Ciolanil derivative and application thereof

Abstract

The invention provides a siolanib derivative and application thereof. Specifically, the invention provides a compound as shown in a general formula (I), an isomer, a crystal form, a pharmaceutically acceptable salt, a hydrate or a solvate of the compound, and application of the compound in preparation of antitumor drugs. # imgabs0 #

Description

Technical Field

[0001] The present invention relates to the field of medicinal chemistry; in particular, to a ceorone derivative and a use thereof. Background Art

[0002] Protein kinases are a class of enzymes that catalyze protein phosphorylation, particularly catalyzing the phosphorylation of hydroxyl groups on specific tyrosine, serine and threonine residues in proteins. A partial non-limiting list of such kinases includes PDGFRα, AuroraB, MAPKAPK2, FRK, CaMK1β, DDR1, FLT1, FGFR1, AuroraC, KIT, VEGFR, etc. Protein kinases have been shown to be key regulators of most cell functions, including cell signaling, proliferation, differentiation, metabolism, survival, apoptosis, movement, DNA damage repair, etc. Protein kinases are generally divided into two categories, namely protein tyrosine kinases (PTKs) and serine-threonine kinases (STKs).

[0003] Protein tyrosine kinases (PTKs) can be divided into two categories, namely non-transmembrane tyrosine kinases and transmembrane growth factor receptor tyrosine kinases (RTKs). Currently, many different subfamilies of transmembrane growth factor receptor tyrosine kinases (RTKs) have been identified, such as epidermal growth factor receptor (EGFR), vascular endothelial growth factor receptor (VEGFR), platelet-derived growth factor receptor (PDGFR) and fibroblast growth factor receptor (FGFR).

[0004] Serine-threonine kinases (STKs) are mainly found in cells, and they perform their functions in the cytoplasm rather than in organelles and the cytoskeleton in the cytoplasm, such as glycogen synthase kinase-3 (GSK-3), Aurora kinase, and cell cycle-dependent kinases (CDKs).

[0005] Inhibition of protein kinases, thereby inhibiting phosphorylation of substrate peptides or proteins, has been shown to be effective in the treatment of a variety of diseases. Given the large number of kinases and related diseases, and the inevitable emergence of drug resistance, which ultimately leads to disease progression, there is a constant need for new inhibitors that are selective for multiple kinases and effective in the treatment of related diseases. Summary of the invention

[0006] The present invention has discovered a class of naphthamide compounds which have an inhibitory effect on various protein kinases such as vascular endothelial growth factor receptor (VEGFR) and Aurora.

[0007] The present invention provides a compound or isomer represented by general formula (I), a crystal form, a pharmaceutically acceptable salt, a hydrate or a solvate:

[0008]

[0009] Among them, X1, X2, X3, X4, X5, X6, X7, X8, X9, X 10 , X 11 , X 12 , X 13 , X 14 , X 15 , X 16 , X 17 and X 18 are each independently selected from hydrogen or deuterium, and X1, X2, X3, X4, X5, X6, X7, X8, X9, X 10 , X 11 , X 12 , X 13 , X 14 , X 15 , X 16 , X 17 or X 18 At least one of them is selected from deuterium.

[0010] In some embodiments, the compound represented by general formula (I) has one of the following structures:

[0011]

[0012]

[0013]

[0014] In some embodiments, the compound represented by general formula (I) has one of the following structures:

[0015]

[0016] In some embodiments, the compound represented by general formula (I) has one of the following structures:

[0017]

[0018]

[0019]

[0020] In some embodiments, the compound represented by general formula (I) has one of the following structures:

[0021]

[0022] Any atom of the compound of the present invention, unless otherwise specified, refers to the isotope of its stable atom. Unless otherwise specified, when a site on the molecular structure is selected as "H" or "hydrogen", the site should be understood to have the natural abundance of hydrogen isotopes. Similarly, if not otherwise specified, when a site is selected as "D" or "deuterium", the site should be understood to have a deuterium isotope abundance of at least 3000 times its natural abundance (the natural abundance of deuterium isotopes is 0.015%), i.e., 45% deuterium atom enrichment. Preferably, the deuterium atom abundance of each deuterated site of the deuterated compound of the present invention is at least 3500 times its natural abundance (52.2% deuterium atom enrichment). More preferably, at least 4500 times (67.5% deuterium atom enrichment). More preferably, at least 5000 times (75% deuterium atom enrichment). More preferably, it is at least 6000 times (90% deuterium atom enrichment). More preferably, it is at least 6333 times (95% deuterium atom enrichment). More preferably, it is at least 6466.7 times (97% deuterium atom enrichment). More preferably, it is at least 6600 times (99% deuterium atom enrichment). More preferably, it is at least 6633.3 times (99.5% deuterium atom enrichment).

[0023] The preparation of the compound described in the general formula (I) of the present invention can be completed by the following exemplary methods and examples, but these methods and examples should not be considered in any way to limit the scope of the present invention. The compound described in the present invention can also be synthesized by synthetic techniques known to those skilled in the art, or a combination of synthetic methods known in the art and the method described in the present invention. The product obtained by each step of the reaction is obtained by separation techniques known in the art, including but not limited to extraction, filtration, distillation, crystallization, chromatographic separation, etc. The starting materials and chemical reagents required for the synthesis can be conventionally synthesized according to the literature (such as provided by Scifinder) or purchased.

[0024] The following synthetic route describes the preparation method of the compound of formula (I) of the present invention, and the raw materials, reagents, catalysts, solvents, etc. used in the following synthetic schematic diagram can be prepared by methods well known to those of ordinary skill in the field of organic chemistry or can be commercially available. All final derivatives of the present invention can be prepared by the methods described in the schematic diagram or similar methods thereof, which are all well known to those of ordinary skill in the field of organic chemistry. All variable factors used in these schematic diagrams are defined as above and below.

[0025] The definitions of the following variables are as described above, and the definitions of new variables are as described in this section. In addition, the compounds described in the general formula (I) and the intermediates involved can be purified by common separation methods, such as extraction, recrystallization and silica gel column chromatography. The 200-300 mesh silica gel and thin layer chromatography silica gel plates used are all produced by Qingdao Ocean Chemical Plant. The chemical reagents used are analytically pure or chemically pure commercial products of general reagents and are not further purified when used.

[0026] The present invention provides a method for preparing a compound represented by general formula (I), comprising the following steps:

[0027]

[0028] 1) The compound represented by formula (Ia) is reacted in a first solvent with a compound represented by formula (Ib) through a nucleophilic substitution reaction after being catalyzed by a first base to obtain a compound represented by formula (Ic); or the compounds represented by formula (Ia) and formula (Ib) are reacted through a Mitsunobu reaction in the presence of triphenylphosphine (PPh3) / diethyl azodiacid (DEAD) or triphenylphosphine (PPh3) / diisopropyl azodiacid (DIAD) to obtain a compound represented by formula (Ic);

[0029] 2) the compound represented by formula (Ic) reacts with the compound represented by formula (Id) in a second solvent under the action of a second base to obtain a compound represented by formula (Ie) through a nucleophilic substitution reaction;

[0030] 3) the compound represented by formula (Ie) and the compound represented by (If) undergo condensation reaction in a third solvent under the action of a condensing agent and a third base to obtain a compound represented by formula (I);

[0031] in:

[0032] X1, X2, X3, X4, X5, X6, X7, X8, X9, X 10 , X 11 , X 12 , X 13 , X 14 , X 15 , X 16 , X 17 and X 18 The definition of is as mentioned above;

[0033] M is selected from bromine, iodine, -OMs, -OTf, -OTs, -OH.

[0034] In some embodiments, in step 1), the first solvent is selected from dichloromethane (DCM), 1,2-dichloroethane, 1,4-dioxane, tetrahydrofuran (THF), acetonitrile (MeCN), N,N'-dimethylformamide (DMF), N-methylpyrrolidone (NMP) or a combination thereof.

[0035] In some embodiments, in step 1), the first base is selected from potassium carbonate (K2CO3), cesium carbonate (Cs2CO3) or a combination thereof.

[0036] In some embodiments, in step 2), the second solvent is selected from 1,2-dichloroethane, 1,4-dioxane, tetrahydrofuran (THF), acetonitrile (MeCN), N,N'-dimethylformamide (DMF), N-methylpyrrolidone (NMP), isopropanol, N,N'-dimethyl sulfoxide (DMSO) or a combination thereof.

[0037] In some embodiments, in step 2), the second base is selected from potassium carbonate (K2CO3), cesium carbonate (Cs2CO3), diisopropylethylamine (DIEA) or a combination thereof.

[0038] In some embodiments, in step 3), the condensing agent is selected from 1-(3-dimethylaminopropyl)-3-ethylcarbodiimide hydrochloride (EDCI·HCl), 1-hydroxybenzotriazole (HOBt), 2-(7-azobenzotriazole)-N,N,N',N'-tetramethyluronium hexafluorophosphate (HATU), N,N,N',N'-tetramethylchloroformamidine hexafluorophosphate (TCFH) and N-methylimidazole, among which TCFH is particularly suitable for the condensation of sterically hindered larger amines, 1H-benzotriazol-1-yloxytripyrrolidino hexafluorophosphate (PyBOP) or a combination thereof.

[0039] In some embodiments, in step 3), the third solvent is selected from dichloromethane (DCM), 1,4-dioxane (1,4-dioxane), tetrahydrofuran (THF), acetonitrile (MeCN), N,N'-dimethylformamide (DMF) or a combination thereof.

[0040] In some embodiments, the third base is selected from triethylamine (TEA) and diisopropylethylamine (DIEA).

[0041] The compounds provided by the present invention can all be prepared by the above general preparation method using one-to-one corresponding raw materials.

[0042] The present invention also provides a pharmaceutical composition, which contains the compound described in the first aspect above, its isomers, crystal forms, pharmaceutically acceptable salts, hydrates or solvates, and optional pharmaceutical carriers and / or adjuvants and / or diluents.

[0043] Methods for preparing various pharmaceutical compositions containing a certain amount of active ingredients are known or will be apparent to those skilled in the art based on the disclosure of the present invention. As described in REMINGTON'S PHARMACEUTICAL SCIENCES, Martin, EW, ed., Mack Publishing Company, 19th ed. (1995), the method for preparing the pharmaceutical composition includes incorporating appropriate pharmaceutical excipients, carriers, diluents, etc.

[0044] The present invention also provides the use of the compound described in the first aspect, its isomer, crystal form, pharmaceutically acceptable salt, hydrate or solvate, or the pharmaceutical composition described in the second aspect in the preparation of drugs for diseases related to abnormal protein kinase activity.

[0045] In some embodiments, the protein kinase is VEGFR 2 and / or Aurora B.

[0046] In some embodiments, the disease is an autoimmune disease, cancer, a neurological disease and a neurodegenerative disease, an allergy, asthma, a cardiovascular disease and a metabolic disease, or a hormone-related disease.

[0047] In some embodiments, the cancer is a hematological cancer and a solid tumor.

[0048] Beneficial effects of the present invention:

[0049] The compounds of the present invention (such as representative compounds I-1, I-2 and I-3) have excellent inhibitory effects on protein kinases (such as VEGFR2 and Aurora B), and can be used as drugs for treating diseases related to the activity of kinases such as VEGFR2 and Aurora B. Furthermore, the pharmacokinetic experiments in mice show that after oral administration, the compounds of the present invention (such as representative compounds I-2 and I-3) have a significantly lower plasma exposure (AUC) and a lower half-life (T 1 / 2 ) and mean residence time (MRT) were higher than those of sioronib, indicating that after oral administration, the compound of the present invention can significantly prolong the half-life in vivo, improve the metabolic stability of the drug in vivo, increase the drug exposure level in vivo, and has the potential to bring unexpected anti-tumor activity.

[0050] Technical terms of the present invention:

[0051] In the following description, certain specific details are set forth to provide a thorough understanding of different embodiments. However, it will be appreciated by those skilled in the art that the present invention can be implemented without these details. In other cases, known structures are not shown or described in detail to avoid making the description of the embodiments unclear unnecessarily. In addition, the titles provided herein are merely for convenience and are not intended to explain the scope or meaning of the invention for which protection is sought.

[0052] References throughout this specification to "some embodiments" or "an embodiment" mean that a particular feature, structure, or characteristic described with respect to that embodiment is included in at least one embodiment. Therefore, the phrases "in some embodiments" or "in an embodiment" appearing in different places throughout the specification do not necessarily all refer to the same embodiment. In addition, the particular features, structures, or characteristics may be combined in any suitable manner in one or more embodiments. Moreover, as used in this specification and the appended claims, the singular forms "a" and "an" include plural referents unless the context clearly dictates otherwise. It should also be noted that the term "or" is generally used in a sense that includes "and / or" unless the context clearly dictates otherwise.

[0053] The term "pharmaceutically acceptable salt" refers to a salt of a compound of the present invention, prepared from a compound having a specific substituent discovered by the present invention and a relatively non-toxic acid or base. When the compound of the present invention contains a relatively acidic functional group, a base addition salt can be obtained by contacting such compound with a sufficient amount of a base in a pure solution or a suitable inert solvent. When the compound of the present invention contains a relatively basic functional group, an acid addition salt can be obtained by contacting such compound with a sufficient amount of an acid in a pure solution or a suitable inert solvent.

[0054] The term "isomer" refers to any tautomer, stereoisomer, isotope isomer, enantiomer or diastereomer of any compound of the present invention. Among them, isotope isomers refer to different molecules that differ only in isotopes but have the same other structures. Specific implementation plan

[0055] Unless otherwise defined, all technical and scientific terms in the present invention have the same meaning as those commonly understood by those skilled in the art. Other various forms of modification, substitution or change may be made according to common technical knowledge and customary means in the art without departing from the above basic technical concept of the present invention.

[0056] The present invention is described in detail below by examples, but it is not intended to limit the present invention in any adverse way. The compounds of the present invention can be prepared by a variety of synthesis methods well known to those skilled in the art, including the specific embodiments listed below, the embodiments formed by the combination of the embodiments with other chemical synthesis methods, and equivalent substitutions well known to those skilled in the art, and preferred embodiments include but are not limited to the embodiments of the present invention. It will be obvious to those skilled in the art that various changes and improvements are made to the specific embodiments of the present invention without departing from the spirit and scope of the present invention, and they should also be regarded as the protection scope of the present invention.

[0057] LC-MS analysis method:

[0058] Mass spectrometry conditions: instrument Thermo ISQ EC; ion source ESI (EA+EA-); source temperature 300°C; sheath gas pressure 50.0 psi; auxiliary gas pressure 5.0 psi; purge gas pressure 0.5 psi; vaporization chamber temperature 300°C.

[0059] Chromatographic conditions: instrument Thermo U3000; detector DAD-3000 (RS) (diode array detector); chromatographic column Pheromone Titank C18 3μm 4.6×50mm; flow rate 2.0mL / min, split; column temperature 35°C; mobile phase A contained 0.05% formic acid and 5% acetonitrile in water; mobile phase B contained 0.05% formic acid in acetonitrile; elution method was linear elution from 100% to 5% of phase A within 0 to 1.0min, and then 5% of phase A was maintained for 1.2min.

[0060] HPLC analysis method:

[0061] Instrument: Thermo U3000; detector: VWD-3×00(RS) (ultraviolet detector); wavelength: 254nm; chromatographic column: Shimadzu inertsil 3μm 4.6×150mm; flow rate: 0.8mL / min; column temperature: 35°C; mobile phase A: water containing 0.05% formic acid and 5% acetonitrile; mobile phase B: acetonitrile containing 0.05% formic acid; elution method: first maintain 100% phase A for 1.0min, then linearly elute phase A from 100% to 5% within 1.0-8.0min, and finally maintain 5% phase A for 4.0min.

[0062] 1 H-NMR analysis method:

[0063] 1H-NMR was measured at room temperature using a BRUKER AVANCE-400 MHz NMR spectrometer in DMSO-d6 or CDCl3 with TMS as the internal standard. Signal peaks were expressed as s (singlet), d (doublet), t (triplet), q (quartet), m (multiplet), dd (double of doublets), tt (double of triplets). The unit of coupling constant (J) is Hertz (Hz).

[0064] According to the above-described method, the present invention prepared representative compounds I-1 to I-3 (see Table 1), and detected and recorded their purities.

[0065] Table 1: Representative compounds

[0066]

[0067]

[0068] The present invention is further described below in conjunction with specific examples, but the protection scope of the present invention is not limited to these examples. The percentages described in the present invention are all weight percentages unless otherwise specified. The numerical ranges described in the specification, such as units of measurement, reaction conditions, physical states of compounds or percentages, are all for providing unambiguous written references. When implementing the present invention, those skilled in the art use temperatures, concentrations, quantities, carbon atoms, etc. outside this range or different from a single numerical value, and it is still possible to obtain the expected results. In addition, the raw materials in the following examples, if not otherwise specified, can all be commercially available, for example, can be purchased from Shanghai Bid Pharmaceutical Technology Co., Ltd., Jiangsu Aikang Biopharmaceutical R&D Co., Ltd., Nanjing Yaoshi Technology Co., Ltd., Shanghai Shaoyuan Reagent Co., Ltd., and Hechun Biotechnology (Shanghai) Co., Ltd.

[0069] Intermediate: Preparation of I-1c

[0070]

[0071] The commercially available I-1a (1.80 g, 10.02 mmol, 1.0 eq) was dissolved in DMF (30 mL), Cs2CO3 (9.79 g, 30.06 mmol, 3.0 eq) and I-1b (4.27 g, 30.06 mmol, 3.0 eq) were added, and the reaction solution was stirred at 50 ° C for 3 hours, then water (50 mL) was added to the reaction solution for dilution, extracted with ethyl acetate (80 mL), washed once with saturated brine (50 mL), dried over anhydrous Na2SO4, and concentrated to obtain a crude product. The crude product was purified by silica gel column chromatography (PE / EA (v / v) = 2 / 1) to obtain a light yellow solid I-1c. (1.77 g, yield 91.2%). LC-MS MS-ESI (m / z) 194.0 [M+H]+ .

[0072] Intermediate: Preparation of I-1e

[0073]

[0074] The homemade I-1c (1.00 g, 5.16 mmol, 1.0 eq) was dissolved in DMSO (50 mL), Cs2CO3 (5.55 g, 17.03 mmol, 3.3 eq) and commercially available I-1d (971.11 mg, 5.16 mmol, 1.0 eq) were added, and the reaction solution was stirred at 140 ° C for 4 hours, then the reaction solution was cooled to room temperature, diluted with water (100 mL), and then the pH was adjusted to neutral, the solid was precipitated, filtered, and dried to obtain a yellow solid I-1e. (1.41 g, yield 79.2%). LC-MS MS-ESI (m / z) 346.1 [M+H] + .

[0075] Intermediate: Preparation of I-2c

[0076]

[0077] The light yellow solid I-2c was prepared from I-1a (1.80 g, 10.02 mmol, 1.0 eq) and commercially available I-2b (2.00 g, 13.79 mmol, 1.38 eq) and K2CO3 (3.20 g, 23.15 mmol, 2.3 eq) according to the similar steps in intermediate I-1c. (1.60 g, yield 81.2%). LC-MS MS-ESI (m / z) 197.1 [M+H] + .

[0078] Intermediate: Preparation of I-2e

[0079]

[0080] Yellow solid I-2e was prepared from homemade I-2c (1.60 g, 8.14 mmol, 1.0 eq), K2CO3 (3.40 g, 24.60 mmol, 3.0 eq) and commercially available I-1d (1.50 g, 8.14 mmol, 1.0 eq) according to the similar steps of intermediate I-1e. (0.92 g, yield 32.4%). LC-MS MS-ESI (m / z) 349.1 [M+H] + .

[0081] Example 1: Preparation of Compound I-1

[0082]

[0083] I-1e (150.00 mg, 0.43 mmol, 1.0 eq) and I-1f (70.00 mg, 0.62 mmol, 1.4 eq) were suspended in THF (20 mL) and NMP (5 mL), and N-methylimidazole (NMI, 162.00 mg, 1.97 mmol, 4.5 eq) and TCFH (280.00 mg, 1.00 mmol, 2.3 eq) were added, and the resulting solution was stirred at room temperature for 2 h. The reaction solution was concentrated to remove THF, and the residue was diluted with water (50 mL). The precipitated solid was collected by filtration and rinsed with water (3 mL) 3 times. The crude product was dried and separated by preparative TLC (DCM / MeOH (v / v) = 15 / 1) to obtain a yellow solid I-1. (104.0 mg, yield 53.5%). LC-MS MS-ESI (m / z) 440.3 [M+H] +.

[0084] 1 H-NMR(400MHz,DMSO-d6)δppm 9.88(s,1H),8.65(d,J=5.2Hz,1H),8.43(d,J=9.2Hz,1H),8.25(d,J=9.2Hz,1H),8.08(d,J=8.3Hz,1H),7.87-7.92(m,2H),7.63-7.69(m, 1H),7.58(dd,J=9.4,2.4Hz,1H),7.45(d,J=2.6Hz,1H),7.32(dd,J=9.2,2.6Hz,1H),6.60(d,J=4.8Hz,1H),4.99(s,2H),3.95ppm(s,3H).

[0085] Example 2: Preparation of Compound I-2

[0086]

[0087] I-2e (175.00 mg, 0.50 mmol, 1.0 eq) and I-2f (108.00 mg, 1.00 mmol, 2.0 eq) were suspended in DMF (10 mL), Et3N (310.00 mg, 3.06 mmol, 6.1 eq) and PyBOP (520.00 mg, 1.00 mmol, 2.0 eq) were added, and the resulting solution was stirred at room temperature for 2 h. The reaction solution was diluted with water (50 mL), and the precipitated solid was collected by filtration. The crude product was dried and separated by preparative TLC (DCM / MeOH (v / v) = 15 / 1) to obtain a yellow solid I-2. (78.0 mg, yield 34.8%). LC-MS MS-ESI (m / z) 439.4 [M+H] + .

[0088] 1 H-NMR(400MHz,DMSO-d6)δppm 9.88(s,1H),8.64(d,J=5.2Hz,1H),8.43(d,J=9.2Hz,1H),8.25(d,J=9.2Hz,1H),8.08(d,J=8 .3Hz,1H),7.89(s,1H),7.89(d,J=11.5Hz,1H),7.66(t,J=7.2Hz,1H),7.58(dd,J=9.2,2.6Hz ,1H),7.44(d,J=2.6Hz,1H),7.37(d,J=8.2Hz,1H),7.32(dd,J=9.2,2.6Hz,1H),6.98(t,J=7. 1Hz, 1H), 6.82 (d, J = 8.3Hz, 1H), 6.64 (t, J = 7.0Hz, 1H), 6.59 (d, J = 5.2Hz, 1H), 5.00ppm (s, 2H).

[0089] Example 3: Preparation of Compound I-3

[0090]

[0091] Yellow compound I-3 was prepared from I-2e (175.00 mg, 0.50 mmol, 1.0 eq) and I-1f (112.00 mg, 1.00 mmol, 2.0 eq), Et3N (310.00 mg, 3.06 mmol, 6.1 eq) and PyBOP (520.00 mg, 1.00 mmol, 2.0 eq) according to the similar steps in compound I-2. (85.7 mg, yield 38.3%). LC-MS MS-ESI (m / z) 443.3 [M+H] + .

[0092] 1 H-NMR(400MHz,DMSO-d6)δppm 9.88(s,1H),8.64(d,J=5.2Hz,1H),8.43(d,J=9.6Hz,1H),8.25(d,J=9.2Hz,1H),8.08(d,J=8.3Hz,1H),7.84-7.94(m,2H),7.66(t,J =7.7Hz,1H),7.58(dd,J=9.2,2.6Hz,1H),7.44(d,J=2.6Hz,1H),7.32(dd,J=9.2,2.6Hz,1H),6.59(d,J=5.2Hz,1H),4.99ppm(s,2H).

[0093] In vitro biological evaluation

[0094] Example A Test of the Inhibitory Activity of the Compounds of the Invention on Aurora B and VEGFR2 Kinases

[0095] Prepare DMSO working solutions of compound gradient concentrations: prepare the compound into 0.2 mM DMSO solution, and dilute it into 10 working solutions of gradient concentrations with DMSO (corresponding to final concentrations of the compound in the reaction system of 1000 nM, 300 nM, 100 nM, 30 nM, 10 nM, 3 nM, 1 nM, 0.3 nM, 0.1 nM, and 0.03 nM).

[0096] Aurora B kinase and VEGFR2 inhibitory activity test: The half inhibitory concentration (IC50) of the compound on Aurora B kinase and VEGFR2 was tested by HTRF KinEASE-TK kit (Cisbio, Cat#62TK0PEC) and HTRF KinEASE-STK kit (Cisbio, Cat#61ST2BLE), respectively. 50 ). According to the instructions of the kit, prepare 2×kinase&Metal solution (Aurora B / Cat#05-102 and VEGFR2 / Cat#08-191 are purchased from Carna) and 2×Substrate&ATP solution (substrates S2 and TK are used for Aurora B and VEGFR2, respectively). Add 25nL of working solution of the compound to be tested at each gradient concentration and 2.5μL of 2×kinase&Metal solution to each well of the 384-well plate, seal the plate with a sealing film, mix well, centrifuge, and incubate at 25°C for 10 minutes. Then add 2.5μL of 2×Substrate&ATP solution to each well and incubate again at 25°C for 30 minutes. Finally, add 5μL of 2×XL665&Antibody kinase detection reaction solution to each well, incubate at 25°C for 60 minutes, and read the fluorescence signals of each well at 665nm and 620nm wavelengths using an enzyme reader.

[0097] The 665 nm / 620 nm fluorescence signal ratio (X) of each well was calculated, and the well without compound was used as the negative reference (Y) and the well without enzyme was used as the positive reference (Z). The inhibition rate of the compound in each test well on the corresponding kinase was calculated by the formula: Inhibition (%) = 100% - (XZ) / (YZ) × 100%, and the half inhibitory concentration (IC) of each compound on the corresponding kinase was calculated by GraphPad 7.0 software. 50 ). The specific data are shown in Table 2 below:

[0098] Table 2: Inhibitory activity of the compounds of the present invention on AuroraB and VEGFR kinases

[0099]

[0100] The results show that the representative compounds I-1, I-2 and I-3 prepared in the present invention have excellent VEGFR2 and Aurora B kinase inhibitory activities.

[0101] Example B Pharmacokinetics of the compounds of the present invention in mice

[0102] Twenty-four female BALB / c mice of about 8 weeks of age (purchased from Zhuhai Baishitong Biotechnology Co., Ltd.) were randomly divided into four groups, with 6 mice in each group. The test compound was prepared in an aqueous solvent containing 0.2% CMC-Na, and the suspension was gavaged with the compound at 5 mg / kg. The single dose was blood drawn alternately through the fundus venous plexus before and 15 min, 30 min, 1 h, 2 h, 4 h, 8 h and 24 h after administration. About 0.06 mL of blood was collected in a centrifuge tube (heparin sodium anticoagulation), centrifuged at 4 ° C, 3200 g for 10 minutes, the supernatant plasma was separated, and frozen at -20 ° C for testing. Several plasma samples were accurately taken, and after extraction with a certain proportion of acetonitrile, the drug concentration in plasma was determined by liquid chromatography-mass spectrometry (LC-MS / MS). The pharmacokinetic parameters were calculated using Phoenix WinNonlin 7.0 based on the average blood drug concentration data at different time points. The experimental data are summarized in Table 3:

[0103] Table 3 Pharmacokinetic parameters of the test compounds in mice after oral administration

[0104]

[0105] From the above results, it can be seen that after oral administration, the representative compounds I-2 and I-3 of the present invention have a significant effect on the plasma exposure (AUC) and half-life (T 1 / 2 ) and mean residence time (MRT) are higher than those of sioronib. This indicates that the representative compounds I-2 and I-3 of the present invention can significantly prolong the half-life in vivo, improve the metabolic stability of the drug in vivo, increase the drug exposure level in vivo, and have the potential to bring unexpected anti-tumor activity after oral administration.

[0106] Industrial Applicability

[0107] The compound of the invention has excellent VEGFR2, Aurora B and other kinase inhibitory activities and can be used as a drug for treating diseases related to the activity of VEGFR2, Aurora B and other kinases.

[0108] The above is only a preferred embodiment of the present invention. It should be pointed out that for ordinary technicians in this technical field, several improvements and modifications can be made without departing from the principle of the present invention. These improvements and modifications should also be regarded as the scope of protection of the present invention.

Claims

1. A compound represented by general formula (I), its isomer, crystal form, pharmaceutically acceptable salt, hydrate or solvate: in, X1, X2, X3, X4, X5, X6, X7, X8, X9, X 10 , X 11 , X 12 , X 13 , X 14 , X 15 , X 16 , X 17 and X 18 are each independently selected from hydrogen or deuterium, and X1, X2, X3, X4, X5, X6, X7, X8, X9, X 10 , X 11 , X 12 , X 13 , X 14 , X 15 , X 16 , X 17 or X 18 At least one of them is selected from deuterium.

2. The compound according to claim 1, its isomer, crystalline form, pharmaceutically acceptable salt, hydrate or solvate, wherein the compound has one of the following structures:

3. The compound according to claim 1, its isomer, crystalline form, pharmaceutically acceptable salt, hydrate or solvate, wherein the compound has one of the following structures:

4. The compound according to claim 1, its isomer, crystalline form, pharmaceutically acceptable salt, hydrate or solvate, wherein the compound has one of the following structures:

5. The compound according to claim 1, its isomer, crystalline form, pharmaceutically acceptable salt, hydrate or solvate, wherein the compound has one of the following structures:

6. A pharmaceutical composition, characterized in that Contains the compound according to any one of claims 1 to 5, its isomer, crystal form, pharmaceutically acceptable salt, hydrate or solvate, and a pharmaceutically acceptable excipient or carrier.

7. Use of the compound according to any one of claims 1 to 5, its isomer, crystal form, pharmaceutically acceptable salt, hydrate or solvate, or the pharmaceutical composition according to claim 6 in the preparation of a medicament for a disease associated with abnormal protein kinase activity.

8. The use according to claim 7, characterized in that The protein kinase is VEGFR 2 and / or Aurora B.

9. The use according to claim 7, characterized in that The diseases are inflammation, autoimmune diseases, cancer, neurological and neurodegenerative diseases, allergies, asthma, cardiovascular and metabolic diseases, or hormone-related diseases.

10. The use according to claim 9, characterized in that The cancers are blood cancers and solid tumors.

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