CN110204532A - A kind of compound and its preparation method and application of targeting EGFR protein degradation - Google Patents

A kind of compound and its preparation method and application of targeting EGFR protein degradation Download PDF

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CN110204532A
CN110204532A CN201910403317.2A CN201910403317A CN110204532A CN 110204532 A CN110204532 A CN 110204532A CN 201910403317 A CN201910403317 A CN 201910403317A CN 110204532 A CN110204532 A CN 110204532A
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preparation
formula
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cancer
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CN110204532B (en
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张兴贤
何凯伦
王文冰
王孝举
郑晓亮
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Zhejiang University of Technology ZJUT
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    • A61K31/00—Medicinal preparations containing organic active ingredients
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    • A61K31/395—Heterocyclic compounds having nitrogen as a ring hetero atom, e.g. guanethidine or rifamycins
    • A61K31/495—Heterocyclic compounds having nitrogen as a ring hetero atom, e.g. guanethidine or rifamycins having six-membered rings with two or more nitrogen atoms as the only ring heteroatoms, e.g. piperazine or tetrazines
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    • A61K31/506—Pyrimidines; Hydrogenated pyrimidines, e.g. trimethoprim not condensed and containing further heterocyclic rings
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    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
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    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61P—SPECIFIC THERAPEUTIC ACTIVITY OF CHEMICAL COMPOUNDS OR MEDICINAL PREPARATIONS
    • A61P35/00—Antineoplastic agents
    • A61P35/02—Antineoplastic agents specific for leukemia
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    • C07—ORGANIC CHEMISTRY
    • C07D—HETEROCYCLIC COMPOUNDS
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    • C07D401/14—Heterocyclic compounds containing two or more hetero rings, having nitrogen atoms as the only ring hetero atoms, at least one ring being a six-membered ring with only one nitrogen atom containing three or more hetero rings

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Abstract

The invention discloses a kind of compound or its pharmaceutically acceptable salt with the protein degradation of structure targeting EGFR shown in following formula, in formula, R C1~C6Alkyl, the integer that m is 1~7, the integer that n is 1~6.The invention also discloses the preparation method of above compound and preparation made of the pharmaceutical composition including the compound or its pharmaceutically acceptable salt and pharmaceutically acceptable carrier and the medical compounds.Such compound shows excellent EGFR degradation and has good anti-tumor activity, and therefore, the invention also discloses application of the compound in preparation prevention or/and treating cancer, has huge application prospect in medical field.

Description

A kind of compound and its preparation method and application of targeting EGFR protein degradation
Technical field
The invention belongs to compound synthesis technical fields, a kind of compound more particularly to targeting EGFR protein degradation and Preparation method and application.
Background technique
Proteolysis targeted chimeric molecules (Proteolysis Targeting Chimeras, PROTACs) are to utilize one The difunctional small molecule of kind furthers target protein and intracellular E3, so as to cause the degradation of target protein.PROTACs includes Three parts functional structure: (1) part that can be combined with protein substrate；(2) part that can be combined with E3；(3) two before Partial connection chain.Intracellular PROTACs can be simultaneously in conjunction with target protein and E3, and making originally cannot be with the target protein in conjunction with E3 Ubiquitination, and then identified by proteasome and degrade (Angew.Chem.Int.Ed.Engl, 2016,55 (6), 1996-1973).
Research has shown that following formula: compound A analog can be CUL4-RBX1- with CRBN protein binding, CRBN albumen DDB1-Cereblon(CRL4CRBN) E3 ubiquitin ligase complex substrate receptor albumen, pass through uiquitin-protease in conjunction with after Body approach carries out proteolysis (Nature, 2014,512 (7512): 49-53)；
The proliferation of nowadays, the high expression in many entity tumors there are EGFR or unconventionality expression, EGFR and tumour cell, Angiogenesis, tumor invasion, transfer and the inhibition of Apoptosis are related.The lifting in the evolution of malignant tumour of the overexpression of EGFR It acts on, there is the overexpression of EGFR in the tissue such as spongiocyte, kidney, lung cancer, prostate cancer, cancer of pancreas, breast cancer.EGFR Effective antitumour target spot is had become, multiple EGFR inhibitors have listed the treatment for kinds cancer.But it is presently used EGFR inhibitor inhibitory effect it is limited, and therefore existing certain resistance problems develop a kind of degradable EGFR albumen Compound with extremely important pharmacy be worth.
Summary of the invention
The object of the present invention is to provide a kind of compound and its preparation method and application of targeting EGFR protein degradation, The compound not only has the excellent effect of EGFR protein degradation and anti-tumor activity, small to human body toxic side effect, can be used for making Standby anti-tumor drug.
The object of the invention is also to provide the preparation methods of above compound.
Another object of the present invention is to provide above compounds or its pharmaceutically acceptable salt and hydrate to make Application in the drug of standby prevention or/and treating cancer.
In order to achieve the above-mentioned object of the invention, the present invention provides with targeting EGFR protein degradation shown in formula I or formula II Compound or its pharmaceutically acceptable salt:
In formula I and formula II, R C1~C6Alkyl, the integer that m is 1~7, the integer that n is 1~6.
Compound of the present invention is a kind of proteolysis targeted chimeric molecules (PROTACs) comprising Ke Yiyu Part that EGFR protein substrate combines, can in conjunction with E3 part and connection chain, can using this difunctional small molecule Target protein and intracellular E3 to further, so that making originally cannot be with the EGFR target protein ubiquitination in conjunction with E3, and then quilt Proteasome is identified and is degraded.
Preferably, R is methyl, and the integer that m 1, n are 2~5, preferred degradation EGFR albumen effect is more preferable, Anti-tumor activity is also stronger.
The invention also discloses the preparation method of the compound of above-mentioned targeting EGFR protein degradation, the formula I and formula II are changed The preparation process for closing object is as follows:
The preparation process of the type I compound is as follows:
It specifically includes: formula III compound is dissolved in organic solvent, formula IV compound and diisopropyl are added at room temperature Ethamine is heated to 80~90 DEG C of 10~14h of reaction and obtains Formulas I compound represented after extraction, washing, drying.
The preparation process of the Formula II compound is as follows:
It specifically includes: formula III compound is dissolved in organic solvent, Formula V compound and diisopropyl are added at room temperature Ethamine is heated to 80~90 DEG C of 10~14h of reaction and obtains Formula II compound represented after extraction, washing, drying.
The invention also discloses a kind of pharmaceutical compositions, including above compound or its pharmaceutically acceptable salt, with medicine Acceptable excipient on.
The pharmaceutically acceptable excipient refers to any diluent that can be used for drug field, adjuvant and/or load Body.
Pharmaceutical composition of the invention can be made into various preparations, comprising: be added in pharmaceutical composition pharmaceutically acceptable Auxiliary material clinically acceptable dosage form is made.The clinically acceptable dosage form is oral preparation, ejection preparation or part Preparation.
Wherein, oral preparation can be tablet, capsule, solution or suspension；Ejection preparation can be injection solution or suspension The dried powder of agent or injectable, water for injection is added before the injection to be used immediately；Topical formulations can be ointment or solution.
Wherein, pharmaceutically acceptable auxiliary material include: the adhesive of oral preparation, lubricant, disintegrating agent, cosolvent, Diluent, stabilizer, suspending agent, pigment or corrigent；Preservative, solubilizer or the stabilizer of ejection preparation；Topical formulations Matrix, diluent, lubricant, preservative etc..Pharmaceutical preparation can by oral administration or parenteral (as it is intravenous, subcutaneous, In peritonaeum or part) administration, if some drugs be under the conditions of stomach it is unstable, enteric coated tablets can be configured to.
The compounds of this invention or its pharmaceutically acceptable salt can be used for preparing treatment and EGFR proteinase activity exception table Up to the application of the drug of relevant disease, screened by external activity, it has been found that the compounds of this invention or its can pharmaceutically connect The salt received has anti-tumor activity, therefore the invention also discloses above compounds or its pharmaceutically acceptable salt to prepare in advance Application in the anti-or/and drug for the treatment of cancer.
The cancer is breast cancer, colon cancer, prostate cancer, cancer of pancreas, thyroid papillary carcinoma, oophoroma, black Plain tumor, leukaemia or non-small cell lung cancer.Preferably non-small cell carcinoma, this is because experiment shows the anti-non-small cell of the compound Cancer activity is high.
Compared with the prior art, the present invention has the following beneficial effects:
The present invention selects Compound A structure analog as the position being combined in PROTACs with E3 ligase, selects It is connected by suitable connection chain with the structure with EGFR protease inhibitory activity constructs PROTACs.Extracorporeal anti-tumor Active testing and external EGFR protein degradation activity show that the compound shows good anti-tumor activity, and show Excellent EGFR protein degradation effect out, can be used for preventing or/and treating kinds cancer, have huge application in field of medicaments Prospect.
Detailed description of the invention
Fig. 1 is the nuclear magnetic spectrogram for the compound P2 that embodiment 5 obtains；
Fig. 2 is the nuclear magnetic spectrogram for the compound P5 that embodiment 8 obtains.
Specific embodiment
Examples provided hereinafter and preparation example further elucidate and illustrate this compound and preparation method thereof, answer Work as understanding, the range of following embodiments and preparation example does not limit the scope of the invention in any way.
Abbreviation meaning involved in the present invention are as follows: Boc is tertbutyloxycarbonyl, and DMF is dimethylformamide, and DIPEA is Diisopropylethylamine, TBAB are tetrabutylammonium bromide, and EA is ethyl acetate, and Et is ethyl, and Me is methyl, and i.e. mole, PE is stone to M Oily ether, pH are phenyl, and THF is tetrahydrofuran, and TFA is trifluoroacetic acid, and TLC is thin-layer chromatography, and NSCLC is non-small cell lung cancer, WT is wild type.
(1) based on-iodine substitution " design the synthesis for synthesizing E3 small molecule (compound 7):
(2) EGFR targets the synthesis of small molecule (compound III):
(3) synthetic method of PROTACs compound (Formulas I and Formula II)
In targeting proteins embeding technique, the selection of linking group is had very important effect, presently mainly its The screening of length and type.The present invention screens the length of linking group, and having synthesized following 9 has different length connection PROTACs small molecule compound, specific structure is as shown in table 1 below:
Table 1
The synthesis of embodiment 1:E3 small molecule (compound 7)
(1) preparation of compound 2
The preparation of compound 2a:
Diethylene glycol (DEG) (7.0g, 66.0mmol) is dissolved in 10mL dichloromethane solution, Et is added3N(13.3g, 132.2mmol), the dichloromethane solution of TsCl (15.1g, 79.1mmol) is added dropwise under ice bath.Room temperature is transferred to after being added dropwise Under be stirred overnight.After 6mol/L HCl adjusting pH value of solution to 7,10mL water is added, is extracted 3 times with 30mL methylene chloride, merges Organic phase, anhydrous sodium sulfate are dry.Column chromatographs (EA) and obtains 10g yellow liquid product, yield 60%.
1H NMR (500MHz, Chloroform-d) δ 7.75 (d, J=8.4Hz, 2H), 7.31 (d, J=8.0Hz, 2H), 4.19-4.09 (m, 2H), 3.62 (ddd, J=12.1,5.4,4.1Hz, 4H), 3.48 (dd, J=5.3,3.8Hz, 2H), 2.40 (s,3H).
The preparation of compound 2b:
Compound triethylene-glycol (5.0g, 33.3mmol) is dissolved in 10mL dichloromethane solution, Et is added3N (6.7g, 66.6mmol) dichloromethane solution of TsCl (8.9g, 46.6mmol) is added dropwise under ice bath.It is transferred to after being added dropwise It is stirred to react at room temperature 12 hours.10mL water is added after 6MHCl adjusting solution PH to 7, is extracted 3 times with 30mL methylene chloride, Merge organic phase, anhydrous sodium sulfate is dry.Column chromatographs (EA) and obtains 7.5g yellow liquid product, yield 74%.
1H NMR (500MHz, Chloroform-d) δ 7.79 (d, J=8.4Hz, 2H), 7.35 (d, J=7.7Hz, 2H), 4.24–4.07(m,2H),3.72–3.66(m,4H),3.65–3.51(m,6H),2.44(s,3H).
The preparation of compound 2c:
Compound tetraethylene-glycol (5.0g, 25.8mmol) is dissolved in 20mL methylene chloride, triethylamine is added dropwise (3.9g, 38.7mmol), while TsCl is configured to dichloromethane solution, it is slowly added dropwise in solution with dropping funel, at room temperature Stir 15h.Thin-layer chromatography (EA) monitoring reaction, compound TsCl some residual.Solution PH=7 are adjusted with 4M HCl, are added 10mL water is extracted 3 times with 30mL methylene chloride, merges organic phase, and anhydrous sodium sulfate is dry.Column chromatography (EA) obtains product 6.8g yield 76%.
1H NMR (500MHz, Chloroform-d) δ 7.74 (d, J=8.4Hz, 2H), 7.48-7.16 (m, 2H), 4.31- 3.96(m,2H),3.70–3.61(m,4H),3.60–3.58(m,4H),3.56–3.51(m,6H),2.39(s,3H).
The preparation of compound 2d:
Four contracting five ethylene glycol (6.0g, 25.2mmol) of compound is dissolved in 20mL methylene chloride, triethylamine is added dropwise (4.9g, 48,4mmol), while TsCl is configured to dichloromethane solution, it is slowly added dropwise in solution with dropping funel, at room temperature Stir 15h.Thin-layer chromatography (EA) monitoring reaction, compound TsCl some residual.Solution PH=7 are adjusted with 4M HCl, are added 10mL water is extracted 3 times with 30mL methylene chloride, merges organic phase, and anhydrous sodium sulfate is dry.Column chromatography (EA) obtains product 6.52g, yield 66%.
1H NMR (500MHz, Chloroform-d) δ 7.79 (d, J=8.3Hz, 2H), 7.33 (d, J=8.0Hz, 2H), 4.18-4.11(m,2H),3.75-3.65(m,13H),3.61-3.56(m,7H),2.44(s,3H).
(2) preparation of compound 3
The preparation of compound 3a:
Compound 2a (2.954g, 11.3mmol) is dissolved in 10mL toluene solution, at room temperature be added TBAB (0.181g, 0.56mmol), KOH (1.01g, 18.1mmol), bromo-acetic acid tert-butyl (2.7g, 13.6mmol).It is stirred to react at room temperature overnight. (PE:EA=1:2) is detected through thin-layer chromatography, feedstock portions are remaining.It is concentrated under reduced pressure and removes toluene, 5mL water is added, with 15mL dichloro Methane extracts 3 times, merges organic phase, and anhydrous sodium sulfate is dry.Column chromatographs (EA) and obtains 3.4g product, yield 83.9%.
1H NMR (500MHz, Chloroform-d) δ 7.80 (d, J=8.4Hz, 2H), 7.37-7.31 (m, 2H), 4.19- 4.14(m,2H),3.98(s,1H),3.72–3.67(m,2H),3.67–3.59(m,4H),2.44(s,3H),1.47(s,9H).
The preparation of compound 3b:
Compound 2b (3.73g, 12.3mmol) is dissolved in 10mL toluene solution, at room temperature be added TBAB (0.198g, 0.6mmol), KOH (1.1g, 20.0mmol), bromo-acetic acid tert-butyl (2.9g, 14.7mmol).It is stirred to react at room temperature 12 hours. (PE:EA=1:2) is detected through thin-layer chromatography, feedstock portions are remaining.It is concentrated under reduced pressure and removes toluene, 5mL water is added, with 15mL dichloro Methane extracts 3 times, merges organic phase, and anhydrous sodium sulfate is dry.Column chromatographs (EA) and obtains 4.2g product, white solid, yield 81.5%.
1H NMR (500MHz, CDCl3) δ 7.80 (d, J=8.4Hz, 2H), 7.58-7.07 (m, 3H), 4.19-4.12 (m, 2H),4.01(s,2H),3.73-3.63(m,6H),3.60(s,4H),2.45(s,3H),1.47(s,9H).
The preparation of compound 3c:
Compound 2c (5.0g, 25.8mmol) is dissolved in 20mL methylene chloride, dropwise addition triethylamine (3.9g, 38.7mmol), while by TsCl it is configured to dichloromethane solution, is slowly added dropwise in solution with dropping funel, stirred at room temperature 15h.Thin-layer chromatography (EA) monitoring reaction, compound TsCl some residual.PH value of solution=7 are adjusted with 4N HCl, 10mL water is added, It is extracted 3 times with 30mL methylene chloride, merges organic phase, anhydrous sodium sulfate is dry.Column chromatographs (EA), and obtaining product is 6.8g, yield It is 74%.
1H NMR(500MHz,CDCl3): δ 7.73 (d, J=8.3Hz, 2H), 7.30-7.27 (m, 2H), 4.10-4.08 (m, 2H),3.96(s,2H),3.66-3.62(m,6H),3.61-3.60(m,4H),3.52-3.50(m,4H),2.39(s,3H), 1.41(s,9H).
The preparation of compound 3d:
Compound 2d (5.0g, 12.8mmol) is dissolved in 20mL methylene chloride, dropwise addition triethylamine (3.9g, 38.7mmol), while by TsCl it is configured to dichloromethane solution, is slowly added dropwise in solution with dropping funel, stirred at room temperature 15h.Thin-layer chromatography (EA) monitoring reaction, compound TsCl some residual.PH value of solution=7 are adjusted with 4N HCl, 10mL water is added, It is extracted 3 times with 30mL methylene chloride, merges organic phase, anhydrous sodium sulfate is dry.Column chromatographs (EA), and obtaining product is 4.5g, yield It is 70%.
1H NMR(500MHz,CDCl3): δ 7.80 (d, J=8.3Hz, 2H), 7.40-7.31 (m, 2H), 4.15 (d, J= 5.0Hz, 2H), 4.02 (s, 2H), 3.73-3.67 (m, 6H), 3.67-3.63 (m, 4H), 3.62 (q, J=1.9Hz, 4H), 3.58 (s,4H),2.45(s,3H),1.47(s,9H).
(3) preparation of compound 4
The preparation of compound 4a:
Compound 3a (2.271g, 6.1mmol) is dissolved in 5mL acetone, NaI (1.28g, 8.5mmol) is added at room temperature. Reflux at 70 DEG C is transferred to after being added dropwise to be stirred to react 6 hours.It is concentrated under reduced pressure.15mL water is added, is extracted with 20mL ethyl acetate It takes 3 times, anhydrous sodium sulfate is dry, merges organic phase.Obtain crude yellow liquid 1.41g, yield 70.3%.1H NMR(500MHz, Chloroform-d) δ 4.04 (s, 2H), 3.77 (d, J=7.1Hz, 2H), 3.75-3.68 (m, 4H), 3.28 (t, J=6.9Hz, 2H),1.48(s,9H).
The preparation of compound 4b:
Compound 3b (0.30g, 0.72mmol) is dissolved in 5mL acetone, NaI (0.15g, 1.0mmol) is added at room temperature. Reflux at 70 DEG C is transferred to after being added dropwise to be stirred to react 6 hours.It is concentrated under reduced pressure.5mL water is added, is extracted with 10mL ethyl acetate 3 times, anhydrous sodium sulfate is dry, merges organic phase.Obtain crude yellow liquid 221mg, thick yield 95%.1H NMR(500MHz, Chloroform-d) δ 4.03 (s, 2H), 3.76 (dd, J=7.3,6.5Hz, 2H), 3.72-3.69 (m, 4H), 3.67 (s, 4H), 3.26 (dd, J=7.3,6.5Hz, 2H), 1.47 (s, 9H)
The preparation of compound 4c
Compound 3c (1.0g, 2.2mmol) is placed in 5mL acetone, NaI (0.462g, 3.3mmol) is added at room temperature. It heats after being added dropwise and is stirred to react 6 hours at 80 DEG C.It is concentrated under reduced pressure, 5mL water is added, extracted 3 times with 10mL ethyl acetate, satisfied With brine It 3 times, anhydrous sodium sulfate is dry, merges organic phase.Obtain 0.871g product, yield 94.7%.
1H NMR (500MHz, Chloroform-d) δ 4.02 (s, 2H), 3.76 (dd, J=7.3,6.5Hz, 2H), 3.74- 3.68 (m, 4H), 3.67 (d, J=4.3Hz, 8H), 3.27 (dd, J=7.3,6.5Hz, 2H), 1.48 (s, 9H)
The preparation of compound 4d
Compound 3d (1.2g, 2.6mmol) is placed in 5mL acetone, NaI (0.562g, 3.7mmol) is added at room temperature. It heats after being added dropwise and is stirred to react 6 hours at 80 DEG C.It is concentrated under reduced pressure, 5mL water is added, extracted 3 times with 10mL ethyl acetate, satisfied With brine It 3 times, anhydrous sodium sulfate is dry, merges organic phase.Obtain 0.971g product, yield 89.2%.
1H NMR(500MHz,Chloroform-d)δ4.01(s,2H),3.78-3.72(m,2H),3.72-3.67(m, 4H), 3.65 (d, J=1.9Hz, 12H), 3.25 (dd, J=7.4,6.5Hz, 2H), 1.46 (s, 9H)
(4) preparation of compound 5
The preparation of compound 5a:
Compound 4a (1.252g, 3.8mmol) is placed in 3mL methylene chloride, is added dropwise TFA (8.65g, 75.8mmol), It is transferred to and is stirred to react at room temperature 2 hours after being added dropwise.It is detected through thin-layer chromatography (PE:EA=1:1), raw material reacts completely. It is concentrated under reduced pressure, 5mL water is added, adjust PH=12 with 1M NaOH solution, water phase is washed 2 times with 10mL methylene chloride, with 6M HCl Water phase pH=2 is adjusted, is extracted 3 times after adjusting with 15mL ethyl acetate, organic phase is merged, anhydrous sodium sulfate is dry.? 0.843g crude product, yield 81.1%.The preparation of compound 5b:
Compound 4b (0.229g, 0.6mmol) is placed in 3mL methylene chloride, is added dropwise TFA (1.7g, 12.2mmol), drop It is transferred to and is stirred to react at room temperature 2 hours after adding.It is detected through thin-layer chromatography (PE:EA=1:2), raw material reacts completely.Subtract Pressure concentration, is added 5mL water, adjusts pH=10 with 1M NaOH solution, water phase is washed 2 times with 10mL methylene chloride, with 6M HCl tune Water-saving phase PH=3 is extracted 3 times after adjusting with 15mL ethyl acetate, merges organic phase, and anhydrous sodium sulfate is dry.? 0.190g crude product, yield 97.5%.
The preparation of compound 5c:
Compound 4c (0.871g, 2.08mmol) is placed in 6mL methylene chloride, at room temperature be added TFA (4.7g, 41.6mmol,3mL).It is stirred overnight at room temperature after being added dropwise.It is concentrated under reduced pressure, 5mL water is added, adjust pH=with 1M NaOH solution 10, it is washed 2 times with 10mL methylene chloride, water phase adjusts pH=2 with 6M HCl solution, is extracted 3 times with 10mL methylene chloride, anhydrous Sodium sulphate is dry, merges organic phase.Obtain 0.66g product, yield 87.7%.
The preparation of compound 5d:
Compound 4d (0.971g, 2.10mmol) is placed in 6mL methylene chloride, at room temperature be added TFA (4.7g, 41.6mmol,3mL).It is stirred overnight at room temperature after being added dropwise.It is concentrated under reduced pressure, 5mL water is added, adjust pH=with 1M NaOH solution 10, it is washed 2 times with 10mL methylene chloride, water phase adjusts pH=2 with 6M HCl solution, is extracted 3 times with 10mL methylene chloride, anhydrous Sodium sulphate is dry, merges organic phase.Obtain 0.785g product, yield 92.1%
(5) preparation of compound 7
The preparation of compound 7a:
Compound 5a (0.600g, 2.2mmol) is placed in 25mL round-bottomed flask, displaced air, Ar protection, is dripped under ice bath Add (COCl)2(1.53g,10.9mmol).It is transferred to and is stirred to react at room temperature 2 hours after being added dropwise.It is concentrated under reduced pressure, is added dropwise 3mL DMF is added pomalidomide (0.54g, 1.91mmol), stirs at room temperature, detects (PE:EA=1:2) through thin-layer chromatography, former Expect end of reaction.10mL water is added, is extracted 3 times with 20mL ethyl acetate, saturated common salt water washing 3 times, anhydrous sodium sulfate is dry, Merge organic phase, obtains 0.245g crude product.Column chromatographs (EA), obtains 0.254g white solid product, yield 62%.
1H NMR (500MHz, Chloroform-d) δ 10.48 (s, 1H), 8.88 (d, J=8.2Hz, 1H), 8.26 (s, 1H), 7.74 (dd, J=8.5,7.3Hz, 1H), 7.60 (dd, J=7.3,0.8Hz, 1H), 4.99 (dt, J=12.6,5.8Hz, 1H), 4.27-4.19 (m, 2H), 3.83 (dd, J=8.7,3.8Hz, 6H), 3.74-3.60 (m, 1H), 3.30 (dd, J=7.4, 6.6Hz, 1H), 3.02-2.89 (m, 1H), 2.89-2.71 (m, 2H), 2.18 (qd, J=6.5,5.8,2.0Hz, 1H)
The preparation of compound 7b
Compound 5b (0.228g, 0.72mmol) is placed in 25mL round-bottomed flask, displaced air, Ar protection, under ice bath It is added dropwise (COCl)2(0.502g,3.58mmol).It is transferred to and is stirred to react at room temperature 2 hours after being added dropwise.It is concentrated under reduced pressure, drop Add 3mL DMF, be added pomalidomide (0.197g, 0.72mmol), heat 40 DEG C of reaction 0.5h, detects (EA) through thin-layer chromatography, Raw material end of reaction.5mL water is added, is extracted 3 times with 10mL ethyl acetate, saturated common salt water washing 3 times, anhydrous sodium sulfate is dry It is dry, merge organic phase, obtains 0.245g crude product.Column chromatographs (EA), obtains 0.253g white solid product, yield 61%.
1H NMR (500MHz, Chloroform-d) δ 10.47 (s, 1H), 8.87 (dd, J=8.5,0.8Hz, 1H), 8.41 (s, 1H), 7.74 (dd, J=8.5,7.3Hz, 1H), 7.59 (dd, J=7.3,0.8Hz, 1H), 5.03-4.87 (m, 1H), 4.23 (s, 2H), 3.84 (dq, J=3.0,1.4Hz, 4H), 3.79-3.64 (m, 6H), 3.31-3.22 (m, 2H), 2.97-2.89 (m, 1H),2.89–2.72(m,2H),2.22–2.14(m,1H).
The preparation of compound 7c
Compound 5c (307mg, 0.84mmol) is set in a round bottom flask, Ar is replaced, injects (COCl) under ice bath2, to After injection, it is transferred to room temperature reaction 1h, is concentrated under reduced pressure, addition DMF solution, addition pomalidomide (218mg, 0.84mmol), 2h is reacted at room temperature, and 5mL water is added, three times with the extraction of 20mL ethyl acetate, saturated common salt water washing, anhydrous sulphur Sour sodium is dry, merges.Column chromatography, obtains white solid 340mg, yield 67%.1H NMR(500MHz,Chloroform-d)δ 10.47 (s, 1H), 8.67 (s, 1H), 7.74 (dd, J=8.5,7.3Hz, 1H), 7.59 (dd, J=7.3,0.8Hz, 1H), 4.96 (dd, J=12.2,5.4Hz, 1H), 4.21 (d, J=3.1Hz, 2H), 3.83 (d, J=1.3Hz, 4H), 3.76 (td, J=6.8, 1.5Hz, 2H), 3.74-3.69 (m, 2H), 3.70-3.62 (m, 7H), 3.27 (t, J=6.9Hz, 2H), 2.94-2.84 (m, 1H),2.84–2.71(m,2H),2.17-2.15(m,1H).
The preparation of compound 7d
Compound 5d (376mg, 0.93mmol) is set in a round bottom flask, Ar is replaced, injects (COCl) under ice bath2, to After injection, it is transferred to room temperature reaction 1h, is concentrated under reduced pressure, addition DMF solution, addition pomalidomide (258mg, 0.99mmol), 2h is reacted at room temperature, and 5mL water is added, three times with the extraction of 20mL ethyl acetate, saturated common salt water washing, anhydrous sulphur Sour sodium is dry, merges.Column chromatography, obtains white solid 379.5mg, yield 62%.1H NMR(500MHz,Chloroform-d)δ 10.46 (s, 1H), 8.94 (s, 1H), 8.83 (dd, J=8.5,0.7Hz, 1H), 7.71 (dd, J=8.5,7.3Hz, 1H), 7.56 (dd, J=7.3,0.8Hz, 1H), 4.95 (dd, J=12.1,5.4Hz, 1H), 4.19 (d, J=1.8Hz, 2H), 3.80 (d, J= 1.9Hz, 4H), 3.75-3.67 (m, 4H), 3.64 (s, 10H), 3.23 (t, J=6.9Hz, 2H), 2.90-2.71 (m, 3H), 2.18-2.10(m,1H).
The synthesis of embodiment 2:EGFR targeting small molecule:
(1) preparation of compound 11:
3- (2- chlorine pyrimidine-4-yl) -1- methyl indol 9 (5g, 20.5mmol) is placed in Isosorbide-5-Nitrae-dioxane, at room temperature Compound 10 (3.8g, 20.5mmol) is added, p-methyl benzenesulfonic acid monohydrate (4.7g, 24.6mmol) heats after being added dropwise It is stirred to react at 105 DEG C 2 hours.It is down to room temperature after completion of the reaction, solid is precipitated in solution, filters, and ethyl acetate washs filter cake, dries It is dry, obtain yellow solid 8.0g, yield 100%.1H NMR(500MHz,DMSO-d6) d 8.86 (s, 1H), 8.77 (d, J= 8.2Hz, 1H), 8.55-8.44 (m, 2H), 7.95 (s, 1H), 7.57 (d, J=7.2Hz, 1H), 7.41 (d, J=13.3Hz, 1H),7.34-7.20(m,3H),3.99(s,3H),2.30(s,3H).
(2) preparation of compound 13
Compound 11 (6.8g, 17.3mmol) is dissolved in 30mL DMSO, at room temperature be added 12 (3.911g, 20.8mmol)、K2CO3(3.6g, 26mmol), is added dropwise, and is heated to 90 DEG C of reaction 10h.TLC detection (DCM:MeOH=80: 1) 10mL water is added in raw material fully reacting, three times with the extraction of 30mL methylene chloride, saturated common salt water washing 6 times, anhydrous sodium sulfate It is dry, merge, column chromatographs (DCM:MeOH=80:1), obtains orange color solid 6.473g, yield 66.8%.1H NMR(500MHz, Chloroform-d) δ 9.52 (s, 1H), 8.34 (d, J=5.3Hz, 1H), 8.18 (s, 1H), 8.16-8.09 (m, 1H), 7.51 (s, 1H), 7.39-7.31 (m, 1H), 7.32-7.22 (m, 2H), 7.12 (d, J=5.3Hz, 1H), 6.69 (s, 1H), 3.97 (s, 3H), 3.87 (s, 3H), 3.47 (d, J=7.5Hz, 2H), 2.96-2.77 (m, 8H), 1.45 (s, 9H)
(3) preparation of compound 14
Compound 13 (6.5g, 11.6mmol) is dissolved in ethyl alcohol: in water (V/V=3:1) solution, iron powder being added at room temperature (3.9g, 69.4mmol), ammonium chloride (0.46g, 8.7mmol), is added dropwise, heating reflux reaction 4h.TLC detection (DCM: MeOH=60:1) raw material fully reacting is cooled to room temperature, and is filtered, and is merged.Column chromatographs (DCM:MeOH=30:1), obtains yellow green Paste 5.4g, yield 87.5%.Bright brown paste；1H NMR(500MHz,Chloroform-d)δ8.60–8.43(m,1H), 8.34 (d, J=5.3Hz, 1H), 8.18 (s, 1H), 7.79 (s, 1H), 7.60 (s, 1H), 7.44-7.36 (m, 1H), 7.36- 7.20 (m, 2H), 7.02 (d, J=5.3Hz, 1H), 6.74-6.65 (m, 1H), 3.87 (s, 6H), 3.37 (s, 2H), 3.02 (s, 2H), 2.88 (d, J=14.6Hz, 3H), 2.70 (s, 3H), 1.48 (s, 9H)
(4) preparation of compound 15
Compound 14 (3.735g, 7.0mmol) is dissolved in 20mL methylene chloride, ice bath bet chlorpromazine chloride (1.07g, 8.4mmol), it is added dropwise and is transferred to room temperature reaction 1h.TLC detects (DCM:MeOH=60:1) raw material fully reacting, depressurizes dense Contracting, with saturated common salt water washing, anhydrous sodium sulfate is dry, thick yield 78%.Green paste；1H NMR(500MHz, Chloroform-d) δ 9.92 (s, 1H), 9.02 (s, 2H), 7.97-7.77 (m, 2H), 7.43 (d, J=7.9Hz, 1H), 7.36- (m, 2H), 7.20-7.10 (m, 1H), 3.98 (s, 6H), 3.93 (t, J=6.0Hz, 2H), 3.75 (t, J=6.7Hz, 2H), 3.45 (s, 2H), 3.05 (d, J=22.7Hz, 2H), 2.90 (s, 5H), 2.82 (t, J=6.6Hz, 3H), 1.46 (s, 9H)
(5) preparation of compound 16
Compound 15 is dissolved in acetonitrile, triethylamine (2.5g, 21.3mmol) is added at room temperature, is heated to 80 DEG C of reactions 6h, TLC detect (DCM:MeOH=60:1), raw material fully reacting.It is concentrated under reduced pressure, with saturated common salt water washing, anhydrous sodium sulfate It is dry.Column chromatographs (DCM:MeOH=60:1), obtains yellow paste product 3.45g, yield 85%.1H NMR(500MHz, Chloroform-d)δ9.87(s,1H),9.74(s,1H),9.07(s,1H),8.95(s,1H),8.39–8.38(m,1H), 8.09-8.07 (m, 1H), 7.75 (d, J=7.4Hz, 1H), 7.41-7.40 (m, 1H), 7.32-7.28 (m, 2H), 7.22-7.21 (m, 1H), 6.79 (d, J=3.5Hz, 1H), 6.47 (d, J=5.0Hz, 1H), 5.78 (d, J=5.0Hz, 1H), 3.99 (s, 3H), 3.90 (s, 3H), 3.37 (d, J=7.6Hz, 2H), 3.00 (s, 2H), 2.86 (s, 3H), 2.69 (s, 3H), 1.47 (s, 9H).
(6) preparation of compound III (m=1, R are methyl)
Compound 16 (0.808g, 1.4mmol) is dissolved in 4mL methylene chloride, is added TFA (2.08g, 24.8mmol), It is stirred to react 3h.TLC detects (DCM:MeOH=10:1) raw material fully reacting, is concentrated under reduced pressure.With methylene chloride band it is extra three Fluoroacetic acid.Column chromatographs (DCM:MeOH=60:1), obtains yellow solid 0.640g, yield 94.1%.1H NMR(500MHz,DMSO- d6) δ 9.38 (s, 1H), 8.69 (s, 1H), 8.60 (s, 1H), 8.29 (d, J=6.0Hz, 1H), 7.57 (dd, J=8.3, 4.4Hz, 1H), 7.35-7.31 (m, 1H), 7.31-7.26 (m, 1H), 7.18 (t, J=7.7Hz, 1H), 7.02 (d, J= 12.0Hz, 1H), 6.75 (dd, J=17.0,10.2Hz, 1H), 6.29 (dd, J=16.9,2.0Hz, 1H), 5.79 (dd, J= 10.2,2.0Hz, 1H), 3.93 (s, 3H), 3.85 (s, 3H), 3.27 (d, J=5.9Hz, 2H), 3.16 (d, J=6.0Hz, 2H), 2.64(s,6H).
Embodiment 3: the synthesis of compound 17:
The preparation of compound 17a
Pomalidomide (0.2mg, 0.73mmol) is dissolved in 10mL THF, at room temperature be added chlorpromazine chloride (0.102mg, 0.8mmol), 60 DEG C of back flow reactions 2h, TLC are heated and detect EA starting material left.It is cooled to room temperature, is concentrated under reduced pressure, is analysed with petroleum ether White crystal out filters, drying, obtains 0.254g white solid, and thick yield is 95.8%.White solid；1H NMR(500MHz, DMSO-d6) δ 11.16 (s, 1H), 9.90 (s, 1H), 8.44 (d, J=8.4Hz, 1H), 7.85 (dd, J=8.4,7.3Hz, 1H), 7.65 (d, J=7.2Hz, 1H), 5.16 (dd, J=12.9,5.4Hz, 1H), 3.90 (t, J=6.2Hz, 2H), 3.01 (t, J= 6.2Hz, 2H), 2.91 (ddd, J=17.1,13.9,5.4Hz, 1H), 2.62 (dt, J=17.0,3.0Hz, 1H), 2.58-2.47 (m,2H),2.13–2.01(m,1H).
The preparation of compound 17b
Pomalidomide (200mg, 0.73mmol) is dissolved in tetrahydrofuran, at room temperature inject 4- chlorobutanoylchloride (112mg, 0.8mmol), heating reflux reaction 4h after injection.It is concentrated under reduced pressure, washs solid with ether, filter, wash filter with ether Cake, obtains white solid 220mg, and thick yield is 80%.White solid；1H NMR(500MHz,DMSO-d6)δ11.16(s,1H), 9.81 (s, 1H), 8.41 (d, J=8.3Hz, 1H), 7.84 (dd, J=8.4,7.3Hz, 1H), 7.63 (dd, J=7.3,0.8Hz, 1H), 5.15 (dd, J=12.8,5.4Hz, 1H), 3.73 (t, J=6.6Hz, 2H), 2.69-2.57 (m, 3H), 2.56-2.48 (m,2H),2.10–2.04(m,3H).
The preparation of compound 17c
Pomalidomide (200mg, 0.73mmol) is dissolved in tetrahydrofuran, injects 5- chloro valeric chloride at room temperature (125mg, 0.8mmol), heating reflux reaction 4h after injection.It is concentrated under reduced pressure, washs solid with ether, filter, use ether Filter cake is washed, white solid 228mg is obtained, thick yield is 79.7%.White solid；1H NMR(500MHz,Chloroform-d)δ 9.43 (s, 1H), 8.82 (d, J=8.5Hz, 1H), 8.54 (s, 1H), 7.72 (dd, J=8.5,7.3Hz, 1H), 7.56 (d, J= 7.3Hz, 1H), 4.97 (dd, J=12.4,5.3Hz, 1H), 3.59 (t, J=6.0Hz, 2H), 3.02-2.86 (m, 1H), 2.86- 2.74 (m, 2H), 2.52 (t, J=6.9Hz, 2H), 2.22-2.11 (m, 1H), 1.90 (qd, J=8.9,8.0,4.2Hz, 4H)
The preparation of compound 17d
Pomalidomide (200mg, 0.73mmol) is dissolved in 15mL tetrahydrofuran, injects 6- bromo valeric chloride at room temperature (187mg, 0.88mmol), heating reflux reaction 4h after injection.It is concentrated under reduced pressure, washs solid with ether, filter, use ether Filter cake is washed, white solid 276mg is obtained, thick yield is 83.9%.White solid；1H NMR(500MHz,Chloroform-d)δ 9.42 (s, 1H), 8.83 (d, J=8.5Hz, 1H), 8.52 (s, 1H), 7.72 (dd, J=8.5,7.3Hz, 1H), 7.56 (d, J= 7.2Hz, 1H), 5.08-4.85 (m, 1H), 3.43 (t, J=6.7Hz, 2H), 2.99-2.86 (m, 1H), 2.85-2.72 (m, 2H), 2.50 (t, J=7.5Hz, 2H), 2.24-2.12 (m, 1H), 1.914.89 (m, 2H), 1.81.76 (m 2H), 1.63- 1.50(m,2H).
The preparation of embodiment 4:PROTACs compound P1
Compound 7a (140mg, 0.26mmol) made from embodiment 1 is dissolved in 5mL DMF, be added embodiment 2 be made Compound III (141mg, 0.29mmol), DIPEA (101mg, 0.78mmmol), heat 80 DEG C of reaction 12h.10mL is added Water, three times with the extraction of 20mL methylene chloride, saturated common salt water washing, anhydrous sodium sulfate is dry, merges organic phase, and column chromatography obtains Yellow solid 135mg, yield 57%.1H NMR(500MHz,Chloroform-d)δ10.40(s,1H),9.79(s,1H), 9.60 (s, 1H), 9.05 (s, 1H), 8.78 (dd, J=8.5,0.7Hz, 1H), 8.39 (d, J=5.2Hz, 1H), 8.14-7.96 (m, 1H), 7.72 (s, 1H), 7.66 (dd, J=8.5,7.3Hz, 1H), 7.53 (dd, J=7.3,0.8Hz, 1H), 7.43-7.36 (m, 1H), 7.28-7.23 (m, 2H), 7.20 (d, J=5.3Hz, 1H), 6.77 (s, 1H), 6.49-6.37 (m, 1H), 5.80- 5.62 (m, 1H), 4.95-4.81 (m, 1H), 4.12 (d, J=3.6Hz, 2H), 3.99 (s, 3H), 3.86 (s, 3H), 3.78- 3.66 (m, 5H), 3.62 (s, 2H), 2.94 (s, 2H), 2.86-2.77 (m, 1H), 2.69 (d, J=8.9Hz, 7H), 2.48 (s, 2H),2.33(s,3H),2.14–2.10(m,1H),1.88–1.84(m,1H).
The preparation of embodiment 5:PROTACs compound P2
Compound 7b (200mg, 0.35mmol) made from embodiment 1 is dissolved in 5mL DMF, be added embodiment 2 be made Compound III (186mg, 0.38mmol), DIPEA (136mg, 1.05mmmol), heat 80 DEG C of reaction 12h.10mL is added Water, three times with the extraction of 20mL methylene chloride, saturated common salt water washing, anhydrous sodium sulfate is dry, merges organic phase, and column chromatography obtains Yellow solid 181mg, yield 55.3%.1H NMR(500MHz,Chloroform-d)δ10.41(s,1H),9.80(s,1H), 9.58 (s, 1H), 9.03 (s, 1H), 8.81 (d, J=8.4Hz, 1H), 8.38 (d, J=5.3Hz, 1H), 8.05 (dd, J=7.0, 2.0Hz, 1H), 7.77 (s, 1H), 7.68 (dd, J=8.5,7.3Hz, 1H), 7.57-7.50 (m, 1H), 7.40-7.35 (m, 1H), 7.28-7.24 (m, 2H), 7.19 (d, J=5.3Hz, 1H), 6.77 (s, 1H), 6.45 (dd, J=16.7,1.9Hz, 1H), 5.78-5.64 (m, 1H), 4.89 (dd, J=12.3,5.3Hz, 1H), 4.12 (d, J=3.6Hz, 2H), 3.97 (s, 3H), 3.85 (s,3H),3.71(s,4H),3.65–3.49(m,6H),2.96(s,2H),2.86–2.58(m,9H),2.57–2.42(m,2H), 2.42–2.19(m,3H),2.11–2.07(m,1H).
The preparation of embodiment 6:PROTACs compound P3
Compound 7c (153mg, 0.25mmol) made from embodiment 1 is dissolved in 5mL DMF, be added embodiment 2 be made Compound III (132mg, 0.27mmol), DIPEA (104mg, 0.8mmol), heat 80 DEG C of reaction 12h.10mL water is added, Three times with the extraction of 20mL methylene chloride, saturated common salt water washing, anhydrous sodium sulfate is dry, merges organic phase, and column chromatography obtains yellow Solid 118mg, yield 50%.1H NMR(500MHz,Chloroform-d)δ10.45(s,1H),9.82(s,1H),9.69(s, 1H), 9.06 (s, 1H), 8.83 (d, J=8.4Hz, 1H), 8.40 (d, J=5.3Hz, 1H), 8.11-7.99 (m, 1H), 7.78 (s, 1H), 7.73-7.65 (m, 1H), 7.55 (d, J=7.3Hz, 1H), 7.44-7.35 (m, 1H), 7.29-7.23 (m, 3H), 7.20 (d, J=5.3Hz, 1H), 6.78 (s, 1H), 6.46 (dd, J=16.5,2.0Hz, 1H), 5.75-5.59 (m, 1H), 4.90 (dd, J=12.3,5.4Hz, 1H), 4.15 (d, J=3.6Hz, 2H), 3.99 (s, 3H), 3.87 (s, 3H), 3.75 (p, J= 2.2,1.7Hz,4H),3.64–3.52(m,10H),2.95–2.94(m,2H),2.86–2.64(m,8H),2.48(s,2H), 2.33(s,3H),2.13–2.10(m,2H).
The preparation of embodiment 7:PROTACs compound P4
Compound 7d (150mg, 0.23mmol) made from embodiment 1 is dissolved in 5mL DMF, embodiment is added at room temperature Compound III (110mg, 0.23mmol) made from 2, DIPEA (89mg, 0.69mmol), finish, and are slowly increased to 80 DEG C of reactions 12h.TLC detects (DCM:MeOH=10:1), starting material left.10mL water is added, methylene chloride extracts (20mL × 3), saturation food Salt water washing, anhydrous sodium sulfate is dry, and concentration, column chromatographs (DCM:MeOH=10:1), obtains yellow solid 120mg, yield 53%.1H NMR (500MHz, Chloroform-d) δ 9.79 (s, 1H), 9.59 (s, 1H), 8.98 (d, J=17.9Hz, 2H), 8.38 (d, J =5.3Hz, 1H), 8.12-8.01 (m, 1H), 7.79 (s, 1H), 7.75-7.67 (m, 2H), 7.44 (t, J=7.8Hz, 1H), 7.41-7.35 (m, 1H), 7.29-7.25 (m, 3H), 7.18 (d, J=5.3Hz, 1H), 6.76 (s, 1H), 6.42 (dd, J= 16.6,1.9Hz, 1H), 5.73-5.65 (m, 1H), 5.16 (dd, J=13.3,5.2Hz, 1H), 4.39 (s, 2H), 4.08 (d, J =8.5Hz, 2H), 3.96 (s, 3H), 3.85 (s, 3H), 3.67 (dt, J=5.3,2.6Hz, 2H), 3.61-3.49 (m, 5H), 3.42 (dt, J=16.3,5.6Hz, 5H), 2.91 (s, 3H), 2.83-2.71 (m, 2H), 2.63 (s, 3H), 2.52 (s, 2H), 2.29–2.26(m,2H),2.20–2.06(m,1H).
The preparation of embodiment 8:PROTACs compound P5
Compound 17a (257mg, 0.6mmol) made from embodiment 3 is dissolved in 5mL DMF, be added embodiment 2 be made Compound III (320mg, 0.66mmol), DIPEA (233mg, 1.8mmmol), heat 80 DEG C of reaction 8h.10mL water is added, Three times with the extraction of 20mL methylene chloride, saturated common salt water washing, anhydrous sodium sulfate is dry, merges organic phase, and column chromatography obtains yellow Solid 258mg, yield 51%.1H NMR(500MHz,Chloroform-d)δ10.07(s,1H),9.64(s,1H),9.22(s, 1H), 8.94 (s, 1H), 8.72 (d, J=8.5Hz, 1H), 8.38 (d, J=5.3Hz, 1H), 8.07-7.98 (m, 1H), 7.79 (d, J=14.0Hz, 1H), 7.61-7.52 (m, 1H), 7.42-7.31 (m, 2H), 7.27-7.24 (m, 2H), 7.17 (d, J= 5.4Hz, 1H), 6.74 (s, 1H), 6.38-6.24 (m, 2H), 5.66 (dd, J=8.5,3.3Hz, 1H), 4.79 (dd, J= 12.3,5.7Hz,1H),3.91(s,3H),3.80(s,3H),2.96–2.93(m,2H),2.83–2.78(m,2H),2.74– 2.56 (m, 7H), 2.48-2.44 (m, 2H), 2.54-2.51 (m, 2H), 2.33 (d, J=8.9Hz, 3H), 2.12-2.00 (m, 1H).
The preparation of embodiment 9:PROTACs compound P6
Compound 17b (254mg, 0.54mmol) made from embodiment 3 is dissolved in 5mL DMF, be added embodiment 2 be made Compound III (287mg, 0.59mmol), DIPEA (210mg, 1.6mmmol), heat 80 DEG C of reaction 8h.10mL water is added, Three times with the extraction of 20mL methylene chloride, saturated common salt water washing, anhydrous sodium sulfate is dry, merges organic phase, and column chromatography obtains yellow Solid 232mg, yield 52.2%.1H NMR(500MHz,Chloroform-d)δ9.79(s,1H),9.58(s,1H),9.37 (s, 1H), 9.01 (s, 1H), 8.70 (d, J=8.3Hz, 1H), 8.37 (dd, J=5.3,2.6Hz, 1H), 8.03 (s, 1H), 7.73 (d, J=11.0Hz, 1H), 7.68-7.54 (m, 1H), 7.47-7.45 (m, 1H), 7.42-7.35 (m, 1H), 7.27- 7.22 (m, 2H), 7.19-7.13 (m, 1H), 6.76 (d, J=10.4Hz, 1H), 6.40 (d, J=15.1Hz, 2H), 5.75- 5.61 (m, 1H), 4.85 (td, J=12.4,5.3Hz, 1H), 3.96 (s, 3H), 3.87 (s, 3H), 2.95 (s, 2H), 2.84- 2.74(m,2H),2.73–2.60(m,6H),2.59–2.43(m,7H),2.35–2.39(m,4H).
The preparation of embodiment 10:PROTACs compound P7
Compound 17c (249mg, 0.52mmol) made from embodiment 3 is dissolved in 10mL DMF, the system of embodiment 2 is added Compound III (275mg, 0.57mmol), the DIPEA (202mg, 1.6mmmol) obtained, heats 80 DEG C of reaction 9h.10mL is added Water, three times with the extraction of 20mL methylene chloride, saturated common salt water washing, anhydrous sodium sulfate is dry, merges organic phase, and column chromatography obtains Yellow solid 215mg, yield 49.2%.1H NMR(500MHz,Chloroform-d)δ9.77(s,1H),9.66(s,1H), 9.32 (s, 1H), 9.00 (s, 1H), 8.70 (d, J=8.5Hz, 1H), 8.37 (d, J=5.3Hz, 1H), 8.08-7.96 (m, 1H), 7.79 (s, 1H), 7.60 (dd, J=8.5,7.3Hz, 1H), 7.44 (dd, J=7.3,0.9Hz, 1H), 7.41-7.32 (m, 1H), 7.27-7.21 (m, 2H), 7.16 (d, J=5.4Hz, 1H), 6.75 (s, 1H), 6.43 (s, 1H), 5.83-5.56 (m, 1H), 4.86 (dd, J=12.3,5.5Hz, 1H), 3.94 (s, 3H), 3.84 (s, 3H), 2.92 (t, J=5.8Hz, 2H), 2.81- 2.74 (m, 1H), 2.74-2.63 (m, 6H), 2.44 (dt, J=23.0,7.8Hz, 6H), 2.32-2.25 (m, 3H), 2.04 (s, 2H), 1.71 (t, J=7.7Hz, 2H), 1.63-1.51 (m, 2H)
The preparation of embodiment 11:PROTACs compound P8
Compound 17d (306mg, 0.61mmol) made from embodiment 3 is dissolved in 10mL DMF, the system of embodiment 2 is added Compound III (329mg, 0.67mmol), the DIPEA (261mg, 1.8mmmol) obtained, heats 80 DEG C of reaction 6h.10mL is added Water, three times with the extraction of 20mL methylene chloride, saturated common salt water washing, anhydrous sodium sulfate is dry, merges organic phase, and column chromatography obtains Yellow solid 224mg, yield 43%.1H NMR(500MHz,Chloroform-d)δ9.76(s,1H),9.71(s,1H),9.31 (d, J=4.8Hz, 1H), 9.00 (s, 1H), 8.68 (dd, J=8.6,6.1Hz, 1H), 8.36 (dd, J=5.3,2.4Hz, 1H), 8.08-7.96 (m, 1H), 7.79 (d, J=9.1Hz, 1H), 7.63-7.54 (m, 1H), 7.46-7.39 (m, 1H), 7.39-7.30 (m, 1H), 7.24-7.21 (m, 2H), 7.14 (d, J=5.3Hz, 1H), 6.73 (d, J=9.4Hz, 1H), 6.42 (d, J= 15.4Hz, 1H), 5.78-5.61 (m, 1H), 4.86 (dd, J=12.2,5.6Hz, 1H), 3.94 (s, 3H), 3.84 (s, 3H), 2.91–2.88(m,2H),2.80–2.62(m,8H),2.44–2.36(m,6H),2.27(s,3H),2.12–2.05(m,1H), 1.72–1.68(m,2H),1.54–1.49(m,2H),1.38–1.33(m,2H).
Performance measurement:
(1) mtt assay measures PROTACs compound on tumor cell proliferation inhibition activity
The research of this screening active ingredients relates generally to following several non-small cell lung cancer cell strains, title and feature such as following table Shown in 2:
2 non-small cell lung cancer of table (NSCLC) cell line type and mutational site
By the tumour cell (H1975, PC9, PC9-IR, HCC827) in logarithmic growth phase respectively with 5 × 103、5× 103、5×103、1.5×103A cell inoculation is cultivated for 24 hours in 96 orifice plates, after various concentration PROTACs compound is added, cell In 37 DEG C, 5%CO2Under the conditions of continue culture 72 hours, every hole is added 20uL MTT (5mg/mL) solution and continues culture 4 hours, It is dissolving crystallized with DMSO, its OD value is measured at 490nm wavelength with enzyme-linked immunosorbent assay instrument and calculates IC50, concrete outcome is as follows Shown in table 3.
Table 3
(2) Western Blot measures the effect of EGFR protein degradation
It after PROTACs drug-treated PC9 or PC9-IR cell 48h, is washed 2 times with the PBS of pre-cooling, is added and contains 10% in right amount The RIPA lysate of PMSF and 5%cOmplete collects cell, on ice after lytic cell 30min, 4 DEG C, 12000r/min, 30min centrifugation, takes supernatant, i.e. total protein of cell.Protein content is detected with BCA standard measure, dilutes egg with 5 × albumen sample-loading buffer 100 DEG C of denaturation 5min after white.Albumen is separated by electrophoresis in SDS-PAGE, transferring film, closes 2h, 4 DEG C of primary antibody overnight incubations.TBST is washed Film, secondary antibody 1:2000 are incubated for 2h, wash film, develop after chemiluminescence.
Table 4
By table 3 and table 4 it is found that compound provided by the present invention all has preferable EGFR protein degradation effect, and to more Kind tumour has good inhibiting effect.

Claims (8)

1. compound or its pharmaceutically acceptable salt with targeting EGFR protein degradation shown in formula I or formula II:
In formula I and formula II, R C1~C6Alkyl, the integer that m is 1~7, the integer that n is 1~6.
2. the compound or its pharmaceutically acceptable salt of targeting EGFR protein degradation according to claim 1, feature It is, R is methyl, the integer that m 1, n are 2~5.
3. the preparation method of the compound of targeting EGFR protein degradation according to claim 1 or 2, which is characterized in that institute The preparation process for the type I compound stated is as follows:
It specifically includes: formula III compound is dissolved in organic solvent, formula IV compound and diisopropyl second are added at room temperature Amine is heated to 80~90 DEG C of 10~14h of reaction and obtains Formulas I compound represented after extraction, washing, drying.
4. the preparation method of the compound of targeting EGFR protein degradation according to claim 1 or 2, which is characterized in that institute The preparation process for the Formula II compound stated is as follows:
It specifically includes: formula III compound is dissolved in organic solvent, Formula V compound and diisopropylethylamine are added at room temperature, It is heated to 80~90 DEG C of 10~14h of reaction and obtains Formula II compound represented after extraction, washing, drying.
5. a kind of pharmaceutical composition, which is characterized in that including compound of any of claims 1 or 2 or its is pharmaceutically acceptable Salt and pharmaceutically acceptable excipient.
6. a kind of compound according to claim 1 or 2 or its pharmaceutically acceptable salt are in preparation prevention or/and treatment It is applied in the drug of cancer.
7. application according to claim 6, which is characterized in that the cancer be breast cancer, colon cancer, prostate cancer, Cancer of pancreas, thyroid papillary carcinoma, oophoroma, melanoma, leukaemia or non-small cell lung cancer.
8. application according to claim 6, which is characterized in that the cancer is non-small cell lung cancer.
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