CN109400597A - A kind of protein degradation targeting chimera and preparation method and application based on VEGFR-2 inhibitor ABT-869 - Google Patents
A kind of protein degradation targeting chimera and preparation method and application based on VEGFR-2 inhibitor ABT-869 Download PDFInfo
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Abstract
A kind of protein degradation targeting chimera and preparation method and application based on VEGFR-2 inhibitor ABT-869, alkyl dicarboxylic aid and 1- (4- (3- amino-1 h-indazole -4- base) phenyl) -3- (the fluoro- 5- aminomethyl phenyl of 2-) urea obtain the intermediate product with monocarboxylic acid under the condensation of PyBop；2) intermediate product and (2S with monocarboxylic acid; 4R) -1- ((S) -2- amino -3; 3- dimethylbutanoyl) -4- hydroxy-n-(4- (4- methylthiazol -5- base) benzyl) pyrrolidines -2- formamide is under the condensation of HATU; obtain protein degradation targeting chimera; it is simple that protein degradation of the invention targets chimeric preparation; it is easily achieved; and yield is higher; it can be applied to the preparation of anti-tumor drug, in particular for preparing using VEGFR-2 kinases as the anti-tumor drug of target spot.
Description
Technical field
The present invention relates to a kind of, and the protein degradation based on VEGFR-2 inhibitor ABT-869 targets chimera and preparation method
And application.
Background technique
Protein degradation targets chimera (PROTACs), has to corresponding target proteins (such as VEGFR-2 albumen) and polypeptide
The function of ubiquitination and degradation.For target protein ligand moiety in conjunction with target protein, E3 ubiquitin ligase ligand moiety and E3 are general
Plain ligase combines, and two parts are connected by linker, the ubiquitin of activation is transferred on target protein by E3 ubiquitin ligase,
The selective ubiquitination to target protein is realized, the target protein of final ubiquitination is identified and degraded by proteasome.
Specifically, target proteins are degraded protein degradation targeting chimera (PROTACs) ubiquitination and and polypeptide is related to
A kind of intracorporal proteolytic pathway of biology, i.e. Ubiquitin-proteasome system.Ubiquitin-proteasome system is intracellular egg
The main path of white matter degradation, participates in the degradation of intracellular 80% or more protein.Ubiquitin-proteasome system protein degradation mistake
Cheng Shi: there are three the ubiquitinations that enzyme takes part in target protein.That is E1: ubiquitin kinase；E2: ubiquitin binding enzyme；E3: ubiquitinbond
Enzyme.Ubiquitin is then passed to E2 by ubiquitin activating by E1 first, and under the action of E3, ubiquitin molecule is transferred to target proteins
On, realize the ubiquitination of albumen.It is finally identified by proteasome and is degraded to the certain peptide fragment of length.
It can treat various diseases using protein degradation targeting chimera, mode and traditional small molecule compound are completely not
Together.Protein degradation targets chimera by identification and ubiquitination target proteins, is then degraded by proteasome, so as to select
The level for reducing target proteins in Patient cells of selecting property, to treat some diseases.
Vascular endothelial growth factor receptor (vascular endothelial growth factor receptor) be by
The memebrane protein of VEGF gene expression belongs to tyrosine family protein, is the high molecular weight protein closely related with malignant tumour.Blood vessel
Endothelial growth factors and its receptor have overexpression in a series of tumour cells, and this receptor family includes three hypotypes:
VEGFR-1,VEGFR-2,VEGFR-3.Wherein VEGFR-2 is primarily involved in the proliferation of vascular endothelial cell, is distributed in cancer cell
Also most extensively.A series of research confirms that it can be used as effective drug targets.
Since 2008, E3 ubiquitin ligase MDM2 (mouse double minute was reported in succession
2homologue), clAP1 (cellulr inhibitor of apoptosis), CRBN (cereblon) and VHL (von
Hippel-Lindau the small molecule PROTACs, the more outstanding E3 of one of effect that ligand and protein ligands) is constituted
Ubiquitin ligase ligand is VHL (von Hippel-Lindau) ligand.
Summary of the invention
The purpose of the present invention is to provide a kind of, and the protein degradation based on VEGFR-2 inhibitor ABT-869 targets chimera
And preparation method and application.
In order to achieve the above objectives, the invention adopts the following technical scheme:
A kind of protein degradation targeting chimera based on VEGFR-2 inhibitor ABT-869, structural formula are as follows:
Wherein, integer of the n between 1-20.
A further improvement of the present invention lies in that n 3,8 or 12.
A kind of preparation method of the protein degradation targeting chimera based on VEGFR-2 inhibitor ABT-869, including following step
It is rapid:
1) alkyl dicarboxylic aid and 1- (4- (3- amino-1 h-indazole -4- base) phenyl) -3- (the fluoro- 5- aminomethyl phenyl of 2-) urea exist
Under the condensation of PyBop, the intermediate product with monocarboxylic acid is obtained；
2) intermediate product and (2S, 4R) -1- ((S) -2- amino -3,3- dimethylbutanoyl) -4- hydroxyl with monocarboxylic acid
Base-N- (4- (4- methylthiazol -5- base) benzyl) pyrrolidines -2- formamide under the condensation of HATU, is based on
The protein degradation of VEGFR-2 inhibitor ABT-869 targets chimera, and structural formula is as follows:
Wherein, integer of the n between 1-20.
A further improvement of the present invention lies in that the detailed process of the step 1) are as follows: alkyl dicarboxylic aid and PyBop to be dissolved in
In methylene chloride, triethylamine is added dropwise, after mixing evenly, 1- (4- (3- amino-1 h-indazole -4- base) phenyl) is added, and (2- is fluoro- by -3-
5- aminomethyl phenyl) urea, it is handled after 12h is stirred at room temperature, obtains the intermediate product with monocarboxylic acid.
A further improvement of the present invention lies in that pimelic acid 3.99mmol and PyBop 3.19mmol are dissolved in 20mL dichloromethane
In alkane, triethylamine 4.79mmol is added dropwise, after mixing evenly, 1- (4- (3- amino-1 h-indazole -4- base) phenyl) -3- (2- is added
Fluoro- 5- aminomethyl phenyl) urea 0.799mmol, it is handled after 12h is stirred at room temperature, obtains the intermediate product with monocarboxylic acid.
A further improvement of the present invention lies in that the detailed process of the step 2) are as follows: have single carboxylic for what step 1) obtained
The intermediate product of acid is dissolved in methylene chloride, and (2S, 4R) -1- ((S) -2- amino -3,3- dimethylbutanoyl) -4- hydroxyl is added
Base-N- (4- (4- methylthiazol -5- base) benzyl) pyrrolidines -2- formamide is stirred evenly under ice bath, is then added dropwise
DIPEA is stirred evenly, and HATU is added, stirs 12h at 25 DEG C, is handled after reaction, obtains inhibiting based on VEGFR-2
The protein degradation of agent ABT-869 targets chimera.
A further improvement of the present invention lies in that by 7- (3- amino -4- (4- (3- (the fluoro- 5- aminomethyl phenyl of 2-) urea groups) benzene
Base) -1H- indazole -1- base) -7- oxo-heptanoic acid 0.193mmol is dissolved in 20mL methylene chloride, (2S, 4R) -1- ((S) -2- is added
Amino -3,3- dimethylbutanoyl) -4- hydroxy-n-(4- (4- methylthiazol -5- base) benzyl) pyrrolidines -2- formamide
0.193mmol is stirred evenly under ice bath, DIPEA0.773mmol is then added dropwise, stirs evenly, and HATU is added
0.29mmol is handled after stirring 12h at 25 DEG C, and it is embedding to obtain the protein degradation targeting based on VEGFR-2 inhibitor ABT-869
It is fit.
It is a kind of based on VEGFR-2 inhibitor ABT-869 protein degradation targeting chimera preparation treat or prevent cancer
Drug in application.
A further improvement of the present invention lies in that the protein degradation targeting chimera based on VEGFR-2 inhibitor ABT-869 exists
Preparation is using VEGFR-2 kinases as the application in the anti-tumor drug of target spot.
Compared with prior art, the invention has the following advantages:
By the present invention in that with alkyl dicarboxylic aid that biphenyl ureas VEGFR-2 protein inhibitor and E3 ubiquitin ligase is compound
The connection of von Rippel-Lindau (VHL) protein ligands obtains protein degradation and targets chimera in body.Protein degradation targeting is embedding
Fit (PROTACs) is capable of the degradation of selective induction VEGFR-2 albumen.Protein degradation of the invention targets chimera preparation side
Method is simple, it is easy to accomplish, and yield is higher.
Small molecular protein degradation targeting chimera in the present invention can carry out ubiquitination label to VEGFR-2 albumen, lure
Protein degradation is led, antitumous effect is better than VEGFR-2 protein inhibitor.VEGFR-2 albumen is inhibited to generally require drug is long-term
Maintain higher concentration, it is possible to cause serious side effect；And inducible protein degradation only needs a small amount of compound, this
Process is similar to catalysis reaction, does not need the drug of equimolar amounts, so degrading targeting chimera using small molecular protein can
To reduce drug dosage, mitigate toxic side effect.Protein degradation targeting chimera of the invention has antitumor work in vitro
Property, it can be applied to the preparation of anti-tumor drug, particularly for preparing using VEGFR-2 kinases as the anti-tumor drug of target spot.
Detailed description of the invention
Fig. 1 is the synthetic route of small molecular protein degradation targeting chimera with anti-tumor activity provided by the invention
Figure；
Wherein, compound 1 is 1- (4- (3- amino-1 h-indazole -4- base) phenyl) -3- (the fluoro- 5- aminomethyl phenyl of 2-) urea,
Compound 2 is alkyl dicarboxylic aid, and compound 3 is the intermediate product with monocarboxylic acid, and compound 4 is (2S, 4R) -1- ((S) -2-
Amino -3,3- dimethylbutanoyl) -4- hydroxy-n-(4- (4- methylthiazol -5- base) benzyl) pyrrolidines -2- formamide, chemical combination
Object (X) is small molecular protein with anti-tumor activity degradation targeting chimera.
It is marked in figure specifically:
a.PyBop,TEA,CH2Cl2,rt；b.HATU,DIPEA,CH2Cl2,rt。
Specific embodiment
The present invention is described in further detail with specific embodiment with reference to the accompanying drawing, and described is to of the invention
It explains rather than limits.
Protein degradation targeting chimera (PROTACs) of the present invention is capable of the drop of selective induction VEGFR-2 albumen
Solution.By the present invention in that will be in biphenyl ureas VEGFR-2 protein inhibitor and E3 ubiquitin ligase complex with alkyl dicarboxylic aid
The connection of von Rippel-Lindau (VHL) protein ligands obtains protein degradation and targets chimera.These compounds have induction
The function of VEGFR-2 protein degradation can be used for preparing new type antineoplastic medicine.
The present invention provides a kind of small molecular protein degradation targeting chimera with anti-tumor activity, the protein degradation targets
To chimera there is anti-tumor activity in vitro, can be applied to the preparation of anti-tumor drug.
The chemical structural formula of small molecular protein degradation targeting chimera with anti-tumor activity provided by the invention is specific
It is as follows:
Wherein, n is selected from the integer between 1-20；Preferably, 3,8 or 12 n.
Protein degradation of the present invention targets chimera comprising:
(2S, 4R) -1- ((S) -2- (7- (3- amino -4- (4- (3- (the fluoro- 5- aminomethyl phenyl of 2-) urea groups) phenyl) -1H- Yin
Azoles -1- base) -7- oxo heptanamido) -3,3- dimethylbutanoyl) -4- hydroxy-n-(4- (4- methylthiazol -5- base) benzyl)
Pyrrolidines -2- formamide；
(2S, 4R) -1- ((S) -2- (12- (3- amino -4- (4- (3- (the fluoro- 5- aminomethyl phenyl of 2-) urea groups) phenyl) -1H-
Indazole -1- base) -12- oxododecanoyl amido) -3,3- dimethylbutanoyl) -4- hydroxy-n-(4- (4- methylthiazol -5-
Base) benzyl) pyrrolidines -2- formamide；
(2S, 4R) -1- ((S) -2- (16- (3- amino -4- (4- (3- (the fluoro- 5- aminomethyl phenyl of 2-) urea groups) phenyl) -1H-
Indazole -1- base) -16- oxo hexadecanoyl amido) -3,3- dimethylbutanoyl) -4- hydroxy-n-(4- (4- methylthiazol -5-
Base) benzyl) pyrrolidines -2- formamide；
Carry out the tool that the present invention will be described in detail provides below with reference to synthetic route shown in Fig. 1 and specific synthetic example
There are preparation and the method for screening active ingredients of the drug candidate small molecular protein degradation targeting chimera of anti-tumor activity.
Referring to Fig. 1, a kind of preparation method of the protein degradation targeting chimera based on VEGFR-2 inhibitor ABT-869, packet
Include following steps:
1) alkyl dicarboxylic aid and biphenyl ureas VEGFR-2 protein ligands 1- (4- (3- amino-1 h-indazole -4- base) phenyl) -
3- (the fluoro- 5- aminomethyl phenyl of 2-) urea obtains the intermediate product with monocarboxylic acid under the condensation of PyBop condensing agent；
2) this intermediate product and E3 ubiquitin ligase ligand (2S, 4R) -1- ((S) -2- amino -3,3- with monocarboxylic acid
Dimethylbutanoyl) -4- hydroxy-n-(4- (4- methylthiazol -5- base) benzyl) pyrrolidines -2- formamide is in HATU condensing agent
The compound that logical formula (X) indicates is obtained under condensation；
The concrete operations of the step 1) are as follows: by alkyl dicarboxylic aid, PyBop is dissolved in methylene chloride, and three second are slowly added dropwise
Amine, after stirring 3min, sampling.VEGFR-2 protein ligands 1- (4- (3- amino-1 h-indazole -4- base) phenyl) -3- (2- is added
Fluoro- 5- aminomethyl phenyl) urea, it is stirred overnight at room temperature, after reaction, low pressure rotation removes organic solvent, suitable quantity of water is added, with acetic acid second
Ester extraction, the organic phase of extraction is washed, it is dry after decompression boil off solvent, obtain crude product, with chromatography post separation crude product, had
The intermediate product of monocarboxylic acid.
The concrete operations of the step 2) are as follows: the intermediate product with monocarboxylic acid for obtaining step 1) is dissolved in methylene chloride
In, (2S, 4R) -1- ((S) -2- amino -3,3- dimethylbutanoyl) -4- hydroxy-n-(4- (4- methylthiazol -5- base) is added
Benzyl) pyrrolidines -2- formamide, 5min is stirred under ice bath, DIPEA is then added dropwise, stirs 5min, is added HATU, 25 DEG C
Lower stirring 12h, after reaction, low pressure rotation remove organic solvent, and suitable quantity of water is added, is extracted with ethyl acetate, the organic phase of extraction
Decompression boils off solvent after washed, dry, obtains crude product, with chromatography post separation crude product, obtains the compound that logical formula (X) indicates.
The small molecular protein with anti-tumor activity degradation targeting chimera is in preparation using VEGFR-2 kinases as target
Application in the anti-tumor drug of point.
Embodiment 1
In the structural formula of the small molecular protein degradation targeting chimera with anti-tumor activity, n 3 passes through following step
Rapid preparation (referring to Fig. 1):
1) pimelic acid (compound 2) and VEGFR-2 protein ligands 1- (4- (3- amino-1 h-indazole -4- base) phenyl) -3-
(the fluoro- 5- aminomethyl phenyl of 2-) urea (compound 1) obtains 7- (3- amino -4- (4- (3- under the condensation of PyBop condensing agent
(the fluoro- 5- aminomethyl phenyl of 2-) urea groups) phenyl) -1H- indazole -1- base) -7- oxo-heptanoic acid (compound 3)；Detailed process is as follows:
By pimelic acid (0.64g, 3.99mmol), PyBop (1.66g, 3.19mmol) is dissolved in 20mL methylene chloride, slowly
It is added dropwise triethylamine (665 μ L, 4.79mmol), after stirring 3min, sampling.1- (4- (3- amino-1 h-indazole -4- base) benzene is added
Base) -3- (the fluoro- 5- aminomethyl phenyl of 2-) urea (0.3g, 0.799mmol), it is stirred overnight at room temperature, then low pressure rotation removes organic solvent,
Suitable quantity of water is added, is extracted with ethyl acetate, anhydrous sodium sulfate is dry, and decompression rotation removes organic solvent, and residue passes through silicagel column color
Chromatographic purifying is composed, affords white solid, weight 0.28g, yield using petrol ether/ethyl acetate (V/V=6/1-3/1)
67.7%.
LCMS(ESI,m/z):518.20[M-H]-
2) 7- (3- amino -4- (4- (3- (the fluoro- 5- aminomethyl phenyl of 2-) urea groups) phenyl) -1H- indazole -1- base) -7- oxo
Enanthic acid (compound 3) and E3 ubiquitin ligase ligand (2S, 4R) -1- ((S) -2- amino -3,3- dimethylbutanoyl) -4- hydroxyl
Condensation of base-N- (4- (4- methylthiazol -5- base) benzyl) pyrrolidines -2- formamide (compound 4) in HATU condensing agent
Under obtain (2S, 4R) -1- ((S) -2- (7- (3- amino -4- (4- (3- (the fluoro- 5- aminomethyl phenyl of 2-) urea groups) phenyl) -1H- Yin
Azoles -1- base) -7- oxo heptanamido) -3,3- dimethylbutanoyl) -4- hydroxy-n-(4- (4- methylthiazol -5- base) benzyl)
Pyrrolidines -2- formamide (compound X)；Detailed process is as follows:
By 7- (3- amino -4- (4- (3- (the fluoro- 5- aminomethyl phenyl of 2-) urea groups) phenyl) -1H- indazole -1- base) -7- oxo
Enanthic acid (0.1g, 0.193mmol) is dissolved in 20mL methylene chloride, and (2S, 4R) -1- ((S) -2- amino -3,3- dimethyl butyrate is added
Acyl group) -4- hydroxy-n-(4- (4- methylthiazol -5- base) benzyl) pyrrolidines -2- formamide (0.084g, 0.193mmol), ice
Bath is lower to stir 5min, is then added dropwise DIPEA (132 μ L, 0.773mmol), stirring 5min, addition HATU (0.11g,
0.29mmol), 12h is stirred at 25 DEG C, then low pressure rotation removes organic solvent, and suitable quantity of water is added, is extracted with ethyl acetate, anhydrous sulphur
Sour sodium is dry, and decompression rotation removes organic solvent, and residue is purified by silica gel column chromatography, uses petrol ether/ethyl acetate (V/
V=1/1-0/1 target compound, weight 0.11g, yield 61.22%) are afforded.
The structure of gained target compound is as follows:
Hydrogen composes nuclear magnetic resonance data are as follows:1H NMR(400MHz,DMSO-D6)δ9.28(s,1H),δ8.98(s,1H),δ
8.57(s,2H),δ8.31-8.33(d,1H),δ8.00-8.02(d,1H),δ7.87-7.89(d,1H),δ7.58-7.64(m,
3H),δ7.37-7.43(m,6H),δ7.10-7.19(m,2H),δ6.81-6.84(m,1H),δ5.21(s,2H),δ5.14(d,
1H),δ4.53-4.56(d,1H),δ4.41-4.47(t,2H),δ4.35(s,1H),δ4.19-4.25(q,1H),δ3.63-3.70
(t,2H),δ2.45(s,3H),δ2.29(s,3H),δ2.12-2.19(m,1H),δ2.02-2.06(m,1H),δ1.87-1.94
(m,1H),δ1.67-1.75(m,3H),δ1.50-1.61(m,2H),δ1.34-1.39(m,2H),δ1.24(m,2H),δ0.91
(s,9H).
LCMS(ESI,m/z):930.45[M-H]-
Embodiment 2
In the structural formula of the small molecular protein degradation targeting chimera with anti-tumor activity, n 8.
Synthesis step is the same as embodiment 1.
The structure of gained target compound is as follows:
Hydrogen composes nuclear magnetic resonance data are as follows:1H NMR(400MHz,DMSO-D6)δ9.28(s,1H),δ8.99(s,1H),δ
8.58(s,2H),δ8.31-8.33(d,1H),δ8.01-8.03(d,1H),δ7.85-7.87(d,1H),δ7.58-7.64(m,
3H),δ7.40-7.43(m,6H),δ7.10-7.19(m,2H),δ6.83(m,1H),δ5.21(s,2H),δ5.14(s,1H),δ
4.54-4.56(d,1H),δ4.43-4.45(t,2H),δ4.36(s,1H),δ4.21-4.25(q,1H),δ3.63-3.70(t,
2H),δ2.45(s,3H),δ2.29(s,3H),δ2.00-2.12(m,3H),δ1.91-1.92(m,1H),δ1.66-1.71(m,
2H),δ1.43-1.55(m,2H),δ1.17-1.36(m,14H),δ0.93(s,9H).LCMS(ESI,m/z):1000.55[M-
H]-
Embodiment 3
In the structural formula of the small molecular protein degradation targeting chimera with anti-tumor activity, n 12.
Synthesis step is the same as embodiment 1
The structure of gained target compound is as follows:
Hydrogen composes nuclear magnetic resonance data are as follows:1H NMR(400MHz,DMSO-D6)δ9.27(s,1H),δ8.98(s,1H),δ
8.57(s,2H),δ8.30-8.32(d,1H),δ8.00-8.02(d,1H),δ7.83-7.86(d,1H),δ7.57-7.64(m,
3H),δ7.37-7.43(m,6H),δ7.10-7.19(m,2H),δ6.82(m,1H),δ5.19(s,2H),δ5.13-5.14(d,
1H),δ4.53-4.56(d,1H),δ4.41-4.47(t,2H),δ4.35(s,1H),δ4.19-4.24(q,1H),δ3.63-3.70
(t,2H),δ2.45(s,3H),δ2.29(s,3H),δ1.99-2.12(m,2H),δ1.87-1.94(m,1H),δ1.67-1.71
(m,2H),δ1.23-1.49(m,25H),δ0.94(s,9H).
LCMS(ESI,m/z):1056.60[M-H]-
Embodiment 4
Protein degradation targets chimera and screens to the inhibitory activity of VEGFR-2 kinases.
Using ADP-Glo luminescent method measurement protein degradation targeting chimera to the inhibitory activity of VEGFR-2 kinases.
It is 250 with Buffer (Tris 80mM, MgCl2 20mM, BSA 0.2mg/mL, DTT 2mM) dilution ATP (10mM)
μM；ATP and substrate Poly (4:1Glu, Tyr) Peptide is made into ATP (125 μM)-Poly (4:1Glu, Tyr) by volume 1:1
Peptide (0.5 μ g/ μ L) mixed solution；It is 1.5ng/ μ L with Buffer dilution kinases.Untested compound is made into 6 concentration
The solution of gradient, in sequentially adding on 384 orifice plates, 2 μ L ATP-Poly (4:1Glu, Tyr) Peptide solution, 1 μ L sample are molten
Liquid, the starting reaction of 2 μ L enzyme solutions.After 30 DEG C of incubation 60min, 5 μ L of ADP-Glo reagent is added and terminates reaction.Add Kinase
ADP is converted ATP by 10 μ L of Detection reagent, in 25 DEG C of incubation 30min, uses PerkinElmer multi-function microplate reader
Chemiluminescence module measures luminous value, calculates inhibiting rate.
Numerical value processing: inhibiting rate=(positive value-administration class value)/(positive value-feminine gender value) × 100%；
The experimental result of compound is shown in Table 1:
1 protein degradation of table targets chimera to the inhibitory activity result of VEGFR-2 kinases.(60nM)
As it can be seen from table 1 the protein degradation targeting prepared by the present invention based on VEGFR-2 inhibitor ABT-869 is chimeric
Body has preferable inhibitory activity.
Embodiment 5
Protein degradation targets the determination of activity of chimera cellular level.
The Activity determination that protein degradation targets chimera cellular level uses MTT detection method.Increased logarithmic phase will be in
Single cell suspension is made with 0.25% trypsin digestion in EA.hy926 cell or SMMC-7721 cell, is inoculated in 96 orifice plates
(2×104A/hole), every 180 μ L of hole.37 DEG C are put into, 5%CO2It is cultivated in constant incubator, for 24 hours the dosing after cell is adherent afterwards.
20 hole μ L/ serum free mediums are added in 3 multiple holes of every group of setting, negative control group, and 20 μ of drug of various concentration is added in experimental group
The hole L/ (dilutes drug with serum free medium), is put into 37 DEG C, 5%CO2Continue to cultivate in constant incubator.Drug effect 72h
Afterwards, careful inhale abandons supernatant, and the 200 μ L/ of MTT solution (final concentration of 0.5mg/mL) that serum free medium dilutes 10 times is added
Hole, after 37 DEG C of incubation 4-6h, careful inhale abandons supernatant, and 150 hole μ L/ DMSO, shake well 15min on decolorization swinging table is added.With
Enzyme-linked immunosorbent assay instrument measures each hole absorbance (OD) value under 490nm wavelength.
Numerical value processing: inhibiting rate=(ODNegative group-ODAdministration group)/(ODNegative group-ODBlank group) × 100%；
The experimental result of part of compounds is shown in Table 2:
Inhibitory activity (100nM, 72h) of 2 preferred compound of table to EA.hy926 cell and SMMC-7721 cell
From table 2 it can be seen that chimera prepared by the present invention has preferably EA.hy926 cell and SMMC-7721 cell
Inhibitory activity.
Claims (9)
1. a kind of protein degradation based on VEGFR-2 inhibitor ABT-869 targets chimera, which is characterized in that structural formula is as follows:
Wherein, integer of the n between 1-20.
2. a kind of protein degradation based on VEGFR-2 inhibitor ABT-869 according to claim 1 targets chimera,
It is characterized in that, n 3,8 or 12.
3. a kind of preparation method of the protein degradation targeting chimera based on VEGFR-2 inhibitor ABT-869, which is characterized in that
The following steps are included:
1) alkyl dicarboxylic aid and 1- (4- (3- amino-1 h-indazole -4- base) phenyl) -3- (the fluoro- 5- aminomethyl phenyl of 2-) urea exist
Under the condensation of PyBop, the intermediate product with monocarboxylic acid is obtained；
2) intermediate product and (2S, 4R) -1- ((S) -2- amino -3,3- dimethylbutanoyl) -4- hydroxy-n-with monocarboxylic acid
(4- (4- methylthiazol -5- base) benzyl) pyrrolidines -2- formamide obtains pressing down based on VEGFR-2 under the condensation of HATU
The protein degradation of preparation ABT-869 targets chimera, and structural formula is as follows:
Wherein, integer of the n between 1-20.
4. a kind of system of protein degradation targeting chimera based on VEGFR-2 inhibitor ABT-869 according to claim 3
Preparation Method, which is characterized in that the detailed process of the step 1) are as follows: alkyl dicarboxylic aid and PyBop are dissolved in methylene chloride, drip
Add triethylamine, after mixing evenly, 1- (4- (3- amino-1 h-indazole -4- base) phenyl) -3- (the fluoro- 5- aminomethyl phenyl of 2-) is added
Urea is handled after 12h is stirred at room temperature, and obtains the intermediate product with monocarboxylic acid.
5. a kind of system of protein degradation targeting chimera based on VEGFR-2 inhibitor ABT-869 according to claim 4
Preparation Method, which is characterized in that pimelic acid 3.99mmol and PyBop 3.19mmol are dissolved in 20mL methylene chloride, three second are added dropwise
1- (4- (3- amino-1 h-indazole -4- base) phenyl) -3- (the fluoro- 5- aminomethyl phenyl of 2-) is added after mixing evenly in amine 4.79mmol
Urea 0.799mmol is handled after 12h is stirred at room temperature, and obtains the intermediate product with monocarboxylic acid.
6. a kind of system of protein degradation targeting chimera based on VEGFR-2 inhibitor ABT-869 according to claim 3
Preparation Method, which is characterized in that the detailed process of the step 2) are as follows: the intermediate product with monocarboxylic acid for obtaining step 1) is molten
In methylene chloride, (2S, 4R) -1- ((S) -2- amino -3,3- dimethylbutanoyl) -4- hydroxy-n-(4- (4- methyl is added
Thiazole -5- base) benzyl) pyrrolidines -2- formamide, it stirs evenly under ice bath, DIPEA is then added dropwise, stirs evenly, be added
HATU stirs 12h at 25 DEG C, is handled after reaction, obtain the protein degradation based on VEGFR-2 inhibitor ABT-869
Target chimera.
7. a kind of system of protein degradation targeting chimera based on VEGFR-2 inhibitor ABT-869 according to claim 6
Preparation Method, which is characterized in that by 7- (3- amino -4- (4- (3- (the fluoro- 5- aminomethyl phenyl of 2-) urea groups) phenyl) -1H- indazole -1-
Base) -7- oxo-heptanoic acid 0.193mmol is dissolved in 20mL methylene chloride, (2S, 4R) -1- ((S) -2- amino -3,3- diformazan is added
Base bytyry) -4- hydroxy-n-(4- (4- methylthiazol -5- base) benzyl) pyrrolidines -2- formamide 0.193mmol, is stirred under ice bath
It mixes uniformly, DIPEA0.773mmol is then added dropwise, stirs evenly, HATU 0.29mmol is added, after stirring 12h at 25 DEG C
It is handled, obtains the protein degradation targeting chimera based on VEGFR-2 inhibitor ABT-869.
8. a kind of egg based on VEGFR-2 inhibitor ABT-869 of the method preparation as described in any one of claim 3-7
Application of the white degradation targeting chimera in the drug that preparation treats or prevents cancer.
9. application according to claim 8, which is characterized in that the protein degradation target based on VEGFR-2 inhibitor ABT-869
The application in the anti-tumor drug using VEGFR-2 kinases as target spot is being prepared to chimera.
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