CN116693505B - Compound for targeted degradation of EGFR protein and preparation method and application thereof - Google Patents

Compound for targeted degradation of EGFR protein and preparation method and application thereof Download PDF

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CN116693505B
CN116693505B CN202310935511.1A CN202310935511A CN116693505B CN 116693505 B CN116693505 B CN 116693505B CN 202310935511 A CN202310935511 A CN 202310935511A CN 116693505 B CN116693505 B CN 116693505B
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杨鹏
肖易倍
朱亚胜
郝海平
叶秀全
伍昱星
沈豪
王晓
王丽萍
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China Pharmaceutical University
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Abstract

The invention discloses a compound for targeted degradation of EGFR protein, a preparation method and application thereof, wherein the compound for targeted degradation of EGFR protein has degradation activity on EGFR kinase, can effectively solve the problem of drug resistance caused by target mutation of octenidine, and has strong inhibition activity on various tumor cells when applied to preparation of drugs for preventing or treating cancers or tumor-related diseases.

Description

Compound for targeted degradation of EGFR protein and preparation method and application thereof
Technical Field
The invention relates to a compound for targeted degradation of EGFR protein, and also relates to a preparation method and application of the compound.
Background
EGFR (epidermal growth factor receptor) is an important cell membrane receptor, playing a key role in tumor growth and progression. Abnormal activation of EGFR is closely related to the occurrence and development of various malignant tumors, and thus EGFR becomes an important target for anti-tumor treatment. Normally, EGFR is activated by binding its ligand EGF (epidermal growth factor) or other similar growth factor, thereby initiating a series of signaling pathways that promote cell proliferation, survival and migration. However, in some cases, EGFR may be subject to abnormal changes such as mutation, overexpression or overactivation, resulting in excessive cell proliferation and tumor formation. Thus, findDrugs capable of effectively inhibiting EGFR are one of the important directions for tumor treatment. Ornitinib (Osimertinib) is a third generation EGFR tyrosine kinase inhibitor for use in the treatment of EGFR mutation-positive non-small cell lung cancer (NSCLC) patients. Although the Ornitinib achieves remarkable curative effects in the early stage of treatment, the problem of drug resistance gradually emerges with the prolongation of the treatment time. A number of mechanisms have been discovered which lead to resistance to octreotide. The most common mechanism is EGFR C797S Mutations, EGFR C797S Resulting in failure of the oritinib to bind to EGFR effectively, thereby causing tumor cells to develop resistance to the oritinib. In addition, other mechanisms include amplification of MET, KRAS mutation, HER2 mutation, activation of the cell repair system, and the like. These mechanisms may exist alone or simultaneously, resulting in failure of the treatment with octenib. Therefore, solving the problem of the resistance of the octreotide is an urgent clinical requirement.
Disclosure of Invention
The invention aims to: the invention aims to provide a compound for targeted degradation of EGFR protein, and a second aim is to provide a preparation method and application of the compound.
The technical scheme is as follows: the compound for targeted degradation of EGFR protein is shown as a general formula (I):
；
wherein linker is a linking group, the linking group is a straight chain or branched chain alkylene chain with total length of 1-20 atoms, and the straight chain or branched chain alkylene chain is formed by-CH 2 -, -O-, -CO-; -CONH-; -S-, -N-, and one or more of alkynylene or cycloalkylene.
Preferably, the straight or branched alkylene chain consists of-CH 2 -, -O-, -CO-; -CONH-; -one or more of S-or-N-.
Preferably, the straight or branched alkylene chain consists of-CH 2 -, -O-, -CO-; -one or more of CONH-or-N-.
Further, the compound is selected from C-1 to C-16:
。
the preparation method of the compound comprises the following steps:
and (3) dissolving the A and the B in DMF, adding N, N-diisopropylethylamine or a mixed solution of the N, N-diisopropylethylamine and HATU, and purifying after the reaction is finished to obtain the compound for targeted degradation of EGFR protein.
Preferably, the reaction temperature of the reaction is 20-100 ℃ and the reaction time is 12-24 hours.
The preparation method of the compound further comprises the following steps:
c and D are dissolved in DMF, HATU and N, N-diisopropylethylamine are added, and after the reaction is finished, the compound for targeted degradation of EGFR protein is obtained by purification.
Preferably, the reaction temperature of the reaction is 20-50 ℃ and the reaction time is 12-24 hours.
The pharmaceutically acceptable salts are acid addition salts of the compounds of formula (I), wherein the acid used for salt formation is selected from the group consisting of inorganic acids selected from the group consisting of hydrochloric acid, sulfuric acid, phosphoric acid, and organic acids selected from the group consisting of acetic acid, propionic acid, butyric acid, maleic acid, p-toluenesulfonic acid, malic acid, methanesulfonic acid, malonic acid, trichloroacetic acid, trifluoroacetic acid, fumaric acid, citric acid, digluconic acid, camphoric acid, cinnamic acid, aspartic acid, and tartaric acid.
Further, the dosage form of the medicine is at least one selected from the group consisting of tablets, capsules, granules, injections, powder injections, eye drops, powders, drop pills, transdermal absorption patches, ointments, aerosols, emulsions, smears, suppositories, films, controlled release preparations and nano-preparations.
The invention also provides a pharmaceutical composition, which is the compound for targeted degradation of EGFR protein or pharmaceutically acceptable salt or optical isomer thereof, and a pharmaceutically acceptable carrier.
Pharmaceutically acceptable carrier refers to excipients or diluents that do not cause significant irritation to the organism and do not interfere with the biological activity and properties of the compound being administered. The excipient comprises a flavoring agent, an antioxidant, a flavoring agent, a cosolvent, an emulsifying agent, a solubilizer, a preservative, a binding agent, an osmotic pressure regulator, a disintegrating agent, a colorant filler, a lubricant and the like, and the diluent comprises dextrin, starch, sucrose, physiological saline, lactose and the like.
The application of the compound for targeted degradation of EGFR protein in preparing medicines for preventing or treating cancers or tumor related diseases.
Preferably, the cancer or tumor comprises lung cancer, stomach cancer, prostate cancer, ovarian cancer, testicular cancer, colon cancer, leukemia, breast cancer, multiple myeloma, liver cancer, pancreatic cancer, melanoma, glioma, brain glioma or pituitary tumor.
Further, the tumor is a malignant tumor of EGFR gene mutation.
The principle of the invention: the compounds of the invention, when the PROTAC molecule of EGFR binds to intracellular EGFR, the PROTAC molecule causes binding of EGFR to ubiquitin ligases, resulting in ubiquitination of EGFR, labeled as a target for degradation. Ubiquitinated EGFR is recognized by the proteasome and eventually degraded. Since EGFR is a key factor in tumor development and progression, EGFR function can be effectively inhibited by promoting EGFR degradation, thereby inhibiting tumor growth and spread. Compared with the traditional drug target inhibitor, the PROTAC of EGFR can degrade EGFR protein level more rapidly, can effectively degrade EGFR mutant protein and has obvious killing effect on the Ornitanib drug-resistant cells, and can effectively solve the drug resistance problem caused by the Ornitanib target mutation.
The beneficial effects are that: compared with the prior art, the invention has the following remarkable advantages: the compound for targeted degradation of EGFR protein prepared by the invention has degradation activity on EGFR kinase and inhibition activity on various tumor cells, and can effectively solve the problem of drug resistance caused by target mutation of the Ornitinib.
Drawings
FIG. 1 is a general formula of a compound targeted to degrade EGFR protein;
FIG. 2 is a synthetic route for intermediate A1-4 of example 1;
FIG. 3 is a synthetic route for intermediate B1-13 of example 1;
FIG. 4 is the synthesis of compounds C1-C8;
FIG. 5 shows the synthesis of compounds C9-C16.
Description of the embodiments
The technical scheme of the invention is further described below with reference to the accompanying drawings.
Example 1
Synthesis of intermediate reactants
(1) Synthesis of 4- (1-ethylsulfonyl) -1H-indol-3-yl) -N- (4- (piperazin-1-yl) phenyl) -5-trifluoromethylpyrimidin-2-amine (A-1):
step one, synthesis of 3- (2-chloro-5-trifluoromethyl) pyrimidin-4-yl) -1H-indole (a-1-1): 2, 4-dichloro-5- (trifluoromethyl) pyrimidine (1 eq, 100 mmol) and aluminum trichloride (1.2 eq, 120 mmol) were dissolved in 250 mL 1, 2-dichloroethane, stirred at room temperature for 5 minutes, indole (1.2 eq, 120 mmol) was added, reacted at 80℃for 2 hours, then quenched with ice water, filtered through celite, and the mixture extracted with dichloromethane. The organic phase was washed with saturated sodium chloride solution, dried over anhydrous sodium sulfate and concentrated by evaporation under vacuum to give A-1-1 as a pale yellow solid in 70% yield by rapid preparative liquid phase purification. 1 H NMR (400 MHz, DMSO-d 6 ) δ 12.25 (s, 1H), 9.02 (s, 1H), 8.45 – 8.35 (m, 1H), 8.10 (dd,J= 1.2, 3.2 Hz, 1H), 7.64 – 7.53 (m, 1H), 7.27 (tt,J= 5.5, 7.2 Hz, 2H)。
Step two, 3- (2-chloro-5- (trifluoromethyl) pyrimidin-4-yl) -1- (ethylsulfonyl) -1H-indole (A-1-2)Is synthesized by the following steps: a-1-1 (1 eq, 10 mmol) was dissolved in 25 mL anhydrous tetrahydrofuran and NaH (2 eq, 20 mmol) was added slowly in portions at 0℃and after the addition was complete, stirred further for 1h at room temperature. Ethyl sulfonyl chloride (1.5 eq, 15 mmol) was added and the reaction was continued for 1h, quenched with saturated aqueous ammonium chloride and the mixture extracted with dichloromethane. The organic phase was washed with saturated sodium chloride solution, dried over anhydrous sodium sulfate and concentrated by evaporation under vacuum to give A-1-2 as a pale yellow solid in 91% yield by rapid preparative liquid phase purification. 1 H NMR (400 MHz, DMSO-d 6 ) δ 9.30 (d,J= 0.8 Hz, 1H), 8.05 (dt,J= 1.2, 7.8 Hz, 1H), 8.01 – 7.98 (m, 1H), 7.95 (dt,J= 0.9, 8.4 Hz, 1H), 7.57 – 7.44 (m, 2H), 3.82 (q,J= 7.3 Hz, 2H), 1.10 (t,J= 7.3 Hz, 3H)。
Step three, synthesis of 4- (1-ethylsulfonyl) -1H-indol-3-yl) -N- (4- (piperazin-1-yl) phenyl) -5-trifluoromethyl pyrimidin-2-amine (a-1): a-1-2 (1 eq, 5 mmol) and tert-butyl 4- (4-aminophenyl) piperazine-1-carboxylate (1.1 eq, 5.5 mmol) were dissolved in n-butanol of 20 mL and trifluoroacetic acid (2 eq, 10 mmol) was added and reacted at 120℃for 24 hours, after which the reaction mixture was concentrated and the liquid phase was purified rapidly to give A-1 in 73% yield. 1 H NMR (400 MHz, Chloroform-d) δ 8.71 (s, 1H), 8.08 – 8.00 (m, 1H), 7.96 (d,J= 8.3 Hz, 1H), 7.91 (s, 1H), 7.53 – 7.47 (m, 2H), 7.47 – 7.40 (m, 2H), 7.34 (t,J= 7.9 Hz, 1H), 6.92 (d,J= 8.5 Hz, 2H), 3.39 (q,J= 7.4 Hz, 2H), 3.13 (dd,J= 3.2, 6.5 Hz, 4H), 3.05 (dd,J= 3.3, 6.3 Hz, 4H), 1.25 (t,J= 7.4 Hz, 3H)。
(2) Synthesis of tert-butyl 3- (4- (4- ((4- (1- (ethylsulfonyl) -1H-indol-3-yl) -5- (trifluoromethyl) pyrimidin-2-yl) amino) phenyl) piperazine-1-propanoate (A-2):
a-1 (1 eq, 1 mmol) and tert-butyl bromopropionate (1 eq, 1.2 mmol) were dissolved in 10 mL DMF, N-diisopropylethylamine (3 eq, 15 mmol) was added, the reaction was conducted at 80℃12 h, after the completion of the reaction, the reaction mixture was concentrated, and the rapid preparative liquid phase was purified to give A-2 in 63% yield. 1 H NMR (400 MHz, Chloroform-d) δ 8.71 (s, 1H), 8.06 – 8.00 (m, 1H), 8.00 – 7.94 (m, 1H), 7.91 (s, 1H), 7.53 – 7.47 (m, 2H), 7.47 – 7.30 (m, 3H), 6.91 (d,J= 8.5 Hz, 2H), 3.39 (q,J= 7.4 Hz, 2H), 3.17 (t,J= 5.0 Hz, 4H), 2.73 (t,J= 7.3 Hz, 2H), 2.63 (t,J= 5.0 Hz, 4H), 2.46 (t,J= 7.3 Hz, 2H), 1.46 (s, 9H), 1.26 (d,J= 7.3 Hz, 3H)。
(3) Synthesis of tert-butyl 4- (4- (4- (1-ethylsulfonyl) -1H-indol-3-yl) -5- (trifluoromethyl) pyrimidin-2-ylamino) phenyl) piperazin-1-yl) butyrate (A-3):
referring to the synthesis of A-2, the yield was 65%. 1 H NMR (400 MHz, Chloroform-d) δ 8.71 (s, 1H), 8.04 (dd,J= 1.1, 7.9 Hz, 1H), 7.97 (d,J= 1.0 Hz, 1H), 7.91 (s, 1H), 7.53 – 7.47 (m, 2H), 7.43 (ddd,J= 1.3, 7.2, 8.4 Hz, 1H), 7.35 (d,J= 8.5 Hz, 2H), 6.92 (d,J= 8.5 Hz, 2H), 3.39 (q,J= 7.4 Hz, 2H), 3.18 (t,J= 5.0 Hz, 4H), 2.62 (t,J= 5.0 Hz, 4H), 2.42 (t,J= 7.5 Hz, 2H), 2.28 (t,J= 7.4 Hz, 2H), 1.82 (p,J= 7.5 Hz, 2H), 1.45 (s, 9H), 1.24 (d,J= 7.4 Hz, 3H)。
(4) Synthesis of tert-butyl 2- (2- (4- (4- (4- (4- (4- (1- (ethylsulfonyl) -1H-indol-3-yl) -5- (trifluoromethyl) pyrimidin-2-amino) phenyl) piperazin-1-yl) ethoxy) acetate (A-4):
referring to the synthetic method of A-2, 58% yield was obtained. 1 H NMR (400 MHz, Chloroform-d) δ 8.71 (s, 1H), 8.05 (d,J= 8.2 Hz, 1H), 7.96 (d,J= 8.3 Hz, 1H), 7.91 (s, 1H), 7.50 (d,J= 8.9 Hz, 2H), 7.47 – 7.40 (m, 1H), 7.36 (s, 2H), 6.92 (d,J= 8.4 Hz, 2H), 4.04 (s, 2H), 3.76 – 3.64 (m, 6H), 3.40 (q,J= 7.4 Hz, 2H), 3.20 (t,J= 4.9 Hz, 4H), 2.69 (d,J= 5.8 Hz, 6H), 1.48 (s, 9H), 1.25 (t,J= 7.4 Hz, 3H)。
(5) Synthesis of 2-bromo-N- (2, 6-dioxopiperidin-3-yl) -1-oxoisoindolin-4-yl) acetamide (B-1):
in the future, lenalidomide (1 eq, 5 mmol) was dissolved in DMF of 20 mL, bromoacetyl bromide (3 eq, 15 mmol) was slowly added dropwise under ice bath, after the reaction was completed, 50 mL water was added, and the solid precipitated, suction filtration and drying gave B-1 as a white solid in 75% yield. 1 H NMR (400 MHz, DMSO-d 6 ) δ 11.07 (s, 1H), 10.30 (s, 1H), 7.85 (dd,J= 1.5, 7.5 Hz, 1H), 7.62 – 7.44 (m, 2H), 5.17 (dd,J= 5.1, 13.3 Hz, 1H), 4.44 – 4.29 (m, 2H), 4.12 (s, 2H), 3.02 – 2.84 (m, 1H), 2.61 (dt,J= 3.4, 17.2 Hz, 1H), 2.35 (qd,J= 4.4, 13.2 Hz, 1H), 2.09 – 2.00 (m, 1H)。
(6) Synthesis of 5-bromo-N- (2, 6-dioxopiperidin-3-yl) -1-oxoisoindolin-4-yl) pentanamide (B-2):
the lenalidomide (1 eq, 5 mmol) was dissolved in 20 mL DMF, triethylamine (3 eq, 15 mmol) was added, 5-bromopentanoyl chloride (1.5 eq, 7.5 mmol) was slowly added dropwise under ice bath, reacted at room temperature for 5h, after the reaction was completed, the reaction mixture was concentrated and the liquid phase was rapidly prepared to give B-3 as a white solid in 70% yield. 1 H NMR (400 MHz, DMSO-d 6 ) δ 11.02 (s, 1H), 9.80 (s, 1H), 7.81 (dd,J= 2.0, 7.0 Hz, 1H), 7.55 – 7.44 (m, 2H), 5.15 (dd,J= 5.1, 13.3 Hz, 1H), 4.47 – 4.28 (m, 2H), 3.57 (t,J= 6.5 Hz, 2H), 2.92 (ddd,J= 5.4, 13.6, 17.3 Hz, 1H), 2.71 – 2.54 (m, 1H), 2.45 – 2.25 (m, 3H), 2.10 – 1.98 (m, 1H), 1.91 – 1.63 (m, 4H)。
(7) Synthesis of 6-bromo-N- (2, 6-dioxopiperidin-3-yl) -1-oxoisoindolin-4-yl) hexanamide (B-3):
referring to the synthesis of B-2, a white solid was obtained in 73% yield. 1 H NMR (400 MHz, DMSO-d 6 ) δ 11.02 (s, 1H), 9.79 (s, 1H), 7.81 (dd,J= 1.9, 7.1 Hz, 1H), 7.61 – 7.36 (m, 2H), 5.15 (dd,J= 5.1, 13.3 Hz, 1H), 4.48 – 4.27 (m, 2H), 3.73 – 3.49 (m, 2H), 2.92 (ddd,J= 5.3, 13.6, 17.3 Hz, 1H), 2.61 (dt,J= 3.5, 17.2 Hz, 1H), 2.42 – 2.27 (m, 3H), 2.09 – 1.97 (m, 1H), 1.87 – 1.69 (m, 2H), 1.64 (p,J= 7.4 Hz, 2H), 1.52 – 1.37 (m, 2H)。
(8) Synthesis of 7-bromo-N- (2, 6-dioxopiperidin-3-yl) -1-oxoisoindolin-4-yl) heptanamide (B-4):
referring to the synthesis of B-2, a white solid was obtained in 76% yield. 1 H NMR (400 MHz, DMSO-d 6 ) δ 11.02 (s, 1H), 9.77 (s, 1H), 7.81 (dd,J= 2.0, 6.9 Hz, 1H), 7.56 – 7.43 (m, 2H), 5.15 (dd,J= 5.1, 13.3 Hz, 1H), 4.45 – 4.28 (m, 2H), 3.64 (t,J= 6.6 Hz, 2H), 2.98 – 2.84 (m, 1H), 2.70 – 2.56 (m, 1H), 2.40 – 2.27 (m, 3H), 2.10 – 1.99 (m, 1H), 1.78 – 1.56 (m, 4H), 1.53 – 1.29 (m, 4H)。
(9) Synthesis of 8-bromo-N- (2, 6-dioxopiperidin-3-yl) -1-oxoisoindolin-4-yl) octanamide (B-5):
referring to the synthesis of B-2, a white solid was obtained in 72% yield. 1 H NMR (400 MHz, DMSO-d 6 ) δ 11.02 (s, 1H), 9.76 (s, 1H), 7.81 (dt,J= 1.4, 6.9 Hz, 1H), 7.54 – 7.44 (m, 2H), 5.15 (dd,J= 5.1, 13.3 Hz, 1H), 4.45 – 4.27 (m, 2H), 3.53 (t,J= 6.7 Hz, 2H), 2.92 (ddd,J= 5.4, 13.5, 17.2 Hz, 1H), 2.70 – 2.56 (m, 1H), 2.43 – 2.28 (m, 3H), 2.10 – 1.97 (m, 1H), 1.80 (p,J= 6.8 Hz, 2H), 1.66 – 1.54 (m, 2H), 1.45 – 1.30 (m, 6H)。
(10) Synthesis of 9-bromo-N- (2, 6-dioxopiperidin-3-yl) -1-oxoisoindolin-4-yl) nonanamide (B-6):
referring to the synthesis of B-2, a white solid was obtained in 71% yield. 1 H NMR (400 MHz, DMSO-d 6 ) δ 11.02 (s, 1H), 9.75 (s, 1H), 7.81 (dd,J= 2.0, 6.9 Hz, 1H), 7.60 – 7.42 (m, 2H), 5.15 (dd,J= 5.1, 13.3 Hz, 1H), 4.44 – 4.26 (m, 2H), 3.66 – 3.47 (m, 2H), 2.98 – 2.85 (m, 1H), 2.71 – 2.55 (m, 1H), 2.41 – 2.26 (m, 3H), 2.12 – 1.96 (m, 1H), 1.91 – 1.55 (m, 4H), 1.42 – 1.27 (m, 8H)。
(11) Synthesis of 10-bromo-N- (2, 6-dioxopiperidin-3-yl) -1-oxoisoindolin-4-yl) decanoamide (B-7):
referring to the synthesis of B-2, a white solid was obtained in 71% yield. 1 H NMR (400 MHz, DMSO-d 6 ) δ 11.02 (s, 1H), 9.75 (s, 1H), 7.81 (dd,J= 2.1, 6.9 Hz, 1H), 7.55 – 7.43 (m, 2H), 5.15 (dd,J= 5.1, 13.3 Hz, 1H), 4.44 – 4.28 (m, 2H), 3.52 (t,J= 6.7 Hz, 2H), 2.99 – 2.84 (m, 1H), 2.72 – 2.56 (m, 1H), 2.34 (h,J= 8.2, 8.8 Hz, 3H), 2.10 – 2.00 (m, 1H), 1.78 (p,J= 6.8 Hz, 2H), 1.60 (t,J= 7.3 Hz, 2H), 1.43 – 1.26 (m, 10H)。
(12) Synthesis of 11-bromo-N- (2, 6-dioxopiperidin-3-yl) -1-oxoisoindolin-4-yl) undecanamide (B-8):
referring to the synthesis of B-2, a white solid was obtained in 75% yield. 1 H NMR (400 MHz, DMSO-d 6 ) δ 11.02 (s, 1H), 9.75 (s, 1H), 7.81 (dd,J= 2.0, 6.9 Hz, 1H), 7.59 – 7.39 (m, 2H), 5.15 (dd,J= 5.1, 13.3 Hz, 1H), 4.48 – 4.27 (m, 2H), 3.51 (t,J= 6.7 Hz, 2H), 2.92 (ddd,J= 5.4, 13.6, 17.3 Hz, 1H), 2.73 – 2.55 (m, 1H), 2.35 (t,J= 7.3 Hz, 3H), 2.03 (s, 1H), 1.78 (p,J= 6.8 Hz, 2H), 1.65 – 1.56 (m, 2H), 1.40 – 1.26 (m, 12H)。
(13) Synthesis of 4- (2, 6-dioxopiperidin-3-yl) -1-oxoisoindolin-4-ylamino) -4-oxobutanoic acid (B-9):
the lenalidomide (1 eq, 5 mmol) and the succinic anhydride (1.2 eq, 6 mmol) are dissolved in toluene of 20 mL and heated to reflux, after the reaction is completed, the solid is separated out, filtered by suction, and dried to obtain intermediate B-9 as a white solid with a yield of 81%. 1 H NMR (400 MHz, DMSO-d 6 ) δ 12.18 (s, 1H), 11.02 (s, 1H), 9.87 (s, 1H), 7.82 (dd,J= 2.3, 6.7 Hz, 1H), 7.55 – 7.43 (m, 2H), 5.15 (dd,J= 5.1, 13.3 Hz, 1H), 4.46 – 4.25 (m, 2H), 3.01 – 2.86 (m, 1H), 2.66 – 2.52 (m, 5H), 2.42 – 2.27 (m, 1H), 2.08 – 1.96 (m, 1H)。
(14) Synthesis of 5- (2, 6-dioxopiperidin-3-yl) -1-oxoisoindolin-4-ylamino) -5-oxopentanoic acid (B-10):
referring to the synthesis of B-9, a white solid was obtained in 79% yield. 1 H NMR (400 MHz, DMSO-d 6 ) δ 12.15 (s, 1H), 11.03 (s, 1H), 9.81 (s, 1H), 7.82 – 7.80 (m, 1H), 7.48 (q,J= 6.9 Hz, 2H), 5.17 – 5.12 (m, 1H), 4.47 – 4.24 (m, 2H), 2.89 (t,J= 5.8 Hz, 1H), 2.62 (bs, 1H), 2.40 (t,J= 7.3 Hz, 3H), 2.29 (t,J= 6.9 Hz, 2H), 2.05 – 2.04 (m, 1H), 1.86 – 1.80 (m, 2H)。
(15) Synthesis of tert-butyl 6- (2, 6-dioxopiperidin-3-yl) -1-oxoisoindolin-4-ylamino) -6-oxohexanoate (B-11):
the lenalidomide (1 eq, 5 mmol) and 6- (tert-butoxy) -6-oxohexanoic acid (1.2 eq, 6 mmol) were dissolved in 20 mL DMF and N, N-diisopropylethylamine (3 eq, 15 mmol) and HATU (1.5 eq, 7.5 mmol) were added and reacted at room temperature for 12 hours after completion of the reaction mixture was concentrated and the liquid phase was purified rapidly to give B-11 in 65% yield. 1 H NMR (400 MHz, DMSO-d 6 ) δ 11.06 (s, 1H), 9.81 (s, 1H), 7.81 (dd,J= 1.9, 7.1 Hz, 1H), 7.56 – 7.44 (m, 2H), 5.16 (dd,J= 5.1, 13.3 Hz, 1H), 4.46 – 4.27 (m, 2H), 2.93 (ddd,J= 5.4, 13.6, 17.3 Hz, 1H), 2.67 – 2.55 (m, 1H), 2.43 – 2.29 (m, 3H), 2.23 (t,J= 7.0 Hz, 2H), 2.03 (dtd,J= 1.9, 4.2, 4.6, 12.4 Hz, 1H), 1.65 – 1.47 (m, 4H), 1.39 (s, 9H)。
(16) Synthesis of 7- ((2- (2, 6-dioxopiperidin-3-yl) -1-oxoisoindolin-4-yl) amino) -7-oxoheptanoic acid tert-butyl ester (B-12):
referring to the synthesis of B-11, the yield was 63%. 1 H NMR (300 MHz, DMSO-d 6 ) δ 11.04 (s, 1H), 9.78 (s, 1H), 7.81 (dd,J= 2.4, 6.6 Hz, 1H), 7.57 – 7.42 (m, 2H), 5.15 (dd,J= 5.1, 13.2 Hz, 1H), 4.45 – 4.25 (m, 2H), 2.93 (ddd,J= 5.3, 13.4, 17.9 Hz, 1H), 2.61 (d,J= 16.9 Hz, 1H), 2.45 – 2.27 (m, 3H), 2.20 (t,J= 7.2 Hz, 2H), 2.10 – 1.96 (m, 1H), 1.65 – 1.48 (m, 4H), 1.38 (s, 11H)。
(17) 8- ((2- (2, 6-dioxy)Synthesis of tert-butyl substituted piperidin-3-yl) -1-oxoisoindolin-4-yl amino) -8-oxooctanoate (B-13): referring to the synthesis of B-11, the yield was 69%. 1 H NMR (400 MHz, DMSO-d 6 ) δ 11.04 (s, 1H), 9.78 (s, 1H), 7.81 (dd,J= 2.0, 7.0 Hz, 1H), 7.54 – 7.46 (m, 2H), 5.15 (dd,J= 5.1, 13.3 Hz, 1H), 4.43 – 4.29 (m, 2H), 2.92 (ddd,J= 5.3, 13.6, 17.2 Hz, 1H), 2.66 – 2.57 (m, 1H), 2.43 – 2.28 (m, 3H), 2.18 (t,J= 7.3 Hz, 2H), 2.03 (dtd,J= 2.6, 5.5, 6.4, 13.8 Hz, 1H), 1.64 – 1.45 (m, 4H), 1.39 (s, 9H), 1.37 – 1.27 (m, 4H)。
Example 2
Synthesis of Compound C-1-C-16
Synthesis of N- (2, 6-dioxopiperidin-3-yl) -1-oxoisoindolin-4-yl) -2- (4- (4- (1-ethylsulfonyl) -1H-indol-3-yl) -5- (trifluoromethyl) pyrimidin-2-yl) amino) phenyl) piperazin-1-yl) acetamide (C-1):
a-1 (1 eq, 1 mmol) and B-1 (1 eq, 1.2 mmol) were dissolved in 10 mL DMF and N, N-diisopropylethylamine (3 eq, 15 mmol) was added to react at 80℃12 h, after which the reaction mixture was concentrated and the liquid phase was prepared rapidly to give C-1 as a pale yellow solid in 61% yield. 1 H NMR (400 MHz, DMSO-d 6 ) δ 11.02 (s, 1H), 10.14 (s, 1H), 9.79 (s, 1H), 8.84 (s, 1H), 8.01 (s, 1H), 7.94 – 7.87 (m, 2H), 7.81 (dd,J= 1.5, 7.4 Hz, 1H), 7.62 – 7.47 (m, 5H), 7.39 (s, 1H), 6.92 (s, 2H), 5.13 (dd,J= 5.1, 13.3 Hz, 1H), 4.53 – 4.32 (m, 2H), 3.76 (q,J= 7.2 Hz, 2H), 3.31 – 3.10 (m, 6H), 2.98 – 2.85 (m, 1H), 2.79 – 2.54 (m, 5H), 2.46 – 2.33 (m, 1H), 2.06 – 1.97 (m, 1H), 1.07 (t,J= 7.2 Hz, 3H)。
Synthesis of N- (2, 6-dioxopiperidin-3-yl) -1-oxoisoindolin-4-yl) -3- (4- (4- (4- (1-ethylsulfonyl) -1H-indol-3-yl) -5- (trifluoromethyl) pyrimidin-2-yl) amino) phenyl) piperazin-1-yl) propionamide (C-2):
a-2 (1 eq, 1 mmol) was dissolved in 10 mL DCM/TFA at room temperature1:1) and after 3h of reaction, the solvent was removed by evaporation under reduced pressure. The residue was dissolved in 10 mL DMF, lenalidomide (1.2 eq, 1.2 mmol), HATU (1.5 eq, 1.5 mmol) and DIPEA (3 eq, 3 mmol) were added, reacted at room temperature 10 h, after the reaction was completed, the reaction mixture was concentrated and the rapid preparative liquid phase was purified to give C-2 as a pale yellow solid in 65% yield. 1 H NMR (400 MHz, DMSO-d 6 ) δ 11.02 (s, 1H), 10.14 (s, 1H), 10.02 (s, 1H), 8.84 (s, 1H), 8.09 – 7.85 (m, 4H), 7.76 – 7.31 (m, 6H), 6.90 (s, 2H), 5.20 – 5.07 (m, 1H), 4.46 – 4.30 (m, 2H), 3.76 (q,J= 7.2 Hz, 2H), 3.11 (s, 4H), 2.93 – 2.82 (m, 1H), 2.77 – 2.68 (m, 2H), 2.56 (d,J= 27.9 Hz, 7H), 2.37 – 2.14 (m, 1H), 2.03 – 1.94 (m, 1H), 1.07 (t,J= 7.2 Hz, 3H)。
Synthesis of N- (2, 6-dioxopiperidin-3-yl) -1-oxoisoindolin-4-yl) -4- (4- (4- (1-ethylsulfonyl) -1H-indol-3-yl) -5- (trifluoromethyl) pyrimidin-2-yl) amino) phenyl) piperazin-1-yl) butanamide (C-3:
referring to the synthesis of C-2, a pale yellow solid was obtained in 64% yield. 1 H NMR (400 MHz, Chloroform-d) δ 9.76 (s, 1H), 9.20 (s, 1H), 8.70 (s, 1H), 8.02 (d,J= 8.0 Hz, 1H), 7.96 – 7.82 (m, 3H), 7.65 (t,J= 8.5 Hz, 2H), 7.50 – 7.37 (m, 4H), 7.31 (t,J= 7.8 Hz, 1H), 6.83 (d,J= 8.4 Hz, 2H), 4.97 (dd,J= 5.4, 12.8 Hz, 1H), 4.48 – 4.26 (m, 2H), 3.38 (q,J= 7.3 Hz, 2H), 3.14 (s, 4H), 2.74 – 2.49 (m, 10H), 2.16 – 1.92 (m, 4H), 1.23 (t,J= 7.4 Hz, 3H)。
Synthesis of N- (2, 6-dioxopiperidin-3-yl) -1-oxoisoindolin-4-yl) -5- (4- (4- (1-ethylsulfonyl) -1H-indol-3-yl) -5- (trifluoromethyl) pyrimidin-2-yl) amino) phenyl) piperazin-1-yl) pentanamide (C-4):
referring to the synthesis of C-1, a pale yellow solid was obtained in 66% yield. 1 H NMR (400 MHz, DMSO-d 6 ) δ 11.03 (s, 1H), 10.15 (s, 1H), 9.82 (s, 1H), 8.84 (s, 1H), 8.07 – 7.82 (m, 4H), 7.67 – 7.32 (m, 6H), 6.92 (s, 2H), 5.16 (dd,J= 5.1, 13.3 Hz, 1H), 4.47 – 4.31 (m, 2H), 3.76 (q,J= 7.2 Hz, 2H), 3.27 – 2.79 (m, 6H), 2.67 – 2.55 (m, 2H), 2.49 – 2.21 (m, 5H), 2.08 – 1.99 (m, 1H), 1.65 (s, 4H), 1.47 – 1.10 (m, 2H), 1.08 (t,J= 7.2 Hz, 3H)。
Synthesis of N- (2, 6-dioxopiperidin-3-yl) -1-oxoisoindolin-4-yl) -6- (4- (4- (1-ethylsulfonyl) -1H-indol-3-yl) -5- (trifluoromethyl) pyrimidin-2-amino) phenyl) piperazin-1-yl) hexanamide (C-5):
referring to the synthesis of C-1, as a pale yellow solid, yield 69%. 1 H NMR (400 MHz, DMSO-d 6 ) δ 11.05 (s, 1H), 10.19 (s, 1H), 9.84 (s, 1H), 8.85 (s, 1H), 8.09 – 7.82 (m, 4H), 7.62 (d,J= 8.5 Hz, 2H), 7.54 – 7.46 (m, 3H), 7.39 (t,J= 7.8 Hz, 1H), 6.95 (s, 2H), 5.17 (dd,J= 5.2, 13.3 Hz, 1H), 4.48 – 4.28 (m, 2H), 3.77 (q,J= 7.2 Hz, 2H), 3.22 – 2.57 (m, 9H), 2.49 – 2.26 (m, 4H), 2.08 – 1.99 (m, 1H), 1.66 (p,J= 7.5 Hz, 4H), 1.42 – 1.32 (m, 2H), 1.29 – 1.20 (m, 2H), 1.08 (t,J= 7.2 Hz, 3H)。
Synthesis of N- (2, 6-dioxopiperidin-3-yl) -1-oxoisoindolin-4-yl) -7- (4- (4- (1-ethylsulfonyl) -1H-indol-3-yl) -5- (trifluoromethyl) pyrimidin-2-yl) amino) phenyl) piperazin-1-yl) heptanamide (C-6):
referring to the synthesis of C-1, a pale yellow solid was obtained in 65% yield. 1 H NMR (400 MHz, DMSO-d 6 ) δ 11.03 (s, 1H), 10.12 (s, 1H), 9.77 (s, 1H), 8.83 (s, 1H), 8.01 (s, 1H), 7.92 (d,J= 8.4 Hz, 1H), 7.88 (s, 1H), 7.81 (dd,J= 1.9, 7.1 Hz, 1H), 7.57 (d,J= 8.6 Hz, 2H), 7.53 – 7.46 (m, 3H), 7.38 (t,J= 6.0 Hz, 1H), 6.88 (s, 2H), 5.15 (dd,J= 5.1, 13.3 Hz, 1H), 4.44 – 4.29 (m, 2H), 3.76 (q,J= 7.2 Hz, 2H), 3.16 – 2.85 (m, 5H), 2.67 – 2.58 (m, 1H), 2.48 (s, 4H), 2.41 – 2.27 (m, 5H), 2.07 – 1.99 (m, 1H), 1.62 (p,J= 7.3, 7.8 Hz, 2H), 1.47 (t,J= 8.0 Hz, 2H), 1.38 – 1.29 (m, 4H), 1.07 (t,J= 7.2 Hz, 3H)。
Synthesis of N- (2, 6-dioxopiperidin-3-yl) -1-oxoisoindolin-4-yl) -8- (4- (4- (4- (1-ethylsulfonyl) -1H-indol-3-yl) -5- (trifluoromethyl) pyrimidin-2-yl) amino) phenyl) piperazin-1-yl) octanamide (C-7):
referring to the synthesis of C-1, a pale yellow solid was obtained in 60% yield. 1 H NMR (400 MHz, DMSO-d 6 ) δ 11.04 (s, 1H), 10.13 (s, 1H), 9.78 (s, 1H), 8.84 (s, 1H), 8.01 (s, 1H), 7.96 – 7.87 (m, 2H), 7.82 (dd,J= 1.9, 7.1 Hz, 1H), 7.58 (d,J= 8.5 Hz, 2H), 7.54 – 7.45 (m, 3H), 7.44 – 7.31 (m, 1H), 6.90 (s, 2H), 5.16 (dd,J= 5.1, 13.3 Hz, 1H), 4.49 – 4.28 (m, 2H), 3.76 (q,J= 7.2 Hz, 2H), 3.18 – 2.86 (m, 5H), 2.69 – 2.57 (m, 2H), 2.45 – 2.24 (m, 5H), 2.06 – 1.99 (m, 1H), 1.62 (t,J= 7.1 Hz, 2H), 1.49 (s, 2H), 1.37 – 1.23 (m, 9H), 1.08 (t,J= 7.2 Hz, 3H)。
Synthesis of N- (2, 6-dioxopiperidin-3-yl) -1-oxoisoindolin-4-yl) -9- (4- (4- (1-ethylsulfonyl) -1H-indol-3-yl) -5- (trifluoromethyl) pyrimidin-2-yl) amino) phenyl) piperazin-1-yl) nonanamide (C-8):
referring to the synthesis of C-1, a pale yellow solid was obtained in 68% yield. 1 H NMR (400 MHz, Chloroform-d) δ 8.71 (s, 1H), 8.02 (s, 1H), 7.95 (d,J= 8.3 Hz, 1H), 7.88 (d,J= 13.6 Hz, 2H), 7.71 – 7.60 (m, 3H), 7.51 – 7.40 (m, 4H), 7.33 (t,J= 7.1 Hz, 1H), 6.89 (d,J= 8.5 Hz, 2H), 5.11 (dd,J= 5.5, 13.0 Hz, 1H), 4.35 (q,J= 16.6 Hz, 2H), 3.39 (q,J= 7.3 Hz, 2H), 3.19 (s, 4H), 2.82 – 2.61 (m, 6H), 2.41 (tq,J= 7.3, 12.2 Hz, 4H), 2.23 – 2.09 (m, 2H), 1.77 – 1.68 (m, 2H), 1.58 – 1.47 (m, 2H), 1.38 – 1.29 (m, 8H), 1.23 (t,J= 7.4 Hz, 3H)。
Synthesis of N- (2, 6-dioxopiperidin-3-yl) -1-oxoisoindolin-4-yl) -10- (4- (4- (1-ethylsulfonyl) -1H-indol-3-yl) -5- (trifluoromethyl) pyrimidin-2-amino) phenyl) piperazin-1-yl) decanoamide (C-9):
referring to the synthesis of C-1, a pale yellow solid was obtained in 71% yield. 1 H NMR (400 MHz, DMSO-d 6 ) δ 11.04 (s, 1H), 10.16 (s, 1H), 9.78 (s, 1H), 8.84 (s, 1H), 8.07 – 7.86 (m, 3H), 7.81 (dd,J= 1.9, 7.1 Hz, 1H), 7.60 (d,J= 8.5 Hz, 2H), 7.54 – 7.46 (m, 3H), 7.38 (t,J= 7.3 Hz, 1H), 6.93 (s, 2H), 5.16 (dd,J= 5.1, 13.3 Hz, 1H), 4.47 – 4.27 (m, 2H), 3.76 (q,J= 7.2 Hz, 2H), 3.28 – 2.82 (m, 6H), 2.70 – 2.52 (m, 2H), 2.43 – 2.16 (m, 4H), 2.07 – 1.99 (m, 1H), 1.67 – 1.42 (m, 4H), 1.34 – 1.20 (m, 13H), 1.07 (t,J= 7.2 Hz, 3H)。
Synthesis of N- (2, 6-dioxopiperidin-3-yl) -1-oxoisoindolin-4-yl) -11- (4- (4- (1-ethylsulfonyl) -1H-indol-3-yl) -5- (trifluoromethyl) pyrimidin-2-yl) amino) phenyl) piperazin-1-yl) undecanamide (C-10):
referring to the synthesis of C-1, a pale yellow solid was obtained in 67% yield. 1 H NMR (400 MHz, Chloroform-d) δ 9.84 (s, 1H), 8.71 (s, 1H), 8.03 (d,J= 8.0 Hz, 1H), 7.95 (d,J= 8.3 Hz, 1H), 7.90 (s, 1H), 7.69 (t,J= 8.8 Hz, 2H), 7.63 – 7.55 (m, 2H), 7.51 – 7.40 (m, 4H), 7.33 (t,J= 7.6 Hz, 1H), 6.89 (d,J= 8.5 Hz, 2H), 5.14 (dd,J= 5.3, 13.1 Hz, 1H), 4.45 – 4.29 (m, 2H), 3.39 (q,J= 7.3 Hz, 2H), 3.18 (t,J= 4.9 Hz, 4H), 2.86 – 2.61 (m, 6H), 2.45 – 2.36 (m, 4H), 2.26 – 2.12 (m, 2H), 1.76 – 1.67 (m, 2H), 1.52 (t,J= 7.4 Hz, 2H), 1.33 – 1.21 (m, 15H)。
Synthesis of N- (2, 6-dioxopiperidin-3-yl) -1-oxoisoindolin-4-yl) -4- (4- (4- (1-ethylsulfonyl) -1H-indol-3-yl) -5- (trifluoromethyl) pyrimidin-2-yl) amino) phenyl) piperazin-1-yl) -4-oxobutanamide (C-11):
a-1 (1 eq, 1 mmol) and B-9 (1.2 eq, 1.2 mmol) were dissolved in 10 mL of DMF at room temperature, HATU (1.5 eq, 1.5 mmol) and DIPEA (3 eq, 3 mmol) were added,after the reaction is completed, the reaction mixture is concentrated, and the liquid phase is rapidly prepared and purified to obtain C-2 as a pale yellow solid with the yield of 60 percent. 1 H NMR (400 MHz, DMSO-d 6 ) δ 11.05 (s, 1H), 10.17 (s, 1H), 9.88 (s, 1H), 8.85 (s, 1H), 8.01 (s, 1H), 7.95 – 7.82 (m, 3H), 7.61 (d,J= 8.6 Hz, 2H), 7.53 – 7.45 (m, 3H), 7.40 (d,J= 8.0 Hz, 1H), 6.94 (s, 2H), 5.16 (dd,J= 5.1, 13.3 Hz, 1H), 4.45 – 4.29 (m, 2H), 3.76 (q,J= 7.2 Hz, 2H), 3.62 (s, 4H), 3.08 (d,J= 32.7 Hz, 4H), 2.98 – 2.87 (m, 1H), 2.75 – 2.57 (m, 5H), 2.40 – 2.26 (m, 1H), 2.07 – 1.99 (m, 1H), 1.08 (t,J= 7.2 Hz, 3H)。
Synthesis of N- (2, 6-dioxopiperidin-3-yl) -1-oxoisoindolin-4-yl) -5- (4- (4- (1-ethylsulfonyl) -1H-indol-3-yl) -5- (trifluoromethyl) pyrimidin-2-yl) amino) phenyl) piperazin-1-yl) -5-oxopentanamide (C-12):
referring to the synthesis of C-11, a pale yellow solid was obtained in 64% yield. 1 H NMR (400 MHz, DMSO-d 6 ) δ 11.02 (s, 1H), 10.16 (s, 1H), 9.81 (s, 1H), 8.84 (s, 1H), 8.09 – 7.96 (m, 1H), 7.92 (d,J= 8.4 Hz, 1H), 7.88 (s, 1H), 7.83 (dd,J= 1.9, 7.1 Hz, 1H), 7.60 (d,J= 8.6 Hz, 2H), 7.56 – 7.44 (m, 3H), 7.38 (t,J= 7.7 Hz, 1H), 6.93 (s, 2H), 5.15 (dd,J= 5.1, 13.3 Hz, 1H), 4.46 – 4.30 (m, 2H), 3.76 (q,J= 7.2 Hz, 2H), 3.60 (s, 4H), 3.17 – 3.00 (m, 4H), 2.97 – 2.86 (m, 1H), 2.68 – 2.55 (m, 1H), 2.47 – 2.28 (m, 5H), 2.06 – 1.97 (m, 1H), 1.91 – 1.79 (m, 2H), 1.08 (t,J= 7.2 Hz, 3H)。
Synthesis of N- (2, 6-dioxopiperidin-3-yl) -1-oxoisoindolin-4-yl) -6- (4- (4- (4- (1-ethylsulfonyl) -1H-indol-3-yl) -5- (trifluoromethyl) pyrimidin-2-yl) amino) phenyl) piperazin-1-yl) -6-oxohexanamide (C-13):
b-11 (1 eq, 1 mmol) was dissolved in a mixed solvent of DCM/TFA (1:1) 10 mL at room temperature and after 3h the solvent was removed by evaporation under reduced pressure. The residue was dissolved in 10To mL of DMF was added A-1 (0.9 eq, 0.9 mmol), HATU (1.5 eq, 1.5 mmol) and DIPEA (3 eq, 3 mmol), reacted at room temperature for 10 h, after the reaction was completed, the reaction mixture was concentrated and the rapid preparative liquid phase was purified to give C-13 as a pale yellow solid in 58% yield. 1 H NMR (400 MHz, DMSO-d 6 ) δ 11.02 (s, 1H), 10.15 (s, 1H), 9.80 (s, 1H), 8.84 (s, 1H), 8.01 (s, 1H), 7.95 – 7.86 (m, 2H), 7.80 (dd,J= 1.7, 7.3 Hz, 1H), 7.60 (d,J= 8.7 Hz, 2H), 7.55 – 7.46 (m, 3H), 7.39 (t,J= 7.3 Hz, 1H), 6.92 (s, 2H), 5.15 (dd,J= 5.1, 13.3 Hz, 1H), 4.45 – 4.29 (m, 2H), 3.76 (q,J= 7.2 Hz, 2H), 3.59 (s, 4H), 3.13 – 2.87 (m, 5H), 2.70 – 2.56 (m, 1H), 2.44 – 2.33 (m, 5H), 2.07 – 1.99 (m, 1H), 1.70 – 1.54 (m, 4H), 1.08 (t,J= 7.2 Hz, 3H)。
Synthesis of N- (2, 6-dioxopiperidin-3-yl) -1-oxoisoindolin-4-yl) -7- (4- (4- (1-ethylsulfonyl) -1H-indol-3-yl) -5- (trifluoromethyl) pyrimidin-2-yl) amino) phenyl) piperazin-1-yl) -7-oxoheptanamide (C-14):
referring to the synthesis of C-13, a pale yellow solid was obtained in 60% yield. 1 H NMR (400 MHz, DMSO-d 6 ) δ 11.02 (s, 1H), 10.15 (s, 1H), 9.77 (s, 1H), 8.84 (s, 1H), 8.01 (s, 1H), 7.92 (d,J= 8.4 Hz, 1H), 7.88 (s, 1H), 7.81 (dd,J= 1.7, 7.3 Hz, 1H), 7.60 (d,J= 8.6 Hz, 2H), 7.56 – 7.44 (m, 3H), 7.43 – 7.32 (m, 1H), 6.91 (s, 2H), 5.15 (dd,J= 5.1, 13.3 Hz, 1H), 4.45 – 4.30 (m, 2H), 3.76 (q,J= 7.2 Hz, 2H), 3.58 (s, 4H), 3.15 – 2.86 (m, 5H), 2.69 – 2.57 (m, 1H), 2.36 (t,J= 7.4 Hz, 5H), 2.08 – 1.98 (m, 1H), 1.70 – 1.50 (m, 4H), 1.40 – 1.32 (m, 2H), 1.07 (t,J= 7.2 Hz, 3H)。
Synthesis of N- (2, 6-dioxopiperidin-3-yl) -1-oxoisoindolin-4-yl) -8- (4- (4- (4- (1-ethylsulfonyl) -1H-indol-3-yl) -5- (trifluoromethyl) pyrimidin-2-yl) amino) phenyl) piperazin-1-yl) -8-oxooctanamide (C-15):
referring to the synthesis method of C-13, light yellowColor solid, yield 61%. 1 H NMR (400 MHz, Chloroform-d) δ 9.51 (s, 1H), 8.71 (s, 1H), 8.57 (s, 1H), 8.03 (d,J= 8.0 Hz, 1H), 7.97 – 7.83 (m, 3H), 7.66 (t,J= 8.7 Hz, 2H), 7.53 (d,J= 8.4 Hz, 2H), 7.42 (t,J= 7.8 Hz, 2H), 7.32 (t,J= 7.5 Hz, 1H), 6.87 (d,J= 8.3 Hz, 2H), 5.05 (dd,J= 5.0, 13.3 Hz, 1H), 4.36 (s, 2H), 3.74 (s, 2H), 3.59 (s, 2H), 3.39 (q,J= 7.4 Hz, 2H), 3.08 (d,J= 14.8 Hz, 4H), 2.82 – 2.55 (m, 2H), 2.46 – 2.28 (m, 4H), 2.27 – 2.15 (m, 1H), 2.08 (s, 1H), 1.65 (d,J= 41.5 Hz, 4H), 1.34 (s, 4H), 1.24 (t,J= 7.4 Hz, 3H)。
Synthesis of N- (2, 6-dioxopiperidin-3-yl) -1-oxoisoindolin-4-yl) -2- (2- (2- (4- (4- (4- (1-ethylsulfonyl) -1H-indol-3-yl) -5- (trifluoromethyl) pyrimidin-2-amino) phenyl) piperazin-1-yl) ethoxy) acetamide (C-16):
referring to the synthesis of C-2, a pale yellow solid was obtained in 66% yield. 1 H NMR (400 MHz, Chloroform-d) δ 9.51 (s, 1H), 8.71 (s, 1H), 8.57 (s, 1H), 8.03 (d,J= 8.0 Hz, 1H), 7.97 – 7.83 (m, 3H), 7.66 (t,J= 8.7 Hz, 2H), 7.53 (d,J= 8.4 Hz, 2H), 7.42 (t,J= 7.8 Hz, 2H), 7.32 (t,J= 7.5 Hz, 1H), 6.87 (d,J= 8.3 Hz, 2H), 5.05 (dd,J= 5.0, 13.3 Hz, 1H), 4.36 (s, 2H), 3.74 (s, 2H), 3.59 (s, 2H), 3.39 (q,J= 7.4 Hz, 2H), 3.08 (d,J= 14.8 Hz, 4H), 2.82 – 2.55 (m, 2H), 2.46 – 2.28 (m, 4H), 2.27 – 2.15 (m, 1H), 2.08 (s, 1H), 1.65 (d,J= 41.5 Hz, 4H), 1.34 (s, 4H), 1.24 (t,J= 7.4 Hz, 3H)。
Example 3.
Biological evaluation experiment
(1) EGFR in vitro degradation Activity assay:
the ability of compounds targeted to degrade EGFR protein was evaluated using Western immunoblotting, and Western immunoblotting analysis was performed on various tumor cells, respectively.
Experimental results show that the compound C-1~C-16 prepared in the embodiment of the invention is applied to EGFR L858R/T790M/C797S Has significant degradation activity, and the optimal compound C-6 has significant degradation activity on other EGFR mutant kinases and has high selectivity on EGFR wild type protein, and the results are shown in table 1 and table 2, wherein DC of the compound 50 Classification according to description:
"A" means DC 50 A measured value of 100nM or less; "B" means DC 50 A measured value of 250 or less nM or more than 100nM; "C" means DC 50 The measured value is 500 or less and nM or more and 250 or more and nM or less. "D" means DC 50 The measured value is greater than 500 nM.
TABLE 1 compounds of the invention against EGFR L858R/T790M/C797S Is a degradation activity of (2)
TABLE 2 degradation Activity of Compound C-6 of the invention on EGFR other mutant forms
(2) In vitro cell Activity assay: determining the inhibition of the proliferation of various cancer cells by the compound according to the CCK-8 method, and obtaining the half inhibition concentration IC of the proliferation inhibition activity of the compound 50 Values. Inoculating logarithmic growth phase cells into 96-well plate at 3000-5000 cells/well, placing at 37deg.C, 5% CO 2 Culturing for 24 hours under the condition; 1. Mu.M, 0.5. Mu.M, 0.25. Mu.M, 0.1250. Mu.M, and 100. Mu.L of test compound solutions of different concentrations 62.5 nM,31.25 nM,15.63 nM,7.81 nM,3.91 nM were added to the plates, and the plates were incubated at 37℃with 5% CO 2 Incubating for 72 hours under incubator conditions; before the end of incubation, 4. 4 h, 10. Mu.L of CCK-8 solution (5. 5 mg/mL) was added to each well. After incubation, OD was measured by using an ELISA reader 450 Inhibition = (control OD value-experimental OD value)/control OD value x 100%; after data were obtained, graphPad Prism fit gave IC 50 ；
"+". ++'s representing IC 50 The measured value is not more than 250 nM.
"+". ++'s representing IC 50 The measured value is 500nM or less and more than 250 nM.
"++" means IC 50 The measured value is 1 mu M or less and 500nM or more.
"+" indicates IC 50 The measured value is greater than 1. Mu.M.
"-" means IC 50 Not measured.
TABLE 3 antiproliferative Activity of the inventive Compounds against EGFR 858R/T790M/C797S mutant cell H1975TM
(3) In vivo anti-tumor (H1975 TM (EGFR) L858R/T790M/C797S ) Activity assay):
the drug was the compound (C-6) produced in example 6; the cell strain is a human lung adenocarcinoma H1975TM cell; the test animals were SPF-grade BALB/c nude mice (Kwangsi laboratory animal Co., ltd.); a female; 6 in each group, 18 in total; the drug dosage settings are as in table 4.
Table 4, drug dosage configuration
The experimental method comprises the following steps: taking tumor in vigorous growth phase, inoculating H1975TM cells of human lung adenocarcinoma into 25 BALB/c nude mice under right armpit skin under aseptic condition, and inoculating cell amount of 5×10 6 . The diameter of the transplanted tumor is measured by a vernier caliper for the transplanted tumor of the nude mice, and the tumor grows to 85 mm 3 When the number of the tumor-bearing nude mice is about 18, the nude mice with good growth state and good tumor size uniformity are selected and divided into 3 groups, and 6 nude mice in each group are respectively administered according to a dose setting table, namely (1) a model group, (2) a low-dose group of test drugs and (3) a high-dose group of test drugs. The antitumor effect of the test substance was dynamically observed by using a method for measuring tumor diameter. The number of tumor diameter measurements wasThe nude mice were weighed once every other day while tumor diameters were measured. The nude mice were euthanized 31 days after administration, the tumor mass was removed by surgery and weighed, and the tumor inhibition rate (%) was calculated from the tumor mass weight. Tumor inhibition rate (%) = (model tumor weight-dosing tumor weight)/model tumor weight x 100%.
TABLE 5 influence of test samples on tumor growth in human lung adenocarcinoma H1975TM nude mice transplantations
The experimental results are shown in table 5: the test agent example 6 has obvious inhibition effect on tumor growth, and the inhibition effect of the high-dose group (100 mg/kg) is superior to that of the low-dose group (25 mg/kg), and the tumor inhibition rates of the high-dose group and the low-dose group are 66.4% and 48.1%, respectively. The dosing group had no obvious effect on the body weight of the animals compared to the model control group.

Claims (9)

1. A compound for targeted degradation of EGFR protein, which is characterized in that the compound is shown as a general formula (I):
；
wherein linker is a linking group, the linking group is a straight chain or branched chain alkylene chain with total length of 1-20 atoms, and the straight chain or branched chain alkylene chain is formed by-CH 2 -, -O-, -CO-; -CONH-; -S-, -N-, and one or more of alkynylene or cycloalkylene.
2. The compound of claim 1, wherein the straight or branched alkylene chain consists of-CH 2 -, -O-, -CO-; -CONH-; -one or more of S-or-N-.
3. The compound of claim 1, wherein the straight or branched alkylene chain consists of-CH 2 -, -O-, -CO-; -one or more of CONH-or-N-.
4. The compound of claim 1, wherein the compound is as shown in C-1 to C-16:
。
5. a method for preparing a compound according to claim 1, comprising the steps of: dissolving A and B in N, N-dimethylformamide, adding N, N-diisopropylethylamine or a mixed solution of N, N-diisopropylethylamine and 2- (7-azobenzotriazole) -N, N, N ', N' -tetramethylurea hexafluorophosphate, and purifying after the reaction is finished to obtain a compound for targeted degradation of EGFR protein, wherein R is bromine and carboxyl;
。
6. a process for the preparation of a compound as defined in C-2, C-3, C-16, comprising the steps of:
or (b)
Or (b)
。
7. A pharmaceutical composition comprising a compound of claim 1 or a pharmaceutically acceptable salt thereof, and a pharmaceutically acceptable carrier.
8. Use of a compound that targets degradation of EGFR protein according to claim 1, in the manufacture of a medicament for preventing or treating a tumor-related disease.
9. The use according to claim 8, wherein the tumor is lung cancer, gastric cancer, prostate cancer, ovarian cancer, testicular cancer, colon cancer, leukemia, breast cancer, multiple myeloma, liver cancer, pancreatic cancer, melanoma, glioma, brain glioma or pituitary tumor.
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Citations (4)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
CN109928956A (en) * 2019-02-27 2019-06-25 杭州偶联医药科技有限公司 A kind of compound of targeting ubiquitination degradation EGFR albumen and its pharmaceutical composition and application
CN110753693A (en) * 2016-12-23 2020-02-04 阿尔维纳斯运营股份有限公司 EGFR proteolytic targeting chimeric molecules and related methods of use
CN111574498A (en) * 2020-04-10 2020-08-25 浙江工业大学 Lenalidomide-based targeted degradation EGFR protein small molecule compound and preparation and application thereof
CN114805303A (en) * 2021-01-20 2022-07-29 海思科医药集团股份有限公司 Molecule with double functions of degrading EGFR (epidermal growth factor receptor), composition and pharmaceutical application thereof

Patent Citations (4)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
CN110753693A (en) * 2016-12-23 2020-02-04 阿尔维纳斯运营股份有限公司 EGFR proteolytic targeting chimeric molecules and related methods of use
CN109928956A (en) * 2019-02-27 2019-06-25 杭州偶联医药科技有限公司 A kind of compound of targeting ubiquitination degradation EGFR albumen and its pharmaceutical composition and application
CN111574498A (en) * 2020-04-10 2020-08-25 浙江工业大学 Lenalidomide-based targeted degradation EGFR protein small molecule compound and preparation and application thereof
CN114805303A (en) * 2021-01-20 2022-07-29 海思科医药集团股份有限公司 Molecule with double functions of degrading EGFR (epidermal growth factor receptor), composition and pharmaceutical application thereof

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