ylide)methyl)-3-methoxyphenoxy)-3 -Trifluoromethylbenzonitrile (LS 1-113)

4-(4-((5-fluoro-1-(2-morpholinoethyl)-2-oxoindolin-3-ylidene)methyl)-3-methoxyphenoxy)-3-(trifluoromethyl)benzonitrile

The synthetic method is as in Example 1.

¹ H NMR (400 MHz, CDCl ₃ ): δ 8.78 (d, J=9.2 Hz, 1H), 8.01 (dd, J=8.0, 1.6 Hz, 2H), 7.93 (d, J=11.6 Hz, 2H), 7.80(dd,J＝8.8,2.0Hz,1H),7.75(dd,J＝8.8,2.0Hz,1H),7.71(d,J＝9.2Hz,1H),7.34-7.21(m,3H),7.11 (d, J＝8.4Hz, 2H), 7.05-6.92(m, 3H), 6.79-6.66(m, 4H), 4.19-3.96(m, 4H), 3.95-3.66(m, 14H), 3.06-2.41 (m,12H).

MS(ESI): m/z 568[M+H] ⁺ .

Example 11 4-(4-((4-Bromo-1-(2-ethylmorpholine)-2-indolone-3-ylide)methyl)-3-methoxyphenoxy)-3 -Trifluoromethylbenzonitrile (LS 1-123)

4-(4-((4-bromo-1-(2-morpholinoethyl)-2-oxoindolin-3-ylidene)methyl)-3-methoxyphenoxy)-3-(trifluoromethyl)benzonitrile

The synthetic method is as in Example 1.

¹ H NMR (400 MHz, CDCl ₃ ): δ 8.97 (s, 1H), 8.37 (d, J=9.2 Hz, 1H), 8.07-7.95 (m, 2.2H), 7.80-7.69 (m, 1.6H) ,7.23(d,J＝8.0Hz,2H),7.18-7.08(m,3.2H),6.99(d,J＝8.8Hz,0.6H),6.95-6.79(m,1.6H),6.71-6.56( m, 2.8H), 4.06-3.84(m, 6.2H), 3.83(s, 1.8H), 3.78-3.53(m, 6.4H), 2.77-2.34(m, 9.6H).

MS(ESI): m/z 628[M+H] ⁺ .

Example 12 4-(4-((4-Bromo-1-(2-ethylmorpholine)-2-indolone-3-ylide)methyl)-3-hydroxyphenoxy)-3-tri Fluoromethylbenzonitrile (LS 1-125)

4-(4-((4-bromo-1-(2-morpholinoethyl)-2-oxoindolin-3-ylidene)methyl)-3-hydroxyphenoxy)-3-(trifluoromethyl)benzonitrile

The synthetic method is as in Example 6.

¹ H NMR (400 MHz, CDCl ₃ ): δ 9.95 (s, 1H), 8.88 (s, 1H), 8.00 (d, J=1.6 Hz, 1H), 7.77 (dd, J=8.8, 2.0 Hz, 1H) ),7.56(d,J＝8.4Hz,1H),7.34(dd,J＝8.4,0.8Hz,1H),7.21-7.08(m,2H),6.97(d,J＝7.6Hz,1H),6.82 (d, J=2.4Hz, 1H), 6.76 (dd, J=8.4, 2.4Hz, 1H), 4.02 (t, J=6.8Hz, 2H), 3.66 (t, J=4.4Hz, 4H), 2.66 (t, J=6.4Hz, 2H), 2.53 (t, J=4.4Hz, 4H).

MS(ESI): m/z 612[MH] ^- .

Example 13 4-(4-((4-Fluoro-1-(2-ethylmorpholine)-2-indolone-3-ylide)methyl)-3-methoxyphenoxy)-3 -Trifluoromethylbenzonitrile (LS 1-142)

4-(4-((4-fluoro-1-(2-morpholinoethyl)-2-oxoindolin-3-ylidene)methyl)-3-methoxyphenoxy)-3-(trifluoromethyl)benzonitrile

The synthetic method is as in Example 1.

¹ H NMR (400MHz, CDCl ₃ ): δ 8.56 (d, J=9.2Hz, 0.8H), 8.20 (d, J=3.2Hz, 0.8H), 8.09-7.93 (m, 2.8H), 7.75 ( td, J＝8.8, 2.0Hz, 1.8H), 7.35(dd, J＝9.2, 5.6Hz, 1H), 7.30-7.15(m, 1.8H), 7.11(d, J＝8.8Hz, 0.8H), 7.04(d,J＝8.4Hz,1H),6.79-6.64(m,7.2H),3.97-3.83(m,9H),3.72-3.65(m,7.2H),2.68-2.52(m,10.8H) .

MS(ESI): m/z 568[M+H] ⁺ .

Example 14 4-(4-((4-Fluoro-1-(2-ethylmorpholine)-2-indolone-3-ylide)methyl)-3-hydroxyphenoxy)-3-tri Fluoromethylbenzonitrile (LS 1-143)

4-(4-((4-fluoro-1-(2-morpholinoethyl)-2-oxoindolin-3-ylidene)methyl)-3-hydroxyphenoxy)-3-(trifluoromethyl)benzonitrile

The synthetic method is as in Example 6.

¹ H NMR (400MHz, DMSO-d ₆ ): δ 10.81 (s, 1H), 10.63 (s, 0.5H), 8.81 (d, J=8.8Hz, 1H), 8.45-8.35 (m, 1.5H) ,8.27-8.08(m,2.5H),7.85(s,0.5H),7.41-7.26(m,3H),7.22(d,J=8.4Hz,0.5H),6.99(d,J=8.0Hz, 0.5H),6.95(d,J＝8.0Hz,1H),6.89(dd,J＝10.8,8.4Hz,1H),6.81(dd,J＝10.0,8.4Hz,0.5H),6.73-6.57(m ,3H),3.99-3.79(m,3H),3.64-3.41(m,6H),2.61-2.33(m,9H).

MS(ESI): m/z 552[MH] ^- .

Example 15 4-(4-((5-Fluoro-1-(2-ethylpiperidine)-2-indolone-3-ylide)methyl)-2-methoxyphenoxy)-3 -Trifluoromethylbenzonitrile (LS 1-146)

4-(4-((5-fluoro-2-oxo-1-(2-(piperidin-1-yl)ethyl)indolin-3-ylidene)methyl)-2-methoxyphenoxy)-3-(trifluoromethyl)benzonitrile

The synthetic method is as in Example 1.

¹ H NMR (400MHz, CDCl ₃ ): δ 8.76 (s, 0.8H), 7.99 (dd, J=7.2, 2.0 Hz, 1.8H), 7.86 (s, 1H), 7.73 (dd, J=8.8, 2.0Hz, 1H), 7.68(dd, J＝8.8, 2.0Hz, 0.8H), 7.64(dd, J＝8.4, 2.0Hz, 0.8H), 7.48(s, 1H), 7.41(dd, J＝8.8 ,2.4Hz,1H),7.38-7.24(m,4.2H),7.19(d,J=8.4Hz,0.8H),7.07-6.99(m,1.8H),6.98-6.79(m,3H),4.21 -3.85(m, 6H), 3.81(s, 3H), 2.98-2.24(m, 10.8H), 1.62-1.39(m, 10.8H).

MS(ESI): m/z 566[M+H] ⁺ .

Example 16 4-(4-((5-Fluoro-1-(3-propylpiperidine)-2-indolone-3-ylide)methyl)-2-methoxyphenoxy)-3 -Trifluoromethylbenzonitrile (LS 1-147)

4-(4-((5-fluoro-2-oxo-1-(3-(piperidin-1-yl)propyl)indolin-3-ylidene)methyl)-2-methoxyphenoxy)-3-(trifluoromethyl)benzonitrile

The synthetic method is as in Example 1.

¹ H NMR (400MHz, CDCl ₃ ): δ 8.76 (d, J=1.6Hz, 0.8H), 7.97 (dd, J=7.6, 2.0Hz, 1.8H), 7.83 (s, 1H), 7.71 (dd ,J=8.4,2.0Hz,1H),7.66(dd,J=8.8,2.4Hz,0.8H),7.60(dd,J=8.4,2.0Hz,0.8H),7.46(s,0.8H),7.38 (dd,J=8.8,2.4Hz,1H),7.33-7.20(m,3.6H),7.16(d,J=8.4Hz,0.8H),7.00(td,J=8.4,2.4Hz,2H), 6.95-6.87(m, 1.8H), 6.86-6.78(m, 1.8H), 3.89(s, 2.4H), 3.85(t, J=6.8Hz, 3.6H), 3.79(s, 3H), 2.71- 2.31(m, 10.8H), 2.11-1.88(m, 5.4H), 1.65-1.40(m, 9H).

MS(ESI): m/z 580[M+H] ⁺ .

Example 17 4-(4-((5-Fluoro-1-(3-propylpyrrole)-2-indolone-3-ylide)methyl)-2-methoxyphenoxy)-3- Trifluoromethylbenzonitrile (LS 1-148)

4-(4-((5-fluoro-2-oxo-1-(3-(pyrrolidin-1-yl)propyl)indolin-3-ylidene)methyl)-2-methoxyphenoxy)-3-(trifluoromethyl)benzonitrile

The synthetic method is as in Example 1.

¹ H NMR (400 MHz, CDCl ₃ ): δ 8.68 (d, J=1.6 Hz, 1H), 7.97 (dd, J=6.8, 1.6 Hz, 2H), 7.83 (s, 1H), 7.75-7.60 (m ,3H),7.48(s,1H),7.40(dd,J=8.8,2.4Hz,1H),7.37-7.23(m,3H),7.18(d,J=8.4Hz,1H),7.14-6.96( m,3H),6.95-6.88(m,2H),6.87-6.78(m,2H),3.90(t,J=6.8Hz,4H),3.88(s,3H),3.80(s,3H),3.24 -2.69(m, 12H), 2.27-2.17(m, 4H), 2.07-1.96(m, 8H).

MS(ESI): m/z 566[M+H] ⁺ .

Example 18 4-(4-((5-Fluoro-1-(3-(4-methylpiperazin-1-yl)propyl)-2-indolone-3-ylide)methyl)-2 -Methoxyphenoxy)-3-trifluoromethylbenzonitrile (LS 1-149)

4-(4-((5-fluoro-1-(3-(4-methylpiperazin-1-yl)propyl)-2-oxoindolin-3-ylidene)methyl)-2-methoxyphenoxy)-3-(trifluoromethyl)benzonitrile

The synthetic method is as in Example 1.

¹ H NMR (400 MHz, CDCl ₃ ): δ 8.78 (d, J=2.0 Hz, 1H), 7.97 (dd, J=7.2, 1.6 Hz, 2H), 7.83 (s, 1H), 7.71 (dd, J =8.4,2.0Hz,1H),7.66(dd,J=8.8,2.0Hz,1H),7.60(dd,J=8.4,2.0Hz,1H),7.46(s,1H),7.39(dd,J= 9.2, 2.4Hz, 1H), 7.34-7.27 (m, 2H), 7.25-7.21 (m, 2H), 7.17 (d, J=8.4Hz, 1H), 6.98 (td, J=8.8, 2.0Hz, 2H) ), 6.92-6.77(m, 4H), 3.88(s, 3H), 3.83(t, J=6.8Hz, 4H), 3.79(s, 3H), 2.72-2.24(m, 26H), 1.93-1.86( m,4H).

MS(ESI): m/z 595[M+H] ⁺ .

Example 19 4-(4-((1-(3-(dimethylamino)propyl)-5-fluoro-2-indolone-3-ylide)methyl)-2-methoxyphenoxy yl)-3-trifluoromethylbenzonitrile (LS 1-150)

4-(4-((1-(3-(dimethylamino)propyl)-5-fluoro-2-oxoindolin-3-ylidene)methyl)-2-methoxyphenoxy)-3-(trifluoromethyl)benzonitrile

The synthetic method is as in Example 1.

¹ H NMR (400MHz, CDCl ₃ ): δ 8.76 (d, J=1.6Hz, 0.8H), 7.97 (dd, J=7.6, 1.6Hz, 1.6H), 7.84 (s, 1H), 7.71 (dd , J＝8.8, 2.0Hz, 1H), 7.66(dd, J＝8.8, 2.0Hz, 0.8H), 7.61(dd, J＝8.0, 1.6Hz, 0.8H), 7.46(s, 1H), 7.39( dd,J=8.8,2.4Hz,1H),7.36-7.22(m,3.8H),7.17(d,J=8.4Hz,0.8H),7.10-7.04(m,0.8H),7.00(td,J =8.8,2.4Hz,1.6H),6.95-6.77(m,3H),3.88(m,2.4H),3.85(t,J=6.8Hz,3.6H),3.79(s,3H),2.57-2.36 (m, 3.6H), 2.30(d, J=8.0Hz, 10.8H), 1.97-1.89(m, 3.6H).

MS(ESI): m/z 540[M+H] ⁺ .

Example 20 4-(4-((5-Fluoro-1-(2-ethylmorpholine)-2-indolone-3-ylide)methyl)-2-methoxyphenoxy)-benzene Formonitrile (LS 2-16)

4-(4-((5-fluoro-1-(2-morpholinoethyl)-2-oxoindolin-3-ylidene)methyl)-2-methoxyphenoxy)benzonitrile

The synthetic method is as in Example 1.

¹ H NMR (400MHz, CDCl ₃ ): δ 8.75(d, J=2.0Hz, 0.8H), 7.85(s, 1H), 7.67-7.57(m, 4.4H), 7.47(s, 0.8H), 7.43(dd,J=8.8,2.4Hz,1H),7.32-7.26(m,2.8H),7.17(d,J=8.0Hz,1H),7.10(d,J=8.4Hz,1H),7.05- 6.95(m, 5.4H), 6.86-6.80(m, 1.6H), 3.94(t, J=7.2Hz, 3.6H), 3.93(s, 2.4H), 3.82(s, 3H), 3.76-3.66( m,7.2H),2.71-2.62(m,3.6H),2.61-2.50(m,7.2H).

MS(ESI): m/z 450[M+H] ⁺ .

Example 21 5-Fluoro-3-(3-methoxy-4-(4-nitro-2-(trifluoromethyl)phenoxy)benzylylide)-1-(2-ethylmorpholine ) indol-2-one (LS 2-22)

5-fluoro-3-(3-methoxy-4-(4-nitro-2-(trifluoromethyl)phenoxy)benzylidene)-1-(2-morpholinoethyl)indolin-2-one

The synthetic method is as in Example 1.

¹ H NMR (400 MHz, CDCl ₃ ): δ 8.75 (d, J=2.0 Hz, 1H), 8.58 (dd, J=6.8, 2.8 Hz, 1.9 H), 8.31 (dd, J=9.2, 2.8 Hz, 0.9H), 8.26(dd, J=9.2, 2.8Hz, 1H), 7.84(s, 0.9H), 7.62(dd, J=8.4, 2.0Hz, 1H), 7.46(s, 1H), 7.40(dd , J=8.8, 2.4Hz, 0.9H), 7.32 (dd, J=8.0, 1.2Hz, 1H), 7.30-7.23 (m, 2.8H), 7.19 (d, J=8.4Hz, 0.9H), 7.00 (td, J＝8.4, 2.0Hz, 1.9H), 6.88-6.77(m, 3.8H), 3.91(t, J＝6.8Hz, 3.8H), 3.88(s, 3H), 3.79(s, 2.7H ), 3.69(dd, J=9.6, 6.0Hz, 7.6H), 2.67-2.60(m, 3.8H), 2.59-2.49(m, 7.6H).

MS(ESI): m/z 588[M+H] ⁺ .

Example 225-Fluoro-3-(3-methoxy-4-(2-(trifluoromethyl)phenoxy)benzylylide)-1-(2-ethylmorpholine)indole-2- Ketones (LS 2-24)

5-fluoro-3-(3-methoxy-4-(2-(trifluoromethyl)phenoxy)benzylidene)-1-(2-morpholinoethyl)indolin-2-one

The synthetic method is as in Example 1.

¹ H NMR (400MHz, CDCl ₃ ): δ 8.72 (d, J=1.6Hz, 1H), 7.84 (s, 0.5H), 7.72-7.64 (m, 1.5H), 7.58 (dd, J=8.4, 2.0Hz, 1H), 7.52-7.38(m, 3H), 7.31-7.21(m, 2H), 7.20-7.11(m, 1.5H), 7.10-7.03(m, 0.5H), 7.02-6.93(m, 2.5H), 6.87(d, J＝8.0Hz, 1.5H), 6.84-6.73(m, 1.5H), 3.95(s, 3H), 3.94-3.88(m, 3H), 3.77-3.66(m, 6H) ),3.64(s,1.5H),2.70-2.61(m,3H),2.61-2.48(m,6H).

HRMS(ESI): m/z 543.1883[M+H] ⁺ .

Example 234-(3-Methoxy-4-((1-(2-ethylmorpholine)-2-indolone-3-ylide)methyl)phenoxy)-3-trifluoromethyl Benzonitrile (LS 2-25)

4-(3-methoxy-4-((1-(2-morpholinoethyl)-2-oxoindolin-3-ylidene)methyl)phenoxy)-3-(trifluoromethyl)benzonitrile

The synthetic method is as in Example 1.

¹ H NMR (400MHz, CDCl ₃ ): δ 8.75 (d, J=8.4Hz, 0.5H), 8.00 (d, J=2.0Hz, 1H), 7.98 (d, J=1.6Hz, 0.5H), 7.92(s, 0.5H), 7.89(s, 1H), 7.81-7.70(m, 2.5H), 7.61-7.51(m, 1.5H), 7.31-7.21(m, 1.5H), 7.14-7.02(m ,2H),6.93-6.83(m,2.5H),6.76-6.64(m,3H),3.92(t,J＝7.2Hz,3H),3.90(s,1.5H),3.86(s,3H), 3.77-3.64(m, 6H), 2.68-2.60(m, 3H), 2.60-2.47(m, 6H).

MS(ESI): m/z 550[M+H] ⁺ .

Example 24 4-(3-Hydroxy-4-((1-(2-ethylmorpholine)-2-carbonyl-4-phenylindole-3-ylide)methyl)phenoxy)-3- Trifluoromethylbenzonitrile

4-(3-hydroxy-4-((1-(2-morpholinoethyl)-2-oxo-4-phenylindolin-3-ylidene)methyl)phenoxy)-3-(trifluoromethyl)benzonitrile(LS 2-37)

The synthetic method is as in Example 6.

Further column chromatography separation was obtained:

Isomer A ¹ H NMR (400 MHz, CDCl ₃ ): δ 10.39 (s, 1H), 7.97 (d, J=1.6 Hz, 1H), 7.72 (dd, J=8.8, 2.0 Hz, 1H), 7.52 -7.40(m, 5H), 7.36(t, J=7.6Hz, 1H), 7.15(s, 1H), 7.06(d, J=8.4Hz, 1H), 7.00(d, J=8.0Hz, 2H) ,6.77(d,J＝8.8Hz,1H),6.73(d,J＝2.4Hz,1H),6.54(dd,J＝8.8,2.8Hz,1H),4.05(t,J＝6.8Hz,2H) ,3.70(d,J=4.4Hz,4H),2.71(t,J=6.8Hz,2H),2.65-2.49(m,4H).

MS(ESI): m/z 610[MH] ^- .

Example 25 4-(4-((4-Cyclopropyl-1-(2-ethylmorpholine)-2-indolone-3-ylide)methyl)-3-hydroxyphenoxy)-3 -Trifluoromethylbenzonitrile (LS 2-28A)

4-(4-((4-cyclopropyl-1-(2-morpholinoethyl)-2-oxoindolin-3-ylidene)methyl)-3-hydroxyphenoxy)-3-(trifluoromethyl)benzonitrile

The synthetic method is as in Example 6.

Further column chromatography separation was obtained:

Isomer A ¹ H NMR (400 MHz, CDCl ₃ ): δ 10.19 (s, 1H), 8.43 (s, 1H), 7.99 (d, J=1.6 Hz, 1H), 7.76 (dd, J=8.4, 2.0Hz, 1H), 7.48(d, J＝8.8Hz, 1H), 7.25(t, J＝7.6Hz, 1H), 7.12(d, J＝8.8Hz, 1H), 7.02(d, J＝8.0Hz ,1H),6.88(d,J＝7.6Hz,1H),6.84(d,J＝2.4Hz,1H),6.75(dd,J＝8.8,2.4Hz,1H),4.01(t,J＝6.8Hz ,2H),3.68(t,J＝4.4Hz,4H),2.67(t,J＝6.8Hz,2H),2.55(t,J＝4.0Hz,4H),1.12-1.06(m,2H),0.89 -0.85(m,3H).

MS(ESI): m/z 574[MH] ^- .

Example 264-(3-Hydroxy-4-((1-(2-ethylmorpholine)-2-indolone-3-ylide)methyl)phenoxy)-3-trifluoromethylbenzyl Nitrile (LS 2-28B)

4-(3-hydroxy-4-((1-(2-morpholinoethyl)-2-oxoindolin-3-ylidene)methyl)phenoxy)-3-(trifluoromethyl)benzonitrile

The synthetic method is as in Example 6.

¹ H NMR (400 MHz, CDCl ₃ ): δ 8.05-7.96 (m, 2.5H), 7.76 (dt, J=8.8, 2.0 Hz, 1.5H), 7.72-7.63 (m, 1.5H), 7.60 (d ,J=7.2Hz,0.5H),7.55(d,J=7.6Hz,1H),7.54(d,J=8.8Hz,0.5H),7.37-7.32(m,0.5H),7.31-7.24(m ,1.5H),7.17(td,J＝7.6,0.8Hz,0.5H),7.10(d,J＝8.8Hz,1H),6.98(d,J＝8.0Hz,0.5H),6.92(td,J ＝7.6,0.8Hz,1H),6.87(d,J＝7.6Hz,1H),6.81(d,J＝2.4Hz,0.5H),6.74(dd,J＝8.8,2.8Hz,0.5H),6.67 -6.58(m, 2H), 4.01(t, J=6.8Hz, 1H), 3.93(t, J=7.2Hz, 2H), 3.74-3.64(m, 6H), 2.71-2.63(m, 3H), 2.61-2.48(m,6H).

MS(ESI): m/z 534[MH] ^- .

Example 27 4-(3-Hydroxy-4-((4-methyl-1-(2-ethylmorpholine)-2-indolone-3-ylide)methyl)phenoxy)-3- Trifluoromethylbenzonitrile (LS 2-28C)

4-(3-hydroxy-4-((4-methyl-1-(2-morpholinoethyl)-2-oxoindolin-3-ylidene)methyl)phenoxy)-3-(trifluoromethyl)benzonitrile

The synthetic method is as in Example 6.

Further column chromatography separation was obtained:

Isomer A ¹ H NMR (400 MHz, CDCl ₃ ): δ 9.99 (s, 1H), 7.99 (d, J=2.0 Hz, 1H), 7.80 (s, 1H), 7.75 (dd, J=8.8, 2.0Hz, 1H), 7.54 (d, J=8.8Hz, 1H), 7.23 (t, J=8.0Hz, 1H), 7.11 (d, J=8.8Hz, 1H), 6.95 (d, J=8.0Hz) ,1H),6.86(d,J＝8.0Hz,1H),6.78(d,J＝2.4Hz,1H),6.73(dd,J＝8.4,2.4Hz,1H),4.01(t,J＝6.8Hz ,2H),3.68(t,J＝4.8Hz,4H),2.67(t,J＝6.8Hz,2H),2.63(s,3H),2.59-2.52(m,4H).

MS(ESI): m/z 548[MH] ^- .

Example 28 3-(4-(4-Chloro-2-(trifluoromethyl)phenoxy)-3-methoxybenzylylide)-5-fluoro-1-(2-ethylmorpholine) Indol-2-one (LS 2-54)

The synthetic method is as in Example 1.

Further column chromatography separation was obtained:

Isomer A ¹ H NMR for (400 MHz, CDCl ₃ ): δ 8.73 (d, J=2.0 Hz, 1H), 7.64 (d, J=2.8 Hz, 1H), 7.58 (dd, J=8.4, 2.0 Hz,1H),7.45(s,1H),7.37(dd,J=8.8,2.4Hz,1H),7.30-7.23(m,1H),7.02(d,J=8.0Hz,1H),7.00-6.95 (m,1H),6.82-6.74(m,2H),3.97-3.86(m,5H),3.69(t,J=4.8Hz,4H),2.64(t,J=6.8Hz,2H),2.56( t,J=3.6Hz,4H).

MS(ESI): m/z 577[M+H] ⁺ .

Example 29 4-(4-((5-Fluoro-2-indolone-3-ylide)methyl)-2-methoxyphenoxy)-3-trifluoromethylbenzamide (LS 2 -62)

4-(4-((5-fluoro-2-oxoindolin-3-ylidene)methyl)-2-methoxyphenoxy)-3-(trifluoromethyl)benzamide

The synthetic method is as in Example 1, step 2.

¹ H NMR (400 MHz, CD ₃ OD): δ 8.84 (d, J=1.6 Hz, 1H), 8.26 (dd, J=4.0, 2.0 Hz, 2H), 8.07-7.95 (m, 2H), 7.80 ( d, J=14.0Hz, 2H), 7.73 (dd, J=8.4, 1.6Hz, 1H), 7.50-7.44 (m, 2H), 7.44-7.36 (m, 2H), 7.30 (d, J=8.0Hz) ,1H),7.18(d,J＝8.4Hz,1H),7.06-6.93(m,2H),6.93-6.77(m,4H),3.86(s,3H),3.82(s,3H).

MS(ESI): m/z 473[M+H] ⁺ .

Example 30 3-Chloro-4-(4-((5-fluoro-1-(2-ethylmorpholine)-2-indolone-3-ylide)methyl)-2-methoxyphenoxy base) benzonitrile (LS 2-67)

3-chloro-4-(4-((5-fluoro-1-(2-morpholinoethyl)-2-oxoindolin-3-ylidene)methyl)-2-methoxyphenoxy)benzonitrile

The synthetic method is as in Example 1.

¹ H NMR (400 MHz, CDCl ₃ ): δ 8.75 (d, J=1.6 Hz, 0.8H), 7.85 (s, 1H), 7.76 (dd, J=6.8, 2.0 Hz, 1.8H), 7.61 (dd ,J=8.4,1.6Hz,0.8H),7.50-7.42(m,2H),7.44-7.35(m,1.6H),7.32-7.22(m,3H),7.18(d,J=8.0Hz,1H ),7.10(d,J＝8.0Hz,0.8H),7.01(td,J＝8.8,2.8Hz,1.8H),6.88-6.82(m,1.6H),6.82-6.75(m,1.8H), 4.02-3.87(m, 6H), 3.82(s, 3H), 3.78-3.66(m, 7.2H), 2.75-2.65(m, 3.6H), 2.65-2.52(m, 7.2H).

MS(ESI): m/z 534[M+H] ⁺ .

Example 31 4-(4-((5-Fluoro-1-(2-ethylmorpholine)-2-indolone-3-ylide)methyl)-2-methoxyphenoxy)-3 - Methyl nitrobenzoate (LS 3-53)

4-(4-((5-fluoro-1-(2-morpholinoethyl)-2-oxoindolin-3-ylidene)methyl)-2-methoxyphenoxy)-3-nitrophenyl acetate

The synthetic method is as in Example 1.

¹ H NMR (400MHz, CDCl ₃ ): δ 8.75 (d, J=2.0Hz, 0.8H), 8.66 (d, J=2.0Hz, 1H), 8.65 (d, J=2.4Hz, 0.8H), 8.14(dd,J＝8.8,2.4Hz,1H),8.09(dd,J＝8.8,2.0Hz,0.8H),7.84(s,1H),7.62(dd,J＝8.4,2.0Hz,0.8H) ,7.46(s,0.8H),7.40(dd,J=8.8,2.4Hz,1H),7.35-7.23(m,3.6H),7.19(d,J=8.4Hz,0.8H),7.00(td, J=8.8,2.4Hz,2H),6.92(d,J=8.8Hz,1H),6.89(d,J=8.8Hz,0.8H),6.86-6.77(m,1.8H),4.04-3.94(m ,5.4H),3.92(t,J＝6.4Hz,3.6H),3.88(s,2.4H),3.80(s,3H),3.75-3.64(m,7.2H),2.67-2.60(m,3.6 H),2.60-2.49(m,7.2H).

MS(ESI): m/z 578[M+H] ⁺ .

Example 32 4-(4-((5-Fluoro-2-indolone-3-ylide)methyl)-2-methoxyphenoxy)-2-trifluoromethylbenzonitrile (LS3- 58)

4-(4-((5-fluoro-2-oxoindolin-3-ylidene)methyl)-2-methoxyphenoxy)-2-(trifluoromethyl)benzonitrile

The synthetic method is as in Example 1, step 2.

¹ H NMR (400MHz, DMSO-d ₆ ): δ 10.68(s, 1H), 10.67(s, 0.6H), 8.77(d, J=1.6Hz, 1H), 8.16-8.05(m, 1.6H) ,7.95(s,1H),7.94(dd,J=8.4,1.6Hz,1H),7.73(s,0.6H),7.66(dd,J=8.8,2.4Hz,1H),7.60(d,J= 1.6Hz,0.6H),7.58-7.52(m,1.6H),7.48-7.39(m,1.2H),7.39-7.33(m,1.6H),7.30(dd,J=8.4,2.4Hz,0.6H ),7.25(dd,J＝8.8,2.4Hz,1H),7.12(td,J＝9.2,2.8Hz,0.6H),7.09-7.02(m,1H),6.88(dd,J＝8.4,4.4Hz ,0.6H),6.83(dd,J=8.4,4.4Hz,1H),3.82(s,3H),3.78(s,1.8H).

MS(ESI): m/z 455[M+H] ⁺ .

Example 33 4-(4-((5-Fluoro-2-indolone-3-ylide)methyl)-2-methoxyphenoxy)-3-nitrobenzonitrile (LS3-63)

4-(4-((5-fluoro-2-oxoindolin-3-ylidene)methyl)-2-methoxyphenoxy)-3-nitrobenzonitrile

The synthetic method is as in Example 1, step 2.

¹ H NMR (400 MHz, DMSO-d ₆ ): δ 10.69 (s, 1H), 10.67 (s, 1H), 8.77 (d, J=1.6 Hz, 1H), 8.66 (d, J=2.0 Hz, 2H) ), 8.05(td, J=8.8, 2.0Hz, 2H), 7.94(s, 1H), 7.93(dd, J=8.4, 1.6Hz, 1H), 7.71(s, 1H), 7.66(dd, J= 8.8, 2.4Hz, 1H), 7.60(s, 1H), 7.44(d, J=0.8Hz, 2H), 7.44-7.35(m, 2H), 7.18-7.09(m, 2H), 7.09-7.01(m ,2H),6.88(dd,J=8.4,4.4Hz,1H),6.83(dd,J=8.4,4.4Hz,1H),3.81(s,3H),3.78(s,3H).

MS(ESI): m/z 432[M+H] ⁺ .

Example 34 2-(4-((5-Fluoro-2-indolone-3-ylide)methyl)-2-methoxyphenoxy)-5-trifluoromethylbenzonitrile (LS3- 67)

2-(4-((5-fluoro-2-oxoindolin-3-ylidene)methyl)-2-methoxyphenoxy)-5-(trifluoromethyl)benzonitrile

The synthetic method is as in Example 1, step 2.

¹ H NMR (400MHz, DMSO-d ₆ ): δ 10.70(s, 1H), 10.68(s, 0.8H), 8.78(d, J=2.0Hz, 1H), 8.42(d, J=2.0Hz, 1.8H), 7.99-7.93(m, 3.8H), 7.73(s, 0.8H), 7.66(dd, J=8.8, 2.4Hz, 1H), 7.62(d, J=1.6Hz, 0.8H), 7.51 -7.38(m, 3.4H), 7.12(td, J=9.2, 2.8Hz, 0.8H), 7.09-7.03(m, 1H), 7.01(d, J=9.2Hz, 0.8H), 6.94(d, J＝8.8Hz,1H),6.89(dd,J＝8.4,4.4Hz,0.8H),6.83(dd,J＝8.4,4.4Hz,1H),3.83(s,3H),3.79(s,2.4H) ).

MS(ESI): m/z 455[M+H] ⁺ .

Example 35 4-(4-((5-Fluoro-1-(2-ethylmorpholine)-2-indolone-3-ylide)methyl)-2-methoxyphenoxy)-2 -Trifluoromethylbenzonitrile (LS 3-65)

4-(4-((5-fluoro-1-(2-morpholinoethyl)-2-oxoindolin-3-ylidene)methyl)-2-methoxyphenoxy)-2-(trifluoromethyl)benzonitrile

The synthetic method is as in Example 1.

Further column chromatography separation was obtained:

Isomer A ¹ H NMR (400 MHz, DMSO-d ₆ ): δ 8.75 (d, J=2.0 Hz, 1H), 8.11 (d, J=8.8 Hz, 1H), 8.01 (s, 1H), 7.97 (dd, J=8.4, 2.0Hz, 1H), 7.72 (dd, J=8.8, 2.4Hz, 1H), 7.55 (d, J=2.4Hz, 1H), 7.38 (d, J=8.4Hz, 1H) ,7.25(dd,J＝8.8,2.4Hz,1H),7.16(td,J＝8.8,2.4Hz,1H),7.10(dd,J＝8.8,4.4Hz,1H),3.89(t,J＝6.8 Hz, 2H), 3.82(s, 3H), 3.51(t, J=4.4Hz, 4H), 2.54(t, J=6.4Hz, 2H), 2.48-2.37(m, 4H).

Isomer B ¹ H NMR (400 MHz, DMSO-d ₆ ): δ 8.13 (d, J=8.8 Hz, 1H), 7.82 (s, 1H), 7.62 (d, J=1.6 Hz, 1H), 7.55 (d, J=2.8Hz, 1H), 7.49-7.38 (m, 3H), 7.31 (dd, J=8.8, 2.4Hz, 1H), 7.21 (td, J=8.8, 2.4Hz, 1H), 7.15 ( dd,J＝8.8,4.4Hz,1H),3.89(t,J＝6.4Hz,2H),3.78(s,3H),3.53(t,J＝4.4Hz,4H),2.55(t,J＝6.4 Hz,2H),2.39-2.49(m,4H).

MS(ESI): m/z 568[M+H] ⁺ .

Example 36 4-(4-((5-Fluoro-1-(2-ethylmorpholine)-2-indolone-3-ylide)methyl)-2-methoxyphenoxy)-3 -Nitrobenzonitrile (LS 3-68)

2-(4-((5-fluoro-1-(2-morpholinoethyl)-2-oxoindolin-3-ylidene)methyl)-2-methoxyphenoxy)-5-(trifluoromethyl)benzonitrile

The synthetic method is as in Example 1.

Further column chromatography separation was obtained:

Isomer A ¹ H NMR (400 MHz, DMSO-d ₆ ): δ 8.76 (d, J=2.0 Hz, 1H), 8.43 (d, J=2.0 Hz, 1H), 8.02 (s, 1H), 7.99 (dd, J=8.4, 1.6Hz, 1H), 7.95 (dd, J=9.2, 2.4Hz, 1H), 7.73 (dd, J=8.8, 2.4Hz, 1H), 7.45 (d, J=8.4Hz, 1H), 7.16(td, J＝8.8, 2.8Hz, 1H), 7.10(dd, J＝8.8, 4.4Hz, 1H), 6.95(d, J＝8.8Hz, 1H), 3.90(t, J＝6.4 Hz, 2H), 3.84(s, 3H), 3.52(t, J=4.4Hz, 4H), 2.54(t, J=6.4Hz, 2H), 2.45(s, 4H).

Isomer B ¹ H NMR (400 MHz, DMSO-d ₆ ): δ 8.42 (d, J=2.4 Hz, 1H), 7.97 (dd, J=9.2, 2.0 Hz, 1H), 7.82 (s, 1H) ,7.64(d,J＝1.2Hz,1H),7.53-7.40(m,3H),7.22(td,J＝8.8,2.4Hz,1H),7.15(dd,J＝8.8,4.4Hz,1H), 7.01(d,J＝8.8Hz,1H),3.89(t,J＝6.4Hz,2H),3.80(s,3H),3.53(t,J＝4.8Hz,4H),2.55(t,J＝6.4 Hz,2H),2.48-2.38(m,4H).

MS(ESI): m/z 568[M+H] ⁺ .

Example 37 4-(4-((5-Fluoro-1-(2-ethylmorpholine)-2-indolone-3-ylide)methyl)-2-methoxyphenoxy)-3 -Nitrobenzonitrile (LS 3-69)

4-(4-((5-fluoro-1-(2-morpholinoethyl)-2-oxoindolin-3-ylidene)methyl)-2-methoxyphenoxy)-3-nitrobenzonitrile

The synthetic method is as in Example 1.

Further column chromatography separation was obtained:

Isomer A ¹ H NMR (400 MHz, DMSO-d ₆ ): δ 8.75 (d, J=2.0 Hz, 1H), 8.67 (d, J=2.4 Hz, 1H), 8.04 (dd, J=8.8, 2.0Hz, 1H), 8.01(s, 1H), 7.97(dd, J=8.4, 1.6Hz, 1H), 7.72(dd, J=8.8, 2.4Hz, 1H), 7.40(d, J=8.4Hz, 1H), 7.19-7.12(m, 1H), 7.11-7.07(m, 1H), 7.06(d, J=8.8Hz, 1H), 3.89(t, J=6.4Hz, 2H), 3.82(s, 3H) ), 3.51(t, J=4.4Hz, 4H), 2.54(t, J=6.8Hz, 2H), 2.49-2.39(m, 4H).

Isomer B ¹ H NMR (400 MHz, DMSO-d ₆ ): δ 8.67 (d, J=2.0 Hz, 1H), 8.07 (dd, J=8.8, 2.0 Hz, 1H), 7.80 (s, 1H) ,7.62(s,1H),7.48-7.41(m,3H),7.21(td,J=8.8,2.4Hz,1H),7.17-7.10(m,2H),3.88(t,J=6.4Hz,2H ), 3.78(s, 3H), 3.53(t, J=4.8Hz, 4H), 2.55(t, J=6.4Hz, 2H), 2.48-2.37(m, 4H).

MS(ESI): m/z 545[M+H] ⁺ .

Example 38 4-(4-((5-Fluoro-1-(3-propylmorpholine)-2-indolone-3-ylide)methyl)-2-methoxyphenoxy)-3 -Trifluoromethylbenzonitrile (LS 3-155)

4-(4-((5-fluoro-1-(3-morpholinopropyl)-2-oxoindolin-3-ylidene)methyl)-2-methoxyphenoxy)-3-(trifluoromethyl)benzonitrile

The synthetic method is as in Example 1.

¹ H NMR (400MHz, CDCl ₃ ): δ 8.78(s, 0.8H), 7.98(d, J=7.2Hz, 1.8H), 7.85(s, 1H), 7.78-7.57(m, 2.6H), 7.47(s, 0.8H), 7.44-7.13(m, 6H), 7.00(t, J=8.4Hz, 1.6H), 6.94-6.73(m, 3.4H), 3.99-3.74(m, 9H), 3.70 (d, J=3.6Hz, 7.2H), 2.43 (d, J=6.0Hz, 10.8H), 1.91 (t, J=6.4Hz, 3.6H).

MS(ESI): m/z 582[M+H] ⁺ .

Example 39 4-(4-((5-Fluoro-1-(4-butylmorpholine)-2-indolone-3-ylide)methyl)-2-methoxyphenoxy)-3 -Trifluoromethylbenzonitrile (LS 4-1)

4-(4-((5-fluoro-1-(4-morpholinobutyl)-2-oxoindolin-3-ylidene)methyl)-2-methoxyphenoxy)-3-(trifluoromethyl)benzonitrile

The synthetic method is as in Example 1.

Further column chromatography separation was obtained:

Isomer A ¹ H NMR (400 MHz, CDCl ₃ ): δ 8.76 (s, 1H), 7.95 (s, 1H), 7.65 (d, J=8.8 Hz, 1H), 7.60 (d, J=8.0 Hz ,1H),7.45(s,1H),7.25-7.20(m,1H),7.16(d,J=8.0Hz,1H),6.99(t,J=6.8Hz,1H),6.89-6.70(m, 2H), 3.88(s, 3H), 3.79(t, J=7.2Hz, 2H), 3.69(s, 4H), 2.53-2.29(m, 6H), 1.78-1.67(m, 2H), 1.65-1.50 (m,2H).

MS(ESI): m/z 596[M+H] ⁺ .

Example 40 4-(4-((5-Fluoro-1-(4-(4-methylpiperazin-1-yl)butyl)-2-indolone-3-ylide)methyl)-2 -Methoxyphenoxy)-3-trifluoromethylbenzonitrile (LS 4-3)

4-(4-((5-fluoro-1-(4-(4-methylpiperazin-1-yl)butyl)-2-oxoindolin-3-ylidene)methyl)-2-methoxyphenoxy)-3-(trifluoromethyl)benzonitrile

The synthetic method is as in Example 1.

¹ H NMR (400 MHz, CDCl ₃ ): δ 8.77 (d, J=2.0 Hz, 1H), 7.98 (d, J=1.6 Hz, 0.7H), 7.96 (d, J=2.0 Hz, 1H), 7.84 (s,0.7H),7.71(dd,J＝8.4,2.0Hz,0.7H),7.66(dd,J＝8.4,2.0Hz,1H),7.60(dd,J＝8.4,2.0Hz,1H), 7.46(s, 1H), 7.38(dd, J=8.8, 2.4Hz, 0.7H), 7.32-7.22(m, 3.4H), 7.17(d, J=8.0Hz, 1H), 7.03-6.96(m, 1.7H), 6.86-6.76(m, 3.1H), 3.89(s, 3H), 3.86-3.72(m, 5.5H), 2.85-2.30(m, 17H), 2.28(d, J=2.4Hz, 5.1 H), 1.80-1.68(m, 3.4H), 1.60-1.52(m, 3.4H).

MS(ESI): m/z 609[M+H] ⁺ .

Example 41 3-Bromo-4-(4-((5-fluoro-1-(2-ethylmorpholine)-2-indolone-3-ylide)methyl)-2-methoxyphenoxy base) benzonitrile (LS 4-35)

3-bromo-4-(4-((5-fluoro-1-(2-morpholinoethyl)-2-oxoindolin-3-ylidene)methyl)-2-methoxyphenoxy)benzonitrile

The synthetic method is as in Example 1.

¹ H NMR (400MHz, CDCl ₃ ): δ8.74 (d, J=2.0Hz, 0.8H), 7.91 (d, J=2.0Hz, 1H), 7.89 (d, J=2.0Hz, 0.8H), 7.82(s,1H),7.59(dd,J=8.8,2.0Hz,0.8H),7.49(dd,J=8.4,2.0Hz,1H),7.47-7.41(m,1.8H),7.38(dd, J=8.8, 2.4Hz, 1H), 7.30-7.26(m, 0.8H), 7.26-7.21(m, 1.8H), 7.16(d, J=8.4Hz, 1H), 7.08(d, J=8.4Hz) ,0.8H),7.02-6.92(m,1.8H),6.82-6.76(m,1.8H),6.74(d,J＝8.4Hz,1H),6.72(d,J＝8.4Hz,0.8H), 3.92-3.86(m, 6H), 3.79(s, 3H), 3.70-3.64(m, 7.2H), 2.65-2.58(m, 3.6H), 2.57-2.50(m, 7.2H).

HRMS(ESI): m/z 578.1064[M+H] ⁺ .

Example 42 4-(4-((5-Fluoro-1-(2-ethylmorpholine)-2-indolone-3-ylide)methyl)-2-methoxyphenoxy)-1 - Naphthalonitrile (LS 4-36)

4-(4-((5-fluoro-1-(2-morpholinoethyl)-2-oxoindolin-3-ylidene)methyl)-2-methoxyphenoxy)-1-naphthonitrile

The synthetic method is as in Example 1.

¹ H NMR (400 MHz, CDCl ₃ ): δ 8.79 (d, J=1.2 Hz, 1H), 8.50 (t, J=8.8 Hz, 2H), 8.24 (dd, J=8.0, 4.8 Hz, 2H), 7.87(s, 1H), 7.85-7.72(m, 4H), 7.72-7.56(m, 3H), 7.55-7.39(m, 2H), 7.37-7.19(m, 4H), 7.14(d, J=8.4 Hz,1H),7.06-6.94(m,2H),6.82(td,J=8.8,4.4Hz,2H),6.69(dd,J=8.0,5.6Hz,2H),4.02-3.83(m,7H) ,3.80(s,3H),3.70(dd,J=9.2,5.2Hz,8H),2.73-2.60(m,4H),2.57(s,8H).

HRMS(ESI): m/z 550.2114[M+H] ⁺ .

Example 43 3-(4-(4-Bromo-2-(trifluoromethyl)phenoxy)-3-methoxybenzylylide)-5-fluoro-1-(2-ethylmorpholine) Indol-2-one (LS 4-38)

3-(4-(4-bromo-2-(trifluoromethyl)phenoxy)-3-methoxybenzylidene)-5-fluoro-1-(2-morpholinoethyl)indolin-2-one

The synthetic method is as in Example 1.

¹ H NMR (400 MHz, CDCl ₃ ): δ 8.72 (d, J=2.0 Hz, 1H), 7.83 (s, 1H), 7.78 (dd, J=6.4, 2.4 Hz, 2H), 7.58 (dd, J ＝8.4,1.6Hz,1H),7.54(dd,J＝8.8,2.4Hz,1H),7.50(dd,J＝8.8,2.0Hz,1H),7.46-7.40(m,2H),7.28-7.26( m,1H),7.26-7.22(m,2H),7.11(d,J=8.4Hz,1H),7.03(d,J=8.4Hz,1H),7.01-6.94(m,2H),6.80(td ,J=8.4,4.0Hz,2H),6.71(dd,J=8.8,6.4Hz,2H),3.97-3.87(m,7H),3.82(s,3H),3.69(dd,J=9.2,4.8 Hz, 8H), 2.63(td, J=6.8, 3.6Hz, 4H), 2.60-2.46(m, 8H).

HRMS(ESI): m/z 621.0985[M+H] ⁺ .

Example 44 4-(4-((5-Fluoro-1-(2-ethylmorpholine)-2-indolone-3-ylide)methyl)-2-methoxyphenoxy)-3 - Methyl trifluoromethylbenzoate (LS 4-39)

4-(4-((5-fluoro-1-(2-morpholinoethyl)-2-oxoindolin-3-ylidene)methyl)-2-methoxyphenoxy)-3-(trifluoromethyl)benzoate

The synthetic method is as in Example 1.

¹ H NMR (400 MHz, CDCl ₃ ): δ 8.73 (d, J=1.6 Hz, 1H), 8.36 (d, J=2.0 Hz, 1H), 8.35 (d, J=2.0 Hz, 1H), 8.09 ( dd,J=8.8,2.4Hz,1H),8.05(dd,J=8.8,2.0Hz,1H),7.83(s,1H),7.60(dd,J=8.4,2.0Hz,1H),7.45(s ,1H),7.42(dd,J=8.8,2.4Hz,1H),7.31-7.27(m,1H),7.26-7.22(m,2H),7.20(d,J=8.0Hz,1H),7.12( d, J=8.0Hz, 1H), 7.01-6.94(m, 2H), 6.84-6.75(m, 4H), 3.94-3.87(m, 13H), 3.79(s, 3H), 3.68(dd, J= 9.6,5.6Hz,8H),2.66-2.60(m,4H),2.60-2.50(m,8H).

HRMS(ESI): m/z 601.1945[M+H] ⁺ .

Example 454-(4-((5-Fluoro-1-(2-ethylmorpholine)-2-indolone-3-ylide)methyl)-phenoxy)-3-trifluoromethylbenzene Nitrile (LS 4-46)

4-(4-((5-fluoro-1-(2-morpholinoethyl)-2-oxoindolin-3-ylidene)methyl)phenoxy)-3-(trifluoromethyl)benzonitrile

The synthetic method is as in Example 1.

¹ H NMR (400MHz, CDCl ₃ ): δ 8.45-8.35 (m, 2H), 8.01 (d, J=2.0Hz, 1H), 7.99 (d, J=2.0Hz, 1H), 7.84 (s, 1H) ), 7.78(dd, J=8.8, 2.0Hz, 1H), 7.74(dd, J=8.4, 2.0Hz, 1H), 7.72-7.65(m, 2H), 7.47(s, 1H), 7.33(dd, J=8.8, 2.4Hz, 1H), 7.27-7.22(m, 1H), 7.23-7.16(m, 2H), 7.16-7.11(m, 2H), 7.06(t, J=8.8Hz, 2H), 6.99 (td,J=8.8,2.8Hz,2H),6.80(td,J=8.4,4.0Hz,2H),3.96-3.86(m,4H),3.69(dd,J=9.2,4.8Hz,8H), 2.66-2.58(m, 4H), 2.59-2.49(m, 8H).

HRMS(ESI): m/z 538.1742[M+H] ⁺ .

Example 46 3-Difluoromethyl-4-(4-((5-fluoro-1-(2-ethylmorpholine)-2-indolinone-3-ylide)methyl)-2-methoxy phenoxy) benzonitrile (LS 4-52)

3-(difluoromethyl)-4-(4-((5-fluoro-1-(2-morpholinoethyl)-2-oxoindolin-3-ylidene)methyl)-2-methoxyphenoxy)benzonitrile

The synthetic method is as in Example 1.

¹ H NMR (400 MHz, CDCl ₃ ): δ 8.74 (d, J=1.6 Hz, 1H), 7.96 (d, J=6.0 Hz, 2H), 7.84 (s, 1H), 7.67-7.56 (m, 3H) ),7.46(s,1H),7.40(dd,J=8.8,2.4Hz,1H),7.32-7.26(m,2H),7.26-7.20(m,2.5H),7.16(d,J=8.0Hz ,1H),7.13(d,J＝4.4Hz,1H),7.04-6.96(m,2.5H),6.88-6.72(m,4H),3.92(t,J＝7.2Hz,4H),3.87(s ,3H),3.78(s,3H),3.70(dd,J=9.6,5.2Hz,8H),2.64(td,J=6.8,4.8Hz,4H),2.56(s,8H).

HRMS(ESI): m/z 550.1935[M+H] ⁺ .

Example 47 3-(4-(2-Chloro-5-methoxy-4-nitrophenoxy)-3-methoxybenzylylide)-5-fluoro-1-(2-ethylmorph Lino)indol-2-one (LS 4-83)

3-(4-(2-chloro-5-methoxy-4-nitrophenoxy)-3-methoxybenzylidene)-5-fluoro-1-(2-morpholinoethyl)indolin-2-one

The synthetic method is as in Example 1.

¹ H NMR (400 MHz, CDCl ₃ ): δ 8.82 (d, J=2.0 Hz, 1H), 8.14 (d, J=6.0 Hz, 2H), 7.86 (s, 1H), 7.63 (dd, J=8.4 ,1.6Hz,1H),7.46(s,1H),7.32(dd,J=9.2,2.8Hz,1H),7.30-7.24(m,3H),7.18(d,J=8.0Hz,1H),7.09 (d, J＝8.0Hz, 1H), 7.04-6.95(m, 2H), 6.86-6.77(m, 2H), 6.42(s, 2H), 3.98-3.88(m, 7H), 3.85(s, 3H) ), 3.79(s, 3H), 3.76(s, 3H), 3.74-3.67(m, 8H), 2.65(t, J=6.8Hz, 4H), 2.57(s, 8H).

HRMS(ESI): m/z 584.1573[M+H] ⁺ .

Example 48 3-(4-(2,6-Dichloro-4-nitrophenoxy)-3-methoxybenzylylide)-5-fluoro-1-(2-ethylmorpholine)indone Indol-2-one (LS 4-90)

3-(4-(2,6-dichloro-4-nitrophenoxy)-3-methoxybenzylidene)-5-fluoro-1-(2-morpholinoethyl)indolin-2-one

The synthetic method is as in Example 1.

¹ H NMR (400 MHz, CDCl ₃ ): δ 8.83 (d, J=1.6 Hz, 1H), 8.31 (d, J=7.2 Hz, 4H), 7.80 (s, 1H), 7.50-7.38 (m, 3H) ),7.27(d,J＝2.0Hz,1H),7.23(dd,J＝8.4,2.4Hz,1H),7.15(dd,J＝8.4,1.2Hz,1H),7.02-6.92(m,2H) ,6.80(td,J＝9.2,4.0Hz,2H),6.60(d,J＝8.4Hz,1H),6.52(d,J＝8.4Hz,1H),4.05(s,3H),3.95(s, 3H), 3.91 (td, J=7.2, 2.4Hz, 4H), 3.69 (dd, J=7.6, 3.6Hz, 8H), 2.63 (td, J=6.8, 2.0Hz, 4H), 2.60-2.46 (m ,8H).

HRMS(ESI): m/z 588.1072[M+H] ⁺ .

Example 49 4-(4-((5-Fluoro-1-(2-(4-(4-methylpiperazin-1-yl)piperidin-1-yl)ethyl)-2-indolone -3-ylide)methyl)-2-methoxyphenoxy)-3-trifluoromethylbenzonitrile (LS 4-70)

4-(4-((5-fluoro-1-(2-(4-(4-methylpiperazin-1-yl)piperidin-1-yl)ethyl)-2-oxoindolin-3-ylidene)methyl)-2- methoxyphenoxy)-3-(trifluoromethyl)benzonitrile

The synthetic method is as in Example 1.

HRMS(ESI): m/z 664.2887[M+H] ⁺ .

Example 50 4-(4-((5-Fluoro-2-carbonyl-1-(2-(4-(pyrrol-1-yl)piperidin-1-yl)ethyl)indole-3-ylide) Methyl)-2-methoxyphenoxy)-3-trifluoromethylbenzonitrile (LS 4-77)

4-(4-((5-fluoro-2-oxo-1-(2-(4-(pyrrolidin-1-yl)piperidin-1-yl)ethyl)indolin-3-ylidene)methyl)-2-methoxyphenoxy )-3-(trifluoromethyl)benzonitrile

The synthetic method is as in Example 1.

HRMS(ESI): m/z 635.2609[M+H] ⁺ .

Example 514-((2-(2-Ethylmorpholine)-1-carbonyl-1,2-dihydrobenzo[6,7]oxepino[4,3,2-cd]indol-8-yl) oxy)-3-trifluoromethylbenzonitrile (LS 1-132)

4-((2-(2-morpholinoethyl)-1-oxo-1,2-dihydrobenzo[6,7]oxepino[4,3,2-cd]indol-8-yl)oxy)-3-(trifluoromethyl) benzonitrile

4-(4-((4-Bromo-1-(2-ethylmorpholine)-2-indolone-3-ylide)methyl)-3-hydroxyphenoxy)-3-trifluoromethane benzonitrile (LS 1-125, 24 mg, 0.04 mmol), cesium carbonate (32 mg, 0.1 mmol), cuprous iodide (1 mg, 0.004 mmol), tetramethylethylenediamine (1 drop) in 5 mL N , N-dimethylformamide, under argon protection, heated to 100 degrees Celsius and reacted overnight. The reaction system was cooled, filtered through celite, and the filtrate was extracted three times with dichloromethane. The organic phases were combined, washed with saturated brine, spun dry, and separated by column chromatography to obtain a white solid (7 mg, 32%).

¹ H NMR (400 MHz, CDCl ₃ ): δ 8.00 (d, J=1.6 Hz, 1H), 7.79 (dd, J=8.4, 2.0 Hz, 1H), 7.16-7.01 (m, 3H), 6.88 (s ,1H),6.67(dd,J＝8.4,2.4Hz,1H),6.56(d,J＝2.4Hz,1H),6.43(d,J＝7.6Hz,1H),6.31(d,J＝8.4Hz ,1H),3.81(t,J＝6.4Hz,2H),3.77-3.66(m,4H),2.68-2.45(m,6H).

MS(ESI): m/z 534[M+H] ⁺ .

Example 52 4-(2-((5-Fluoro-2-indolone-3-ylide)methyl)-6-methoxyphenoxy)-3-trifluoromethylbenzonitrile (LS3-80 )

4-(2-((5-fluoro-2-oxoindolin-3-ylidene)methyl)-6-methoxyphenoxy)-3-(trifluoromethyl)benzonitrile

The synthetic method is as in Example 1, step 2.

Further column chromatography separation was obtained:

Isomer A ¹ H NMR (400 MHz, CDCl ₃ ): δ 10.61 (s, 1H), 8.28 (d, J=2.0 Hz, 1H), 7.94 (dd, J=8.8, 2.0 Hz, 1H), 7.68 -7.49(m,1H),7.47-7.31(m,3H),7.14-7.01(m,2H),6.89-6.72(m,2H),3.79(s,3H).

MS(ESI): m/z 455[M+H] ⁺ .

Example 53 4-(2-((5-Fluoro-1-(2-ethylmorpholine)-2-indolone-3-ylide)methyl)-6-methoxyphenoxy)-3 -Trifluoromethylbenzonitrile (LS 3-82)

4-(2-((5-fluoro-1-(2-morpholinoethyl)-2-oxoindolin-3-ylidene)methyl)-6-methoxyphenoxy)-3-(trifluoromethyl)benzonitrile

The synthetic method is as in Example 1.

Further column chromatography separation was obtained:

Isomer A ¹ H NMR (400 MHz, CDCl ₃ ): δ 7.91 (d, J=2.0 Hz, 1H), 7.69 (s, 1H), 7.58 (dd, J=8.8, 2.0 Hz, 1H), 7.39 (t,J=8.0Hz,1H),7.36-7.30(m,1H),7.19(dd,J=8.8,2.8Hz,1H),7.16-7.10(m,1H),6.98(td,J=8.8 ,2.4Hz,1H),6.76(dd,J＝8.8,4.4Hz,1H),6.65(d,J＝8.8Hz,1H),3.84(t,J＝6.8Hz,2H),3.80(s,3H ), 3.65(t, J＝4.4Hz, 4H), 2.59(t, J＝6.8Hz, 2H), 2.51(t, J＝4.4Hz, 4H).

Isomer B ¹ H NMR (400 MHz, CDCl ₃ ): δ 8.28 (dd, J=8.0, 0.8 Hz, 1H), 7.97 (d, J=2.0 Hz, 1H), 7.67-7.60 (m, 2H) ,7.39(t,J＝8.0Hz,1H),7.14(dd,J＝8.4,1.2Hz,1H),7.10(dd,J＝8.0,2.4Hz,1H),6.98(td,J＝8.4,2.4 Hz,1H),6.76(t,J＝4.0Hz,1H),6.72(d,J＝8.8Hz,1H),3.85(t,J＝7.2Hz,2H),3.80(s,3H),3.69( t, J=4.8Hz, 4H), 2.60 (t, J=7.2Hz, 2H), 2.54 (t, J=4.4Hz, 4H).

MS(ESI): m/z 568[M+H] ⁺ .

Example 543-Amino-4-(4-((5-fluoro-1-(2-ethylmorpholine)-2-indolone-3-ylide)methyl)-2-methoxyphenoxy ) benzonitrile (LS 4-93)

3-amino-4-(4-((5-fluoro-1-(2-morpholinoethyl)-2-oxoindolin-3-ylidene)methyl)-2-methoxyphenoxy)benzonitrile

4-(4-((5-Fluoro-1-(2-ethylmorpholine)-2-indolone-3-ylide)methyl)-2-methoxyphenoxy)-3-nitro Benzonitrile (LS 3-69, 55mg, 0.1mmol), reduced iron powder (17mg, 0.3mmol), ammonium chloride (77mg, 1mmol) were dissolved in a mixed solvent of 8 ml of ethanol and 4 ml of water, heated to Reflux until the reaction is complete. After suction filtration through celite, the filtrate was spin-dried and separated by column chromatography to obtain a yellow solid (37 mg, 73%).

¹ H NMR (400 MHz, CDCl ₃ ): δ 8.77 (d, J=1.6 Hz, 1H), 7.84 (s, 1H), 7.55 (dd, J=8.8, 2.0 Hz, 1H), 7.48-7.41 (m ,2H),7.27-7.22(m,3H),7.15-7.03(m,3H),7.04-6.93(m,5H),6.81(td,J=8.4,4.4Hz,2H),6.72(d,J ＝5.2Hz,1H),6.70(d,J＝5.2Hz,1H),4.15(d,J＝10.0Hz,4H),3.99-3.88(m,7H),3.85(s,3H),3.70(dd , J=9.2, 4.8Hz, 8H), 2.64(td, J=7.2, 3.2Hz, 4H), 2.56(s, 8H).

MS(ESI): m/z 515[M+H] ⁺ .

Example 55 Time-resolved-fluorescence resonance energy transfer experiment (TR-FRET test)

The time-resolved fluorescence resonance energy transfer assay (TR-FRET test) was used to investigate the effects of the compounds described in Examples 1-54 on the interaction between the estrogen-related receptor ERRα and the coactivator protein PGC-1α.

Use a commercial screening kit (LanthaScreenTM ^Etrogen Related Receptor alphaTR-FRET CoactivatorAssay, Invitrogen PV4663), which adopts the principle of fluorescence resonance energy transfer. solution (the 2-indolone compounds in this example all exist in the mixed form of cis-trans isomer interconversion in the solution of the activity test), 10 μL of the solutions of each concentration were taken into 384-well black plates (Thermo, #267461), then 5 μL of ERR alpha-LBD (20nM, 4×) solution was added to each well, followed by 5 μL of fluo-rescein-PGC1α (2 μM, 4×) and Tb anti-GST antibody (20nM, 4×) The mixed solution; placed at room temperature, reacted in the dark for 1 hour, detected the emission intensity at 520nm and 495nm and calculated the ratio. The ratio (emission coefficient) is positively correlated with the concentration of the compound, and the _IC50 value is calculated by plotting its relationship with the concentration.

Table 1 lists the compound numbers of the examples and the corresponding TR-FRET activity results.

Table 1

Among them, the literature source of the positive molecule C29 is J.Med.Chem.2011,54,788-808.

It can be seen from the data in Table 1 that the indolinone molecules of the present invention have strong binding ability to ERRα, thereby affecting the interaction between ERRα and coactivator PGC-1α, and the activity of some compounds is better than that of the positive molecule C29.

Example 56 Effect of compound on tumor cell proliferation inhibitory activity

Tumor cells were stored in the corresponding growth medium and passaged for at least two passages before resuscitation for use. An appropriate number of cells in logarithmic growth phase were seeded in a 96-well plate with a volume of 100 μL per well, and cultured overnight in a 37 °C incubator with 5% CO ₂ . Take 2 μL of compound solutions of different concentrations (the 2-indolone compounds in this example are in the form of cis-trans isomer interconversion in the solution of the activity test) and mix them with 998 μL of culture medium, respectively. Add more than 100 μL of culture medium to the corresponding 96-well plate, and use 2 μL DMSO as blank control. After 72 hours of co-incubation, 20 μL of MTT (5 mg/mL) reagent was added, shaken for 10 minutes, and the number of stained cells was read with a Synergy ^™ HT (Bio Tek)/OD570. Data were processed with GraphPad Prism version 4.0 and a nonlinear regression curve was fitted to derive _IC50 values.

Tumor cell lines were SKBr3 (HER2+), BT-474 (HER2+), MDA-MB-231 (TNBC), HS578T, A549, H460, etc.

Table 2 lists the compound numbers of the examples and the corresponding cell proliferation inhibitory activity results.

Table 2

Note: - is not detected.

It can be seen from the data in Table 2 that the indolinone compounds of the present invention have good proliferation inhibitory activity on tumor cells, which is better than that of the positive molecule C29. Among them, the IC ₅₀ values of compound LS 1-86 on the cell proliferation inhibitory activity of breast cancer cells SKBr3 and BT-474 were 5.3 μM and 6.8 μM, respectively.

The effect of the compound of Example 57 on oral maximal glucose tolerance test in BKS-db (DB/DB) mice

6-week-old DB/DB mice (ie, leptin receptor knockout mice, animal source: Nanjing Institute of Model Animals) were selected and raised for one week after purchase to eliminate stress and adapt to the environment. 32 male mice were randomly divided into 4 groups, 8 mice in each group, as LS 1-86 group (15mg/kg, intraperitoneal injection), LS 1-149 group (15mg/kg, intraperitoneal injection), rosiglitazone group (10 mg/kg, oral administration) and negative control group. It was administered once a day for 28 consecutive days. The body weight and feed intake of the mice were monitored daily. On the 28th day, an oral maximum glucose tolerance test (OGTT) was performed. Fasting overnight before the test, glucose (2g/kg) was intragastrically injected, and blood was collected from the tail vein to measure the blood glucose concentration of the mice at 0, 30, 60, 90, 120, and 150 minutes. , and draw a graph.

The test results are shown in Figure 1. It can be seen from the curve in FIG. 1 that the indolinone compounds of the present invention have a greater effect on the down-regulation of blood glucose, indicating that the mice have improved tolerance to glucose after injection of the indolinone compounds of the present invention.

The technical features of the above-described embodiments can be combined arbitrarily. For the sake of brevity, all possible combinations of the technical features in the following embodiments are not described. However, as long as there is no contradiction between the combinations of these technical features, All should be regarded as the scope described in this specification.

The above-mentioned embodiments only represent several embodiments of the present invention, and the descriptions thereof are specific and detailed, but should not be construed as a limitation on the scope of the patent of the present invention. It should be pointed out that for those of ordinary skill in the art, without departing from the concept of the present invention, several modifications and improvements can also be made, which all belong to the protection scope of the present invention. Therefore, the protection scope of the patent of the present invention should be subject to the appended claims.

Claims (18)

1. An indolone compound having a structure shown in formula (I) or a pharmaceutically acceptable salt thereof or a stereoisomer thereof or a prodrug molecule thereof:

wherein,

m is selected from: 0.1 and 2;

ring (C)

Selected from: s number of R₀Substituted or unsubstituted C₆-C₁₀Aryl, s R₀Substituted or unsubstituted C₅-C₁₀A heteroaryl group;

R₁、R₂、R₃、R₄、R₅、R₆、R₇、R₈、R₀each independently selected from: hydrogen, halogen, cyano, nitro, amino, hydroxy, R_fSubstituted or unsubstituted C₁-C₆Alkyl radical, R_fSubstituted or unsubstituted C₃-C₆Cycloalkyl radical, R_fSubstituted or unsubstituted C₁-C₆Alkoxy radical, R_fSubstituted or unsubstituted C₃-C₆Cycloalkoxy, R_fSubstituted or unsubstituted C₂-C₆Alkenyl radical, R_fSubstituted or unsubstituted C₂-C₆Alkynyl, R_fSubstituted or unsubstituted phenyl, - (CH)₂)_pNR_aR_b-C (═ O) OR, -CONHR, OR R₃And R₅Are linked together by a chemical bond to form-O-, -NH-, or- (CH)₂)_n；

R₉Selected from: hydrogen, - (CH)₂)_qNR_dR_e、-(CH₂)_qOR_d、-(CH₂)_qOCR_cR_dR_e、-(CH₂)_qCR_cR_dR_e；

s is selected from: an integer of 0 to 5;

n is selected from: 1 or 2;

p is selected from; an integer of 0 to 6;

q is selected from: 1 to 6;

R_a、R_beach independently selected from: hydrogen, R_fSubstituted or unsubstituted C₁-C₆Alkyl radical, R_fSubstituted or unsubstituted C₁-C₆Heteroalkyl, or R_a、R_bTogether with the N atom to which they are attached form R_fA substituted or unsubstituted heteroatom-containing monocyclic, fused, spiro or bridged ring;

r is selected from: hydrogen, C₁-C₆An alkyl group;

R_cselected from: hydrogen, C₁-C₃An alkyl group;

R_d、R_eeach independently selected from: hydrogen, R_fSubstituted or unsubstituted C₁-C₆Alkyl radical, R_fSubstituted or unsubstituted C₁-C₆Heteroalkyl, or R_d、R_eTogether with the N or C atom to which they are attached form R_fA substituted or unsubstituted heteroatom-containing monocyclic, fused, spiro or bridged ring;

each R_fEach independently selected from: halogen, hydroxy, amino, C₁-C₃Alkyl, hydroxy substituted C₁-C₃Alkyl radical, C₁-C₃Alkoxy, -NH (C)₁-C₃Alkyl), -N (C)₁-C₃Alkyl) (C₁-C₃Alkyl), -C (═ O) (C)₁-C₃Alkyl), R is a substituted or unsubstituted 5-to 7-membered heterocyclic group.

2. The indolone compound or pharmaceutically acceptable salt thereof or stereoisomer thereof or prodrug molecule thereof of claim 1, wherein the ring is

Selected from: a benzene ring, a pyridine ring, a naphthalene ring, a furan ring, a thiophene ring, a pyrrole ring, a pyrazole ring, a thiazole ring, an isothiazole ring, an oxazole ring, an isoxazole ring, a pyrazine ring, a pyridazine ring, an imidazole ring or a pyrimidine ring.

3. The indolone compound of claim 1, or a pharmaceutically acceptable salt thereof, or a stereoisomer thereof, or a prodrug molecule thereof, wherein R is₁、R₂、R₃、R₄、R₅、R₆、R₇、R₈、R₀Each independently selected from: hydrogen, halogen, cyano, nitro, amino, hydroxy, C₁-C₆Alkyl, halo C₁-C₆Alkyl radical, C₃-C₆Cycloalkyl radical, C₁-C₆Alkoxy, halo C₁-C₆Alkoxy radical, C₃-C₆Cycloalkoxy, C₂-C₆Alkenyl radical, C₂-C₆Alkynyl, phenyl, - (CH)₂)_pNR_aR_b、-C(＝O)OR、-CONHROr R is₃And R₅Are linked together by a chemical bond to form-O-;

R_aand R_bEach independently selected from: hydrogen, R_fSubstituted or unsubstituted C₁-C₃Alkyl, or R_a、R_bTogether with the N atom to which they are attached form R_fA substituted or unsubstituted 3-8 membered monocyclic ring containing 1,2 or 3 heteroatoms, or R_a、R_bTogether with the N atom to which they are attached form R_fA substituted or unsubstituted 8-12 membered fused, spiro or bridged ring containing 1,2 or 3 heteroatoms; the heteroatom is selected from: o, N, S are provided.

4. The indolone compound of claim 3, or a pharmaceutically acceptable salt thereof, or a stereoisomer thereof, or a prodrug molecule thereof, wherein R is₁、R₂、R₃、R₄、R₅、R₆、R₇、R₈、R₀Each independently selected from: hydrogen, halogen, cyano, nitro, hydroxy, C₁-C₃Alkyl, halo C₁-C₃Alkyl radical, C₃-C₆Cycloalkyl radical, C₁-C₃Alkoxy, phenyl, -C (═ O) OR, -CONHR, OR R₃And R₅Are linked together by a chemical bond to form-O-; wherein R is selected from: hydrogen, C₁-C₃An alkyl group.

5. The indolone compound of claim 3, or a pharmaceutically acceptable salt thereof, or a stereoisomer thereof, or a prodrug molecule thereof, wherein R is₁、R₂、R₃、R₄、R₅、R₆、R₇、R₈、R₀Each independently selected from: hydrogen, fluorine, chlorine, bromine, iodine, cyano, nitro, amino, hydroxy, methyl, ethyl, isopropyl, propyl, cyclopropyl, butyl, tert-butyl, cyclobutyl, cyclohexyl, trifluoromethyl, difluoromethyl, methoxy, ethoxy, propoxy, trifluoromethoxy, cyclopropyloxy, vinyl, ethynyl, phenyl, - (CH)₂)_p-NH₂、

-C(＝O)OH、-C(＝O)OCH₃、-C(＝O)OCH₂CH₃、-C(＝O)OCH₂(CH₃)CH₃、-CONH₂、-CONHCH₃Or R is₃And R₅Are linked together by a chemical bond to form-O-; wherein p is an integer of 0 to 3.

6. The indolone compound of claim 1, or a pharmaceutically acceptable salt thereof, or a stereoisomer thereof, or a prodrug molecule thereof, wherein R is₉Selected from: hydrogen, - (CH)₂)_qNR_dR_e(ii) a Wherein,

q is selected from: 1.2, 3 or 4;

R_d、R_eeach independently selected from: hydrogen, R_fSubstituted or unsubstituted C₁-C₆Alkyl, or R_a、R_bTogether with the N atom to which they are attached form R_fA substituted or unsubstituted 3-8 membered monocyclic ring containing 1,2 or 3 heteroatoms, or R_a、R_bTogether with the N atom to which they are attached form R_fA substituted or unsubstituted 8-12 membered fused, spiro or bridged ring containing 1,2 or 3 heteroatoms; the heteroatom is selected from: o, N, S are provided.

7. The indolone compound of claim 6, or a pharmaceutically acceptable salt thereof, or a stereoisomer thereof, or a prodrug molecule thereof, wherein R is_d、R_eEach independently selected from: hydrogen, C₁-C₃Alkyl, or R_a、R_bTogether with the N atom to which they are attached form R_fA substituted or unsubstituted 5-7 membered monocyclic ring containing 1,2 or 3 heteroatoms; the heteroatom is selected from: o, N, respectively; r_fSelected from: H. c₁-C₃Alkyl, -NH (C)₁-C₃Alkyl), -N (C)₁-C₃Alkyl) (C₁-C₃Alkyl), 5-to 7-membered heterocyclic group, C₁-C₃An alkyl-substituted 5-to 7-membered heterocyclic group.

8. The indolone compound of claim 6, or a pharmaceutically acceptable salt thereof, or a stereoisomer thereof, or a prodrug molecule thereof, wherein R is₉Selected from: hydrogen, - (CH)₂)_q-NH₂、

-(CH₂)_q-OH、-(CH₂)_q-OH₃、-(CH₂)_q-OCH₂CH₃(ii) a Wherein q is selected from 1,2, 3 or 4.

9. The indolone compound or pharmaceutically acceptable salt thereof or stereoisomer thereof or prodrug molecule thereof of claim 1, wherein the ring is

Selected from: benzene ring, naphthalene ring;

R₁、R₂、R₃、R₄、R₅、R₆、R₇、R₈each independently selected from: hydrogen, methoxy, hydroxy, fluoro, chloro, bromo, methyl, cyclopropyl, phenyl, or R₃And R₅Are linked together by a chemical bond to form-O-;

R₀selected from: hydrogen, cyano, trifluoromethyl, nitro, fluoro, chloro, bromo, carboxamido, formyl, methyl formate, difluoromethyl, methoxy;

R₉selected from: hydrogen, - (CH)₂)_q-NH₂、

Wherein q is selected from 1,2, 3 or 4.

10. The indolone compound of claim 9, or a pharmaceutically acceptable salt thereof, or a stereoisomer thereof, or a prodrug molecule thereof, wherein the ring is

Selected from: benzene ring, naphthalene ring;

R₁、R₂、R₃、R₄、R₅、R₆、R₇、R₈each independently selected from: hydrogen, methoxy, hydroxy, fluoro, chloro, bromo, methyl, cyclopropyl, phenyl, or R₃And R₅Are linked together by a chemical bond to form-O-;

R₀selected from: hydrogen, cyano, trifluoromethyl, nitro, fluoro, chloro, bromo, methyl formate, difluoromethyl, methoxy;

R₉selected from: hydrogen, hydrogen,

Wherein q is selected from 1,2, 3 or 4.

11. The indolone compound or the pharmaceutically acceptable salt thereof, or the stereoisomer thereof, or the prodrug molecule thereof according to any one of claims 1 to 10, wherein the indolone compound has a structure represented by formula (II) or formula (III):

wherein s is selected from: an integer of 0 to 4.

12. The indolone compound of claim 11, or a pharmaceutically acceptable salt thereof, or a stereoisomer thereof, or a prodrug molecule thereof, wherein R is₁、R₂、R₃、R₄、R₆、R₇、R₈Each independently selected from: hydrogen, methoxy, hydroxy, fluoro, chloro, bromo, methyl, cyclopropyl, phenyl;

R₅selected from: hydrogen, phenyl; and when R is₅When it is hydrogen, R₂And R₃Are not hydroxyl;

R₀selected from: hydrogen, cyano, trifluoromethyl, nitro, fluoro, chloro, bromo, methyl formate, difluoromethyl, methoxy;

R₉selected from: hydrogen, hydrogen,

q is selected from: 1.2, 3 or 4.

13. The indolone compound or pharmaceutically acceptable salt thereof or stereoisomer thereof or prodrug molecule thereof of claim 1, wherein the indolone compound or stereoisomer thereof is selected from the following compounds:

14. use of an indolone compound of any one of claims 1-13 or a pharmaceutically acceptable salt or stereoisomer thereof or prodrug molecule thereof in the preparation of an ERR α inverse agonist.

15. Use of the indolone compound or pharmaceutically acceptable salt thereof or stereoisomer thereof or prodrug molecule thereof according to any one of claims 1 to 13 in the preparation of a medicament for preventing and/or treating a disease associated with abnormal expression of ERR α protein activity.

16. The use of claim 15, wherein the disease associated with aberrant expression of ERR α protein activity is: tumors, metabolic diseases.

17. The use of claim 16, wherein the tumor is: non-small cell lung cancer, lung adenocarcinoma, lung squamous carcinoma, pancreatic cancer, breast cancer, prostate cancer, liver cancer, skin cancer, epithelial cell cancer, gastrointestinal stromal tumor, endometrial cancer, histiocytic lymphoma, nasopharyngeal carcinoma, head and neck tumors, colon cancer, rectal cancer, glioma, malignant melanoma, renal cancer, bladder cancer, ovarian cancer, cervical cancer, laryngeal cancer, or multiple myeloma, B lymphoma, leukemia;

the metabolic disease is: hyperglycemia, diabetes, obesity, atherosclerosis, metabolic syndrome, liver fibrosis, non-alcoholic fatty liver disease, gallstone, hyperlipidemia, hypercholesterolemia, hyperlipoproteinemia, hypertriglyceridemia, hypertension, hyperinsulinemia, hyperuricemia, Parkinson's disease, and Alzheimer's disease.

18. A pharmaceutical composition for preventing and/or treating diseases associated with abnormal expression of ERR α protein activity, which is prepared from an active ingredient and pharmaceutically acceptable excipients, wherein the active ingredient comprises the indolone compound or the pharmaceutically acceptable salt thereof or the stereoisomer thereof or the prodrug molecule thereof according to any one of claims 1 to 13.

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