O-aminopyridine formyl hydrazones and methods of making and using the same

Abstract

This disclosure relates to the field of biomedical technology, specifically to o-aminopyridinecarboxyhydrazone compounds, their preparation methods, and uses. This disclosure offers the following advantages: it provides the use of o-aminopyridinecarboxyhydrazone compounds in the preparation of drugs for the prevention or treatment of angiogenesis-related diseases mediated by abnormal VEGFR-2 kinase. This disclosure of o-aminopyridinecarboxyhydrazone compounds holds promise for providing new strategies for the prevention or treatment of angiogenesis-related diseases.

Description

Technical Field

[0001] This disclosure relates to the field of biomedical technology, specifically to o-aminopyridine carboxyhydrazone compounds, their preparation methods, and uses. Background Technology

[0002] Angiogenesis refers to the process by which new blood vessels form from existing ones. This process includes physiological and pathological angiogenesis. Physiological angiogenesis generally occurs during wound healing, tissue repair, and placental development during pregnancy. Pathological angiogenesis, on the other hand, is caused by abnormal expression of pro-angiogenic factors, leading to abnormal blood vessel proliferation and resulting in many diseases, such as tumor growth and metastasis, age-related macular degeneration or neovascular glaucoma, obesity, kidney disease, arthritis, and atherosclerosis.

[0003] Vascular endothelial growth factor receptor (VEGFR) belongs to the receptor tyrosine kinase (RTK) superfamily and includes three subtypes: VEGFR-1 (Flt-1), VEGFR-2 (KDR / FLK-1), and VEGFR-3 (Flt-4). VEGFR-2 is a high-affinity receptor for VEGF (Vascular Endothelial Growth Factor) and plays a crucial role in angiogenesis. VEGFR-2 is mainly distributed in vascular endothelial cells and can selectively bind to growth factors such as VEGF-A, VEGF-C, VEGF-D, and VEGF-E, thereby playing a vital role in increasing vascular permeability, promoting vascular endothelial cell proliferation, and facilitating angiogenesis.

[0004] The specific binding of vascular endothelial growth factor (VEGF) to VEGFR phosphorylates specific tyrosine residues within cells, thereby activating downstream signaling pathways and inducing persistent angiogenesis. Therefore, directly or indirectly inhibiting VEGFR and blocking its expression can be used to prevent or treat VEGF-induced pathological angiogenesis, such as tumor angiogenesis and neovascular fundus diseases. For example, sorafenib and regorafenib inhibit the expression of receptors such as VEGFR-2, thereby blocking tumor angiogenesis and cutting off the nutrient supply to tumor cells, thus inhibiting tumor growth. Ranibizumab and conbercept inhibit VEGF receptor activation, blocking pathological angiogenesis, and are used to treat fundus neovascular diseases such as wet age-related macular degeneration, diabetic macular edema, pathological myopia, and macular edema secondary to retinal vein occlusion.

[0005] Given the crucial role of VEGFR-2 in angiogenesis, research on anti-tumor drugs targeting VEGFR-2 has garnered increasing attention. To date, numerous VEGFR-2 inhibitors have been reported, with some already approved for marketing or in clinical trials. VEGFR-2 inhibitors can be categorized by structure into pyridine, pyrimidine, indoleone, anthranilamide, and other classes. These small-molecule VEGFR-2 inhibitors have all demonstrated effective inhibitory effects against cancer. Among them, sorafenib was approved by the FDA in 2005 for the treatment of advanced renal cell carcinoma; regorafenib was approved by the FDA in 2012 for the treatment of metastatic colorectal cancer; cabozantinib was approved by the FDA in 2012 for the treatment of medullary thyroid carcinoma, prostate cancer, renal cell carcinoma, and glioblastoma; pazopanib was approved by the FDA in 2009 for the treatment of advanced renal cell carcinoma; vandetanib was approved by the FDA in 2011 for the treatment of advanced metastatic thyroid cancer; sunitinib was approved by the FDA in 2006 for the treatment of gastrointestinal stromal tumors and metastatic renal cell carcinoma; nintedanib was approved by the FDA in 2014 for the treatment of non-small cell lung cancer and idiopathic pulmonary fibrosis; and apatinib was approved by the CFDA in 2014 for the treatment of advanced gastric adenocarcinoma.

[0006] From a pharmaceutical perspective, current small-molecule VEGFR-2 kinase inhibitors suffer from drawbacks such as drug resistance, significant toxicity, and low bioavailability, particularly in the treatment of retinal neovascularization. Therefore, developing novel small-molecule drugs targeting VEGFR-2 kinase with good selectivity, high bioavailability, and clinically viable alternatives to drug-resistant formulations is an urgent need in the fields of malignant tumors and retinal neovascularization.

[0007] Therefore, this disclosure provides a novel and highly active o-aminopyridine carboxyhydrazone compound targeting VEGFR-2 kinase, its preparation method, and its uses. Summary of the Invention

[0008] This disclosure aims to develop o-aminopyridine carboxyhydrazone compounds, their preparation methods, and uses. As vascular endothelial growth factor receptor (VEGFR-2) kinase inhibitors, o-aminopyridine carboxyhydrazone compounds can be used to prevent or treat angiogenesis-related diseases mediated by abnormal VEGFR-2 kinases.

[0009] To achieve the above-mentioned technical objectives, the technical solution adopted in this disclosure is as follows:

[0010] On the one hand, this disclosure provides compounds of formula (I) or pharmaceutically acceptable salts, solvates, isotopic variants, tautomers, diastereomers, enantiomers, or cis-trans isomers thereof:

[0011] (I)

[0012] In formula (I),

[0013] Each R is independently absent or selected from halogen, cyano, C1-6 alkyl, C1-6 alkoxy, haloC1-6 alkyl, haloC1-6 alkoxy, or -NR. 1 R 2 ;

[0014] R 1 and R 2 Each is independently selected from hydrogen, halogen, cyano, C 1-6 Alkyl, C 1-6 Alkoxy, halogenated C 1-6 Alkyl, Halogenated C 1-6 Alkoxy, C 3-6 cycloalkyl, or R 1 and R 2 Together with the nitrogen atom attached to it, they form nitrogen-containing 3-6 membered rings;

[0015] The nitrogen-containing 3-6 membered ring optionally contains other nitrogen, sulfur, or oxygen atoms, or contains the group -N(R) 3 )-;

[0016] R 3 It is hydrogen or C 1-6 alkyl;

[0017] The C1-6 alkyl, C1-6 alkoxy, halogenated C1-6 alkyl, halogenated C1-6 alkoxy, or -NR 1 R 2 Optionally, it is affected by one or more halogens, cyano groups, hydroxyl groups, C 1-6 Alkyl, C 1-6 Alkoxy, halogenated C 1-6 Alkyl or halogenated C 1-6 Alkyl substitution;

[0018] n is an integer selected from 0 to 5.

[0019] In some embodiments, the compound represented by formula (I) has the structure represented by formula (II):

[0020] (II)

[0021] In equation (II), R is defined in the same way as in equation (I).

[0022] In some embodiments, the compound represented by formula (I) has the structure represented by formula (III):

[0023] (III)

[0024] In equation (Ⅲ), R is defined in the same way as in equation (I).

[0025] In some embodiments, the compound represented by formula (I) has the structure represented by formula (IV):

[0026] (Ⅳ)

[0027] In equation (Ⅳ), R is defined in the same way as in equation (I).

[0028] In some embodiments, each R is independently absent or selected from cyano, C1-3 alkoxy, halo-C1-3 alkyl, halo-C1-3 alkoxy, or -NR. 1 R 2 ;R 1 and R 2 Each is independently selected from hydrogen or C. 1-3 alkyl.

[0029] In some embodiments, each R is independently absent or selected from cyano, methoxy, ethoxy, trifluoromethyl, difluoromethyl, monofluoromethyl, trifluoromethoxy, difluoromethoxy, monofluoromethoxy, -NH2, -N(H)(CH3), -N(CH3)(CH3), -N(CH3)(CH2CH3), -N(CH2CH3)(CH2CH3), or -N(H)(CH2CH3).

[0030] In some implementations, each R is independently absent or selected from cyano, methoxy, ethoxy, trifluoromethyl, difluoromethyl, monofluoromethyl, trifluoromethoxy, difluoromethoxy, monofluoromethoxy, -NH2, -N(H)(CH3) or -N(CH3)(CH3).

[0031] In some implementations, each R is independently absent or selected from cyano, methoxy, ethoxy, trifluoromethyl, trifluoromethoxy, -NH2, -N(H)(CH3) or -N(CH3)(CH3).

[0032] In some implementations, each R is independently absent or selected from cyano, methoxy, trifluoromethyl, trifluoromethoxy, or -N(CH3)(CH3).

[0033] In some embodiments, each R is independently absent or selected from halogen, cyano, C1-5 alkyl, C1-5 alkoxy, halo-C1-5 alkyl, halo-C1-5 alkoxy, or -NR. 1 R 2 .

[0034] In some embodiments, each R is independently absent or selected from halogen, cyano, C1-3 alkyl, C1-3 alkoxy, haloC1-3 alkyl, haloC1-3 alkoxy, or -NR. 1 R 2 .

[0035] In some embodiments, each R is independently absent or selected from fluorine, chlorine, cyano, methyl, ethyl, methoxy, ethoxy, trifluoromethyl, difluoromethyl, monofluoromethyl, trifluoromethoxy, difluoromethoxy, monofluoromethoxy, -NH2, -N(H)(CH3), -N(CH3)(CH3), -N(CH3)(CH2CH3), -N(CH2CH3)(CH2CH3), or -N(H)(CH2CH3).

[0036] In some implementations, each R is independently absent or selected from fluorine, chlorine, cyano, methyl, ethyl, methoxy, ethoxy, trifluoromethyl, difluoromethyl, monofluoromethyl, trifluoromethoxy, difluoromethoxy, monofluoromethoxy, -NH2, -N(H)(CH3) or -N(CH3)(CH3).

[0037] In some implementations, each R is independently absent or selected from fluorine, chlorine, cyano, methyl, ethyl, methoxy, ethoxy, trifluoromethyl, trifluoromethoxy, -NH2, -N(H)(CH3) or -N(CH3)(CH3).

[0038] In some implementations, each R is independently absent or selected from fluorine, chlorine, cyano, methyl, methoxy, trifluoromethyl, trifluoromethoxy, or -N(CH3)(CH3).

[0039] In some implementation schemes, R 1 and R 2 Each is independently selected from hydrogen, halogen, cyano, C 1-5 Alkyl, C 1-5 Alkoxy, halogenated C 1-5 Alkyl, Halogenated C 1-5 Alkoxy, C 3-6 cycloalkyl, or R 1 and R 2 Together with nitrogen atoms, they form nitrogen-containing 3-6 membered rings;

[0040] The nitrogen-containing 3-6 membered ring optionally includes other nitrogen atoms, or includes the group -N(R) 3 )-.

[0041] In some implementation schemes, R 1 and R 2 Each is independently selected from hydrogen, halogen, cyano, C 1-3 Alkyl, C 1-3 Alkoxy, halogenated C 1-3 Alkyl, Halogenated C 1-3 Alkoxy, C 3-6 cycloalkyl, or R 1 and R 2 Together with nitrogen atoms, they form nitrogen-containing C atoms. 3-6 ring;

[0042] The nitrogen-containing 3-6 membered ring optionally includes other nitrogen atoms, or includes the group -N(R) 3 )-.

[0043] In some implementation schemes, R 1 and R 2 Each is independently selected from hydrogen, halogen, cyano, C 1-3Alkyl, C 1-3 Alkoxy, halogenated C 1-3 Alkyl, Halogenated C 1-3 Alkoxy or C 3-6 Cycloalkyl.

[0044] In some implementation schemes, R 1 and R 2 Each is independently selected from hydrogen and C. 1-3 Alkyl, C 1-3 Alkoxy, halogenated C 1-3 Alkyl, Halogenated C 1-3 Alkoxy or C 3-4 Cycloalkyl.

[0045] In some implementation schemes, R 1 and R 2 Each is independently selected from hydrogen and C. 1-3 Alkyl, C 1-3 alkoxy or halogenated C 1-3 alkyl.

[0046] In some implementation schemes, R 1 and R 2 Each is independently selected from hydrogen and C. 1-3 Alkyl or C 1-3 Alkyl group.

[0047] In some implementation schemes, R 1 and R 2 Each is independently selected from hydrogen or C. 1-5 alkyl.

[0048] In some implementation schemes, R 1 and R 2 Each is independently selected from hydrogen or C. 1-4 alkyl.

[0049] In some implementation schemes, R 1 and R 2 Each is independently selected from hydrogen or C. 1-3 alkyl.

[0050] In some implementation schemes, R 1 and R 2 Each is independently selected from hydrogen, methyl, or ethyl.

[0051] In some implementation schemes, R 1 and R 2 Each is independently selected from hydrogen or methyl.

[0052] In some implementation schemes, R 1 and R 2 Each is independently selected from methyl groups.

[0053] In some implementation schemes, R 3 It is hydrogen or C 1-5 alkyl.

[0054] In some implementation schemes, R 3 It is hydrogen or C 1-4 alkyl.

[0055] In some implementation schemes, R 3 It is hydrogen or C 1-3 alkyl.

[0056] In some implementation schemes, R 3 It can be hydrogen, methyl, or ethyl.

[0057] In some implementation schemes, R 3 It can be hydrogen or methyl.

[0058] In some implementation schemes, R 3 It is hydrogen.

[0059] In some embodiments, the C1-6 alkyl, C1-6 alkoxy, halo-C1-6 alkyl, halo-C1-6 alkoxy, or -NR 1 R 2 Optionally, it is affected by one or more halogens, cyano groups, hydroxyl groups, C 1-3 Alkyl, C 1-3 Alkoxy, halogenated C 1-3 Alkyl or halogenated C 1-3 Alkyl-substituted.

[0060] In some embodiments, the C1-6 alkyl, C1-6 alkoxy, halo-C1-6 alkyl, halo-C1-6 alkoxy, or -NR 1 R 2 Optionally, it is affected by one or more halogens, cyano groups, hydroxyl groups, C 1-3 Alkyl or C 1-3 Alkyl-substituted.

[0061] In some embodiments, the C1-6 alkyl, C1-6 alkoxy, halo-C1-6 alkyl, halo-C1-6 alkoxy, or -NR 1 R 2 It may optionally be substituted with one or more halogens, cyano groups, hydroxyl groups, methyl groups, ethyl groups, or methoxy groups.

[0062] In some embodiments, the C1-6 alkyl, C1-6 alkoxy, halo-C1-6 alkyl, halo-C1-6 alkoxy, or -NR 1 R 2 Optionally substituted with one or more halogens, methyl, ethyl or methoxy groups.

[0063] In some implementations, n is selected from integers between 0 and 4.

[0064] In some implementations, n is selected from integers between 0 and 3.

[0065] In some implementations, n is selected from integers between 0 and 2.

[0066] In some implementations, n is selected from 0 or 1.

[0067] In some implementations, n is 1.

[0068] On the other hand, this disclosure provides the following compounds or their pharmaceutically acceptable salts, solvates, isotopic variants, tautomers, diastereomers, enantiomers, or cis-trans isomers:

[0069] .

[0070] On the other hand, this disclosure provides a method for preparing the compound as described above or its pharmaceutically acceptable salt, solvate, isotopic variant, tautomer, diastereomer, enantiomer, or cis-trans isomer, comprising the following steps:

[0071] <1> Compound C was prepared by reacting compound A and compound B under alkaline conditions via a nucleophilic substitution reaction;

[0072] <2> Compound D was prepared from compound C via hydrazinolysis.

[0073] <3> The compound described above was prepared by condensation reaction of compound D and compound E under acidic conditions;

[0074]

[0075] R and n are defined as previously.

[0076] The compound E is selected from , , , , , , , , , , , , , , , or .

[0077] The compound DE is the compound as described above.

[0078] In some embodiments, compound E is selected from... , or .

[0079] In some embodiments, compound E is selected from... or .

[0080] In some embodiments, the compound DE is selected from the compound represented by formula (I), the structure represented by formula (II), the structure represented by formula (III), or the structure represented by formula (IV).

[0081] In some embodiments, the compound DE is selected from the following compounds:

[0082] .

[0083] In some implementation schemes, the steps <1> In this reaction, the base is selected from sodium hydroxide, potassium hydroxide, anhydrous potassium carbonate, and anhydrous sodium carbonate, and the reaction solvent is N,N-dimethylformamide (DMF), dimethyl sulfoxide (DMSO), acetonitrile, or tetrahydrofuran (THF).

[0084] In some implementation schemes, the steps <1> In this process, the reaction temperature ranges from room temperature to 100°C.

[0085] In some implementation schemes, the steps <2> In the hydrazolysis reaction, the reagent used is an 80% hydrazine hydrate solution.

[0086] In some implementation schemes, the steps <2> In this reaction, the solvent is selected from anhydrous methanol or anhydrous ethanol.

[0087] In some implementation schemes, the steps <2> In this process, the reaction temperature is between 65℃ and 80℃.

[0088] In some implementation schemes, the steps <3> In this context, the acid is selected from formic acid or glacial acetic acid.

[0089] In some implementation schemes, the steps <3> In this reaction, the solvent is anhydrous methanol or anhydrous ethanol.

[0090] In some implementation schemes, the steps <3> In this process, the reaction temperature ranges from room temperature to 80°C.

[0091] On the other hand, this disclosure provides pharmaceutical compositions comprising the compounds as described above or pharmaceutically acceptable salts, solvates, isotopic variants, tautomers, diastereomers, enantiomers, or cis-trans isomers thereof, or compounds obtained by the preparation methods described above or pharmaceutically acceptable salts, solvates, isotopic variants, tautomers, diastereomers, enantiomers, or cis-trans isomers thereof.

[0092] On the other hand, this disclosure provides the use of the compounds as described above, or pharmaceutically acceptable salts, solvates, isotopic variants, tautomers, diastereomers, enantiomers, or cis-trans isomers thereof, or compounds obtained by the methods described above, or pharmaceutically acceptable salts, solvates, isotopic variants, tautomers, diastereomers, enantiomers, or cis-trans isomers thereof, or pharmaceutical compositions as described above, in the preparation of medicaments for the prevention or treatment of diseases related to angiogenesis mediated by abnormal VEGFR-2 kinase.

[0093] In some embodiments, the drug acts as a VEGFR tyrosine kinase inhibitor;

[0094] In some implementations, the drug is a VEGFR-2 tyrosine kinase inhibitor.

[0095] In some implementations, the VEGFR is VEGFR1, VEGFR2, and / or VEGFR3.

[0096] In some implementations, the VEGFR is VEGFR-2.

[0097] In some embodiments, the drug is used in combination with other active ingredients in the preparation of a medicament for the prevention or treatment of diseases related to angiogenesis mediated by abnormal VEGFR-2 kinase.

[0098] In some implementations, abnormal VEGFR-2 kinase-mediated angiogenesis-related diseases include kidney disease, immune system disorders, embolic microvascular syndromes, retinal neovascularization, or malignant tumors.

[0099] In some implementation schemes, ocular neovascularization-related diseases include diabetic retinopathy, wet age-related macular degeneration, corneal vascular eye disease, iris vascular eye disease, pterygium, neovascular glaucoma, and choroidal neovascularization.

[0100] On the other hand, this disclosure provides a method for preventing or treating diseases related to angiogenesis mediated by abnormal VEGFR-2 kinase, comprising administering to a patient in need a compound as described above or a pharmaceutically acceptable salt, solvate, isotopic variant, tautomer, diastereomer, enantiomer, or cis-trans isomer thereof, or a compound obtained by the preparation method described above or a pharmaceutically acceptable salt, solvate, isotopic variant, tautomer, diastereomer, enantiomer, or cis-trans isomer thereof, or a pharmaceutical composition as described above.

[0101] In some implementations, diseases associated with abnormal VEGFR-2 kinase-mediated angiogenesis are selected from non-small cell lung cancer, liver cancer, hemangioma, angiofibroma, malignant melanoma, gastrointestinal stromal tumor, pancreatic cancer, metastatic colorectal cancer, recurrent glioblastoma, biliary tract tumors, soft tissue sarcoma, breast cancer, ovarian cancer, rheumatoid arthritis, kidney diseases such as glomerulonephritis, diabetic nephropathy, malignant nephrosclerosis, embolic microangiitis syndrome, or eye diseases.

[0102] In some implementations, the eye disease includes, but is not limited to, diabetic retinopathy, neovascular glaucoma, or age-related macular degeneration.

[0103] This disclosure has the following advantages:

[0104] (1) This disclosure provides the use of o-aminopyridine carboxyhydrazone compounds or pharmaceutically acceptable salts, solvates, isotopic variants, tautomers, diastereomers, enantiomers or cis-trans isomers of the same in the preparation of medicaments for the prevention or treatment of angiogenesis-related diseases.

[0105] (2) This disclosure provides the use of o-aminopyridine carboxyhydrazone compounds or pharmaceutically acceptable salts, solvates, isotopic variants, tautomers, diastereomers, enantiomers or cis-trans isomers thereof in the preparation of medicaments for the prevention or treatment of diseases related to angiogenesis mediated by abnormal VEGFR-2 kinase.

[0106] (3) The present disclosure provides that o-aminopyridine carboxyhydrazone compounds or their pharmaceutically acceptable salts, solvates, isotopic variants, tautomers, diastereomers, enantiomers or cis-trans isomers have good inhibitory activity against VEGFR-2 kinase.

[0107] (4) The present disclosure provides that o-aminopyridine carboxyhydrazone compounds or pharmaceutically acceptable salts, solvates, isotopic variants, tautomers, diastereomers, enantiomers or cis-trans isomers thereof may be used to treat diseases related to angiogenesis mediated by abnormal VEGFR-2 kinase, including nephropathy, immune system diseases, embolic microvascular syndromes, fundus neovascularization diseases, malignant tumors, etc.

[0108] (5) This disclosure provides o-aminopyridine carboxyhydrazone compounds, which are expected to provide new strategies for the prevention or treatment of angiogenesis-related diseases. Detailed Implementation

[0109] Definitions and Explanations

[0110] To facilitate understanding of this disclosure, certain technical and scientific terms are specifically defined below. In this disclosure, unless otherwise stated, the scientific and technical terms used herein have meanings commonly understood by those skilled in the art. Furthermore, the cell and tissue culture, microbiology-related terms, and laboratory procedures used herein are all widely used terms and routine procedures in their respective fields. Meanwhile, to better understand this disclosure, definitions and explanations of relevant terms are provided below. It should be understood that this disclosure is not limited to specific methods, reagents, compounds, compositions, or biological systems, and variations thereof are certainly possible. It should also be understood that the terminology used in this application is for describing specific embodiments only and is not intended to be limiting.

[0111] Unless otherwise expressly stated, the terms “a,” “an,” and “the” as used in this specification and the appended claims cover one or more types.

[0112] As used herein, the terms “comprising” and “having”, and any variations thereof, are intended to cover non-exclusive inclusion. For example, a process, method, apparatus, product, or device that includes a series of steps is not limited to the steps or modules listed, but may optionally include steps not listed, or may optionally include other steps inherent to such process, method, product, or device.

[0113] In the description herein, references to “some embodiments,” “some implementations,” or “some implementation schemes” describe a subset of all possible embodiments. However, it is understood that “some embodiments” may be the same subset or different subsets of all possible embodiments and may be combined with each other without conflict.

[0114] When listing a range of values, it is assumed that each value and the subranges within that range are included. For example, "C 1-6 Alkyl groups include C1, C2, C3, C4, C5, C6, and C6. 1-6 C 1-5C 1-4 C 1-3 C 1-2 C 2-6 C 2-5 C 2-4 C 2-3 C 3-6 C 3-5 C 3-4 C 4-6 C 4-5 and C 5-6 alkyl.

[0115] As used herein, the term “substitution” means that any one or more hydrogen atoms on a particular atom are replaced by a substituent, which may include deuterium and hydrogen variants, provided that the valence state of the particular atom is normal and the substituted compound is stable. The terms “optional substitution” or “optionally substituted” mean that a substance may or may not be substituted, and unless otherwise specified, the type and number of substituents may be arbitrary on a chemically feasible basis.

[0116] When any variable (e.g., R1, R2) appears more than once in the composition or structure of a compound, its definition is independent in each case. Therefore, for example, if a group is substituted by 0-2 R1s, the group can optionally be substituted by at most two R1s, and R1 has independent options in each case. Furthermore, combinations of substituents and / or their variants are only permitted if such combinations produce a stable compound.

[0117] In any embodiment, any or all hydrogen atoms present in the compound, or hydrogen atoms in a specific group or portion of the compound, may be replaced by deuterium or tritium. One to a maximum number of hydrogen atoms present in the compound may be replaced by deuterium. One to a maximum number of hydrogen atoms present in any group of the general formula compound or a specific compound may be replaced by deuterium. For example, when a group is described as ethyl, the ethyl group may be C2H5 or a C2H5 in which x (1 to 5) hydrogen atoms are replaced by deuterium, such as C2D. x H 5-x When a group is described as a deuterated ethyl group, the deuterated ethyl group can be a C2H5 with x (1 to 5) hydrogen atoms replaced by deuterium, such as C2D. x H 5-x The stable deuterated derivatives described in this disclosure are preferably stable deuterated isotope derivatives obtained by replacing any deuterated hydrogen atom in each formula with 1 to a maximum number (e.g., 1 to 2, 1 to 3, 1 to 4, 1 to 5, 1 to 6, 1 to 7, etc.) of deuterium atoms.

[0118] This disclosure refers to compounds of formula (I), and also includes their tautomers, stereoisomers, mixtures of stereoisomers, solvates or derivatives.

[0119] This disclosure of "compounds" also includes tautomer forms. A tautomer form arises from the exchange of a single bond with an adjacent double bond, accompanied by the migration of a proton. The terms "tautomer" or "tautomer form" refer to isomers of different functional groups in dynamic equilibrium at room temperature that can rapidly interconvert. It refers to one of two or more structural isomers that exist in equilibrium and readily transform from one isomer form to another. This transformation results in the formal migration of a hydrogen atom, accompanied by the conversion of adjacent conjugated double bonds. Tautomers exist as a mixture of tautomer groups in solution. In solutions where tautomerization is possible, chemical equilibrium of the tautomers will be reached. The exact proportions of the tautomers depend on several factors, including temperature, solvent, and pH conditions. The concept of tautomers that can interconvert through tautomerization is called tautomerism.

[0120] When this specification describes a compound that is readily tautomerizable, but only one of its tautomers is described, it should be understood that all tautomers are included as part of the chemical meaning described. It should be understood that when a compound has tautomeric forms, it is intended to include all tautomeric forms, and the naming of the compound does not exclude any tautomeric form.

[0121] Of the various possible types of tautomerism, two are typically observed. In keto-enol tautomerism, both electrons and hydrogen atoms move simultaneously.

[0122] Common tautomer pairs are: keto-enol, amide-nitrile, lactam-lactam, amide-imine tautomer in heterocycles, imine-enamine, and enamine-enamine.

[0123] The term "isomer" refers to different compounds having the same molecular formula but different atomic arrangements and configurations. Depending on their structure, the compounds of this disclosure can exist in different stereoisomeric forms. These forms include configurational isomers or optical conformational isomers (enantiomers and / or diastereomers, including those that are blocked from rotation). Therefore, this disclosure includes enantiomers, diastereomers, and mixtures thereof. This disclosure further includes all mixtures of the above-described stereoisomers, regardless of proportions, including racemic mixtures.

[0124] Based on their structure, the compounds disclosed herein can exist in various stable isotopic forms. These forms include those in which one or more hydrogen atoms have been replaced by deuterium atoms, and those in which one or more nitrogen atoms have been replaced by deuterium atoms. 15Those that are replaced by N atoms, or one or more of the carbon, fluorine, chlorine, bromine, sulfur, or oxygen atoms, have been replaced by stable isotopes of their respective original atoms.

[0125] According to this disclosure, some compounds and salts can exist in different crystalline forms (polymorphs) within the scope of this disclosure.

[0126] The term "alkyl" refers to a chain-like (straight-chain or branched) saturated aliphatic hydrocarbon group. The term "alkyl" can refer to a straight-chain or branched alkyl group containing 1 to 10 carbon atoms. 1-10 Alkyl groups, preferably alkyl groups containing 1 to 6 carbon atoms (C 1-6 Alkyl groups. Non-limiting examples include methyl, ethyl, n-propyl, isopropyl, n-butyl, isobutyl, tert-butyl, sec-butyl, n-pentyl, 1,1-dimethylpropyl, 1,2-dimethylpropyl, 2,2-dimethylpropyl, 1-ethylpropyl, 2-methylbutyl, 3-methylbutyl, n-hexyl, 1-ethyl-2-methylpropyl, 1,1,2-trimethylpropyl, 1,1-dimethylbutyl, 1,2-dimethylbutyl, 2,2-dimethylbutyl, 1,3-dimethylbutyl, 2-ethylbutyl, 2-methylpentyl, 3-methylpentyl, 4-methylpentyl, 2,3-dimethylbutyl, and various branched isomers thereof. More preferably are lower alkyl groups containing 1 to 3 carbon atoms (C... 1-3 Alkyl groups, including methyl, ethyl, n-propyl, isopropyl, etc., in non-limiting embodiments. Alkyl groups may be substituted or unsubstituted, and when substituted, the substituents are preferably one or more groups described in this application.

[0127] The term "aryl" should be understood to preferably refer to a monovalent aromatic or partially aromatic monocyclic, bicyclic, or tricyclic hydrocarbon ring ("C") having 6, 7, 8, 9, or 10 carbon atoms. 6-10 Aryl), particularly a ring with 6 carbon atoms (“C6 aryl”), such as phenyl; or biphenyl, or a ring with 9 carbon atoms (“C9 aryl”), such as indenyl or indenyl, or a ring with 10 carbon atoms (“C9 aryl”). 10 Aryl), such as tetrahydronaphthyl, dihydronaphthyl, or naphthyl. When the C 6-10 When the aryl group is substituted, it can be monosubstituted or polysubstituted. Furthermore, there are no restrictions on the substitution site; for example, it can be ortho, para, or meta substituted.

[0128] The term "heteroaryl" refers to a monocyclic, bicyclic, or tricyclic aromatic ring system having 5, 6, 7, 8, 9, or 10 ring atoms, wherein the ring atoms comprise 1 to 5 heteroatoms independently selected from N, O, and S, and the bicyclic and tricyclic aromatic ring systems can be fused rings, spirocyclic, or bridged rings (5-10 membered heteroaryls). The 5-10 membered heteroaryl group contains 1 to 5 heteroatoms, preferably 1 to 3. Additionally, in each case, the 5-10 membered heteroaryl group can be benzofused. Examples of heteroaryl groups include, but are not limited to: 5-membered rings, such as oxazolyl, pyrazolyl, thiophene, thiazolyl, triazole, imidazolyl, etc.; and 6-membered rings, such as pyridinyl, pyrimidinyl, pyrazinyl, pyridazinyl, etc. The heteroaryl group can be bicyclic, including but not limited to: 5,5-membered rings, such as tetrahydrocyclopentanopyrazole; 5,6-membered rings, such as tetrahydroindole, tetrahydropyrazolopyridine, tetrahydroimidazopyridine, tetrahydrobenzisoxazole, tetrahydrobenzoxazole, tetrahydrobenzothiazole, tetrahydrobenzisoxiazole, dihydrofuranopyrazole, tetrahydrobenzofuran, dihydrobenzofuran, tetrahydrobenzothiophene; 6,6-membered rings, such as tetrahydroquinoline; 5,7-membered rings, such as tetrahydrocycloheptazothiazole, tetrahydrocycloheptazofuran. When the 5-10-membered heteroaryl group is substituted, it can be monosubstituted or polysubstituted. Furthermore, there are no restrictions on the substitution site; for example, the hydrogen atom bonded to a carbon atom on the heteroaryl ring can be substituted, or the hydrogen atom bonded to a heteroatom on the heteroaryl ring can be substituted.

[0129] The term "solvent" as used in this disclosure refers to a complex formed by the compound of this disclosure with a solvent. These complexes either react in the solvent or precipitate or crystallize from the solvent. For example, a complex formed with water is called a "hydrate". Solvents of the compounds represented by formula (I) of this disclosure are within the scope of this disclosure.

[0130] IC 50 (IC50) refers to the concentration of an inhibitor required under specific experimental conditions to reduce a biological activity (such as cell proliferation) to 50% of its original value. 50 IC is an important parameter for measuring drug efficacy. 50 The smaller the value, the stronger the inhibitory effect of the drug at a lower concentration.

[0131] Pterygium is a common ocular proliferative disease characterized by abnormal proliferation of conjunctival tissue that invades the cornea, forming growths.

[0132] As used in this article, the term "hydroxyl group" refers to -OH.

[0133] As used in this article, the term "carboxyl group" refers to -C(=O)OH.

[0134] As used in this article, VEGFR (Vascular Endothelial Growth Factor Receptor) refers to the vascular endothelial growth factor receptor.

[0135] As used in this article, VEGF (Vascular Endothelial Growth Factor) refers to vascular endothelial growth factor.

[0136] As used in this article, VEGFR2 (Vascular Endothelial Growth Factor Receptor-2) refers to vascular endothelial growth factor receptor 2.

[0137] As used herein, "pharmaceuticalally acceptable carrier" refers to any form of substance that is non-toxic, inert, and compatible with the body in a patient (preferably a mammal, more preferably a human), including but not limited to solid, semi-solid, and liquid diluents, encapsulation materials, excipients, or other forms of excipients; its function is to effectively deliver the drug to the target site of action without destroying the activity of the active ingredient.

[0138] As used in this article, "patient" or "subject" refers to an animal, preferably a mammal, and more preferably a human. The term "mammal" refers to warm-blooded vertebrate mammals, including animals such as cats, dogs, rabbits, bears, foxes, wolves, monkeys, deer, rats, pigs, and humans.

[0139] As used in this article, the term "disease" is a state of health in an animal in which the animal is unable to maintain homeostasis and the animal's health continues to deteriorate if the disease does not improve.

[0140] The term "pharmaceutical composition" refers to a mixture of at least one compound that can be used in this disclosure with a pharmaceutically acceptable carrier. This pharmaceutical composition facilitates the administration of the compound to a patient or subject. Various techniques for administering compounds exist in the art, including but not limited to intravenous, oral, aerosol, parenteral, ocular, pulmonary, and topical administration. The terms "pharmaceutical composition," "therapeutic composition," "therapeutic formulation," or "pharmaceuticalally acceptable formulation" can refer to, but are by no means limited to, compositions or formulations that allow for the effective distribution of the pharmaceutical agents provided in this disclosure, in a form suitable for administration to the physical site most optimal for their desired activity, such as systemic administration.

[0141] The o-aminopyridine carboxyhydrazone compounds of formula (I) disclosed herein may exist in various tautomers, diastereomers, enantiomers, or cis-trans isomers, including but not limited to molecular conjugation theory. Some molecules have chiral carbon atoms, and this disclosure includes mixtures of these epimerizations, racemic mixtures, and compounds with a single configuration.

[0142] The o-aminopyridine carboxylhydrazone compounds disclosed herein can form salts, provided they are pharmaceutically permissible. Pharmaceutical salts are defined as follows: Acid-additive salts include, but are not limited to, hydrochlorides, hydrobroms, hydroiodates, sulfates, nitrates, phosphates, perchlorates, borates, tartrates, maleates, citrates, succinates, palmitates, methanesulfonates, benzoates, benzenesulfonates, salicylates, glycerophosphates, ketoglutarate, ascorbic acid salts, etc. Salts formed with bases include those with alkali metal salts such as Li, Na, and K salts; alkaline earth metal salts such as Ca and Mg salts; organic base salts such as various amino acids, guanidines, diethanolamine, choline, etc.; ammonium salts or substituted ammonium salts and aluminum salts. Furthermore, this invention also includes various hydrates, solvates, and polymorphs of the compounds and their salts, as well as pharmaceutically acceptable excipients, diluents, or carriers.

[0143] A typical pharmaceutical composition contains a compound of general formula (I) or a pharmaceutically acceptable salt thereof, and a pharmaceutically acceptable excipient, which may be a carrier or diluent, or diluted by a carrier, or packaged in a carrier, which may be in the form of capsules, sachets, paper, or other containers. When the carrier is used as a diluent, it can be a solid, semi-solid, or liquid substance, and can serve as a carrier, excipient, or medium for the active compound. The active compound can be absorbed in a container such as a sachet as a particulate solid. Some suitable carriers include water, salt solutions, alcohols, polyethylene glycol, polyhydroxyethoxylated castor oil, peanut oil, olive oil, gelatin, lactose, gypsum powder, sucrose, cyclodextrin, amylose, magnesium stearate, talc, agar, pectin, gum arabic, lower alkyl ethers of stearic acid, silicic acid, fatty acids, fatty acid amines, fatty acid monoglycerides, fatty acid diglycerides, quaternary tetraol fatty acid esters, polyoxyethylene, hydroxymethylcellulose, or polyvinylpyrrolidone, etc. Similarly, the carrier or diluent may comprise any sustained-release material in the art, such as glyceryl monostearate or glyceryl distearate. Formulations of this disclosure may be formulated using methods known in the art to provide rapid, sustained, or delayed release of the active ingredient after administration.

[0144] The pharmaceutical compositions disclosed herein may be sterile and, if desired, may be mixed with excipients, emulsifiers, buffers, sweeteners and / or colorants, provided that they do not react with the active compound.

[0145] The pharmaceutical compositions disclosed herein can be administered in any form, as long as they can effectively deliver the active drug to the appropriate or desired site of activity, such as oral, nasal, transdermal, etc.; or parenteral, such as rectal, subcutaneous, intravenous, intramuscular, intranasal, ophthalmic, etc.

[0146] For formulation into eye drops, they may contain sterile aqueous or non-aqueous solutions, suspensions, or emulsions. Aqueous solutions and suspensions may contain physiological saline and distilled water. Non-aqueous solutions and suspensions may contain alcohol solvents.

[0147] Topical ophthalmic formulations may contain one or more pharmaceutically acceptable excipients selected from: stabilizers, surfactants, polymer-based carriers, gelling agents, organic cosolvents, pH-active components, penetration-active components, and with or without preservatives. In some cases, a sustained-release semi-solid formulation, a sustained-release solid formulation, or an ophthalmic implant is injected into the affected eye. In some embodiments, the sustained-release semi-solid formulation, sustained-release solid formulation, or ophthalmic implant further contains pharmaceutically acceptable excipients. The sustained-release semi-solid formulation, sustained-release solid formulation, or ophthalmic implant contains multi-kinase inhibitors, antimetabolites, or combinations thereof; and biodegradable polymers selected from polylactic acid (PLA), polyglycolic acid (PLGA), and polylactic acid-polyglycolic acid copolymers. Excipients for topical ophthalmic formulations are, for example, those described in CN114010787A.

[0148] Example

[0149] To make the objectives, technical solutions, and advantages of the embodiments of this disclosure clearer, the technical solutions of the embodiments of this disclosure will be clearly and completely described below in conjunction with the structural formulas in the embodiments of this disclosure. Obviously, the described embodiments are only some embodiments of this disclosure, and not all embodiments. The following is merely a further description of this disclosure, and the protection scope of this disclosure is not limited thereto.

[0150] The preparation route of the disclosed compound is as follows:

[0151]

[0152] Wherein, compound E and compound DE are as defined in this disclosure.

[0153] The compound E is selected from , , , , , , , , , , , , , , , or .

[0154] The above preparation method includes the following steps:

[0155] <1> Compound C was prepared by reacting compound A and compound B under alkaline conditions via a nucleophilic substitution reaction;

[0156] <2> Compound D was prepared from compound C via hydrazinolysis.

[0157] <3> Compound DE was prepared by condensation reaction of compound D and compound E under acidic conditions (in Examples 1-15, compound DE is compound LHM-01~LHM-13).

[0158] step <1> In the reaction, the base is sodium hydroxide, potassium hydroxide, anhydrous potassium carbonate, or anhydrous sodium carbonate; the reaction solvent is N,N-dimethylformamide (DMF), dimethyl sulfoxide (DMSO), acetonitrile, tetrahydrofuran (THF), etc.; and the reaction temperature is from room temperature to 100℃.

[0159] step <2> In the hydrazolysis reaction, the reagent used is 80% hydrazine hydrate solution, the reaction solvent is anhydrous methanol, anhydrous ethanol, etc., and the reaction temperature is 65℃ to 80℃.

[0160] step <3> In the reaction, the acid is formic acid or glacial acetic acid, the solvent is anhydrous methanol or anhydrous ethanol, and the reaction temperature is from room temperature to 80°C.

[0161] The o-aminopyridine carboxyhydrazone compounds disclosed herein can all be prepared by the above synthetic route, and the corresponding starting materials can be selected according to the different substituents and substitution positions.

[0162] The structural formulas of compounds LHM-01 to LHM-13 are shown below:

[0163] .

[0164] Example 1: Preparation of methyl 2-[(4-pyridinemethyl)amino]nicotinic acid (C)

[0165] 2-Fluoronic acid methyl ester (6.2 g, 40 mmol), 4-pyridinemethylamine (6.48 g, 60 mmol), and potassium carbonate (11.04 g, 80 mmol) were sequentially added to DMF (60 mL), and the mixture was heated to 80 °C and reacted for 4 h. After the reaction was complete, the reaction solution was cooled to room temperature and then poured into a large amount of water, resulting in the precipitation of a solid. The solid was filtered, the filter cake was washed with water, and dried to obtain the crude product. Purification was performed by silica gel column chromatography (petroleum ether:ethyl acetate = 5:1) to give 7.3 g of an off-white solid, with a yield of 75%. mp: 93.2–95.3 °C; MS (ESI) m / z : 244.1; 1 H NMR (400 MHz, DMSO- d 6) δ 8.48 – 8.41 (m, 3H), 8.23 ​​(dd, J= 4.8, 2.0 Hz, 1H), 8.13 (dd, J = 7.8, 2.0 Hz, 1H), 7.30 – 7.25 (m, 2H), 6.66(dd, J = 7.8, 4.7 Hz, 1H), 4.73 (d, J = 6.1 Hz, 2H), 3.85 (s, 3H).

[0166] Example 2: Preparation of 2-[(4-pyridinemethyl)amino]-3-pyridinecarboxylhydrazide (D)

[0167] Intermediate C (4.86 g, 20 mmol) was dissolved in anhydrous ethanol (50 mL), and 80% hydrazine hydrate (6.25 g, 100 mmol) was slowly added. The mixture was then heated to 80 °C and reacted for 4 h. After the reaction was complete, the reaction solution was cooled to room temperature and concentrated under reduced pressure. A white solid precipitated, which was filtered. The filter cake was washed with a small amount of cold anhydrous ethanol and dried to give 4.0 g of a white solid, with a yield of 82.5%. mp: 176–178 °C; MS (ESI) m / z : 244.1; 1 H NMR (400 MHz, DMSO- d 6) δ 9.82 (s, 1H), 8.68 (t, J = 6.1 Hz, 1H), 8.56 – 8.37 (m, 2H), 8.09 (dd, J = 4.8, 1.8 Hz, 1H), 7.88 (dd, J = 7.6, 1.8 Hz, 1H), 7.35 – 7.18 (m, 2H), 6.58 (dd, J = 7.6, 4.8Hz, 1H), 4.67 (d, J = 6.0 Hz, 2H), 4.50 (s, 2H).

[0168] Example 3: Preparation of (E)-N'-benzylidene-2-[(4-pyridinemethyl)amino]-3-pyridinecarboxylhydrazide (LHM-01)

[0169] Intermediate D (0.24 g, 1 mmol) was added to 12 mL of anhydrous ethanol and heated to 80 °C to dissolve gradually. Then, benzaldehyde (1.1 mmol) and 1 drop of glacial acetic acid were added, and the reaction was continued at 80 °C with stirring for 4 h. After the reaction was complete, the reaction solution was cooled to room temperature, and a white solid precipitated. This was filtered, and the filter cake was washed with a small amount of cold anhydrous ethanol and dried to give a white solid in 55% yield. mp: 200–201 °C; MS (ESI) m / z 332.2[M+H] + ; 1 H NMR (400 MHz, DMSO- d 6) δ11.86 (s, 1H), 8.61 – 8.33 (m, 4H), 8.16 (dd, J = 4.9, 1.8 Hz, 1H), 8.03 (d, J =7.6 Hz, 1H), 7.79 – 7.66 (m, 2H), 7.52 – 7.41 (m, 3H), 7.34 – 7.26 (m, 2H), 6.68 (dd, J = 7.6, 4.8 Hz, 1H), 4.69 (d, J = 6.0 Hz, 2H).

[0170] Example 4: Preparation of (E)-N'-(3-fluorobenzyl)-2-[(4-pyridinemethyl)amino]-3-pyridinecarboxylhydrazide (LHM-02)

[0171] The synthesis method is similar to that used for compound LHM-01. It is a white solid with a yield of 58%. mp: 190.7–191.6 °C; MS (ESI) m / z 350.2 [M+H] + ; 1 H NMR (400 MHz, DMSO- d 6) δ 11.98 (s, 1H),8.59 – 8.34 (m, 4H), 8.16 (dd, J = 4.9, 1.7 Hz, 1H), 8.03 (d, J = 7.6 Hz, 1H),7.61 – 7.47 (m, 3H), 7.34 – 7.25 (m, 3H), 6.68 (dd, J = 7.6, 4.8 Hz, 1H), 4.69(d, J = 6.0 Hz, 2H).

[0172] Example 5: Preparation of (E)-N'-(3-chlorobenzyl)-2-[(4-pyridinemethyl)amino]-3-pyridinecarboxylhydrazide (LHM-03)

[0173] The synthesis method is similar to that used for compound LHM-01. It is a white solid with a yield of 50%. mp: 194-196℃; MS (ESI). m / z 366.2 [M+H] + ; 1 H NMR (400 MHz, DMSO- d 6) δ 12.00 (s, 1H), 8.55 –8.44 (m, 3H), 8.38 (s, 1H), 8.16 (dd, J = 4.8, 1.8 Hz, 1H), 8.03 (d, J = 7.7 Hz,1H), 7.80 – 7.66 (m, 2H), 7.53 – 7.47 (m, 2H), 7.32 – 7.27 (m, 2H), 6.68 (dd, J = 7.7, 4.8 Hz, 1H), 4.69 (d, J = 6.0 Hz, 2H).

[0174] Example 6: Preparation of (E)-N'-(3-methylbenzyl)-2-[(4-pyridinemethyl)amino]-3-pyridinecarboxylhydrazide (LHM-04)

[0175] The synthesis method is similar to that used for compound LHM-01. It is a white solid with a yield of 52%. mp: 196-198℃; MS (ESI). m / z 346.2 [M+H] + ; 1 H NMR (400 MHz, DMSO- d 6) δ 11.85 (s, 1H), 8.59 –8.42 (m, 3H), 8.37 (s, 1H), 8.15 (dd, J = 4.8, 1.7 Hz, 1H), 8.02 (d, J = 7.6 Hz,1H), 7.59 – 7.46 (m, 2H), 7.35 (t, J = 7.6 Hz, 1H), 7.32 – 7.29 (m, 2H), 7.26(d, J= 7.6 Hz, 1H), 6.68 (dd, J = 7.6, 4.8 Hz, 1H), 4.69 (d, J = 6.0 Hz, 2H), 2.36(s, 3H).

[0176] Example 7: Preparation of (E)-N'-(3-methoxybenzyl)-2-[(4-pyridinemethyl)amino]-3-pyridinecarboxylhydrazide (LHM-05)

[0177] The synthesis method is similar to that used for compound LHM-01. It is a white solid with a yield of 60%. mp: 194-196℃; MS (ESI). m / z 362.2 [M+H] + ; 1 H NMR (400 MHz, DMSO- d 6) δ 11.88 (s, 1H), 8.60 –8.42 (m, 3H), 8.38 (s, 1H), 8.16 (dd, J = 4.9, 1.8 Hz, 1H), 8.02 (d, J = 7.6 Hz, 1H), 7.38 (t, J = 8.0 Hz, 1H), 7.35 – 7.23 (m, 4H), 7.02 (m, 1H), 6.68 (dd, J =7.6, 4.8 Hz, 1H), 4.69 (d, J = 6.1 Hz, 2H), 3.81 (s, 3H).

[0178] Example 8: Preparation of (E)-N'-(4-methoxybenzyl)-2-[(4-pyridinemethyl)amino]-3-pyridinecarboxylhydrazide (LHM-06)

[0179] The synthesis method is similar to that used for compound LHM-01. It is a white solid with a yield of 62%. mp: 194-196℃; MS (ESI). m / z 362.2 [M+H] + ; 1 H NMR (400 MHz, DMSO- d 6) δ 11.74 (s, 1H), 8.57 –8.41 (m, 3H), 8.35 (s, 1H), 8.14 (dd, J = 5.0, 1.8 Hz, 1H), 8.00 (d,J = 7.6 Hz, 1H), 7.67 (d, J = 8.3 Hz, 2H), 7.33 – 7.27 (m, 2H), 7.07 – 6.99 (m, 2H), 6.67(dd, J = 7.6, 4.8 Hz, 1H), 4.68 (d, J = 6.0 Hz, 2H), 3.81 (s, 3H).

[0180] Example 9: Preparation of (E)-N'-(3-cyanobenzylidene)-2-[(4-pyridinemethyl)amino]-3-pyridinecarboxylhydrazide (LHM-07)

[0181] The synthesis method is similar to that used for compound LHM-01. It is a white solid with a yield of 57%. mp: 190-192℃; MS (ESI). m / z 357.2 [M+H] + ; 1 H NMR (400 MHz, DMSO- d 6) δ 12.08 (s, 1H), 8.59 –8.38 (m, 4H), 8.17 (dd, J = 4.8, 1.8 Hz, 1H), 8.13 (s, 1H), 8.11 – 8.01 (m,2H), 7.90 (dt, J = 7.7, 1.4 Hz, 1H), 7.68 (t, J = 7.8 Hz, 1H), 7.32 – 7.26 (m,2H), 6.69 (dd, J = 7.6, 4.8 Hz, 1H), 4.69 (d, J = 6.0 Hz, 2H).

[0182] Example 10: Preparation of (E)-N'-(4-cyanobenzylidene)-2-[(4-pyridinemethyl)amino]-3-pyridinecarboxylhydrazide (LHM-08)

[0183] The synthesis method is similar to that used for compound LHM-01. It is a white solid with a yield of 58%. mp: 216-218℃; MS (ESI) m / z 366.2 [M+H] + ; 1 H NMR (400 MHz, DMSO- d6) δ 12.09 (s, 1H), 8.62 –8.38 (m, 4H), 8.17 (dd, J = 4.9, 1.7 Hz, 1H), 8.04 (d, J = 7.6 Hz, 1H), 7.98 –7.85 (m, 4H), 7.32 – 7.26 (m, 2H), 6.69 (dd, J = 7.7, 4.8 Hz, 1H), 4.69 (d, J =6.0 Hz, 2H).

[0184] Example 11: Preparation of (E)-N'-(3-trifluoromethylbenzyl)-2-[(4-pyridinemethyl)amino]-3-pyridinecarboxylhydrazide (LHM-09)

[0185] The synthesis method is similar to that used for compound LHM-01. It is a white solid with a yield of 55%. mp: 197-200℃; MS (ESI). m / z 400.2 [M+H] + ; 1 H NMR (400 MHz, DMSO- d 6) δ 12.07 (s, 1H), 8.63 –8.38 (m, 4H), 8.17 (dd, J = 4.8, 1.7 Hz, 1H), 8.08 (s, 1H), 8.06 – 7.98 (m,2H), 7.80 (d, J = 7.8 Hz, 1H), 7.71 (t, J = 7.8 Hz, 1H), 7.34 – 7.26 (m, 2H), 6.69 (dd, J = 7.6, 4.8 Hz, 1H), 4.69 (d, J = 6.0 Hz, 2H).

[0186] Example 12: Preparation of (E)-N'-(4-trifluoromethylbenzyl)-2-[(4-pyridinemethyl)amino]-3-pyridinecarboxylhydrazide (LHM-10)

[0187] The synthesis method is similar to that used for compound LHM-01. It is a white solid with a yield of 58%. mp: 208-210℃; MS (ESI). m / z 400.2 [M+H] + ; 1H NMR (400 MHz, DMSO- d 6) δ 12.06 (s, 1H), 8.61 –8.35 (m, 4H), 8.17 (dd, J = 4.9, 1.7 Hz, 1H), 8.04 (d, J = 7.7 Hz, 1H), 7.99 –7.88 (m, 2H), 7.87 – 7.80 (m, 2H), 7.33 – 7.26 (m, 2H), 6.69 (dd, J = 7.6, 4.8Hz, 1H), 4.69 (d, J = 6.0 Hz, 2H).

[0188] Example 13: Preparation of (E)-N'-(3-trifluoromethoxybenzyl)-2-[(4-pyridinemethyl)amino]-3-pyridinecarboxylhydrazide (LHM-11)

[0189] The synthesis method is similar to that used for compound LHM-01. It is a white solid with a yield of 60%. mp: 190-192℃; MS (ESI). m / z 416.2 [M+H] + ; 1 H NMR (400 MHz, DMSO- d 6) δ 12.03 (s, 1H), 8.62 –8.37 (m, 4H), 8.17 (dd, J = 4.8, 1.8 Hz, 1H), 8.03 (d, J = 7.6 Hz, 1H), 7.79 –7.67 (m, 2H), 7.61 (t, J = 7.9 Hz, 1H), 7.44 (d, J = 8.0 Hz, 1H), 7.33 – 7.26 (m,2H), 6.69 (dd, J = 7.6, 4.8 Hz, 1H), 4.69 (d, J = 6.0 Hz, 2H).

[0190] Example 14: Preparation of (E)-N'-(4-trifluoromethoxybenzyl)-2-[(4-pyridinemethyl)amino]-3-pyridinecarboxylhydrazide (LHM-12)

[0191] The synthesis method is similar to that used for compound LHM-01. It is a white solid with a yield of 62%. mp: 186-188℃; MS (ESI) m / z 416.2 [M+H] + ; 1 H NMR (400 MHz, DMSO- d 6) δ 11.96 (s, 1H), 8.62 –8.35 (m, 4H), 8.16 (dd, J = 4.8, 1.8 Hz, 1H), 8.03 (d, J = 7.6 Hz, 1H), 7.86 (d, J = 8.2 Hz, 2H), 7.46 (d, J = 8.2 Hz, 2H), 7.30 (d, J = 5.8 Hz, 2H), 6.69 (dd, J =7.6, 4.8 Hz, 1H), 4.69 (d, J = 6.0 Hz, 2H).

[0192] Example 15: Preparation of (E)-N'-(4-dimethylaminobenzylidene)-2-[(4-pyridinemethyl)amino]-3-pyridinecarboxylhydrazide (LHM-13)

[0193] The synthesis method is similar to that used for compound LHM-01. It is a yellow solid with a yield of 52%. mp: 230-232℃; MS (ESI). m / z 375.2 [M+H] + ; 1 H NMR (400 MHz, DMSO- d 6) δ 11.57 (s, 1H), 8.63 –8.38 (m, 3H), 8.27 (s, 1H), 8.17 – 8.10 (m, 1H), 7.98 (d, J = 7.5 Hz, 1H), 7.54(d, J = 8.4 Hz, 2H), 7.30 (d, J = 5.2 Hz, 2H), 6.76 (d, J = 8.6 Hz, 2H), 6.66 (dd, J = 7.6, 4.9 Hz, 1H), 4.68 (d, J= 6.0 Hz, 2H), 2.98 (s, 6H).

[0194] Example 16: Evaluation of the in vitro inhibitory activity of the acylhydrazone compounds LHM-01~LHM-13 of this disclosure against VEGFR-2 kinase.

[0195] The in vitro inhibitory activity of the acylhydrazone compounds disclosed herein against VEGFR-2 kinase was investigated, and all compounds were evaluated using the LanthaScreen kinase binding assay. AAL-993 (CAS No. 269390-77-4, purchased from Guangzhou Invevo Chemical Technology Co., Ltd., batch number V209218) was used as the positive control. The inhibition rate was at least the average of two experiments.

[0196] Specific methods:

[0197] (1) Compound preparation

[0198] The initial concentration of the compound for testing on kinase KDR / VEGFR-2 was 0.1 μM, which was then prepared to a 100-fold concentration, i.e., 10 μM. 4995 μL of 100% DMSO was added to an EP tube, followed by 5 μL of the prepared 10 mM compound solution. 50 μL of the 10 μM compound solution was added to the second well of an Echo 384-well plate. 50 μL of 100% DMSO was transferred to two empty wells as controls (no compound and no enzyme). 200 nmL of the compound solution was transferred to the 384-well plate using an Echo 650.

[0199] (2) Preparation of kinase buffer

[0200] 50mM Hepes (pH 7.5), 10mM MgCl2, 0.01% Tween-20, 0.01% BSA, 2mM DTT.

[0201] (3) Kinase response and termination

[0202] 1. Add the kinase to a 1-fold kinase buffer to form a kinase solution; the final concentration of KDR enzyme is 0.05 nM.

[0203] 2. Transfer 10 μL of the above kinase solution to the reaction wells of a 384-well plate, add 1x kinase buffer to the negative control wells, centrifuge at 1000 rpm for 1 minute, and incubate at room temperature for 10 minutes;

[0204] 3. Add the substrate Fluorescein-PolyGT (final concentration 20 nM) and ATP (final concentration 100 μM) to the kinase buffer to form the substrate solution;

[0205] 4. Transfer 10 μL of the above substrate solution into the reaction wells of a 384-well plate and centrifuge at 1000 rpm for 1 minute;

[0206] 5. React at room temperature for 30 minutes;

[0207] 6. Prepare a 2-fold stop reaction solution containing the antibody. Add 20 μL of the above reagent to a 384-well plate and centrifuge at 1000 rpm for 1 minute. Incubate at room temperature for 60 minutes.

[0208] (4) Data reading

[0209] Fluorescence values ​​were read on the Envision 2104 Multilabel Reader.

[0210] (5) Data calculation

[0211] 1. The numerical ratio of replicated fluorescence readings (Lance signal ratio (520nm / 495nm));

[0212] 2. Convert the above data into a suppression percentage using a formula;

[0213] Percent inhibition = (max – Sample signal ratio) / (max – min)*100

[0214] “min” represents the reading of the control wells without enzyme; “max” represents the reading of the control wells with DMSO added.

[0215] 3. Import the data into MS Excel and use XLFit Excel add-in version 5.4.0.8 to perform curve fitting;

[0216] Fitting formula: Y = Bottom + (Top – Bottom) / (1 + (IC) 50 / X)^Hillslope)

[0217] The experimental results are shown in Table 1.

[0218] Example 17: Evaluation of the in vitro cytotoxic activity of the acylhydrazone compounds LHM-01~LHM-13 disclosed herein against human umbilical vein endothelial cells.

[0219] The inhibitory activity of the acylhydrazone compounds disclosed herein against angiogenesis was investigated using the MTT assay to determine the inhibitory activity of all compounds against VEGF-induced proliferation of human umbilical vein endothelial cells (HUVECs). AAL-993 was used as the positive control. Inhibition rates were at least the average of two experiments.

[0220] Specific methods:

[0221] HUVEC cells in the logarithmic growth phase were counted and seeded into 96-well plates (5 × 10⁶ cells / well). 3 Cells were cultured in wells (cells / well) for 24 h until adherence, then different concentrations of the target compound and positive control were added. The plates were then incubated at 37°C with 5% CO2 for 72 h. After 72 h, the liquid in the 96-well plates was discarded, and 200 μL of physiological saline was added to each well for washing. 10 μL of 5 mg / mL MTT solution was added, and the plates were incubated at 37°C with 5% CO2 for 4 h. After 4 h, the suspension was discarded, and 100 μL of DMSO was added to each well to dissolve the blue-purple formazan crystals. The absorbance of the solution at 490 nm was measured using a multi-mode microplate reader. The cell inhibition rate of the test compound was calculated using the formula (Inh% = (negative control OD value – compound OD value) / negative control OD value * 100%), and the IC50 was calculated using GraphPad Prism 6.0. 50 Values. The experimental results are shown in Table 1.

[0222] Table 1. In vitro VEGFR-2 kinase inhibitory activity and HUVEC proliferation inhibitory activity of compounds LHM-01~LHM-13 and AAL-993

[0223]

[0224] a The values ​​represent the IC50 values ​​of the test compound at five different concentrations. 50 value;

[0225] b The values ​​are processed values ​​of three independent tests of the test compound using the MTT assay after 72 hours of incubation.

[0226] Table 1 shows that compounds LHM-01 to LHM-13 disclosed in this paper all exhibit strong inhibitory activity against VEGFR-2 kinase. Among them, compounds LHM-02 and LHM-03 show the highest IC50 values ​​against VEGFR-2 kinase. 50 Values ​​below 5 nM; the IC50 values ​​of compounds LHM-06, LHM-08, LHM-09, LHM-10, LHM-11 and LHM-12 against VEGFR-2 kinase were... 50 The values ​​were all below 10 nM; the IC50 values ​​of compounds LHM-04, LHM-05, and LHM-07 against VEGFR-2 kinase were... 50 All values ​​were below 20 nM; the IC50 values ​​for all compounds against VEGFR-2 kinase were... 50 The values ​​were all lower than the positive control AAL-993 (IC). 50The value was 35 nM. Subsequent evaluation of HUVEC cell proliferation inhibition activity showed that the o-aminopyridine carboxyhydrazone compound disclosed in this paper had a lower IC50 value compared to the positive control AAL-993. 50 The value indicates that it has better anti-angiogenic activity and cell proliferation inhibition activity.

[0227] Examples 16 and 17 above demonstrate that the o-aminopyridine carboxyhydrazone compounds of this disclosure, or their pharmaceutically acceptable salts, solvates, isotopic variants, tautomers, diastereomers, enantiomers, or cis-trans isomers, have good inhibitory activity against VEGFR-2 kinase.

[0228] The method for synthesizing o-aminopyridine carboxylhydrazone compounds disclosed herein is simple, the raw materials are inexpensive and readily available, and the in vitro biological evaluation shows significant activity, indicating good application prospects.

[0229] The foregoing description of specific exemplary embodiments of this disclosure is for illustrative and explanatory purposes. These descriptions are not intended to limit this disclosure to the precise forms disclosed, and it will be apparent that many changes and variations can be made in accordance with the foregoing teachings. The exemplary embodiments were chosen and described in order to explain the specific principles of this disclosure and their practical application, thereby enabling those skilled in the art to implement and utilize various different exemplary embodiments of this disclosure, as well as various different choices and variations. The scope of this disclosure is intended to be defined by the claims and their equivalents.

Claims

1. The compound represented by formula (II) or a pharmaceutically acceptable salt thereof: (II) In formula (II), R is absent or selected from halogen, cyano, C1-3 alkyl, C1-3 alkoxy, halogenated C1-3 alkyl, halogenated C1-3 alkoxy, or -NR. 1 R 2 ;R 1 and R 2 Each is independently selected from hydrogen or C. 1-3 alkyl.

2. The compound according to claim 1 or a pharmaceutically acceptable salt thereof, characterized in that, It has the structure shown in equation (Ⅲ): (Ⅲ) R is defined in the same way as in equation (II).

3. The compound according to claim 1 or a pharmaceutically acceptable salt thereof, characterized in that, It has the structure shown in equation (Ⅳ): (Ⅳ) R is defined in the same way as in equation (II).

4. The compound or a pharmaceutically acceptable salt thereof according to any one of claims 1-3, characterized in that, R is absent or selected from fluorine, chlorine, cyano, methyl, ethyl, methoxy, ethoxy, trifluoromethyl, difluoromethyl, monofluoromethyl, trifluoromethoxy, difluoromethoxy, monofluoromethoxy, -NH2, -N(H)(CH3), -N(CH3)(CH3), -N(CH3)(CH2CH3), -N(CH2CH3)(CH2CH3), or -N(H)(CH2CH3).

5. The compound or a pharmaceutically acceptable salt thereof according to any one of claims 1-3, characterized in that, R is absent or selected from fluorine, chlorine, cyano, methyl, ethyl, methoxy, ethoxy, trifluoromethyl, difluoromethyl, monofluoromethyl, trifluoromethoxy, difluoromethoxy, monofluoromethoxy, -NH2, -N(H)(CH3) or -N(CH3)(CH3).

6. The compound or a pharmaceutically acceptable salt thereof according to any one of claims 1-3, characterized in that, R is absent or selected from fluorine, chlorine, cyano, methyl, ethyl, methoxy, ethoxy, trifluoromethyl, trifluoromethoxy, -NH2, -N(H)(CH3) or -N(CH3)(CH3).

7. The compound or a pharmaceutically acceptable salt thereof according to any one of claims 1-3, characterized in that, R is either absent or selected from fluorine, chlorine, cyano, methyl, methoxy, trifluoromethyl, trifluoromethoxy, or -N(CH3)(CH3).

8. The compound according to claim 1 or a pharmaceutically acceptable salt thereof: 。 9. A method for preparing the compound of any one of claims 1-8 or a pharmaceutically acceptable salt thereof, comprising the following steps: <1> Compound C was prepared by reacting compound A and compound B under alkaline conditions via a nucleophilic substitution reaction; <2> Compound D was prepared from compound C via hydrazinolysis. <3> The compound according to any one of claims 1-8 is prepared by condensation reaction of compound D and compound E under acidic conditions; Wherein, R is defined as in any one of claims 1-8; The compound E is selected from , , , , , , , , , , , , , , or ; The compound DE is the compound according to any one of claims 1-8.

10. A pharmaceutical composition comprising the compound of any one of claims 1-8 or a pharmaceutically acceptable salt thereof, or the compound of the preparation method of claim 9 or a pharmaceutically acceptable salt thereof.

11. Use of the compound of any one of claims 1-8 or a pharmaceutically acceptable salt thereof, or the compound of the preparation method of claim 9 or a pharmaceutically acceptable salt thereof, or the pharmaceutical composition of claim 10, in the preparation of a medicament for the prevention or treatment of diseases related to angiogenesis mediated by abnormal VEGFR-2 kinase.

12. The use according to claim 11, characterized in that, The diseases associated with abnormal VEGFR-2 kinase-mediated angiogenesis are selected from non-small cell lung cancer, liver cancer, hemangioma, angiofibroma, malignant melanoma, gastrointestinal stromal tumor, pancreatic cancer, metastatic colorectal cancer, recurrent glioblastoma, biliary tract tumors, soft tissue sarcoma, breast cancer, ovarian cancer, rheumatoid arthritis, kidney diseases such as glomerulonephritis, diabetic nephropathy, malignant nephrosclerosis, embolic microangiitis syndrome, or eye diseases; The eye disease is selected from diabetic retinopathy, neovascular glaucoma, or age-related macular degeneration.

Images