CN109485695A - A kind of protein degradation targeting chimera and preparation method and application based on VEGFR-2 inhibitor S7 - Google Patents
A kind of protein degradation targeting chimera and preparation method and application based on VEGFR-2 inhibitor S7 Download PDFInfo
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Abstract
A kind of protein degradation targeting chimera and preparation method and application based on VEGFR-2 inhibitor S7, alkyl dicarboxylic aid and 1- (3- (3- amino-1 h-indazole -4- base) phenyl) -3- (3- isopropyl phenyl) urea obtain the intermediate product with monocarboxylic acid under the condensation of PyBop；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 under the condensation of HATU, obtain based on VEGFR-2 inhibitor S7 protein degradation targeting chimera.It is simple that protein degradation of the invention targets chimeric preparation, it is easy to accomplish, and yield is higher, can be used in preparation and treats or prevents in the drug of cancer, particularly for preparing using VEGFR-2 kinases as in the anti-tumor drug of target spot.
Description
Technical field
The protein degradation based on VEGFR-2 inhibitor S7 that the present invention relates to a kind of targets chimera and preparation method and answers
With.
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 S7 targets chimera and preparation
Methods and applications, protein degradation targeting chimera have the function of induction VEGFR-2 protein degradation, can be used for preparing new antitumoral
Drug.
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 S7, 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 S7, comprising the following steps:
1) alkyl dicarboxylic aid and 1- (3- (3- amino-1 h-indazole -4- base) phenyl) -3- (3- isopropyl phenyl) 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 S7 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, is stirring evenly and then adding into 1- (3- (3- amino-1 h-indazole -4- base) phenyl) -3- (3- isopropyl
Base 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 alkyl dicarboxylic aid 0.778mmol and PyBop 0.648mmol are dissolved in 20mL
In methylene chloride, triethylamine 1.55mmol is added dropwise, after mixing evenly, is added 1- (3- (3- amino-1 h-indazole -4- base) phenyl) -
3- (3- isopropyl phenyl) urea 0.259mmol, is handled after 12h is stirred at room temperature, and 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 and is handled after stirring 12h at 25 DEG C, obtains the egg based on VEGFR-2 inhibitor S7
White degradation targets chimera.
A further improvement of the present invention lies in that the intermediate product 0.284mmol with monocarboxylic acid is dissolved in 20mL dichloromethane
In alkane, (2S, 4R) -1- ((S) -2- amino -3,3- dimethylbutanoyl) -4- hydroxy-n-(4- (4- methylthiazol -5- is added
Base) benzyl) pyrrolidines -2- formamide 0.284mmol, it stirs evenly under ice bath, DIPEA 1.137mmol is then added dropwise,
5min is stirred, HATU 0.426mmol is added, stirs 12h at 25 DEG C, obtains the protein degradation target based on VEGFR-2 inhibitor S7
To chimera.
A kind of protein degradation targeting chimera based on VEGFR-2 inhibitor S7 treats or prevents the drug of cancer in preparation
In application.
A further improvement of the present invention lies in that prepared by the protein degradation targeting chimera based on VEGFR-2 inhibitor S7
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
Von Rippel-Lindau (VHL) protein ligands connect in body, obtain protein degradation and target chimera.Protein degradation targeting
Chimera (PROTACs) is capable of the degradation of selective induction VEGFR-2 albumen.Protein degradation targeting chimera preparation of the invention
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.Small molecular protein degradation targeting chimera of the invention can be used in preparing
In the drug for treating or preventing cancer, particularly for preparing using VEGFR-2 kinases as in the anti-tumor drug of target spot.
Detailed description of the invention
Fig. 1 is the synthetic route chart that the protein degradation provided by the invention based on VEGFR-2 inhibitor S7 targets chimera；
Wherein, compound 1 is 1- (3- (3- amino-1 h-indazole -4- base) phenyl) -3- (3- isopropyl phenyl) urea, chemical combination
Object 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- dimethylbutanoyls) -4- hydroxy-n-(4- (4- methylthiazol -5- base) benzyl) pyrrolidines -2- formamide, compound (X)
It degrades for small molecular protein with anti-tumor activity and targets 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.
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.New albumen of the invention
Degrading targeted chimeric molecules can be in the application in treating cancer.Protein degradation of the present invention targets chimera
(PROTACs) it is capable of the degradation of selective induction VEGFR-2 albumen.
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.
Small molecular protein degradation targeting chimera with anti-tumor activity of the present invention, comprising:
(2S, 4R) -1- ((S) -2- (7- (3- amino -4- (3- (3- (3- isopropyl phenyl) urea groups) phenyl) -1H- indazole -
1- yl) -7- oxo heptanamido) -3,3- dimethylbutanoyl) -4- hydroxy-n-(4- (4- methylthiazol -5- base) benzyl) pyrrole
Cough up alkane -2- formamide；
(2S, 4R) -1- ((S) -2- (12- (3- amino -4- (3- (3- (3- isopropyl phenyl) urea groups) phenyl) -1H- Yin
Azoles -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- (3- (3- (3- isopropyl phenyl) urea groups) phenyl) -1H- Yin
Azoles -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 S7, including with
Lower step:
1) alkyl dicarboxylic aid and biphenyl ureas VEGFR-2 protein ligands 1- (3- (3- amino-1 h-indazole -4- base) phenyl) -
3- (3- isopropyl phenyl) 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
Under condensation, the compound that logical formula (X) indicates is obtained；
The concrete operations of the step 1) are as follows: alkyl dicarboxylic aid and PyBop are dissolved in methylene chloride, three second are slowly added dropwise
Amine, after stirring 3min, sampling；VEGFR-2 protein ligands 1- (3- (3- amino-1 h-indazole -4- base) phenyl) is added, and (3- is different by -3-
Propyl phenyl) urea, 12h is stirred at room temperature, after reaction, low pressure rotation removes organic solvent, and suitable quantity of water is added, is extracted with ethyl acetate
Take, the organic phase of extraction is washed, it is dry after decompression boil off solvent, obtain crude product, with chromatography post separation crude product, obtain with single carboxylic
The intermediate product of 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 egg based on VEGFR-2 inhibitor S7
White degradation targeting chimera leads to the compound that formula (X) indicates.
The above-mentioned protein degradation targeting chimera based on VEGFR-2 inhibitor S7 is in preparation using VEGFR-2 kinases as target spot
Anti-tumor drug in application.
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 biphenyl ureas VEGFR-2 protein ligands 1- (3- (3- amino-1 h-indazole -4- base)
Phenyl) -3- (3- isopropyl phenyl) urea (compound 1) under the condensation of PyBop condensing agent, obtains 7- (3- amino -4-
(3- (3- (3- isopropyl phenyl) urea groups) phenyl) -1H- indazole -1- base) -7- oxo-heptanoic acid (compound 3)；Detailed process is such as
Under:
By pimelic acid (0.12g, 0.778mmol), PyBop (0.33g, 0.648mmol) is dissolved in 20mL methylene chloride, is delayed
It is slow that triethylamine (215 μ L, 1.55mmol) is added dropwise, after stirring 3min, sampling.1- (3- (3- amino-1 h-indazole -4- base) benzene is added
Base) -3- (3- isopropyl phenyl) urea (0.1g, 0.259mmol), it is stirred overnight at room temperature, then low pressure rotation removes organic solvent, is added
Suitable quantity of water is extracted with ethyl acetate, and anhydrous sodium sulfate is dry, and decompression rotation removes organic solvent, and residue passes through silica gel column chromatography layer
Analysis purifying affords white solid, weight 0.11g, yield 80.36% using petrol ether/ethyl acetate (V/V=5/1-3/1).
LCMS(ESI,m/z):528.30[M-H]-
2) 7- (3- amino -4- (3- (3- (3- isopropyl phenyl) urea groups) phenyl) -1H- indazole -1- base) -7- oxo-heptanoic acid
(compound 3) and E3 ubiquitin ligase ligand (2S, 4R) -1- ((S) -2- amino -3,3- dimethylbutanoyl) -4- hydroxy-n -
(4- (4- methylthiazol -5- base) benzyl) pyrrolidines -2- formamide (compound 4) obtains under the condensation of HATU condensing agent
(2S, 4R) -1- ((S) -2- (7- (3- amino -4- (3- (3- (3- isopropyl phenyl) urea groups) phenyl) -1H- indazole -1- base) -7-
Oxo heptanamido) -3,3- dimethylbutanoyl) -4- hydroxy-n-(4- (4- methylthiazol -5- base) benzyl) pyrrolidines -2- first
Amide (compound X)；Detailed process is as follows:
By 7- (3- amino -4- (3- (3- (3- isopropyl phenyl) urea groups) phenyl) -1H- indazole -1- base) -7- oxo-heptanoic acid
(0.15g, 0.284mmol) is dissolved in 20mL methylene chloride, and (2S, 4R) -1- ((S) -2- amino -3,3- dimethyl butyryl is added
Base) -4- hydroxy-n-(4- (4- methylthiazol -5- base) benzyl) pyrrolidines -2- formamide (0.13g, 0.284mmol), under ice bath
5min is stirred, is then added dropwise DIPEA (193 μ L, 1.137mmol), stirring 5min, addition HATU (0.16g,
0.426mmol), 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
Sodium sulphate 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 0.12g, yield 44.9% 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)δ8.98(s,1H),δ8.86(s,1H),δ
8.71(s,1H),δ8.55-8.58(t,1H),δ8.34-8.36(d,1H),δ7.87-7.89(d,1H),δ7.68(s,1H),δ
7.60-7.64(t,1H),δ7.35-7.47(m,6H),δ7.17-7.26(m,3H),δ7.07-7.09(d,1H),δ6.85-6.88
(d,2H),δ5.29(s,2H),δ5.13-5.14(d,1H),δ4.53-4.59(t,1H),δ4.41-4.45(t,2H),δ4.35
(s,1H),δ4.19-4.25(q,1H),δ4.01-4.06(q,1H),δ2.96-3.00(t,2H),δ2.79-2.88(m,1H),δ
2.45(s,3H),δ2.12-2.19(t,3H),δ1.36(s,8H),1.18-1.20(d,6H),δ0.93(s,9H).
LCMS(ESI,m/z):940.55[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)δ8.98(s,1H),δ8.86(s,1H),δ
8.71(s,1H),δ8.55-8.58(t,1H),δ8.34-8.36(d,1H),δ7.83-7.86(d,1H),δ7.68(s,1H),δ
7.60-7.64(t,1H),δ7.36-7.47(m,7H),δ7.19-7.25(m,3H),δ7.07-7.09(d,1H),δ6.86-6.88
(d,1H),δ5.28(s,2H),δ5.13-5.13(d,1H),δ4.54-4.56(d,1H),δ4.41-4.47(q,2H),δ4.36
(s,1H),δ4.20-4.25(q,1H),δ4.01-4.06(q,1H),δ2.97-3.00(t,2H),δ2.81-2.88(m,1H),δ
2.45(s,3H),δ2.00-2.14(m,3H),δ1.68-1.72(m,2H),δ1.19-1.50(m,22H),δ0.94(s,9H).
LCMS(ESI,m/z):1010.45[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)δ8.98(s,1H),δ8.86(s,1H),δ
8.70(s,1H),δ8.55-8.58(t,1H),δ8.34-8.36(d,1H),δ7.83-7.86(d,1H),δ7.68(s,1H),δ
7.60-7.64(t,1H),δ7.35-7.47(m,7H),δ7.19-7.26(m,3H),δ7.08-7.09(d,1H),δ6.85-6.87
(d,1H),δ5.27(s,2H),δ5.13-5.14(d,1H),δ4.53-4.56(d,1H),δ4.41-4.47(q,2H),δ4.35
(s,1H),δ4.18-4.24(m,1H),δ4.01-4.06(q,1H),δ2.96-3.00(t,2H),δ2.79-2.87(m,1H),δ
2.45(s,3H),δ1.99-2.13(m,3H),δ1.68-1.71(m,2H),δ1.23-1.49(m,24H),δ1.18-1.20(d,
6H),δ0.94(s,9H).
LCMS(ESI,m/z):1066.60[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.
With Buffer (Tris 80mM, MgCl220mM, BSA 0.2mg/mL, DTT 2mM) dilution ATP (10mM) be 250
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 protein degradation targeting chimera prepared by the present invention has preferable suppression to VEGFR-2 kinases
System 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, continues to cultivate in 5%CO2 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 S7 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 S7 according to claim 1 targets chimera, feature exists
In n 3,8 or 12.
3. it is a kind of based on VEGFR-2 inhibitor S7 protein degradation targeting chimera preparation method, which is characterized in that including with
Lower step:
1) alkyl dicarboxylic aid and 1- (3- (3- amino-1 h-indazole -4- base) phenyl) -3- (3- isopropyl phenyl) urea are PyBop's
Under condensation, 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 S7 targets chimera, and structural formula is as follows:
Wherein, integer of the n between 1-20.
4. a kind of preparation side of protein degradation targeting chimera based on VEGFR-2 inhibitor S7 according to claim 3
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, are added dropwise three
Ethamine, is stirring evenly and then adding into 1- (3- (3- amino-1 h-indazole -4- base) phenyl) -3- (3- isopropyl phenyl) urea, and room temperature is stirred
It is handled after mixing 12h, obtains the intermediate product with monocarboxylic acid.
5. a kind of preparation side of protein degradation targeting chimera based on VEGFR-2 inhibitor S7 according to claim 4
Method, which is characterized in that alkyl dicarboxylic aid 0.778mmol and PyBop 0.648mmol are dissolved in 20mL methylene chloride, are added dropwise three
1- (3- (3- amino-1 h-indazole -4- base) phenyl) -3- (3- isopropyl phenyl) is added after mixing evenly in ethamine 1.55mmol
Urea 0.259mmol is handled after 12h is stirred at room temperature, and obtains the intermediate product with monocarboxylic acid.
6. a kind of preparation side of protein degradation targeting chimera based on VEGFR-2 inhibitor S7 according to claim 3
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 dissolved in two
In chloromethanes, (2S, 4R) -1- ((S) -2- amino -3,3- dimethylbutanoyl) -4- hydroxy-n-(4- (4- methylthiazol-is added
5- yl) benzyl) pyrrolidines -2- formamide, it stirs evenly under ice bath, DIPEA is then added dropwise, stirs evenly, HATU is added,
It after stirring 12h at 25 DEG C, is handled, obtains the protein degradation targeting chimera based on VEGFR-2 inhibitor S7.
7. a kind of preparation side of protein degradation targeting chimera based on VEGFR-2 inhibitor S7 according to claim 6
Method, which is characterized in that the intermediate product 0.284mmol with monocarboxylic acid is dissolved in 20mL methylene chloride, is added (2S, 4R)-
1- ((S) -2- amino -3,3- dimethylbutanoyl) -4- hydroxy-n-(4- (4- methylthiazol -5- base) benzyl) pyrrolidines -2- first
Amide 0.284mmol is stirred evenly under ice bath, and DIPEA 1.137mmol is then added dropwise, and stirs 5min, and HATU is added
0.426mmol stirs 12h at 25 DEG C, obtains the protein degradation targeting chimera based on VEGFR-2 inhibitor S7.
8. a kind of protein degradation based on VEGFR-2 inhibitor S7 such as any one of claim 3-7 method preparation targets
Application of the chimera in the drug that preparation treats or prevents cancer.
9. application according to claim 8, which is characterized in that the protein degradation targeting based on VEGFR-2 inhibitor S7 is embedding
Zoarium is preparing the application in the anti-tumor drug using VEGFR-2 kinases as target spot.
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| CN104725319A (en) * | 2015-03-11 | 2015-06-24 | 西安交通大学 | 1H-indazole-3-aminobphenyl urea compound with anti-tumor activity as well as preparation method and application of 1H-indazole-3-aminobphenyl urea compound |
| CN108366992A (en) * | 2015-11-02 | 2018-08-03 | 耶鲁大学 | Proteolysis targets chimera compound and its methods for making and using same |
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