CN109400597B - VEGFR-2 inhibitor ABT-869-based protein degradation targeting chimera and preparation method and application thereof - Google Patents
VEGFR-2 inhibitor ABT-869-based protein degradation targeting chimera and preparation method and application thereof Download PDFInfo
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Abstract
A protein degradation targeting chimera based on VEGFR-2 inhibitor ABT-869 and a preparation method and application thereof are disclosed, wherein alkyl dicarboxylic acid and 1- (4- (3-amino-1H-indazol-4-yl) phenyl) -3- (2-fluoro-5-methylphenyl) urea are condensed under PyBop to obtain an intermediate product with monocarboxylic acid; 2) the protein degradation targeting chimera is obtained by condensing the intermediate product with monocarboxylic acid and (2S, 4R) -1- ((S) -2-amino-3, 3-dimethylbutyryl) -4-hydroxy-N- (4- (4-methylthiazol-5-yl) benzyl) pyrrolidine-2-formamide under the action of HATU.
Description
Technical Field
The invention relates to a protein degradation targeting chimera based on a VEGFR-2 inhibitor ABT-869, a preparation method and application thereof.
Background
The protein degradation target chimeras (PROTACs) have the functions of ubiquitination and degradation on corresponding target proteins (such as VEGFR-2 proteins) and polypeptides. The target protein ligand part is combined with target protein, the E3 ubiquitin ligase ligand part is combined with E3 ubiquitin ligase, the two parts are connected by a linker, activated ubiquitin is transferred onto the target protein through the E3 ubiquitin ligase, selective ubiquitination of the target protein is realized, and finally the ubiquitinated target protein is recognized and degraded by proteasome.
In particular, protein degradation targeting chimeras (PROTACs) ubiquitinate and degrade target proteins and polypeptides are involved in a protein degradation pathway in vivo, i.e., the ubiquitin-proteasome system. The ubiquitin-proteasome system is the major pathway for intracellular protein degradation, and is involved in the degradation of more than 80% of proteins in cells. The protein degradation process of the ubiquitin-proteasome system is: there are three enzymes involved in ubiquitination of the target protein. Namely E1: ubiquitin activating enzyme; e2: an ubiquitin-binding enzyme; e3: ubiquitin ligase. The ubiquitin is activated by E1 firstly, and then the ubiquitin is transferred to E2, and the ubiquitin molecule is transferred to the target protein under the action of E3, so that the ubiquitination of the protein is realized. Finally recognized by proteasome and degraded into peptide fragments with certain length.
The protein degradation targeting chimera can be used for treating various diseases in a manner completely different from that of the traditional small molecular compound. Protein degradation targeting chimeras can selectively reduce the level of target proteins in patient cells by recognizing and ubiquitinating the target proteins, followed by proteasomal degradation, to treat several diseases.
Vascular endothelial growth factor receptor (vascular endothelial growth factor receptor) is a membrane protein expressed by VEGF gene, belongs to tyrosine family protein, and is a macromolecular protein closely related to malignant tumor. Vascular endothelial growth factor and its receptor are overexpressed in a range of tumor cells, and the receptor family includes three subtypes: VEGFR-1, VEGFR-2, VEGFR-3. Among them, VEGFR-2 is mainly involved in the proliferation of vascular endothelial cells and is most widely distributed among cancer cells. A series of studies have demonstrated that it can be an effective drug target.
From 2008, small molecule PROTACs composed of ligands of E3 ubiquitin ligase MDM2(mouse double minute2homologue), clAP1(cell inhibitor of apoptosis), crbn (cereblon) and VH L (von Hippel-L indou) and protein ligands were successively reported, wherein one E3 ubiquitin ligase ligand having superior effect is VH L (von Hippel-L indou) ligand.
Disclosure of Invention
The invention aims to provide a protein degradation targeting chimera based on a VEGFR-2 inhibitor ABT-869, a preparation method and application thereof.
In order to achieve the purpose, the invention adopts the following technical scheme:
a protein degradation targeting chimera based on a VEGFR-2 inhibitor ABT-869 has a structural formula as follows:
wherein n is an integer between 1 and 20.
In a further development of the invention, n is 3, 8 or 12.
A preparation method of a protein degradation targeting chimera based on a VEGFR-2 inhibitor ABT-869 comprises the following steps:
1) the condensation of alkyl dicarboxylic acid and 1- (4- (3-amino-1H-indazol-4-yl) phenyl) -3- (2-fluoro-5-methylphenyl) urea under PyBop gives an intermediate product with monocarboxylic acid;
2) the intermediate product with monocarboxylic acid and (2S, 4R) -1- ((S) -2-amino-3, 3-dimethylbutyryl) -4-hydroxy-N- (4- (4-methylthiazol-5-yl) benzyl) pyrrolidine-2-carboxamide are condensed in HATU to obtain a protein degradation targeting chimera based on VEGFR-2 inhibitor ABT-869, and the structural formula is as follows:
wherein n is an integer between 1 and 20.
The further improvement of the invention is that the specific process of the step 1) is as follows: dissolving alkyl dicarboxylic acid and PyBop in dichloromethane, dropwise adding triethylamine, stirring uniformly, adding 1- (4- (3-amino-1H-indazol-4-yl) phenyl) -3- (2-fluoro-5-methylphenyl) urea, stirring at room temperature for 12H, and treating to obtain an intermediate product with monocarboxylic acid.
A further improvement of the invention is that 3.99mmol of pimelic acid and 3.19mmol of PyBop are dissolved in 20m L dichloromethane, 4.79mmol of triethylamine is added dropwise, after stirring uniformly, 0.799mmol of 1- (4- (3-amino-1H-indazol-4-yl) phenyl) -3- (2-fluoro-5-methylphenyl) urea is added, and after stirring for 12H at room temperature, the mixture is treated to obtain an intermediate product with monocarboxylic acid.
The further improvement of the invention is that the specific process of the step 2) is as follows: dissolving the intermediate product with the monocarboxylic acid obtained in the step 1) in dichloromethane, adding (2S, 4R) -1- ((S) -2-amino-3, 3-dimethylbutyryl) -4-hydroxy-N- (4- (4-methylthiazol-5-yl) benzyl) pyrrolidine-2-formamide, stirring uniformly in ice bath, then dropwise adding DIPEA, stirring uniformly, adding HATU, stirring for 12 hours at 25 ℃, and treating after the reaction is finished to obtain the protein degradation targeting chimera based on the VEGFR-2 inhibitor ABT-869.
In a further improvement, 0.193mmol of 7- (3-amino-4- (4- (3- (2-fluoro-5-methylphenyl) ureido) phenyl) -1H-indazol-1-yl) -7-oxoheptanoic acid is dissolved in 20m L dichloromethane, 0.193mmol of (2S, 4R) -1- ((S) -2-amino-3, 3-dimethylbutyryl) -4-hydroxy-N- (4- (4-methylthiazol-5-yl) benzyl) pyrrolidine-2-carboxamide is added, stirring is performed uniformly in ice bath, then 0.773mmol of DIPEA is added dropwise, stirring is performed uniformly, HATU0.29mmol is added, and processing is performed after stirring for 12H at 25 ℃ to obtain the protein degradation targeting chimera based on the VEGFR-2 inhibitor ABT-869.
An application of a protein degradation targeting chimera based on a VEGFR-2 inhibitor ABT-869 in preparing a medicine for treating or preventing cancer.
The further improvement of the invention lies in the application of the protein degradation targeting chimera based on the VEGFR-2 inhibitor ABT-869 in the preparation of the anti-tumor drug taking VEGFR-2 kinase as a target spot.
Compared with the prior art, the invention has the following beneficial effects:
the protein degradation target chimera is obtained by connecting the biphenyl urea VEGFR-2 protein inhibitor with von Rippel-L indou (VH L) protein ligand in an E3 ubiquitin ligase complex through alkyl dicarboxylic acid, and the protein degradation target chimera (PROTACs) can selectively induce the degradation of VEGFR-2 protein.
The small molecule protein degradation target chimera can carry out ubiquitination marking on VEGFR-2 protein, induces protein degradation, and has better anti-tumor effect than VEGFR-2 protein inhibitor. Inhibition of VEGFR-2 proteins often requires maintenance of the drug at higher concentrations for long periods of time, potentially causing serious side effects; only a small amount of compounds are needed for inducing protein degradation, the process is similar to catalytic reaction, and equimolar amount of medicine is not needed, so that the dosage of the medicine can be reduced by using the small molecular protein degradation targeting chimera, and toxic and side effects are reduced. The protein degradation targeting chimera has antitumor activity in vitro, can be applied to the preparation of antitumor drugs, and is particularly used for preparing antitumor drugs taking VEGFR-2 kinase as a target spot.
Drawings
FIG. 1 is a synthesis route diagram of a small molecule protein degradation targeting chimera with anti-tumor activity provided by the present invention;
wherein, the compound 1 is 1- (4- (3-amino-1H-indazol-4-yl) phenyl) -3- (2-fluoro-5-methylphenyl) urea, the compound 2 is an alkyl dicarboxylic acid, the compound 3 is an intermediate product with monocarboxylic acid, the compound 4 is (2S, 4R) -1- ((S) -2-amino-3, 3-dimethylbutyryl) -4-hydroxy-N- (4- (4-methylthiazol-5-yl) benzyl) pyrrolidine-2-carboxamide, and the compound (X) is a small molecule protein degradation targeting chimera with antitumor activity.
The specific notations in the figures are:
a.PyBop,TEA,CH2Cl2,rt；b.HATU,DIPEA,CH2Cl2,rt。
Detailed Description
The present invention will now be described in further detail with reference to the following figures and specific examples, which are intended to be illustrative, but not limiting, of the invention.
The invention relates to a protein degradation target chimera (PROTACs) which can selectively induce the degradation of VEGFR-2 protein, and the protein degradation target chimera is obtained by connecting a biphenyl urea VEGFR-2 protein inhibitor and a von Rippel-L indou (VH L) protein ligand in an E3 ubiquitin ligase complex by using alkyl dicarboxylic acid.
The invention provides a small molecular protein degradation targeting chimera with anti-tumor activity, which has anti-tumor activity in vitro and can be applied to the preparation of anti-tumor drugs.
The chemical structural formula of the micromolecular protein degradation targeting chimera with the anti-tumor activity provided by the invention is as follows:
wherein n is an integer from 1 to 20; preferably, n is 3, 8 or 12.
The invention relates to a protein degradation targeting chimera, which comprises the following components:
(2S, 4R) -1- ((S) -2- (7- (3-amino-4- (4- (3- (2-fluoro-5-methylphenyl) ureido) phenyl) -1H-indazol-1-yl) -7-oxoheptanamido) -3, 3-dimethylbutyryl) -4-hydroxy-N- (4- (4-methylthiazol-5-yl) benzyl) pyrrolidine-2-carboxamide;
(2S, 4R) -1- ((S) -2- (12- (3-amino-4- (4- (3- (2-fluoro-5-methylphenyl) ureido) phenyl) -1H-indazol-1-yl) -12-oxododecanoylamino) -3, 3-dimethylbutyryl) -4-hydroxy-N- (4- (4-methylthiazol-5-yl) benzyl) pyrrolidine-2-carboxamide;
(2S, 4R) -1- ((S) -2- (16- (3-amino-4- (4- (3- (2-fluoro-5-methylphenyl) ureido) phenyl) -1H-indazol-1-yl) -16-oxohexadecanoylamino) -3, 3-dimethylbutyryl) -4-hydroxy-N- (4- (4-methylthiazol-5-yl) benzyl) pyrrolidine-2-carboxamide;
the preparation and activity screening method of the candidate drug small molecule protein degradation targeting chimera with antitumor activity provided by the invention is described in detail below with reference to the synthetic route and the specific synthetic example shown in fig. 1.
Referring to fig. 1, a preparation method of a protein degradation targeting chimera based on a VEGFR-2 inhibitor ABT-869, comprising the following steps:
1) the alkyl dicarboxylic acid and biphenyl urea VEGFR-2 protein ligand 1- (4- (3-amino-1H-indazol-4-yl) phenyl) -3- (2-fluoro-5-methylphenyl) urea are condensed by a PyBop condensing agent to obtain an intermediate product with monocarboxylic acid;
2) condensing the intermediate product with monocarboxylic acid and E3 ubiquitin ligase ligand (2S, 4R) -1- ((S) -2-amino-3, 3-dimethylbutyryl) -4-hydroxy-N- (4- (4-methylthiazol-5-yl) benzyl) pyrrolidine-2-carboxamide in HATU condensing agent to obtain a compound represented by a general formula (X);
the specific operation of the step 1) is as follows: alkyl dicarboxylic acid and PyBop are dissolved in dichloromethane, triethylamine is slowly dropped, and after stirring for 3min, a sample is taken. Adding VEGFR-2 protein ligand 1- (4- (3-amino-1H-indazole-4-yl) phenyl) -3- (2-fluoro-5-methylphenyl) urea, stirring at room temperature overnight, after the reaction is finished, removing the organic solvent at low pressure, adding a proper amount of water, extracting with ethyl acetate, washing and drying the extracted organic phase, evaporating the solvent at reduced pressure to obtain a crude product, and separating the crude product by using a chromatographic column to obtain an intermediate product with monocarboxylic acid.
The specific operation of the step 2) is as follows: dissolving the intermediate product with the monocarboxylic acid obtained in the step 1) in dichloromethane, adding (2S, 4R) -1- ((S) -2-amino-3, 3-dimethylbutyryl) -4-hydroxy-N- (4- (4-methylthiazol-5-yl) benzyl) pyrrolidine-2-carboxamide, stirring for 5min in ice bath, then dropwise adding DIPEA, stirring for 5min, adding HATU, stirring for 12h at 25 ℃, after the reaction is finished, removing the organic solvent by low-pressure rotation, adding a proper amount of water, extracting with ethyl acetate, washing and drying the extracted organic phase, evaporating the solvent under reduced pressure to obtain a crude product, and separating the crude product by using a chromatographic column to obtain the compound represented by the general formula (X).
The micromolecular protein degradation targeting chimera with the anti-tumor activity is applied to preparing anti-tumor drugs taking VEGFR-2 kinase as a target spot.
Example 1
In the structural formula of the small molecule protein degradation targeting chimera with the anti-tumor activity, n is 3, and the small molecule protein degradation targeting chimera is prepared through the following steps (see figure 1):
1) the condensation of pimelic acid (compound 2) with the VEGFR-2 protein ligand 1- (4- (3-amino-1H-indazol-4-yl) phenyl) -3- (2-fluoro-5-methylphenyl) urea (compound 1) in a PyBop condensing agent gives 7- (3-amino-4- (4- (3- (2-fluoro-5-methylphenyl) ureido) phenyl) -1H-indazol-1-yl) -7-oxoheptanoic acid (compound 3); the specific process is as follows:
pimelic acid (0.64g,3.99mmol), PyBop (1.66g,3.19mmol) were dissolved in 20m L dichloromethane, triethylamine (665 μ L, 4.79mmol) was slowly added dropwise, stirring was carried out for 3min, then a sample was taken, 1- (4- (3-amino-1H-indazol-4-yl) phenyl) -3- (2-fluoro-5-methylphenyl) urea (0.3g, 0.799mmol) was added, stirring was carried out overnight at room temperature, then the organic solvent was removed by low pressure rotation, an appropriate amount of water was added, extraction was carried out with ethyl acetate, drying was carried out over anhydrous sodium sulfate, the organic solvent was removed under reduced pressure, and the residue was purified by silica gel column chromatography, eluting with petroleum ether/ethyl acetate (V/V ═ 6/1-3/1) to give a white solid weighing 0.28g, yield 67.7%.
LCMS(ESI,m/z):518.20[M-H]-
2)7- (3-amino-4- (4- (3- (2-fluoro-5-methylphenyl) ureido) phenyl) -1H-indazol-1-yl) -7-oxoheptanoic acid (Compound 3) with E3 ubiquitin ligase ligand (2S, 4R) -1- ((S) -2-amino-3, 3-dimethylbutyryl) -4-hydroxy-N- (4- (4-methylthiazol-5-yl) benzyl) pyrrolidine-2-carboxamide (Compound 4) by condensation with HATU condensing agent to give (2S, 4R) -1- ((S) -2- (7- (3-amino-4- (4- (3- (2-fluoro-5-methylphenyl) urea) Yl) phenyl) -1H-indazol-1-yl) -7-oxoheptanamido) -3, 3-dimethylbutyryl) -4-hydroxy-N- (4- (4-methylthiazol-5-yl) benzyl) pyrrolidine-2-carboxamide (compound X); the specific process is as follows:
7- (3-amino-4- (4- (3- (2-fluoro-5-methylphenyl) ureido) phenyl) -1H-indazol-1-yl) -7-oxoheptanoic acid (0.1g, 0.193mmol) was dissolved in 20m L dichloromethane, (2S, 4R) -1- ((S) -2-amino-3, 3-dimethylbutyryl) -4-hydroxy-N- (4- (4-methylthiazol-5-yl) benzyl) pyrrolidine-2-carboxamide (0.084g, 0.193mmol) was added, stirred under ice bath for 5min, then DIPEA (132 μ L, 0.773mmol) was added dropwise, stirred for 5min, HATU (0.11g, 0.29mmol) was added, stirred at 25 ℃ for 12H, then organic solvent was removed under low pressure, appropriate water was added, silica gel column was extracted with ethyl acetate, dried over anhydrous sodium sulfate, organic solvent was removed under reduced pressure, the residue was purified by chromatography, eluting with petroleum ether/ethyl acetate (V/v. 1/1-0/1 g), the target compound was obtained in 0.22 g.
The structure of the obtained target compound is as follows:
the hydrogen spectrum nuclear magnetic resonance data is as follows:1H NMR(400MHz,DMSO-D6)9.28(s,1H),8.98(s,1H),8.57(s,2H),8.31-8.33(d,1H),8.00-8.02(d,1H),7.87-7.89(d,1H),7.58-7.64(m,3H),7.37-7.43(m,6H),7.10-7.19(m,2H),6.81-6.84(m,1H),5.21(s,2H),5.14(d,1H),4.53-4.56(d,1H),4.41-4.47(t,2H),4.35(s,1H),4.19-4.25(q,1H),3.63-3.70(t,2H),2.45(s,3H),2.29(s,3H),2.12-2.19(m,1H),2.02-2.06(m,1H),1.87-1.94(m,1H),1.67-1.75(m,3H),1.50-1.61(m,2H),1.34-1.39(m,2H),1.24(m,2H),0.91(s,9H).
LCMS(ESI,m/z):930.45[M-H]-
example 2
In the structural formula of the micromolecular protein degradation targeting chimera with the anti-tumor activity, n is 8.
The synthesis procedure was the same as in example 1.
The structure of the obtained target compound is as follows:
the hydrogen spectrum nuclear magnetic resonance data is as follows:1H NMR(400MHz,DMSO-D6)9.28(s,1H),8.99(s,1H),8.58(s,2H),8.31-8.33(d,1H),8.01-8.03(d,1H),7.85-7.87(d,1H),7.58-7.64(m,3H),7.40-7.43(m,6H),7.10-7.19(m,2H),6.83(m,1H),5.21(s,2H),5.14(s,1H),4.54-4.56(d,1H),4.43-4.45(t,2H),4.36(s,1H),4.21-4.25(q,1H),3.63-3.70(t,2H),2.45(s,3H),2.29(s,3H),2.00-2.12(m,3H),1.91-1.92(m,1H),1.66-1.71(m,2H),1.43-1.55(m,2H),1.17-1.36(m,14H),0.93(s,9H).LCMS(ESI,m/z):1000.55[M-H]-
example 3
In the structural formula of the micromolecular protein degradation targeting chimera with the anti-tumor activity, n is 12.
The synthesis procedure is as in example 1
The structure of the obtained target compound is as follows:
the hydrogen spectrum nuclear magnetic resonance data is as follows:1H NMR(400MHz,DMSO-D6)9.27(s,1H),8.98(s,1H),8.57(s,2H),8.30-8.32(d,1H),8.00-8.02(d,1H),7.83-7.86(d,1H),7.57-7.64(m,3H),7.37-7.43(m,6H),7.10-7.19(m,2H),6.82(m,1H),5.19(s,2H),5.13-5.14(d,1H),4.53-4.56(d,1H),4.41-4.47(t,2H),4.35(s,1H),4.19-4.24(q,1H),3.63-3.70(t,2H),2.45(s,3H),2.29(s,3H),1.99-2.12(m,2H),1.87-1.94(m,1H),1.67-1.71(m,2H),1.23-1.49(m,25H),0.94(s,9H).
LCMS(ESI,m/z):1056.60[M-H]-
example 4
And (3) screening the inhibitory activity of the protein degradation targeting chimera on VEGFR-2 kinase.
An ADP-Glo luminescence method is adopted to determine the inhibitory activity of the protein degradation targeting chimera on VEGFR-2 kinase.
ATP (10mM) is diluted to 250 mu M by Buffer (Tris 80mM, MgCl 220 mM, BSA 0.2mg/M L mM), ATP and a substrate Poly (4:1Glu, Tyr) Peptide are prepared into ATP (125 mu M) -Poly (4:1Glu, Tyr) Peptide (0.5 mu g/mu L) mixed solution according to the volume of 1:1, kinase is diluted to 1.5 ng/mu L, a compound to be tested is prepared into 6 concentration gradient solutions, 2 mu L ATP-Poly (4:1Glu, Tyr) Peptide solution, 1 mu L sample solution and 2 mu L enzyme solution are added to start reaction, an ADP-Glo reagent is added to stop the reaction after incubation for 60min at 30 ℃, a KinaseSeDetection reagent of 10 mu L is added to convert ADP into ATP, and the ADP is incubated for 30min at 25 ℃, a chemiluminescence luminescence module of a PerkinElkin multifunctional enzyme analyzer is used for measuring the luminescence inhibition rate.
Numerical treatment, inhibition rate (positive value-administration group value)/(positive value-negative value) × 100%;
the results of the compound experiments are shown in table 1:
table 1 results of inhibitory activity of protein degradation targeting chimeras on VEGFR-2 kinase. (60nM)
As can be seen from Table 1, the protein degradation targeting chimera based on the VEGFR-2 inhibitor ABT-869, prepared by the invention, has better inhibitory activity.
Example 5
And (3) measuring the activity of the protein degradation target chimeric cell level.
The activity of protein degradation targeting chimera cell level is detected by MTT detection method, EA.hy926 cell or SMMC-7721 cell in logarithmic growth period is digested with 0.25% trypsin to prepare single cell suspension, which is inoculated into 96-well plate (2 × 10)4One/well), 180. mu. L per well, 37 ℃ and 5% CO2Culturing in a constant temperature incubator, adding medicine after cell adherence after 24h, arranging 3 multiple wells for each group, adding 20 μ L/well serum-free culture medium for negative control group, adding 20 μ L/well of medicine with different concentrations for experimental group (diluting medicine with serum-free culture medium), placing at 37 deg.C and 5% CO2After the drugs act for 72 hours, carefully sucking and removing the supernatant, adding a 10-fold diluted MTT solution (the final concentration is 0.5mg/m L) with a serum-free culture medium into 200 mu L/hole, incubating at 37 ℃ for 4-6 hours, carefully sucking and removing the supernatant, adding DMSO into 150 mu L/hole, fully shaking on a decoloration shaking table for 15min, and measuring the absorbance (OD) value of each hole at 490nm by using an enzyme-linked immunosorbent assay instrument.
Numerical value processing: inhibition rate (OD)Negative group-ODAdministration set)/(ODNegative group-ODBlank group)×100％；
The results of the experiments for some of the compounds are shown in table 2:
TABLE 2 inhibitory Activity of preferred Compounds on EA.hy926 cells and SMMC-7721 cells (100nM,72h)
As can be seen from Table 2, the chimeras prepared by the present invention have good inhibitory activity against EA.hy926 cells and SMMC-7721 cells.
Claims (7)
1. A protein degradation targeting chimera based on a VEGFR-2 inhibitor ABT-869 is characterized in that the structural formula is as follows:
wherein n is an integer between 1 and 20.
2. The VEGFR-2 inhibitor ABT-869-based proteolytic targeting chimera of claim 1, wherein n is 3, 8 or 12.
3. A preparation method of a protein degradation targeting chimera based on a VEGFR-2 inhibitor ABT-869 is characterized by comprising the following steps:
1) the condensation of alkyl dicarboxylic acid and 1- (4- (3-amino-1H-indazol-4-yl) phenyl) -3- (2-fluoro-5-methylphenyl) urea under PyBop gives an intermediate product with monocarboxylic acid; the specific process is as follows: dissolving alkyl dicarboxylic acid and PyBop in dichloromethane, dropwise adding triethylamine, stirring uniformly, adding 1- (4- (3-amino-1H-indazol-4-yl) phenyl) -3- (2-fluoro-5-methylphenyl) urea, stirring at room temperature for 12H, and treating to obtain an intermediate product with monocarboxylic acid;
2) the intermediate product with monocarboxylic acid and (2S, 4R) -1- ((S) -2-amino-3, 3-dimethylbutyryl) -4-hydroxy-N- (4- (4-methylthiazol-5-yl) benzyl) pyrrolidine-2-carboxamide are condensed under HATU to obtain the protein degradation targeting chimera based on VEGFR-2 inhibitor ABT-869, and the specific process is as follows: dissolving the intermediate product with the monocarboxylic acid obtained in the step 1) in dichloromethane, adding (2S, 4R) -1- ((S) -2-amino-3, 3-dimethylbutyryl) -4-hydroxy-N- (4- (4-methylthiazol-5-yl) benzyl) pyrrolidine-2-formamide, stirring uniformly in ice bath, then dropwise adding DIPEA, stirring uniformly, adding HATU, stirring for 12 hours at 25 ℃, and treating after the reaction is finished to obtain a protein degradation targeting chimera based on a VEGFR-2 inhibitor ABT-869;
the structural formula of the protein degradation targeting chimera is as follows:
wherein n is an integer between 1 and 20.
4. The preparation method of the VEGFR-2 inhibitor ABT-869-based protein degradation targeting chimera according to claim 3, wherein 3.99mmol of pimelic acid and 3.19mmol of PyBop are dissolved in 20m L dichloromethane, 4.79mmol of triethylamine is added dropwise, after stirring uniformly, 0.799mmol of 1- (4- (3-amino-1H-indazol-4-yl) phenyl) -3- (2-fluoro-5-methylphenyl) urea is added, and after stirring for 12H at room temperature, the mixture is treated to obtain an intermediate product with monocarboxylic acid.
5. The preparation method of the VEGFR-2 inhibitor ABT-869-based protein degradation targeting chimera according to claim 3, wherein 0.193mmol of 7- (3-amino-4- (4- (3- (2-fluoro-5-methylphenyl) ureido) phenyl) -1H-indazol-1-yl) -7-oxoheptanoic acid is dissolved in 20m L dichloromethane, 0.193mmol of (2S, 4R) -1- ((S) -2-amino-3, 3-dimethylbutyryl) -4-hydroxy-N- (4- (4-methylthiazol-5-yl) benzyl) pyrrolidine-2-carboxamide is added and stirred uniformly under ice bath, then DIPEA0.773mmol is added dropwise and stirred uniformly, HATU0.29mmol is added, and after stirring at 25 ℃ for 12H, the protein degradation targeting chimera based on the VEGFR-2 inhibitor ABT-869 is obtained.
6. Use of a protein degradation targeting chimera based on the VEGFR-2 inhibitor ABT-869, prepared according to the method of any one of claims 3-5, in the preparation of a medicament for treating or preventing cancer.
7. The use of claim 6, wherein the use of a protein degradation targeting chimera based on the VEGFR-2 inhibitor ABT-869 in the preparation of a VEGFR-2 kinase targeted anti-tumor medicament.
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