GSPT1 degrader-antibody conjugate

Abstract

Disclosed is a GSPT1 degrader-antibody conjugate, and specifically provided are a compound as represented by formula (I), a stereoisomer or a pharmaceutically acceptable salt thereof, a pharmaceutical composition containing same, and a method for using the compound in the treatment of cell proliferative disorders, such as cancer.

Description

GSPT1 degrader antibody conjugate

[0001] This application claims priority to Chinese patent application No. 2024107419077, filed on June 7, 2024, Chinese patent application No. 2024116143437, filed on November 12, 2024, Chinese patent application No. 2025101144499, filed on January 23, 2025, and Chinese patent application No. 2025107154992, filed on May 29, 2025. This application incorporates the entire contents of the above-mentioned Chinese patent applications. TECHNICAL FIELD

[0002] The present application belongs to the field of medicine, and specifically relates to a GSPT1 degrader antibody conjugate, a pharmaceutical composition containing the compound, and a method for treating cell proliferative diseases, such as cancer, using the compound of the present application. BACKGROUND

[0003] Protein mutations, expression imbalances, conformational changes, and functional abnormalities can cause many diseases. Protein synthesis and cell growth and proliferation are strictly regulated processes in space and time. Abnormal regulation of these processes can lead to uncontrolled cell growth, proliferation, and migration, resulting in diseases such as cancer, aging, and viral infection.

[0004] GSPT1 (G1 To S Phase Transition 1) is a translation termination factor that coordinates the completion of protein translation in the body, and down-regulation of GSPT1 can cause abnormal protein expression, thereby inhibiting the proliferation and survival of various tumor cells.

[0005] The antibody conjugate protein degrader ORM-6151 of Orum Therapeutics combines the dual advantages of ADC drugs and PROTAC drugs. This drug uses a CD33 monoclonal antibody as a warhead, conjugated with a GSPT1 protein degrader, so that the protein degrader can be combined with antibody-mediated precise delivery, which can not only accurately target tumor tissue and eliminate the damage of PROTAC to normal cells, but also target and degrade undruggable proteins. It is currently in the clinical stage. SUMMARY

[0006] The present disclosure provides a GSPT1 degrader, specifically, a compound according to the general formula (I), a stereoisomer thereof, or a pharmaceutically acceptable salt thereof:

[0007] wherein ring A is selected from

[0008] R 1 selected from H or halogen;

[0009] R2 -(CH2) m -O-(CH2) n -NR 2a R 2b -(CH2) m -O-C 3-6 cycloalkyl-NR 2a R 2b -(CH2) m -O-3 to 6 membered heterocyclyl, -C(O)NR 2a -(CH2) n -NR 2a R 2b -C 2-6 alkynyl-NR 2a R 2b -O-C 2-6 alkynyl-NR 2a R 2b -C 2-6 alkenyl-NR 2a R 2b -O-C 2-6 alkenyl-NR 2a R 2b -NR 2a C(O)NR 2a (CH2) n -NR 2a R 2b -3 to 6 membered heterocyclyl-(CH2) n -NR 2a R 2b , (CH2) m -SO2-(CH2) n -NR 2a R 2b -(CH2) m -SO2-(CH2) n -3 to 8 membered heterocyclyl, -(CH2) m -COO-3 to 8 membered heterocyclyl, -(CH2) m -COO-C 3-6 cycloalkyl-NR 2a R 2b -(CH2) m -O-(CH2) n -NR 2a CO-C 1-6 alkyl, and said C 1-6 alkyl is optionally further substituted by one or more substituents selected from the group consisting of OH, NH2, -CONH2, -COOH, -NHC(=NH)NH2;

[0010] m, n are each independently selected from 0, 1, 2, 3, 4 or 5;

[0011] R 2a , R 2b are each independently selected from H, C 1-4 alkyl, C 2-4 alkenyl, C 2-4 alkynyl, C 1-4 alkoxy, -C(=NH)-NH2, -C(=NH)-NHCH3, -C(=NH)-R 2h , -CO-R y , C 3-6 cycloalkyl, -CH2C(O)NH2or 5- to 12-membered heterocyclyl, said C 1-4 alkyl can be further substituted by one or more substituents selected from halogen, CN or OH;

[0012] R y is selected from C 1-6 alkyl or 5- to 12-membered heterocyclyl, and said C 1-6 alkyl or 5- to 12-membered heterocyclyl can be further substituted by one or more substituents selected from OH, NH2, -CONH2, -S-CH3, phenyl, 4- to 10-membered heterocyclyl or 7- to 10-membered heteroaryl, said phenyl can be further substituted by OH, NH2, CN or halogen;

[0013] or R 2a , R 2b together with the nitrogen atom to which they are attached form a 3- to 8-membered heterocyclyl;

[0014] R 2c , R 2d are each independently selected from H or C 1-4 alkyl;

[0015] or R 2c , R 2d together with the carbon atom to which they are attached form a C 3-6 cycloalkyl;

[0016] R 2e , R 2f are each independently selected from H or C 1-4 alkyl;

[0017] or R 2e , R 2a together with the atoms to which they are attached form a 3- to 8-membered heterocyclyl;

[0018] or R 2e , R 2b together with the atoms to which they are attached form a 3- to 8-membered heterocyclyl;

[0019] R 2h selected from H, C 1-4 alkyl, C 2-4 alkenyl or C 3-6 cycloalkyl, said C 1-4 alkyl or C 3-6 cycloalkyl can be further substituted by one or more substituents selected from halogen, CN or OH;

[0020] R 3 , R 4 and R 5 are each independently selected from H or halogen;

[0021] with the proviso that the compound of formula (I) is not

[0022] The present disclosure provides a GSPT1 degrader, which is a compound of general formula (I), a stereoisomer thereof or a pharmaceutically acceptable salt thereof:

[0023] wherein ring A is selected from

[0024] R 1 is selected from H or halogen;

[0025] R 2 is selected from -(CH2) m -O-(CH2) n -NR 2a R 2b , -(CH2) m -O-C 3-6 cycloalkyl-NR 2a R 2b , -(CH2) m -O-3 to 6 membered heterocyclyl, -C(O)NR 2a -O-(CH2) n -NR 2a R 2b , -C 2-6 alkenyl-NR 2a R 2b , -O-C 2-6 alkenyl-NR 2a R 2b , -C 2-6 alkenyl-NR 2a R 2b , -O-C 2-6 alkenyl-NR 2a R 2b , -NR 2a C(O)NR 2a (CH2) n -NR2a R 2b , -3 to 6 membered heterocyclyl-(CH2) n -NR 2a R 2b , -(CH2) m -SO2-(CH2) n -NR 2a R 2b , -(CH2) m -SO2-(CH2) n -3 to 8 membered heterocyclyl, -(CH2) m -COO-3 to 8 membered heterocyclyl, -(CH2) m -COO-C 3-6 cycloalkyl-NR 2a R 2b , -(CH2) m -O-(CH2) n -NR 2a CO-C 1-6 alkyl, and said C 1-6 alkyl is optionally further substituted by one or more substituents selected from the group consisting of OH, NH2, -CONH2, -COOH, -NHC(=NH)NH2;

[0026] m, n are each independently selected from 0, 1, 2, 3, 4 or 5;

[0027] R 2a , R 2b are each independently selected from H, C 1-4 alkyl, C 2-4 alkenyl, C 2-4 alkynyl, C 1-4 alkoxy, -C(=NH)-NH2, -C(=NH)-NHCH3, -C(=NH)-R 2h , -CO-R y , C 3-6 cycloalkyl or 5- to 12-membered heterocyclyl, said C 1-4 alkyl can be further substituted by one or more substituents selected from halogen, CN or OH;

[0028] R y is selected from C 1-6 alkyl, and said C 1-6 alkyl can be further substituted by one or more substituents selected from OH, NH2, -CONH2, -S-CH3, phenyl, 4- to 10-membered heterocyclyl or 7- to 10-membered heteroaryl, said phenyl can be further substituted by OH, NH2, CN or halogen;

[0029] or R2a R 2b together with the nitrogen atom to which they are attached form a 3- to 8-membered heterocyclyl group;

[0030] R 2c R 2d are each independently selected from H or C 1-4 alkyl;

[0031] R 2c R 2d together with the carbon atom to which they are attached form a C 3-6 cycloalkyl group;

[0032] R 2e R 2f are each independently selected from H or C 1-4 alkyl;

[0033] R 2e R 2a together with the atom to which they are attached form a 3- to 8-membered heterocyclyl group;

[0034] R 2e R 2b together with the atom to which they are attached form a 3- to 8-membered heterocyclyl group;

[0035] R 2h is selected from H, C 1-4 alkyl, C 2-4 alkynyl or C 3-6 cycloalkyl, which C 1-4 alkyl or C 3-6 cycloalkyl can be further substituted by one or more substituents selected from halogen, CN or OH;

[0036] R 3 R 4 and R 5 are each independently selected from H or halogen;

[0037] with the proviso that the compound of formula (I) is not

[0038] In some embodiments, the compound of formula (I), stereoisomer thereof, or pharmaceutically acceptable salt thereof, is selected from:

[0039] wherein ring A is selected from

[0040] R 1 is selected from H or halogen;

[0041] R 2 is selected from -(CH2) m -O-(CH2) n -NR 2a R2b -(CH2) m -O-C 3-6 cycloalkyl-NR 2a R 2b -(CH2) m -O-3 to 6 membered heterocyclyl, -C(O)NR 2a -(CH2) n -NR 2a R 2b -C 2-6 alkynyl-NR 2a R 2b -O-C 2-6 alkynyl-NR 2a R 2b -C 2-6 alkenyl-NR 2a R 2b -O-C 2-6 alkenyl-NR 2a R 2b -NR 2a C(O)NR 2a (CH2) n -NR 2a R 2b -3 to 6 membered heterocyclyl-(CH2) n -NR 2a R 2b , -(CH2) m -SO2-(CH2) n -NR 2a R 2b -(CH2) m -SO2-(CH2) n -3 to 8 membered heterocyclyl, -(CH2) m -COO-3 to 8 membered heterocyclyl, -(CH2) m -C 3-6 cycloalkyl-NR 2a R 2b -(CH2) m -O-(CH2) n -NR 2a CO-C 1-6 alkyl, and said C 1-6 alkyl is optionally further substituted by one or more substituents selected from the group consisting of OH, NH2, -CONH2, -COOH, -NHC(=NH)NH2;

[0042] m, n are each independently selected from 0, 1, 2, 3, 4 or 5;

[0043] R 2aR 2b each independently selected from H, C 1-4 alkyl, C 2-4 alkenyl, C 2-4 alkynyl, C 1-4 alkoxy, -C(=NH)-NH2, -C(=NH)-NHCH3, -C(=NH)-R 2h , C 3-6 cycloalkyl or 5-12 membered heterocyclyl, said C 1-4 alkyl can be further substituted by one or more substituents selected from halogen, CN or OH;

[0044] or R 2a , R 2b together with the nitrogen atom to which they are attached form a 3- to 8-membered heterocyclyl;

[0045] R 2c , R 2d each independently selected from H or C 1-4 alkyl;

[0046] or, R 2c , R 2d together with the carbon atom to which they are attached form a C 3-6 cycloalkyl;

[0047] R 2e , R 2f each independently selected from H or C 1-4 alkyl;

[0048] or, R 2e , R 2a together with the atom to which they are attached form a 3- to 8-membered heterocyclyl;

[0049] or, R 2e , R 2b together with the atom to which they are attached form a 3- to 8-membered heterocyclyl;

[0050] R 2h selected from H, C 1-4 alkyl, C 2-4 alkynyl or C 3-6 cycloalkyl, said C 1-4 alkyl or C 3-6 cycloalkyl can be further substituted by one or more substituents selected from halogen, CN or OH;

[0051] R 3 , R 4 and R 5 each independently selected from H or halogen;

[0052] with the proviso that the compound of formula (I) is not

[0053] In some embodiments, said ring A is selected from

[0054] R 1 is selected from H or halogen;

[0055] R 2 is selected from -(CH2) m -O-(CH2)n-NR 2a R 2b -(CH2) m -O-C 3-6 cycloalkyl-NR 2a R 2b -(CH2) m -O-3 to 6 membered heterocyclyl, -C(O)NR 2a -O-(CH2)n-NR 2a R 2b -C 2-6 alkynyl-NR 2a R 2b -O-C 2-6 alkynyl-NR 2a R 2b -C 2-6 alkenyl-NR 2a R 2b -O-C 2-6 alkenyl-NR 2a R 2b -NR 2a C(O)NR 2a (CH2)n-NR 2a R 2b -3 to 6 membered heterocyclyl-(CH2) n -NR 2a R 2b or

[0056] m, n are each independently selected from 0, 1, 2, 3, 4 or 5;

[0057] R 2a , R 2b are each independently selected from H, C 1-4 alkyl, C 2-4 alkenyl, C 2-4 alkynyl, C 1-4 alkoxy, -C(=NH)-NH2, -C(=NH)-NHCH3, C 3-6 cycloalkyl, 5-12 membered heterocyclyl, said C 1-4 alkyl can be further substituted with one or more substituents selected from halogen, CN or OH;

[0058] R2c R 2d each independently selected from H or C 1-4 alkyl;

[0059] or, R 2c R 2d together with the carbon atom to which they are attached form a C 3-6 cycloalkyl;

[0060] R 2e R 2f each independently selected from H or C 1-4 alkyl;

[0061] R 3 R 4 and R 5 each independently selected from H or halogen;

[0062] with the proviso that the compound of formula (I) is not

[0063] In some embodiments, ring A is

[0064] R 1 is halogen;

[0065] R 2 is selected from -(CH2) m -O-(CH2) n -NR 2a R 2b -C 2-6 alkynyl-NR 2a R 2b or preferably -(CH2) m -O-(CH2) n -NR 2a R 2b ;

[0066] m and n are 2;

[0067] R 2a R 2b each independently selected from H, C 1-4 alkyl, C 2-4 alkenyl, C 2-4 alkynyl, C 1-4 alkoxy, -C(=NH)-NH2, -C(=NH)-NHCH3, -CH2C(O)NH2, -CO-R y -C(=NH)-R 2h or C 3-6 cycloalkyl, said C 1-4alkyl can be further substituted by one or more substituents selected from the group consisting of halogen or CN;

[0068] R y selected from the group consisting of C 1-6 alkyl or 5- to 12-membered heterocyclyl, and said C 1-6 alkyl or 5- to 12-membered heterocyclyl can be further substituted by one or more substituents selected from the group consisting of OH, NH2, -CONH2, -S-CH3, phenyl or 7- to 10-membered heteroaryl, said phenyl can be further substituted by OH or NH2;

[0069] R 2c , R 2d are each independently selected from the group consisting of H or C 1-4 alkyl;

[0070] or, R 2c , R 2d together with the carbon atom to which they are attached form a C 3-6 cycloalkyl;

[0071] R 2e , R 2f are each independently selected from the group consisting of H or C 1-4 alkyl;

[0072] R 2h selected from the group consisting of C 1-4 alkyl or C 3-6 cycloalkyl, said C 1-4 alkyl can be further substituted by CN;

[0073] R 3 , R 4 and R 5 are each independently selected from H;

[0074] with the proviso that the compound of formula (I) is not

[0075] In some embodiments, said R 1 is halogen; preferably, R 1 is F or Cl, more preferably Cl.

[0076] In some embodiments, R 2 is selected from the group consisting of -(CH2) m -O-(CH2) n -NR 2a R 2b , -(CH2) m -O-cyclobutyl-NR 2a R 2b , -(CH2) m -O-4- to 6-membered heterocyclyl, -C(O)NR 2a-O-(CH2) n -NR 2a R 2b , -C 3-5 alkynyl-NR 2a R 2b , -O-C 3-5 alkynyl-NR 2a R 2b , -4-membered heterocyclyl-(CH2) n -NR 2a R 2b , -(CH2) m -SO2-(CH2) n -NR 2a R 2b , -(CH2) m -SO2-(CH2) n -4- to 6-membered heterocyclyl, -(CH2) m -COO-4- to 6-membered heterocyclyl, -(CH2) m -COO-C 3-6 cycloalkyl-NR 2a R 2b , -(CH2) m -O-(CH2) n -NR 2a CO-C 1- 6alkyl, and said C 1-6 alkyl is optionally further substituted by one or more substituents selected from the group consisting of OH, NH2, -CONH2, -COOH, -NHC(=NH)NH2;

[0077] m and n are each independently selected from 0, 1 or 2;

[0078] R 2a , R 2b are each independently selected from the group consisting of H, methyl, ethyl, propenyl, ethynyl, propynyl, -CH2CF3, -CH2CN, -CH2C(O)NH2, methoxy, cyclopropyl, -CO-R y , -C(=NH)-NH2, -C(=NH)-NHCH3, -C(=NH)-R 2h or 5-membered heterocyclyl;

[0079] R y is selected from C 1-6 alkyl or and said C 1-6 alkyl can be further substituted by one or more substituents selected from the group consisting of OH, NH2, -CONH2, -S-CH3, phenyl or substituted by OH, NH2, CN or halogen;

[0080] or R 2a , R 2b together with the nitrogen atom to which they are attached form a 5- to 6- membered heterocyclyl ring;

[0081] R 2c , R 2d are each independently selected from H or methyl;

[0082] or R 2c , R 2d together with the carbon atom to which they are attached form a cyclopropyl group;

[0083] R 2e , R 2f are each independently selected from H or methyl;

[0084] or R 2e , R 2a together with the atoms to which they are attached form a 5- to 6- membered heterocyclyl ring;

[0085] or R 2e , R 2b together with the atoms to which they are attached form a 5- to 6- membered heterocyclyl ring;

[0086] R 2h is selected from H, methyl, -CH2CN or cyclopropyl.

[0087] In some embodiments, R 2 is selected from -(CH2) m -O-(CH2) n -NR 2a R 2b , -(CH2) m -O-cyclobutyl-NR 2a R 2b , -(CH2) m -O-4- to 6-membered heterocyclyl, -C(O)NR 2a -O-(CH2) n -NR 2a R 2b , -C 3-5 alkynyl-NR 2a R 2b , -O-C 3-5 alkynyl-NR 2a R 2b , -4-membered heterocyclyl-(CH2) n -NR 2a R 2b , -(CH2) m -SO2-(CH2) n -NR 2a R 2b 、-(CH2) m -SO2-(CH2) n -4 to 6 membered heterocyclyl, -(CH2) m -COO-4 to 6 membered heterocyclyl, -(CH2) m -COO-C 3-6 cycloalkyl-NR 2a R 2b 、-(CH2) m -O-(CH2) n -NR 2a CO-C 1-6 alkyl, and said C 1-6 alkyl is optionally further substituted by one or more substituents selected from the group consisting of OH, NH2, -CONH2, -COOH, -NHC(=NH)NH2;

[0088] m and n are each independently selected from 0, 1 or 2;

[0089] R 2a , R 2b are each independently selected from the group consisting of H, methyl, ethyl, propenyl, ethynyl, propynyl, -CH2CF3, -CH2CN, methoxy, cyclopropyl, -CO-R y , -C(=NH)-NH2, -C(=NH)-NHCH3, -C(=NH)-R 2h or 5-membered heterocyclyl;

[0090] R y is selected from the group consisting of C 1-6 alkyl, and said C 1-6 alkyl is optionally further substituted by one or more substituents selected from the group consisting of OH, NH2, -CONH2, -S-CH3, phenyl, which phenyl is optionally further substituted by OH, NH2, CN or halogen; preferably R y is selected from the group consisting of C 1-6 alkyl, and said C 1-6 alkyl is optionally further substituted by one or more substituents selected from the group consisting of OH, NH2, -CONH2, -S-CH3, phenyl or which phenyl is optionally further substituted by OH, NH2, CN or halogen;

[0091] or R 2a , R 2b form together with the nitrogen atom to which they are attached a

[0092] R 2c , R 2d each independently is selected from H or methyl;

[0093] or, R 2c , R 2d together with the carbon atom to which they are attached form a cyclopropyl group;

[0094] R 2e , R 2f each independently is selected from H or methyl;

[0095] or, R 2e , R 2a together with the atom to which they are attached form

[0096] or, R 2e , R 2b together with the atom to which they are attached form

[0097] R 2h is selected from H, methyl, -CH2CN or cyclopropyl.

[0098] In some embodiments, the R y is selected from

[0099] In some embodiments, the R 2 is selected from -(CH2) m -O-(CH2) n -NR 2a R 2b , -(CH2) m -O-cyclobutyl-NR 2a R 2b , -(CH2) m -O-4 to 6 membered heterocyclyl, -C(O)NR 2a -O-(CH2) n -NR 2a R 2b , -C 3-5 alkynyl-NR 2a R 2b , -O-C 3-5 alkynyl-NR 2a R 2b , -4 membered heterocyclyl-(CH2) n -NR 2a R 2b , -(CH2) m -SO2-(CH2) n -NR 2a R2b -(CH2) m -SO2-(CH2) n -4 to 6 membered heterocyclyl, -(CH2) m -COO-4 to 6 membered heterocyclyl, -(CH2) m -COO-C 3-6 cycloalkyl-NR 2a R 2b -(CH2) m -O-(CH2) n -NR 2a CO-C 1-6 alkyl, and said C 1-6 alkyl is optionally further substituted by one or more substituents selected from the group consisting of OH, NH2, -CONH2, -COOH, -NHC(=NH)NH2.

[0100] m and n are each independently selected from 0, 1 or 2;

[0101] R 2a , R 2b are each independently selected from the group consisting of H, methyl, ethyl, propenyl, ethynyl, propynyl, -CH2CF3, -CH2CN, methoxy, cyclopropyl, -C(=NH)-NH2, -C(=NH)-NHCH3, -C(=NH)-R 2h or 5 membered heterocyclyl;

[0102] or R 2a , R 2b form together with the nitrogen atom to which they are attached a

[0103] R 2c , R 2d are each independently selected from H or methyl;

[0104] or, R 2c , R 2d form together with the carbon atom to which they are attached a cyclopropyl group;

[0105] R 2e , R 2f are each independently selected from H or methyl;

[0106] or, R 2e , R 2a form together with the atoms to which they are attached a

[0107] or, R 2e , R 2b form together with the atoms to which they are attached a

[0108] R2h is selected from H, methyl, -CH2CN or cyclopropyl.

[0109] In some embodiments, the R 2 is selected from -(CH2)m-O-(CH2)n-NR 2a R 2b -(CH2)m-O-cyclobutyl-NR 2a R 2b -(CH2)m-O-4 to 6 membered heterocyclyl, -C(O)NR 2a -O-(CH2)n-NR 2a R 2b -C(O)NR 3-5 alkynyl-NR 2a R 2b -O-C 3-5 alkynyl-NR 2a R 2b -4 membered heterocyclyl-(CH2)n-NR 2a R 2b or

[0110] m and n are each independently 2;

[0111] R 2a R 2b are each independently selected from H, methyl, propenyl, ethynyl, -CH2CF3, -CH2CN, methoxy, cyclopropyl, -C(=NH)-NH2, -C(=NH)-NHCH3 or 5 membered heterocyclyl;

[0112] R 2c R 2d are each independently selected from H or methyl;

[0113] Alternatively, R 2c R 2d and the carbon atom to which they are attached together form a cyclopropyl group;

[0114] R 2e R 2f are each independently selected from H or methyl.

[0115] In some embodiments, the R 2 is selected from -(CH2) m -O-(CH2) n -NR 2a R 2b -C(O)NR 2-6 alkynyl-NR 2a R 2b or -(CH2) m -O-(CH2)n -NR 2a R 2b ;

[0116] m and n are 2;

[0117] R 2a , R 2b each independently is selected from H, C 1-4 alkyl, C 2-4 alkenyl, C 2-4 alkynyl, C 1-4 alkoxy, -C(=NH)-NH2, -C(=NH)-NHCH3, -CH2C(O)NH2, -CO-R y , -C(=NH)-R 2h or C 3-6 cycloalkyl, said C 1-4 alkyl can be further substituted with one or more substituents selected from halogen or CN;

[0118] R y is selected from C 1-6 alkyl or 5-12 membered heterocyclyl, and said C 1-6 alkyl or 5-12 membered heterocyclyl can be further substituted with one or more substituents selected from OH, NH2, -CONH2, -S-CH3, phenyl or 7-10 membered heteroaryl, said phenyl can be further substituted with OH or NH2;

[0119] R 2c , R 2d each independently is selected from H or C 1-4 alkyl;

[0120] Alternatively, R 2c , R 2d together with the carbon atom to which they are attached form a C 3-6 cycloalkyl;

[0121] R 2e , R 2f each independently is selected from H or C 1-4 alkyl;

[0122] R 2h is selected from C 1-4 alkyl or C 3-6 cycloalkyl, said C 1-4 alkyl can be further substituted with CN.

[0123] In some embodiments, said R 2 is selected from

[0124] In some embodiments, said R 2selected from the group consisting of

[0125] In some embodiments, the R 2 selected from the group consisting of

[0126] In some embodiments, the R 2 selected from the group consisting of

[0127] In some embodiments, the R 2 selected from the group consisting of

[0128] In some embodiments, the R 2 -(CH2) m -O-(CH2) n -NR 2a R 2b -(CH2) m -O-(CH2) n -NR 2a CO-C 1-6 alkyl, and said C 1-6 alkyl is optionally further substituted with one or more substituents selected from the group consisting of OH or NH2;

[0129] R 2a , R 2b are each independently selected from the group consisting of H, C 1-4 alkyl, C 2-4 alkynyl, or -C(=NH)-NH2;

[0130] m and n are each independently selected from 2.

[0131] In some embodiments, the R 2 selected from the group consisting of

[0132] In some embodiments, the R 2 -(CH2) m -O-(CH2) n -NR 2a R 2b -(CH2) m -O-(CH2) n -NR 2a CO-C 1-6 alkyl, and said C 1-6alkyl is optionally further substituted with one or more substituents selected from OH or NH2;

[0133] R 2a , R 2b each is independently selected from H, methyl, ethynyl, or -C(=NH)-NH2;

[0134] m and n are each independently selected from 2.

[0135] In some embodiments, the R 2 is selected from

[0136] In some embodiments, the R 3 and R 5 are each independently H.

[0137] In some embodiments, the R 4 is selected from H or F.

[0138] In some embodiments, the compound of Formula (I), stereoisomer thereof, or pharmaceutically acceptable salt thereof, is selected from:

[0139] wherein, the R 1 , R 2 and R 4 are as defined in Formula (I).

[0140] In some embodiments, the compound of Formula (I), stereoisomer thereof, or pharmaceutically acceptable salt thereof, is selected from:

[0141] wherein, the R 1 is halogen; preferably, the R 1 is Cl;

[0142] R 2 is selected from

[0143] preferably, the R 2 is selected from

[0144] In some embodiments, the compound of Formula (I) is selected from any one of the following compounds:

[0145] The present disclosure provides a protein degrader antibody conjugate, a stereoisomer thereof, or a pharmaceutically acceptable salt thereof, selected from the structure represented by formula (A):

[0146] wherein Ab is an antibody or an antigen binding fragment;

[0147] L is a linker group;

[0148] s is selected from a number from 1 to 10;

[0149] R 2x is R 2 minus H;

[0150] said R 1 , R 2 , and ring A are as defined in formula (I).

[0151] The present disclosure provides a protein degrader antibody conjugate, a stereoisomer thereof, or a pharmaceutically acceptable salt thereof, selected from the structure represented by formula (A-1):

[0152] wherein Ab is an antibody or an antigen binding fragment;

[0153] L is a linker group;

[0154] t is selected from a number from 1 to 10;

[0155] R 2x is R 2 minus H;

[0156] said R 1 , R 2 , and ring A are as defined in formula (I).

[0157] In some embodiments, Ab is an anti-CD33 antibody; preferably, Ab is Gemtuzumab.

[0158] In some embodiments, s, t are each independently selected from a number from 2 to 8; preferably from 3 to 5.

[0159] In some embodiments, R 2x is R 2 minus H from -NH- or -NH2.

[0160] In some embodiments, the protein degrader antibody conjugate, a stereoisomer thereof, or a pharmaceutically acceptable salt thereof, wherein L is selected from

[0161] In some embodiments, the L is selected from

[0162] In some embodiments, the L is

[0163] In some embodiments, the L is selected from

[0164] In some embodiments, the R 2x is selected from wherein R 2x is attached to L at the N-containing end.

[0165] In some embodiments, the R 2x is selected from wherein R 2x is attached to L at the N-containing end.

[0166] In some embodiments, the R 2x is selected from wherein R 2x is attached to L at the N-containing end.

[0167] In some embodiments, the R 2x is selected from wherein R 2x is attached to L at the N-containing end.

[0168] In some embodiments, the protein degrader antibody conjugate, stereoisomer thereof, or pharmaceutically acceptable salt thereof, is selected from the structure represented by Formula (A-2):

[0169] wherein, Ab is an antibody or antigen binding fragment; preferably, Ab is an anti-CD33 antibody; further preferably, Ab is Gemtuzumab;

[0170] L is selected from

[0171] t is selected from a number from 2 to 8; preferably, a number from 3 to 5;

[0172] R 2x is selected from R 2x is attached to L at the N-containing end.

[0173] The present disclosure also provides a protein degrader linker, a stereoisomer thereof, or a pharmaceutically acceptable salt thereof, selected from a structure represented by Formula (B):

[0174] wherein L a is selected from

[0175] R 2x is R 2 minus H; preferably, R 2 is a divalent structure formed by removing H from -NH- or -NH2; for example, R 2x as defined in Formula (A);

[0176] R 1 , R 2 , and ring A are as defined in Formula (I).

[0177] The present disclosure also provides a protein degrader linker, a stereoisomer thereof, or a pharmaceutically acceptable salt thereof, selected from a structure represented by Formula (B):

[0178] wherein L a is selected from

[0179] R 2x is R 2 minus H;

[0180] R 1 , R 2 , and ring A are as defined in Formula (I).

[0181] In some embodiments, the protein degrader linker, a stereoisomer thereof, or a pharmaceutically acceptable salt thereof, is selected from a structure represented by Formula (B-1):

[0182] L a is selected from

[0183] R 2x is selected from R 2x is attached to L a .

[0184] In some embodiments, the protein degrader linker represented by Formula (B), a stereoisomer thereof, or a pharmaceutically acceptable salt thereof, is selected from any one of:

[0185] In some embodiments, the protein degrader antibody conjugate of Formula (A), stereoisomer thereof, or pharmaceutically acceptable salt thereof, is selected from any one of:

[0186] wherein the Ab is selected from an antibody or antigenic fragment; preferably, Ab is an anti-CD33 antibody; further preferably, Ab is Gemtuzumab; t is a number from 1-10, preferably, t is selected from a number from 2-8; for example, a number from 3-5.

[0187] In some embodiments, the protein degrader antibody conjugate of Formula (A) is selected from any one of:

[0188] The present disclosure also provides a pharmaceutical composition comprising a compound of Formula (I), Formula (I-a), Formula (I-b), stereoisomer thereof, or pharmaceutically acceptable salt thereof, a protein degrader antibody conjugate of Formula (A), Formula (A-1), Formula (A-2), stereoisomer thereof, or pharmaceutically acceptable salt thereof, a protein degrader linker of Formula (B), Formula (B-1), stereoisomer thereof, or pharmaceutically acceptable salt thereof, and a pharmaceutically acceptable carrier.

[0189] The present disclosure also provides use of a compound of Formula (I), Formula (I-a), Formula (I-b), stereoisomer thereof, or pharmaceutically acceptable salt thereof, a protein degrader antibody conjugate of Formula (A), Formula (A-1), Formula (A-2), stereoisomer thereof, or pharmaceutically acceptable salt thereof, a protein degrader linker of Formula (B), Formula (B-1), stereoisomer thereof, or pharmaceutically acceptable salt thereof, or a pharmaceutical composition thereof, for the manufacture of a medicament for treating cancer.

[0190] In some embodiments, the cancer is selected from esophageal cancer, brain cancer, lung cancer, squamous cell carcinoma, bladder cancer, gastric cancer, ovarian cancer, peritoneal cancer, pancreatic cancer, breast cancer, head and neck cancer, cervical cancer, endometrial cancer, colorectal cancer, liver cancer, kidney cancer, urothelial cancer, non-Hodgkin lymphoma, central nervous system tumor, prostate cancer or thyroid cancer, acute myeloid leukemia, or myelodysplastic syndrome.

[0191] In some embodiments, the pharmaceutical composition comprises the compound, stereoisomer thereof, or pharmaceutically acceptable salt thereof in an amount selected from the group consisting of 0.1 mg - 1000 mg.

[0192] In some embodiments, the pharmaceutical composition comprises the compound, stereoisomer thereof, or pharmaceutically acceptable salt thereof in an amount selected from the group consisting of 0.1 mg - 1000 mg.

[0193] In some embodiments, the pharmaceutical composition comprises the compound, stereoisomer thereof, or pharmaceutically acceptable salt thereof in an amount selected from the group consisting of 0.1 mg - 1000 mg.

[0194] Terminology

[0195] The following terms and phrases used herein are intended to have the following meanings unless otherwise indicated. A particular term or phrase should not be construed as indefinite or unclear in the absence of a specific definition, but should be understood according to the ordinary meaning.

[0196] The term "antibody" refers to an immunoglobulin, which is a four polypeptide chain structure connected by interchain disulfide bonds, either two identical heavy chains or two identical light chains;

[0197] The term "pharmaceutically acceptable" means those compounds, materials, compositions, and / or dosage forms which are, within the scope of sound medical judgment, suitable for use in contact with the tissues of human beings and animals without excessive toxicity, irritation, allergic response, or other problem or complication, commensurate with a reasonable benefit / risk ratio.

[0198] The term "pharmaceutically acceptable salt" means a derivative of a compound of the present application produced by reaction with a relatively nontoxic acid or base. These salts can be prepared in situ during the synthesis, isolation, and purification of the compound, or by separately reacting a purified compound with a suitable acid or base. When a compound contains relatively acidic functionalities, base addition salts can be prepared by reaction of the compound with a suitable base, including alkali metal, alkaline earth metal, and organic bases. When a compound contains relatively basic functionalities, acid addition salts can be prepared by reaction of the compound with a suitable acid. Compounds of the present application exist in isomeric forms, such as cis and trans isomers, enantiomers, diastereomers, as well as racemic mixtures and other mixtures thereof, all of which are intended to be within the scope of the present application.

[0199] The term "enantiomer" refers to a stereoisomer which is a mirror image of the other.

[0200] The term "diastereomer" refers to a stereoisomer which has two or more chiral centers and which is not a mirror image of the other.

[0201] The term "cis-trans isomer" refers to the configuration of a molecule in which a double bond or a ring-forming carbon atom single bond cannot freely rotate.

[0202] Unless otherwise indicated, the use of a dashed wedge bond and a dashed wedge bond represents the absolute configuration of a stereocenter.

[0203] Stereoisomers of the compounds of the present application can be prepared by chiral synthesis or chiral reagents or other conventional techniques. For example, one enantiomer of a compound of the present application can be prepared by asymmetric catalytic techniques or by chiral auxiliary derivatization techniques. Alternatively, a single stereoisomer can be obtained from a mixture by chiral resolution techniques. Or directly prepared using chiral starting materials. Isolation of optically pure compounds of the present application is typically accomplished using preparative chromatography on chiral columns to achieve separation of the chiral compounds.

[0204] The term "pharmaceutically acceptable carrier" means a medium generally accepted in the art for the delivery of biologically active agents to animals, particularly mammals, and more particularly humans, and includes adjuvants, excipients or vehicles, such as diluents, preservatives, fillers, flow conditioners, disintegrants, wetting agents, emulsifiers, suspending agents, sweetening agents, flavoring agents, perfuming agents, antibacterial agents, antifungal agents, lubricating agents, and dispersing agents, depending on the nature of the dosage form and the dosage form's intended mode of administration. Pharmaceutically acceptable carriers are formulated in accordance with known methods and are within the scope of those ordinarily skilled in the art. They include, but are not limited to: the type and nature of the active pharmaceutical agent being formulated, the subject to whom the composition containing the agent is to be administered, the intended route of administration of the composition, and the therapeutic indication for which the agent is intended. Pharmaceutically acceptable carriers include both aqueous and nonaqueous media, and a variety of solid and semi-solid dosage forms. Such carriers include a number of different ingredients and additives, and additional ingredients included in the formulation for a variety of reasons (e.g., to stabilize the active pharmaceutical agent, to bind the agents, etc.) are well known to those of ordinary skill in the art. The term "effective prophylactic or therapeutic amount" means a sufficient amount of a compound of the present application or a pharmaceutically acceptable salt thereof to treat a disorder at a reasonable benefit / risk ratio applicable to any medical treatment and / or prophylaxis. It will be appreciated, however, that the appropriate amount of a compound of Formula (I) or a pharmaceutically acceptable salt thereof and compositions of the present application to be administered to a patient will be decided by the attending physician within the scope of sound medical judgment. The specific therapeutically effective dose level for any particular patient will depend upon a variety of factors including the disorder being treated and the severity of the disorder; the activity of the specific compound employed; the specific composition employed; the age, body weight, general health, sex, and diet of the patient; the time of administration, route of administration, and rate of excretion of the specific compound employed; the duration of the treatment; drugs used in combination or coincidental with the specific compound employed; and like factors well known in the medical arts.

[0205] The term "optionally substituted" as used herein means that the one or more hydrogen atoms of the group can be "replaced" by one or more substituents or "not replaced" by one or more substituents.

[0206] Unless otherwise specified, the term "ring" means saturated, partially saturated, or unsaturated monocyclic and polycyclic rings. The term "heterocyclyl" as used herein includes saturated or partially saturated monocyclic and polycyclic heterocyclyl groups; the heterocyclyl groups are independent of the position of attachment (i.e., can be bound via a carbon atom or a heteroatom). The polycyclic heterocyclyl groups include fused heterocyclyl groups, spiro heterocyclyl groups, bridged heterocyclyl groups.

[0207] The term "fused heterocyclyl group" as used herein means a cyclic structure formed by two or more rings sharing two adjacent ring atoms, and at least one of the rings is a heterocyclic ring; the fused heterocyclyl group includes a cyclic structure formed by a monocyclic heterocyclyl group fused with a monocyclic heterocyclyl group or a cycloalkyl group or an aryl group or a heteroaryl group, and also includes a cyclic structure formed by a heteroaryl group fused with a cycloalkyl group or a monocyclic heterocyclyl group.

[0208] The term "spiro heterocyclyl group" as used herein means a cyclic structure formed by two or more rings sharing one ring atom, and at least one of the rings is a heterocyclic ring.

[0209] The term "bridged heterocyclyl group" as used herein means a cyclic structure formed by two or more rings sharing non-adjacent ring atoms, and at least one of the rings is a heterocyclic ring.

[0210] The heterocyclyl group as used herein is preferably a 4-10 membered monocyclic heterocyclyl group, a 5-10 membered fused heterocyclyl group, a 5-10 membered spiro heterocyclyl group, or a 5-10 membered bridged heterocyclyl group; further, the heterocyclyl group is preferably a 4-6 membered monocyclic heterocyclyl group, an 8-10 membered fused heterocyclyl group, an 8-10 membered spiro heterocyclyl group, or a 7-9 membered bridged heterocyclyl group.

[0211] Unless otherwise specified, the term "heterocyclyl" as used herein means a saturated cyclic group derived by replacing one or more ring carbon atoms in a cycloalkyl group with a heteroatom and / or a heteroatom group. The heteroatom and / or heteroatom group is generally selected from N, O, S, NO, SO, S(O)2, P(O), and NR, wherein the carbon atoms in the heterocycle are optionally replaced by an oxo group, i.e., -C(=O); the carbon atoms in the heterocycle are optionally replaced by =NH, i.e., -C(=NH); the heterocyclyl group includes "3-8 membered heterocyclyl", "3-6 membered heterocyclyl", "3-5 membered heterocyclyl", "4-6 membered heterocyclyl", "5-6 membered heterocyclyl". Specific examples include, but are not limited to, azetidinyl, pyrrolidinyl, tetrahydrofuranyl, piperidinyl, piperazinyl, tetrahydropyranyl, morpholinyl, , and the like.

[0212] Unless otherwise specified, the term "aryl" means an unsaturated, typically aromatic, hydrocarbon group that can be a single ring or multiple rings that are fused together. Examples of 6-10 membered aryl groups include, but are not limited to, phenyl, naphthyl.

[0213] Unless otherwise specified, the term "heteroaryl" means a stable, monocyclic or polycyclic aromatic hydrocarbon, preferably containing carbon atoms and 1, 2, 3, or 4 ring heteroatoms independently selected from N, O, and S. Preferred are 5-12 membered heteroaryl groups, more preferred are 5-10 membered heteroaryl groups.

[0214] Unless otherwise specified, "cycloalkyl" means a saturated monocyclic or polycyclic hydrocarbon group. Cycloalkyl groups are preferably C 3-8 cycloalkyl groups, more preferably C 3-6 cycloalkyl groups. Examples of cycloalkyl groups include, but are not limited to, cyclopropyl, cyclobutyl, cyclopentyl, cyclohexyl, cycloheptyl, and the like.

[0215] Unless otherwise specified, the term "alkyl" is used to denote straight-chain or branched-chain saturated hydrocarbon groups. Preferred are C 1-6 alkyl groups, more preferably C 1-4 alkyl groups. Examples of alkyl groups include, but are not limited to, methyl, ethyl, propyl, isopropyl, butyl, isobutyl, t-butyl, pentyl, isopentyl, neopentyl, n-hexyl, and the like.

[0216] Unless otherwise specified, the term "alkoxy" means an alkyl group as defined herein attached through an oxygen atom to the remainder of the molecule, i.e., "-O-alkyl". Included are "C 1-6 alkoxy" groups (of the structure -O-C 1-6 alkyl) and "C 1-4 alkoxy" groups. Specific examples include, but are not limited to, methoxy, ethoxy, propoxy, 1-methylethoxy, butoxy, 2-methylbutoxy, 3-methylbutoxy, 1,1-dimethylpropoxy, 1,2-dimethylpropoxy, and the like; preferably, "alkoxy" groups according to the present disclosure are preferably C 1-4 alkoxy groups, more preferably C 1-3 alkoxy groups. Unless otherwise specified, the term "halogen" denotes a fluorine, chlorine, bromine, or iodine atom.

[0217] Unless otherwise specified, the term "alkenyl" means a straight-chain or branched-chain alkene (containing at least one carbon-carbon double bond) from which one hydrogen atom has been removed. Included are "C 2-6 alkenyl" groups, "C 2-5 alkenyl" groups, "C 2-4 alkenyl" groups, "C 2-3 alkenyl" groups. Specific examples include, but are not limited to: -CH=CH2, -CH=CHCH3, -C(CH2)=CH2, -CH=CHCH2CH3, -CH2CH=CHCH3, and the like.

[0218] Unless otherwise specified, the term "alkynyl" means a straight-chain or branched- chain alkyne (containing at least one carbon-carbon triple bond) from which one hydrogen atom has been removed, including "C 2-5 alkynyl", "C 2-4 alkynyl", "C 2-3 alkynyl", specific examples of which include, but are not limited to: -C≡CH, -C≡C-CH3, -CH2C≡CH, HC≡C-C≡C-, etc.

[0219] It is specifically stated that all combinations of substituents and / or variants thereof herein are only allowed if such combinations will result in stable compounds.

[0220] The "R 2x of the upper N-containing end is connected to L" means that the R 2x of the upper N-containing end is connected to L. For example, when R 2x is selected from , the 2 end of the wavy line (i.e. ) in R 2x is connected to L, and the 1 end of the wavy line is connected to the phenyl group in general formula (A),

[0221] The conjugate generally contains an average of 1-10 cytotoxic compounds or GSPT1 degradation agents (payloads) connected to the targeting agent, which is the degree of grafting or drug-to-antibody ratio, i.e. the DAR value. As s, t outside the brackets of the general formula (A) of the disclosure, when s or t is selected from a number from 1 to 10, it represents the DAR value as an integer or a decimal number from 1 to 10, preferably s or t is selected from 2 to 8; further preferably, s or t is selected from 3 to 5. Preferably, the DAR value described in the disclosure is 3.6, 3.7, 3.8, 3.9, 4.0, 4.1, 4.2, 4.3, 4.4, 4.5, 4.6, 4.7, 4.8, 4.9 or 5.0; further preferably, the DAR value described in the disclosure is 3.8, 3.9, 4.0, 4.1, 4.2, 4.3 or 4.4. DETAILED DESCRIPTION

[0222] In the embodiments of the disclosure, if there is a discrepancy between the compound name and the compound structure, the compound structure given shall prevail if it cannot be confirmed by other means. The preparation method of some compounds in the present application refers to the preparation method of the aforementioned similar compounds. Those skilled in the art should know that when using or referring to using the preparation method cited, the feeding ratio of reactants, reaction solvent, reaction temperature, etc. can be appropriately adjusted according to the different reactants.

[0223] The compounds of the present application can be prepared by a variety of synthetic methods well known to those skilled in the art, including the specific embodiments set forth below, embodiments formed by a combination of the specific embodiments set forth below with other chemical synthetic methods well known in the art, and equivalents thereof as appreciated by those skilled in the art. Preferred embodiments include, but are not limited to, the examples of the present application.

[0224] Summary of experimental instruments:

[0225] The structure of the compounds of the present application is determined by nuclear magnetic resonance (NMR) or / and liquid chromatography-mass spectrometry (LC-MS), or ultra-performance liquid chromatography-mass spectrometry (UPLC-MS). NMR chemical shifts (δ) are given in parts per million (ppm). NMR measurements are determined by Bruker Neo 400M or Bruker Ascend 400 nuclear magnetic instrument, with deuterated dimethyl sulfoxide (DMSO-d6), deuterated methanol (CD3OD) and deuterated chloroform (CDCl3), heavy water (D2O) as the determination solvent, and tetramethylsilane (TMS) as the internal standard.

[0226] Liquid chromatography-mass spectrometry (LC-MS) measurements are determined by Agilent 1260-6125B single quadrupole mass spectrometer, with Welch Biomate column (C18, 2.7 μm, 4.6 x 50 mm) or waters H-Class SQD2, with Welch Ultimate column (XB-C18, 1.8 μm, 2.1 x 50 mm) mass spectrometer (ion source is electrospray ionization).

[0227] Ultra-performance liquid chromatography-mass spectrometry (UPLC-MS) measurements are determined by Waters UPLC H-class SQD mass spectrometer (ion source is electrospray ionization).

[0228] High-performance liquid chromatography (HPLC) measurements use Waters e2695-2998 or Waters ARC and Agilent 1260 or Agilent Poroshell HPH high-performance liquid chromatography.

[0229] Preparative HPLC uses Waters 2555-2489 (10 μm, ODS250 cm x 5 cm) or GILSON Trilution LC, with Welch XB-C18 column (5 μm, 21.2 x 150 mm).

[0230] Thin layer chromatography silica gel plate uses Yantai Jiangyou Silica Gel Development Co., Ltd. GF254 silica gel plate or Lushan City Shangbang New Material Co., Ltd. GF254 silica gel plate, the specification of TLC is 0.15mm-0.20mm, and the preparation type is 20*20cm. Column chromatography generally uses 200-300 mesh silica gel as a carrier.

[0231] The starting materials in the embodiments of the present disclosure are known and commercially available, or can be synthesized by using or according to the methods known in the art. Unless otherwise specified, all reactions of the present application are carried out under continuous magnetic stirring under a dry nitrogen or argon atmosphere, the solvent is a dry solvent, and the reaction temperature unit is Celsius or ℃.

[0232] If TFA (trifluoroacetic acid) is used in the reaction steps of the embodiments of the present disclosure, most of the obtained products are in the form of trifluoroacetate salt. In the route map or the structure of the compound, the structure of the free base form is represented, and the trifluoroacetic acid is not drawn again. Whether it is in the form of free base or salt, it does not affect its biological activity.

[0233] Example 1:

[0234] Preparation of 4-(((S)-2-((R)-2-(6-(2,5-dioxo-2,5-dihydro-1H-pyrrol-1-yl)hexanoylamido)-3- methylbutanamido)-5-ureidopentanamido)benzyl(2-(3-(2-chloro-4-(3-((2-(2,6-dioxopiperidin-3- yl)-1-oxoisoindolin-5-yl)methyl)ureido)phenyl)ureido)ethyl)(methyl)carbamate.

[0235] Reaction Scheme:

[0236] Operation Steps:

[0237] Step A: Compound 1-1 (4g, 12.38mmol) was added to a 250mL single-neck flask, followed by the addition of N,N-dimethylformamide (60mL), Pd(PPh3)4 (1.43g, 1.24mmol) and Zn(CN)2 (1.02g, 8.67mmol). The reaction system was reacted at 100℃ for 3 hours under nitrogen protection. After the reaction was completed by LCMS monitoring, the reaction liquid was poured into water, extracted with ethyl acetate, and the organic phase was washed with saturated sodium chloride solution, dried over anhydrous sodium sulfate, filtered, and the filtrate was concentrated. The obtained crude product was purified by silica gel column (dichloromethane / methanol = 10:1) to obtain compound 1-2 (3.1g, yield 93.01%).

[0238] LCMS (ESI) M / Z: 268.1 [M-H]+ .

[0239] 1 H NMR (400 MHz, DMSO-d6) δ 11.03 (s, 1H), 8.18 (d, J = 10.5 Hz, 1H), 7.98 (dt, J = 10.2, 5.1 Hz, 1H), 7.92 (d, J = 7.8 Hz, 1H), 5.16 (dd, J = 13.3, 5.1 Hz, 1H), 4.56 (d, J = 17.9 Hz, 1H), 4.43 (d, J = 17.9 Hz, 1H), 2.99 - 2.86 (m, 1H), 2.61 (d, J = 17.4 Hz, 1H), 2.49 - 2.36 (m, 1H), 2.09 - 1.99 (m, 1H).

[0240] Step B: Compound 1-2 (1 g, 3.71 mmol) was added to a 100 mL single-necked flask, followed by methanol (15 mL), PtO2(252.74 mg, 1.11 mmol) and concentrated hydrochloric acid (1.2 mL). The reaction system was protected by hydrogen and reacted at room temperature for 15 hours.

[0241] LCMS monitoring showed that the reaction was completed, methanol (250 ml) was added for dilution, filtered with diatomite, the filtrate was rotary evaporated, the residue was slurried with dichloromethane / methanol = 3:1 (20 mL) to obtain compound 1-3 (820 mg, yield 71.39%).

[0242] LCMS (ESI) M / Z: 274.1 [M+H] + .

[0243] 1 H NMR (400 MHz, DMSO-d6) δ 11.00 (s, 1H), 8.64 (s, 3H), 7.81 - 7.72 (m, 2H), 7.65 (d, J = 7.8 Hz, 1H), 5.13 (dd, J = 13.3, 5.1 Hz, 1H), 4.48 (d, J = 17.5 Hz, 1H), 4.34 (d, J = 17.5 Hz, 1H), 4.14 (d, J = 5.5 Hz, 2H), 2.99 - 2.86 (m, 1H), 2.61 (d, J = 17.3 Hz, 1H), 2.45 - 2.34 (m, 1H), 2.07 - 1.96 (m, 1H).

[0244] Step C: Compound 1-4 (1 g, 5.79 mmol) was added to a 100 mL single-necked flask, followed by the addition of N,N-dimethylformamide (15 mL), N,N'- carbonyldiimidazole (1.88 g, 11.58 mmol), triethylamine (1.76 g, 17.37 mmol) and compound 1-5 (1 g, 5.79 mmol). The reaction system was protected by nitrogen and reacted at room temperature for 3 hours.

[0245] After the reaction was completed by LCMS monitoring, compound 1-6 (500 mg, yield 23.15%) was obtained by column chromatography (PE / EA = 70 / 30).

[0246] LCMS (ESI) M / Z: 373.1 [M+H] + .

[0247] Step D: Compound 1-6 (500 mg, 1.34 mmol) was added to a 25 mL single-necked flask, followed by the addition of N,N-dimethylformamide (10 mL), diboronic acid (600 mg, 6.70 mmol) and 4,4'-dipyridine (42 mg, 0.27 mmol), and stirred at room temperature for 10 minutes. After the reaction was completed by LCMS monitoring, compound 1-7 (290 mg, yield 63.07%) was obtained by high performance liquid chromatography preparation purification (acetonitrile / 0.1% formic acid aqueous solution).

[0248] LCMS (ESI) M / Z: 343.2 [M+H] + .

[0249] Step E: Compound 1-7 (50 mg, 0.15 mmol) was added to a 25 mL single-necked flask, followed by the addition of N,N-dimethylformamide (5 mL) for dissolution, compound 1-3 (41 mg, 0.15 mmol), N,N'-carbonyldiimidazole (49 mg, 0.30 mmol) and triethylamine (50.59 mg, 0.50 mmol). The reaction system was protected by nitrogen and reacted at room temperature for 3 hours. After the reaction was completed by LCMS monitoring, compound 1-8 (30 mg, yield 32.03%) was obtained by high performance liquid chromatography preparation purification (acetonitrile / 0.1% formic acid aqueous solution).

[0250] LCMS (ESI) M / Z: 642.2 [M+H] + .

[0251] Step F: Compound 1-8 (30 mg, 0.05 mmol) was taken in 25 mL vial, dichloromethane (5 mL) and TFA (0.5 mL) were added and the reaction mixture was allowed to react at room temperature for 1 h under nitrogen atmosphere. After completion of the reaction as monitored by LCMS, the reaction mixture was concentrated to get compound 1-9 (25 mg, yield 98.73%, crude).

[0252] LCMS (ESI) M / Z: 542.2 [M+H] + .

[0253] Step G: Compound 1-9 (25 mg, 0.046 mmol) was taken in 25 mL vial, N,N- dimethylformamide (2 mL), commercial compound (CAS No. 159857-81-5) 1-10 (33.94 mg, 0.046 mmol), HOBT (3.11 mg, 0.023 mmol) and 2,6-dimethylpyridine (9.86 mg, 0.092 mmol) were added sequentially and the reaction mixture was allowed to react at room temperature for 6 h under nitrogen atmosphere.

[0254] After completion of the reaction as monitored by LCMS, the product was purified by preparative high-performance liquid chromatography to get compound 1 (5.13 mg, yield 9.75%).

[0255] LCMS (ESI) M / Z: 1140.5 [M+H] + .

[0256] 1H NMR (400 MHz, DMSO-d6) δ 10.98 (s, 1H), 9.99 (s, 1H), 8.73 (d, J = 9.1 Hz, 1H), 8.09 (d, J = 7.5 Hz, 1H), 7.98 (s, 1H), 7.85 (d, J = 8.9 Hz, 1H), 7.81 (d, J = 8.6 Hz, 1H), 7.73 - 7.68 (m, 2H), 7.57 (t, J = 6.2 Hz, 2H), 7.52 (s, 1H), 7.45 (d, J = 7.9 Hz, 1H), 7.37 (t, J = 5.9 Hz, 1H), 7.28 (d, J = 8.1 Hz, 2H), 7.07 (dd, J = 8.9, 2.5 Hz, 1H), 7.00 (s, 2H), 6.39 (d, J = 22.4 Hz, 1H), 5.99 (t, J = 5.8 Hz, 1H), 5.41 (s, 2H), 5.11 (dd, J = 13.3, 5.1 Hz, 1H), 4.98 (s, 2H), 4.50 - 4.40 (m, 3H), 4.38 (d, J = 6.2 Hz, 1H), 4.32 (d, J = 17.3 Hz, 1H), 4.19 (dd, J = 8.7, 6.8 Hz, 1H), 3.26 - 3.16 (m, 4H), 3.05 - 2.89 (m, 4H), 2.87 (d, J = 5.1 Hz, 3H), 2.68 - 2.56 (m, 1H), 2.42 - 2.30 (m, 1H), 2.20 - 2.06 (m, 2H), 2.04 - 1.91 (m, 3H), 1.69 (s, 2H), 1.51 - 1.43 (m, 4H), 1.42 - 1.31 (m, 2H), 1.21 - 1.13 (m, 2H), 0.85 (d, J = 6.7 Hz, 3H), 0.82 (d, J = 6.7 Hz, 3H).

[0257] Example 2:

[0258] Preparation of 4-(((S)-2-((R)-2-(6-(2,5-dioxo-2,5-dihydro-1H-pyrrol-1- yl)hexanamido)-3-methylbutanamido)-5-ureidopentanamido)benzyl 3-(2-chloro-4- (3-((2-(2,6-dioxopiperidin-3-yl)-1-oxoisoindolin-5-yl)methyl)ureido)phenethyl- oxy)azetidine-1-carboxylate.

[0259] Reaction Scheme:

[0260] Procedure:

[0261] Step A: Compound 2-1 (500 mg, 2.32 mmol) was added to a 100 mL single-neck flask, and borane dimethyl sulfide (264.38 mg, 3.48 mmol) was added. The reaction system was reacted at room temperature for 8 hours under nitrogen protection. After the reaction was completed by LCMS monitoring, water (20 mL) was added to quench the reaction, and ethyl acetate (50 mL) was added for dilution. After liquid separation and extraction, the crude product 2-2 (450 mg, yield 96.24%) was obtained after drying and concentration.

[0262] LCMS (ESI) M / Z: 202.1 [M+H] + .

[0263] Step B: Compound 2-2 (450 mg, 2.23 mmol) was added to a 25 mL single-neck flask, and dichloromethane (20 mL), NBS (440 mg, 2.45 mmol), and triphenylphosphine (640 mg, 2.45 mmol) were added in sequence. The reaction was stirred at room temperature for 10 minutes. After the reaction was completed by LCMS monitoring, the reaction solution was concentrated, and compound 5-3 (386 mg, yield 65.38%) was obtained by column chromatography purification (PE / EA = 4:1).

[0264] Step C: Compound 2-3 (384 mg, 1.45 mmol) was added to a 25 mL single-neck flask, and tetrahydrofuran (15 mL) was added for dissolution. Compound 2-4 (1.26 g, 7.25 mmol) and 60% sodium hydride (52 mg, 2.17 mmol) were added, and the reaction system was reacted at room temperature for 2 hours under nitrogen protection. After the reaction was completed by LCMS monitoring, the reaction was quenched with methanol (10 mL), and compound 2-5 (64 mg, yield 12.36%) was obtained by column chromatography purification (PE / EA = 2:1) after concentration.

[0265] LCMS (ESI) M / Z: 257.2 [M-BOC+H] + .

[0266] Step D: Compound 2-5 (64 mg, 0.18 mmol) was added to a 25 mL single-neck flask, and N,N-dimethylformamide (3 mL), diboronic acid (80.69 mg, 0.90 mmol), and 4,4'-dipyridine (5.62 mg, 0.036 mmol) were added in sequence. The reaction system was reacted at room temperature for 10 minutes under nitrogen protection. After the reaction was completed by LCMS monitoring, the reaction solution was diluted with ethyl acetate (20 mL), and water (20 mL) was added for liquid separation. After extraction of the oil layer, compound 2-6 (60 mg, yield 102.35%, crude) was obtained after concentration and drying.

[0267] LCMS (ESI) M / Z: 327.1 [M+H] + .

[0268] Step E: Compound 2-6 (60 mg, 0.18 mmol) was added into a 25 mL vial, followed by the addition of N,N-dimethylformamide (2 mL), triethylamine (36.43 mg, 0.36 mmol) and N,N'-carbonyldiimidazole (43.78 mg, 0.27 mmol). The reaction was stirred at room temperature for 1 h under nitrogen. Then compound 1-3 (147.58 mg, 0.54 mmol) and tetrahydrofuran (3 mL) were added. The reaction was stirred at room temperature for 3 h under nitrogen. After the reaction was completed by LCMS, compound 2-7 (24 mg, yield 20.88%) was obtained by preparative high performance liquid chromatography (acetonitrile / 0.1% formic acid in water).

[0269] LCMS (ESI) M / Z: 626.2 [M+H] + .

[0270] Step F: Compound 2-7 (24 mg, 0.038 mmol) was added into a 25 mL vial, followed by the addition of dichloromethane (5 mL) and TFA (0.5 mL). The reaction was stirred at room temperature for 1 h under nitrogen. After the reaction was completed by LCMS, compound 2-8 (20 mg, yield 99.20%) was obtained by concentration.

[0271] LCMS (ESI) M / Z: 526.2 [M+H] + .

[0272] Step G: Compound 2-8 (20 mg, 0.038 mmol) was added into a 25 mL vial, followed by the addition of N,N-dimethylformamide (2 mL), compound 1-10 (28.03 mg, 0.038 mmol), HOBT (5.13 mg, 0.038 mmol) and 2,6-dimethylpyridine (8.14 mg, 0.076 mmol). The reaction was stirred at room temperature for 6 h under nitrogen. After the reaction was completed by LCMS, compound 2 (25.44 mg, yield 90.74%) was obtained by preparative high performance liquid chromatography.

[0273] LCMS (ESI) M / Z: 1124.5 [M+H] + .

[0274] 1H NMR (400 MHz, DMSO-d6) δ 10.98 (s, 1H), 10.00 (s, 1H), 8.88 (s, 1H), 8.11 - 8.07 (m, 2H), 7.80 (d, J = 8.6 Hz, 1H), 7.71 - 7.65 (m, 2H), 7.58 (d, J = 8.3 Hz, 2H), 7.51 (s, 1H), 7.44 (d, J = 7.9 Hz, 1H), 7.27 (d, J = 8.4 Hz, 2H), 7.24 - 7.19 (m, 1H), 7.19 - 7.13 (m, 1H), 6.91 (t, J = 6.0 Hz, 1H), 6.86 - 6.76 (m, 2H), 5.98 (t, J = 5.9 Hz, 1H), 5.41 (s, 2H), 5.10 (dd, J = 13.3, 5.1 Hz, 1H), 4.95 (s, 2H), 4.47 - 4.35 (m, 4H), 4.34 - 4.26 (m, 2H), 4.19 (dd, J = 8.7, 6.8 Hz, 1H), 4.08 (t, J = 7.7 Hz, 2H), 3.70 (s, 2H), 3.50 (t, J = 6.3 Hz, 2H), 3.36 (t, J = 6.8 Hz, 2H), 3.08 - 2.97 (m, 2H), 2.96 - 2.88 (m, 2H), 2.84 (t, J = 7.3 Hz, 2H), 2.69 - 2.56 (m, 1H), 2.41 - 2.32 (m, 1H), 2.23 - 2.07 (m, 2H), 2.04 - 1.92 (m, 2H), 1.74 - 1.64 (m, 1H), 1.65 - 1.53 (m, 1H), 1.53 - 1.44 (m, 4H), 1.21 - 1.14 (m, 2H), 0.88 - 0.79 (m, 6H).

[0275] Example 3:

[0276] Preparation of 2-(methylamino)ethyl-2-(2-chloro-4-(3-((2-(2,6-dioxopiperidin-3-yl)-1- oxoisoindolin-5-yl)methyl)ureido)phenyl)-2-methylpropanoic acid

[0277] Reaction Scheme:

[0278] Procedure:

[0279] Step A: Compound 2-1 (2 g, 9.28 mmol) was added to a 100 mL single-necked flask, dichloromethane (40 mL), compound 3-2 (2.44 g, 13.92 mmol), DCC (2.87 g, 13.92 mmol) and DMAP (1.13 g, 9.28 mmol) were added in turn, and the reaction system was protected by nitrogen, and reacted at room temperature for 12 hours. After the reaction was completed by LCMS monitoring, the reaction liquid was poured into water, extracted with ethyl acetate, and the organic phase was washed with saturated sodium chloride solution. The organic phase was dried over anhydrous sodium sulfate, filtered, and the filtrate was concentrated. The obtained crude product was purified by silica gel column (PE / EA = 10:1) to obtain compound 3-3 (3.3 g, yield 95.42%).

[0280] LCMS (ESI) M / Z: 395.2 [M+Na] + .

[0281] 1 H NMR (400 MHz, DMSO-d6) δ 8.31 (d, J = 2.3 Hz, 1H), 8.20 (dd, J = 8.4, 2.4 Hz, 1H), 7.74 (d, J = 8.4 Hz, 1H), 4.18 (t, J = 5.2 Hz, 2H), 4.08 - 3.93 (m, 2H), 3.39 (t, J = 5.5 Hz, 2H), 2.74 (s, 3H), 1.39 (s, 9H).

[0282] Step B: Compound 3-3 (1.6 g, 4.29 mmol) was added to a 100 mL single-necked flask, and N,N-dimethylformamide (20 mL) was added, and then 60% sodium hydride (343.2 mg, 8.58 mmol) was added at 0°C. After stirring at 0°C for 30 min, iodomethane (0.8 mL, 12.87 mmol) was added, and the reaction system was protected by nitrogen and reacted at room temperature for 2 hours. After the reaction was completed by LCMS monitoring, water (10 mL) was added to quench the reaction, extracted with ethyl acetate, and the organic phase was washed with saturated sodium chloride solution. The organic phase was dried over anhydrous sodium sulfate, filtered, and the filtrate was concentrated. The obtained crude product was purified by silica gel column (PE / EA = 6:1) to obtain compound 3-4 (1.3 g, yield 75.56%).

[0283] LCMS (ESI) M / Z: 423.1 [M+Na] + .

[0284] 1H NMR (400 MHz, DMSO-d6) δ 8.33 - 8.16 (m, 2H), 7.84 (d, J = 8.6 Hz, 1H), 4.13 (t, J = 5.1 Hz, 2H), 3.34 (d, J = 5.7 Hz, 2H), 2.56 (s, 3H), 1.59 (s, 6H), 1.36 (d, J = 7.7 Hz, 9H).

[0285] Step C: Tetrahydroxydiboron (1.16 g, 12.96 mmol) was added to a 50 mL vial, dry N,N-dimethylformamide (25 mL) was added, then 4,4'-dipyridyl (506 mg, 3.24 mmol) and compound 3-4 (1.3 g, 3.24 mmol) were added quickly in turn. The reaction system was protected by nitrogen, and reacted at room temperature for 5 min. After the reaction was completed by LCMS monitoring, a small amount of methanol was added, filtered, and the filtrate was purified by column chromatography (acetonitrile / 0.1% formic acid aqueous solution) to obtain compound 3-5 (1.06 g, yield 88.13%).

[0286] LCMS (ESI) M / Z: 393.2 [M+Na] + .

[0287] 1 H NMR (400 MHz, DMSO-d6) δ 7.10 (d, J = 8.4 Hz, 1H), 6.58 (d, J = 2.2 Hz, 1H), 6.48 (dd, J = 8.5, 2.2 Hz, 1H), 5.26 (s, 2H), 4.07 (t, J = 5.1 Hz, 2H), 3.32 (d, J = 4.7 Hz, 2H), 2.61 (s, 3H), 1.44 (s, 6H), 1.37 (d, J = 2.5 Hz, 9H).

[0288] Step D: Compound 3-5 (200 mg, 0.54 mmol) was added to a 25 mL vial, dry THF (6 mL) was added, then bis(trichloromethyl) carbonate (0.13 mL, 1.08 mmol) was added slowly dropwise. The reaction system was protected by nitrogen, and reacted at room temperature for 1 h. After the reaction was completed by LCMS monitoring, the solvent was rotary evaporated under reduced pressure at low temperature, and then dissolved in dry N,N-dimethylformamide (3 mL). A solution of compound 1-3 (201.33 mg, 0.65 mmol) and triethylamine (0.37 mL, 2.7 mmol) in N,N-dimethylformamide (5 mL) was added slowly dropwise. The reaction system was protected by nitrogen, and reacted at room temperature for 1 h. After the reaction was completed by LCMS monitoring, it was purified by column chromatography (acetonitrile / 0.1% formic acid aqueous solution) to obtain compound 3-6 (287 mg, yield 79.41%).

[0289] LCMS (ESI) M / Z: 570.4 [M-BOC+H]+ .

[0290] 1 H NMR (400 MHz, DMSO-d6) δ 10.98 (s, 1H), 8.89 (s, 1H), 7.68 (t, J = 12.7 Hz, 2H), 7.52 (s, 1H), 7.45 (d, J = 7.9 Hz, 1H), 7.35 (d, J = 8.5 Hz, 1H), 7.24 (t, J = 8.3 Hz, 1H), 6.87 (t, J = 5.9 Hz, 1H), 5.11 (dd, J = 13.3, 5.1 Hz, 1H), 4.44 (t, J = 12.0 Hz, 3H), 4.32 (d, J = 17.3 Hz, 1H), 4.09 (t, J = 5.0 Hz, 2H), 3.33 - 3.31 (m, 2H), 2.98 - 2.85 (m, 1H), 2.65 - 2.55 (m, 4H), 2.46 - 2.31 (m, 1H), 2.04 - 1.95 (m, 1H), 1.50 (s, 6H), 1.37 (s, 9H).

[0291] Step E: Compound 3-6 (250 mg, 0.37 mmol) was added to a 50 mL single-necked flask, dichloromethane (20 mL) was added, TFA (4 mL) was added at 0 °C, the reaction system was protected by nitrogen, and reacted at room temperature for 1 hour. After the reaction was completed by LCMS monitoring, the reaction solution was concentrated, and the crude product was purified by high performance liquid chromatography to obtain compound 3 (180 mg, yield 84.64%).

[0292] LCMS (ESI) M / Z: 570.2 [M+H] + .

[0293] 1 H NMR (400 MHz, DMSO-d6) δ 10.98 (s, 1H), 9.16 (s, 1H), 8.27 (s, 1H), 7.68 (t, J = 5.3 Hz, 2H), 7.51 (s, 1H), 7.44 (d, J = 7.9 Hz, 1H), 7.35 (d, J = 8.7 Hz, 1H), 7.26 (dd, J = 8.7, 2.2 Hz, 1H), 7.17 (t, J = 5.9 Hz, 1H), 5.11 (dd, J = 13.3, 5.1 Hz, 1H), 4.43 (dd, J = 13.8, 12.0 Hz, 3H), 4.31 (d, J = 17.3 Hz, 1H), 4.16 (t, J = 5.7 Hz, 2H), 2.98 - 2.85 (m, 3H), 2.65 - 2.55 (m, 1H), 2.45 - 2.32 (m, 4H), 2.05 - 1.93 (m, 1H), 1.52 (s, 6H).

[0294] Example 4:

[0295] 2-((((4-((S)-2-(S)-2-[(6-(2,5-dioxo-2,5-dihydro-1H-pyrrol-1-yl)hexanoylamido)-3- methylbutanoylamido)-5-ureidopentanoylamido]benzyl)oxy)carbonyl)(methyl)amino)- 2-(2-chloro-4-(3-(((2-(6-dioxopiperidin-3-yl)-1-oxoisoindolin-5-yl)methyl)ureido) phenyl)-2-methylpropanoic acid ethyl ester.

[0296] Reaction Scheme:

[0297] Operation Steps:

[0298] Step A: Compound 3 (60 mg, 0.11 mmol) was added to a 25 mL single-neck flask, followed by the addition of N,N-dimethylformamide (7 mL), compound 1-10 (89.27 mg, 0.12 mmol), HOBT (7.43 mg, 0.06 mmol), and 2,4-dimethylpyridine (23.57 mg, 0.22 mmol). The reaction system was reacted at 40 °C for 12 hours under nitrogen protection. After the reaction was completed by LCMS monitoring, compound 4 (40 mg, yield 32.52%) was obtained by preparative purification by high performance liquid chromatography.

[0299] LCMS (ESI) M / Z: 1168.4 [M+H] + .

[0300] 1H NMR (400 MHz, DMSO-d6) δ 10.98 (s, 1H), 9.98 (s, 1H), 8.85 (s, 1H), 8.08 (d, J = 7.1 Hz, 1H), 7.79 (d, J = 8.7 Hz, 1H), 7.74 - 7.55 (m, 4H), 7.51 (s, 1H), 7.44 (d, J = 7.8 Hz, 1H), 7.29 (dd, J = 21.4, 9.0 Hz, 4H), 6.99 (s, 2H), 6.89 - 6.80 (m, 1H), 5.97 (t, J = 5.7 Hz, 1H), 5.40 (s, 2H), 5.10 (dd, J = 13.3, 5.1 Hz, 1H), 4.93 (d, J = 21.9 Hz, 2H), 4.49 - 4.34 (m, 4H), 4.30 (d, J = 17.3 Hz, 1H), 4.23 - 4.15 (m, 1H), 4.11 (d, J = 5.2 Hz, 2H), 3.44 - 3.35 (m, 4H), 3.07 - 2.85 (m, 3H), 2.71 - 2.55 (m, 4H), 2.44 - 2.31 (m, 1H), 2.23 - 2.07 (m, 2H), 2.04 - 1.90 (m, 2H), 1.76 - 1.56 (m, 2H), 1.54 - 1.30 (m, 12H), 1.23 - 1.13 (m, 2H), 0.83 (dd, J = 12.3, 6.8 Hz, 6H).

[0301] Example 5:

[0302] Preparation of 2-(methylamino)ethyl-1-(2-chloro-4-(3-((2-(2,6-dioxopiperidin-3-yl)-1- oxoisoindol-5-yl)methyl)ureido)phenyl)cyclopropane-1-carboxylate.

[0303] Reaction Scheme:

[0304] Operation Steps:

[0305] Step A: Compound 3-3 (1.7 g, 4.56 mmol) was added to a 50 mL single-neck flask, followed by dimethyl sulfoxide (20 mL) and compound 5-2 (2.48 g, 6.84 mmol), and then DBU (2 mL, 13.68 mmol) was added after stirring for 5 min. The reaction system was protected by nitrogen, and reacted at room temperature for 12 hours. After the reaction was completed by LCMS monitoring, the reaction liquid was poured into water, extracted with ethyl acetate, and then the organic phase was washed with saturated sodium chloride solution. After drying the organic phase with anhydrous sodium sulfate, it was filtered, and the filtrate was concentrated. The obtained crude product was purified by silica gel column (PE / EA = 6:1) to obtain compound 5-3 (1.5 g, yield 82.47%).

[0306] LCMS (ESI) M / Z: 421.0 [M+Na] + .

[0307] 1 H NMR (400 MHz, DMSO-d6) δ 8.36 (d, J = 2.3 Hz, 1H), 8.22 (dd, J = 8.5, 2.3 Hz, 1H), 7.79 (t, J = 11.4 Hz, 1H), 4.16 (t, J = 5.3 Hz, 2H), 3.35 (t, J = 5.3 Hz, 2H), 2.62 (d, J = 13.9 Hz, 3H), 1.74 (d, J = 2.9 Hz, 2H), 1.50 - 1.35 (m, 11H).

[0308] Step B: Tetrahydroxydiboron (1.26 g, 14.04 mmol) was added to a 50 mL single neck flask, dry N,N-dimethylformamide (25 mL) was added, then 4,4'-dipyridyl (548.19 mg, 3.51 mmol) and compound 5-3 (1.4 g, 3.51 mmol) were added quickly in turn. The reaction system was protected by nitrogen, and reacted at room temperature for 5 minutes. After the reaction was completed by LCMS monitoring, a small amount of methanol was added, filtered, and the filtrate was purified by column chromatography (acetonitrile / 0.1% formic acid aqueous solution) to obtain compound 5-4 (1.1 g, yield 84.96%).

[0309] LCMS (ESI) M / Z: 391.1 [M+Na] + .

[0310] 1 H NMR (400 MHz, DMSO-d6) δ 6.98 (d, J = 8.2 Hz, 1H), 6.61 (d, J = 2.2 Hz, 1H), 6.43 (dd, J = 8.3, 2.2 Hz, 1H), 5.33 (s, 2H), 4.04 (t, J = 5.2 Hz, 2H), 3.28 (t, J = 5.3 Hz, 2H), 2.61 (d, J = 12.2 Hz, 3H), 1.50 (d, J = 2.9 Hz, 2H), 1.39 (d, J = 5.4 Hz, 9H), 1.10 (s, 2H).

[0311] Step C: Compound 5-4 (200 mg, 0.54 mmol) was added to a 25 mL vial, after adding dry THF (6 mL), slowly droped bis(trichloromethyl) carbonate (0.13 mL, 1.08 mmol), the reaction system was protected by nitrogen, and reacted at room temperature for 1 hour. After the reaction was completed by LCMS monitoring, the reaction solution was purified by column chromatography (acetonitrile / 0.1% formic acid aqueous solution) to obtain compound 5-5 (300 mg, yield 82.81%).

[0312] LCMS (ESI) M / Z: 568.1 [M-Boc+H] + .

[0313] 1 H NMR (400 MHz, DMSO-d6) δ 10.98 (s, 1H), 8.87 (s, 1H), 7.69 (t, J = 5.2 Hz, 2H), 7.52 (s, 1H), 7.45 (d, J = 7.9 Hz, 1H), 7.21 (q, J = 8.3 Hz, 2H), 6.85 (t, J = 6.0 Hz, 1H), 5.11 (dd, J = 13.3, 5.1 Hz, 1H), 4.44 (t, J = 12.0 Hz, 3H), 4.32 (d, J = 17.4 Hz, 1H), 4.06 (t, J = 5.3 Hz, 2H), 3.29 (t, J = 5.3 Hz, 2H), 2.98 - 2.85 (m, 1H), 2.60 (dd, J = 13.1, 6.2 Hz, 4H), 2.46 - 2.32 (m, 1H), 2.06 - 1.95 (m, 1H), 1.56 (d, J = 2.7 Hz, 2H), 1.38 (d, J = 4.9 Hz, 9H), 1.17 (s, 2H).

[0314] Step D: Compound 5-5 (250 mg, 0.37 mmol) was added to a 50 mL vial, dichloromethane (20 mL) was added, TFA (4 mL) was added at 0°C, the reaction system was protected by nitrogen, and reacted at room temperature for 1 hour. After the reaction was completed by LCMS monitoring, the solvent was evaporated, and the crude product was purified by high performance liquid chromatography to obtain compound 5 (200 mg, yield 94.10%).

[0315] LCMS (ESI) M / Z: 568.2 [M+H] + .

[0316] 1 H NMR (400 MHz, DMSO-d6) δ 10.98 (s, 1H), 9.22 (s, 1H), 8.29 (s, 1H), 7.76 - 7.62 (m, 2H), 7.51 (s, 1H), 7.44 (d, J = 8.0 Hz, 1H), 7.30 - 7.16 (m, 3H), 5.11 (dd, J = 13.3, 5.1 Hz, 1H), 4.43 (dd, J = 14.4, 11.8 Hz, 3H), 4.31 (d, J = 17.4 Hz, 1H), 4.11 (t, J = 5.6 Hz, 2H), 2.96 - 2.84 (m, 3H), 2.64 - 2.56 (m, 1H), 2.44 - 2.32 (m, 4H), 2.05 - 1.95 (m, 1H), 1.69 - 1.56 (m, 2H), 1.26 - 1.13 (m, 2H).

[0317] Example 6:

[0318] Preparation of 2-((((4-((S)-2-(S)-2-[(6-(2,5-dioxo-2,5-dihydro-1H-pyrrol-1- yl)hexanoylamido)-3-methylbutanoylamido)-5-ureidopentanoylamido)benzyl)oxy)carbonyl)(methyl)amino)ethyl-1-(2-chloro-4-(3-(((2-(6-dioxopiperidin-3-yl)-1- oxoisoindolin-5-yl)methyl)ureido)phenyl)cyclopropane-1-carboxylate.

[0319] Reaction Scheme:

[0320] Procedure:

[0321] Step A: Compound 5 (60 mg, 0.11 mmol) was added to a 25 mL single-neck flask, followed by the addition of N,N-dimethylformamide (7 mL), compound 1-10 (89.27 mg, 0.12 mmol), HOBT (7.43 mg, 0.06 mmol) and 2,4-dimethylpyridine (23.57 mg, 0.22 mmol). The reaction system was reacted at 40 °C for 12 hours under nitrogen protection. After the reaction was completed by LCMS monitoring, compound 6 (40 mg, yield 32.52%) was obtained by preparative purification by high performance liquid chromatography.

[0322] LCMS (ESI) M / Z: 1166.3 [M+H] + .

[0323] 1H NMR (400 MHz, DMSO-d6) δ 10.98 (s, 1H), 9.99 (s, 1H), 8.85 (s, 1H), 8.09 (t, J = 8.1 Hz, 1H), 7.79 (d, J = 8.6 Hz, 1H), 7.69 (d, J = 8.2 Hz, 2H), 7.64 - 7.55 (m, 2H), 7.51 (s, 1H), 7.44 (d, J = 7.8 Hz, 1H), 7.27 (t, J = 8.8 Hz, 2H), 7.24 - 7.12 (m, 2H), 6.99 (s, 2H), 6.91 - 6.80 (m, 1H), 5.97 (t, J = 5.4 Hz, 1H), 5.46 (d, J = 46.2 Hz, 2H), 5.10 (dd, J = 13.2, 4.9 Hz, 1H), 4.96 (d, J = 15.0 Hz, 2H), 4.50 - 4.34 (m, 4H), 4.30 (d, J = 17.2 Hz, 1H), 4.19 (t, J = 7.7 Hz, 1H), 4.07 (d, J = 4.0 Hz, 2H), 3.36 (t, J = 6.9 Hz, 4H), 3.08 - 2.85 (m, 3H), 2.63 (t, J = 18.4 Hz, 4H), 2.46 - 2.30 (m, 1H), 2.25 - 2.06 (m, 2H), 2.05 - 1.89 (m, 2H), 1.75 - 1.31 (m, 10H), 1.25 - 1.10 (m, 4H), 0.83 (dd, J = 12.1, 6.7 Hz, 6H).

[0324] Example 7:

[0325] Preparation of 1-(methylamino)propan-2-yl 2-(2-chloro-4-(3-((2-(2,6-dioxopiperidin-3-yl)-1- oxoisoindolin-5-yl)methyl)ureido)phenyl)carbamate.

[0326] Reaction Scheme:

[0327] Procedure:

[0328] Step A: Compound 7-1 (1.2 g, 13.46 mmol) was dissolved in THF (20 mL), TEA (1.87 mL, 13.46 mmol) and Boc20 (3.09 mL, 13.46 mmol) were added successively. The reaction system was protected by nitrogen, and reacted at room temperature for 2 hours. LCMS was used to monitor the completion of the reaction. The reaction liquid was concentrated under reduced pressure, and the obtained crude product was purified by silica gel column (PE / EA = 15:1-3:1) to obtain compound 7-2 (2.2 g, yield 86.35%).

[0329] 1H NMR (400 MHz, DMSO-d6) δ 4.62 (d, J = 12.3 Hz, 1H), 3.76 (dq, J = 11.8, 5.9 Hz, 1H), 3.14 (dd, J = 13.8, 5.3 Hz, 1H), 2.98 (dd, J = 13.8, 6.9 Hz, 1H), 2.82 (d, J = 9.2 Hz, 3H), 1.38 (s, 9H), 1.00 (d, J = 6.2 Hz, 3H).

[0330] Step B: Compound 7-2 (990 mg, 5.23 mmol) was dissolved in dry dichloromethane (30 mL), 2-(2-chloro-4-nitrophenyl)acetic acid (940 mg, 4.36 mmol), DCC (1349 mg, 6.54 mmol) and DMAP (266 mg, 2.18 mmol) were added, the reaction system was protected by nitrogen, and reacted at room temperature for 2 hours. After the reaction was completed by LCMS monitoring, the reaction solution was filtered, and the filtrate was concentrated. The obtained crude product was purified by column chromatography (acetonitrile / 0.1% aqueous ammonium bicarbonate solution) to finally obtain compound 7-3 (1.5 g, yield 88.93%).

[0331] Step C: Tetrahydroxyboron (929 mg, 10.36 mmol) was dissolved in dry N,N- dimethylformamide (25 mL), and then 4,4'-dipyridine (404 mg, 2.59 mmol) and compound 7-3 (1.0 g, 2.59 mmol) were added quickly in turn. The reaction system was protected by nitrogen, and reacted at room temperature for 5 minutes. After the reaction was completed by LCMS monitoring, a small amount of methanol was added for quenching, filtered, and the filtrate was purified by column chromatography (acetonitrile / 0.1% aqueous ammonium bicarbonate solution) to finally obtain compound 7-4 (800 mg, yield 86.72%).

[0332] LCMS (ESI) M / Z: 357.2 [M+H] + .

[0333] 1 H NMR (400 MHz, DMSO-d6) δ 6.97 (d, J = 8.2 Hz, 1H), 6.61 (d, J = 2.2 Hz, 1H), 6.46 (dd, J = 8.2, 2.3 Hz, 1H), 5.30 (s, 2H), 5.00 (dd, J = 9.0, 4.2 Hz, 1H), 3.54 (s, 2H), 3.26 (t, J = 5.4 Hz, 2H), 2.72 (d, J = 6.3 Hz, 3H), 1.37 (d, J = 12.0 Hz, 9H), 1.11 (d, J = 6.3 Hz, 3H).

[0334] Step D: Compound 7-4 (525 mg, 1.47 mmol) was dissolved in dry THF (15 mL), then diphosgene (0.34 mL, 2.79 mmol) was added dropwise slowly. The reaction was stirred at room temperature for 1 h under nitrogen. The solvent was removed by rotary evaporation under reduced pressure at low temperature. The residue was dissolved in dry N,N-dimethylformamide (15 mL), then a solution of compound 1-3 (402 mg, 1.47 mmol) and triethylamine (0.41 mL, 2.94 mmol) in N,N-dimethylformamide (5 mL) was added dropwise slowly. The reaction was stirred at room temperature for 1 h under nitrogen. After the reaction was completed by LCMS monitoring, purification was performed by column chromatography (acetonitrile / 0.1% formic acid in water) to give compound 7-5 (287 mg, yield 69.41%).

[0335] LCMS (ESI) M / Z: 555.9 [M-Boc+H] + .

[0336] 1 H NMR (400 MHz, DMSO-d6) δ 10.98 (s, 1H), 8.86 (s, 1H), 7.70 (d, J = 7.9 Hz, 2H), 7.52 (s, 1H), 7.45 (d, J = 7.9 Hz, 1H), 7.27 - 7.15 (m, 2H), 6.86 (t, J = 6.0 Hz, 1H), 5.19 - 4.96 (m, 2H), 4.52 - 4.27 (m, 4H), 3.68 (s, 2H), 3.29 - 3.24 (m, 2H), 2.98 - 2.90 (m, 1H), 2.73 (d, J = 5.9 Hz, 3H), 2.65 - 2.56 (m, 1H), 2.39 (qd, J = 13.2, 4.3 Hz, 1H), 2.01 (ddd, J = 10.1, 5.1, 3.0 Hz, 1H), 1.39 (s, 9H), 1.13 (d, J = 6.3 Hz, 3H).

[0337] Step E: Compound 7-5 (60 mg, 0.091 mmol) was dissolved in dichloromethane (4 mL), then TFA (2 mL) was added dropwise. The reaction was stirred at room temperature for 1 h under nitrogen. After the reaction was completed by LCMS monitoring, the solvent was removed by rotary evaporation. Purification was performed by high performance liquid chromatography to give compound 7 (30 mg, yield 59%).

[0338] LCMS (ESI) M / Z: 556.2 [M+H] + .

[0339] 1H NMR (400 MHz, DMSO-d6) δ 10.98 (s, 1H), 9.19 (s, 1H), 8.27 (s, 1H), 7.75 - 7.64 (m, 2H), 7.51 (s, 1H), 7.44 (d, J = 7.9 Hz, 1H), 7.27 - 7.17 (m, 3H), 5.10 (dd, J = 13.3, 5.1 Hz, 1H), 5.00 - 4.91 (m, 1H), 4.48 - 4.39 (m, 3H), 4.31 (d, J = 17.3 Hz, 1H), 3.69 (d, J = 1.6 Hz, 2H), 2.96 - 2.86 (m, 1H), 2.82 - 2.69 (m, 2H), 2.65 - 2.54 (m, 1H), 2.41 - 2.33 (m, 4H), 2.08 - 1.92 (m, 1H), 1.17 (d, J = 6.3 Hz, 3H).

[0340] Example 8:

[0341] Preparation of 1-((((4-((S)-2-(S)-2-[(6-(2,5-dioxo-2,5-dihydro-1H-pyrrol-1- yl)hexanoylamido)-3-methylbutanoylamido)-5-ureidopentanoylamido)benzyl)oxy)carbonyl)(methyl)amino)propan-2-yl-2-(2-chloro-4-(3-((2-(2,6-dioxopiperidin-3-yl)-1- oxoisoindolin-5-yl)methyl)ureido)phenyl)acetate.

[0342] Reaction Scheme:

[0343] Procedure:

[0344] Step A: Compound 7 (30 mg, 0.054 mmol) was dissolved in N,N-dimethylformamide (4 mL), compound 1-10 (41.8 mg, 0.057 mmol), HOBT (3.7 mg, 0.027 mmol) and 2,4-dimethylpyridine (11.57 mg, 0.11 mmol), the reaction system was protected by nitrogen, and reacted at 40 °C for 12 hours. After the reaction was completed by LCMS monitoring, the reaction solution was purified by high performance liquid chromatography to obtain compound 8 (10 mg, yield 16.05%).

[0345] LCMS (ESI) M / Z: 1154.4 [M+H] + .

[0346] 1H NMR (400 MHz, DMSO-d6) δ 10.97 (s, 1H), 10.01 (d, J = 23.0 Hz, 1H), 8.89 (s, 1H), 8.08 (dd, J = 7.0, 2.2 Hz, 2H), 8.01 (d, J = 8.4 Hz, 1H), 7.79 (d, J = 8.6 Hz, 1H), 7.72 - 7.65 (m, 1H), 7.59 (d, J = 8.5 Hz, 2H), 7.57 - 7.47 (m, 2H), 7.46 - 7.34 (m, 2H), 7.28 (d, J = 8.6 Hz, 1H), 7.19 (s, 1H), 5.97 (t, J = 5.4 Hz, 1H), 5.42 (t, J = 39.4 Hz, 3H), 5.20 - 4.87 (m, 2H), 4.52 - 4.27 (m, 3H), 4.25 - 4.06 (m, 1H), 3.63 (s, 1H), 3.40 - 3.23 (m, 8H), 3.08 - 2.87 (m, 3H), 2.79 (d, J = 10.9 Hz, 1H), 2.59 (d, J = 17.9 Hz, 1H), 2.45 - 2.26 (m, 1H), 2.15 (dtd, J = 20.7, 13.9, 7.0 Hz, 2H), 1.96 (dt, J = 13.7, 7.8 Hz, 2H), 1.68 (s, 1H), 1.63 - 1.29 (m, 8H), 1.28 - 1.03 (m, 5H), 0.88 - 0.78 (m, 6H).

[0347] Example 9:

[0348] Preparation of 2-(methylamino)ethyl (2-chloro-4-(3-((2-(2,6-dioxopiperidin-3-yl)-1- oxoisoindolin-5-yl)methyl)ureido)phenyl)carbamate.

[0349] Reaction Scheme:

[0350] Procedure:

[0351] Step A: Compound 9-1 (1 g, 5.79 mmol) was dissolved in dry THF (30 mL), then bis(trichloromethyl) carbonate (1.33 mL, 11.0 mmol) was added dropwise slowly. The reaction was stirred at room temperature for 1 h under nitrogen. The solvent was removed by rotary evaporation under reduced pressure. The residue was dissolved in dry N,N-dimethylformamide (30 mL), then a solution of compound 1-3 (1.58 g, 1.47 mmol) and triethylamine (1.61 mL, 11.58 mmol) in N,N-dimethylformamide (5 mL) was added dropwise slowly. The reaction was stirred at room temperature for 1 h under nitrogen. After the reaction was completed by LCMS monitoring, the product was purified by column chromatography (acetonitrile / 0.1% formic acid in water) to give compound 9-2 (2.3 g, 84.12% yield).

[0352] LCMS (ESI) M / Z: 470.0 [M-H] - .

[0353] Step B: Tetrahydroxydiboron (600 mg, 6.8 mmol) was dissolved in dry N,N- dimethylformamide (2 mL), then 4,4'-dipyridine (540 mg, 3.4 mmol) and compound 9-2 (800 mg, 1.7 mmol) were added quickly in turn. The reaction was stirred at room temperature for 5 min under nitrogen. After the reaction was completed by LCMS monitoring, a small amount of methanol was added to quench the reaction. The mixture was filtered, and the filtrate was purified by column chromatography (acetonitrile / 0.1% ammonium bicarbonate in water) to give compound 9-3 (500 mg, 62.50% yield).

[0354] LCMS (ESI) M / Z: 442.0 [M+H] + .

[0355] Step C: Compound 9-4 (145 mg, 1 mmol) was dissolved in THF (2 mL) and dichloromethane (1 mL), then compound 9-5 (335 mg, 2 mmol) and pyridine (0.27 mL, 4 mmol) were added. The reaction was stirred at room temperature for 2 h under nitrogen. After the reaction was completed by LCMS monitoring, the solvent was removed by rotary evaporation to give compound 9-6 (250 mg) as a crude product, which was used directly in the next step.

[0356] LCMS (ESI) M / Z: 363.2 [M+Na] + .

[0357] Step D: Compound 9-6 (250 mg, 0.74 mmol) was dissolved in super dry MeCN (5 mL), compound 9-3 (300 mg, 0.68 mmol), TEA (0.5 mL) and DMAP (150 mg, 1.22 mmol) were added, the reaction was stirred at room temperature for 2 hours under nitrogen protection. After the reaction was completed by LCMS monitoring, the reaction solution was filtered, the filtrate was concentrated, and the obtained crude product was purified by column chromatography (acetonitrile / 0.1% aqueous ammonium bicarbonate solution) to finally obtain compound 9-7 (80 mg, yield 32.12%).

[0358] LCMS (ESI) M / Z: 641.3 [M-H] - .

[0359] Step E: Compound 9-7 (80 mg, 0.125 mmol) was dissolved in dichloromethane (4 mL), and then TFA (2 mL) was added dropwise, and the reaction was stirred at room temperature for 1 hour under nitrogen protection. After the reaction was completed by LCMS monitoring, the solvent was rotary evaporated, and the crude product was purified by high performance liquid chromatography to obtain compound 9 (30 mg, yield 49%).

[0360] LCMS (ESI) M / Z: 543.1 [M+H] + .

[0361] 1 H NMR (400 MHz, DMSO-d6) δ 10.97 (s, 1H), 8.99 (d, J = 2.0 Hz, 1H), 8.88 (s, 1H), 8.60 (s, 2H), 7.72 (dd, J = 22.0, 5.1 Hz, 2H), 7.51 (s, 1H), 7.42 (dd, J = 12.5, 8.5 Hz, 2H), 7.22 (dd, J = 8.8, 2.4 Hz, 1H), 7.09 - 6.90 (m, 1H), 5.21 - 4.95 (m, 1H), 4.57 - 4.21 (m, 6H), 3.22 (s, 2H), 2.99 - 2.83 (m, 1H), 2.68 - 2.55 (m, 4H), 2.45 - 2.31 (m, 1H), 2.08 - 1.92 (m, 1H).

[0362] Example 10:

[0363] Preparation of 4-((S)-2-(S)-2-(6-(2,5-dioxo-2,5-dihydro-1H-pyrrol-1-yl)hexanamido)-3- methylbutanamido)-5-ureidopentanamido)benzyl (2-(2-chloro-4-(3-((2-(2,6-dioxopiperidin-3- yl)-1-oxoisoindolin-5-yl)methyl)ureido)phenyl)carbamoyl)oxy)ethyl)(methyl)carbamate.

[0364] Reaction Scheme:

[0365] Operation Steps:

[0366] Step A: Compound 9 (20 mg, 0.037 mmol) was dissolved in N,N-dimethylformamide (4 mL), compound 1-10 (30 mg, 0.037 mmol), HOBT (3.7 mg, 0.027 mmol) and 2,4-dimethylpyridine (11.57 mg, 0.11 mmol), the reaction system was protected by nitrogen, and reacted at 40 °C for 12 hours. After the reaction was completed by LCMS monitoring, compound 10 (10 mg, yield 32.16%) was obtained by high performance liquid chromatography preparation purification.

[0367] LCMS (ESI) M / Z: 1141.3 [M+H] + .

[0368] 1 H NMR (400 MHz, DMSO-d6) δ 10.98 (s, 1H), 9.99 (s, 1H), 8.92 (s, 1H), 8.85 (s, 1H), 8.08 (d, J = 7.5 Hz, 1H), 7.80 (d, J = 8.6 Hz, 1H), 7.71 (dd, J = 15.2, 5.1 Hz, 2H), 7.64 - 7.53 (m, 2H), 7.52 (s, 1H), 7.44 (d, J = 7.9 Hz, 1H), 7.35 - 7.22 (m, 3H), 7.18 (d, J = 8.4 Hz, 1H), 7.00 (s, 2H), 6.84 (t, J = 5.9 Hz, 1H), 5.97 (t, J = 5.7 Hz, 1H), 5.41 (s, 2H), 5.10 (dd, J = 13.3, 5.1 Hz, 1H), 4.99 (s, 2H), 4.52 - 4.28 (m, 5H), 4.23 - 4.04 (m, 3H), 3.47 (s, 2H), 3.36 (t, J = 7.1 Hz, 2H), 3.09 - 2.79 (m, 6H), 2.69 - 2.55 (m, 1H), 2.45 - 2.32 (m, 1H), 2.23 - 2.06 (m, 2H), 2.03 - 1.89 (m, 2H), 1.74 - 1.56 (m, 2H), 1.51 - 1.31 (m, 6H), 1.23 - 1.10 (m, 2H), 0.83 (dd, J = 12.5, 6.8 Hz, 6H).

[0369] Example 11:

[0370] Preparation of 1-(4-(2-(2-(allylamine)ethoxy)ethyl)-3-chlorophenyl)-3-((2-(2,6-dioxopiperidin-3-yl)-1-oxoisoindolin-5-yl)methyl)urea.

[0371] Reaction Scheme:

[0372] Procedure:

[0373] Step A: Compound 11-1 (5 g, 19.31 mmol) was taken in 100 mL single neck flask, SOCl2 (30 mL) was added and heated to 75 °C for 1 h. Most of the SOCl2 was removed by distillation under reduced pressure, after cooling to room temperature, ammonia in isopropanol (5.72 mL, 20 mmol) was added slowly and allowed to react for 2 h at room temperature. After completion of the reaction as monitored by LCMS, the compound 11-2 (2.8 g, yield 56%) was purified by column chromatography (acetonitrile / 0.1% formic acid in water).

[0374] LCMS (ESI) M / Z: 300.0 [M+ACN+H] + .

[0375] Step B: Compound 11-2 (2.8 g, 10.85 mmol) was taken in 100 mL single neck flask, dissolved in THF (20 mL), borane tetrahydrofuran complex (27 mL, 27.13 mmol) was added slowly at room temperature followed by heating to 70 °C for 2 h. After completion of the reaction as monitored by LCMS, the reaction was quenched by drop wise addition of methanol, solvent was removed under reduced pressure, the crude was purified by column chromatography (acetonitrile / 0.1% formic acid in water) to get compound 11-3 (2.38 g, yield 90%).

[0376] LCMS (ESI) M / Z: 245.0 [M+H] + .

[0377] Step C: Compound 11-3 (2.38 g, 9.75 mmol) was taken in 100 mL single neck flask, MeCN (25 mL), N,N-diisopropylethylamine (3 g, 24.38 mmol) and allyl bromide (1.76 g, 14.63 mmol) were added and allowed to react for 1 h at room temperature, then Boc20 (4.27 g, 19.5 mmol) was added and stirring was continued for 1 h at room temperature. Solvent was removed under reduced pressure, the residue was purified by column chromatography (acetonitrile / 0.1% formic acid in water) to get compound 11-4 (480 mg, yield 12%).

[0378] LCMS (ESI) M / Z: 285.2 [M-BOC+H] + .

[0379] Step D: Compound 11-4 (0.48 g, 1.25 mmol) was taken in 25 mL vial, dissolved in N,N-dimethylformamide (5 mL), B2(OH)4(0.445 g, 5 mmol) and 4,4’-dipyridine (18 mg, 0.125 mmol) were added and stirred, reaction was carried out at room temperature for half an hour. After completion of reaction as monitored by TLC, the reaction mixture was purified by column chromatography (acetonitrile / 0.1% aqueous ammonium bicarbonate solution) to get compound 11-5 (0.342 g, yield 77.6%).

[0380] LCMS (ESI) M / Z: 255.2 [M-BOC+H] + .

[0381] Step E: Compound 11-5 (0.342 g, 0.97 mmol) was taken in 50 mL vial, dissolved in THF (5 mL), followed by addition of TEA (0.97 g, 9.7 mmol) and diphosgene (0.384 g, 1.94 mmol), reaction was carried out at room temperature for half an hour, followed by concentration under reduced pressure, the residue was dissolved in N,N-dimethylformamide (5 mL), followed by addition of compound 1-3 (0.32 g, 1.16 mmol), reaction was carried out at room temperature for 1 hour. After completion of reaction as monitored by TLC, the reaction mixture was purified by column chromatography (acetonitrile / 0.1% aqueous formic acid) to get compound 11-6 (0.37 g, yield 59.45%).

[0382] LCMS (ESI) M / Z: 554.3 [M-BOC+H] + .

[0383] Step F: Compound 11-6 (0.37 g, 0.57 mmol) was taken in 50 mL vial, dissolved in dichloromethane (10 mL), trifluoroacetic acid (4 mL) was added and stirred, reaction was carried out at room temperature for 2 hours. After completion of reaction as monitored by LCMS, the reaction mixture was purified by column chromatography to get compound 11 (90 mg, yield 29%).

[0384] LCMS (ESI) M / Z: 554.3 [M+H] + .

[0385] 1H NMR (400 MHz, DMSO-d6) δ 11.01 (br s, 1H), 9.34 (s, 1H), 8.36 (s, 1H), 7.69 (s, 2H), 7.58 - 7.35 (m, 3H), 7.21 (s, 2H), 5.85 (s, 1H), 5.43 - 5.25 (m, 2H), 5.09 (s, 1H), 4.52 - 4.23 (m, 4H), 3.59 (s, 4H), 3.42 (s, 2H), 3.02 - 2.75 (m, 5H), 2.67 - 2.54 (m, 1H), 2.44 - 2.28 (m, 1H), 1.99 (s, 1H).

[0386] Example 12:

[0387] Preparation of 4-((S)-2-(S)-2-(6-(2,5-dioxo-2,5-dihydro-1H-pyrrol-1-yl)hexanamido)-3- methylbutanamido)-5-ureidopentyl)phenylpropenyl(2-(2-(2,6-dioxo-3-yl)-1-oxoisoindol- 5-yl)methyl)ureidophenoxy)carbamate.

[0388] Reaction Scheme:

[0389] Procedure:

[0390] Step A: Compound 11 (160 mg, 0.29 mmol) was added to a 25 mL single-neck flask, followed by the addition of N,N-dimethylformamide (7 mL), compound 1-10 (240 mg, 0.33 mmol), HOBT (10 mg, 0.08 mmol) and 2,4-dimethylpyridine (35 mg, 0.33 mmol). The reaction system was reacted at 40 °C for 12 hours under nitrogen protection. After the reaction was completed by LCMS monitoring, compound 12 (140 mg, yield 42.04%) was obtained by preparative purification by high performance liquid chromatography.

[0391] LCMS (ESI) M / Z: 1153.6 [M+H] + .

[0392] 1H NMR (400 MHz, DMSO-d6) δ 10.98 (s, 1H), 9.99 (s, 1H), 8.85 (s, 1H), 8.09 (t, J = 8.1 Hz, 1H), 7.79 (d, J = 8.6 Hz, 1H), 7.69 (d, J = 8.2 Hz, 2H), 7.64 - 7.55 (m, 2H), 7.51 (s, 1H), 7.44 (d, J = 7.8 Hz, 1H), 7.27 (t, J = 8.8 Hz, 2H), 7.24 - 7.12 (m, 2H), 6.99 (s, 2H), 6.91 - 6.80 (m, 1H), 5.97 (t, J = 5.4 Hz, 1H), 5.46 (d, J = 46.2 Hz, 2H), 5.10 (dd, J = 13.2, 4.9 Hz, 1H), 4.96 (d, J = 15.0 Hz, 2H), 4.50 - 4.34 (m, 4H), 4.30 (d, J = 17.2 Hz, 1H), 4.19 (t, J = 7.7 Hz, 1H), 4.07 (d, J = 4.0 Hz, 2H), 3.36 (t, J = 6.9 Hz, 4H), 3.08 - 2.85 (m, 3H), 2.63 (t, J = 18.4 Hz, 4H), 2.46 - 2.30 (m, 1H), 2.25 - 2.06 (m, 2H), 2.05 - 1.89 (m, 2H), 1.75 - 1.31 (m, 10H), 1.25 - 1.10 (m, 4H), 0.83 (dd, J = 12.1, 6.7 Hz, 6H).

[0393] Example 13:

[0394] Preparation of 1-(4-(2-(2-(propargylamino)ethoxy)ethyl)-3-chlorophenyl)-3-((2-(2,6-dioxopiperidin-3-yl)-1-oxoisoindolin-5-yl)methyl)urea.

[0395] Reaction Scheme:

[0396] Procedure:

[0397] Step A: Compound 13-1 (1.9 g, 7.8 mmol) was taken in a 100 mL single necked flask, MeCN (20 mL), N,N-diisopropylethylamine (2.4 g, 19.50 mmol) and propargyl bromide (1.41 g, 11.70 mmol) were added, reaction was carried out at room temperature for 1 h, Boc20 (3.42 g, 15.6 mmol) was added and reaction was continued at room temperature for 1 h. The reaction mass was concentrated under reduced pressure to remove the solvent, the crude obtained was purified by column chromatography (acetonitrile / 0.1% formic acid in water) to get compound 13-2 (370 mg, yield 12.41%).

[0398] LCMS (ESI) M / Z: 283.0 [M-BOC+H] + .

[0399] Step B: Compound 13-2 (0.37 g, 0.96 mmol) was taken in 25 mL single necked flask, dissolved in N,N-dimethylformamide (5 mL), diboronic acid (0.445 g, 5 mmol) and 4,4’-dipyridine (18 mg, 0.125 mmol) were added and stirred for half an hour at room temperature. After completion of reaction monitored by TLC, the reaction mixture was purified by column chromatography (acetonitrile / 0.1% aqueous ammonium bicarbonate solution) to get compound 13-3 (0.314 g, yield 92.6%).

[0400] LCMS (ESI) M / Z: 253.2 [M-BOC+H] + .

[0401] Step C: Compound 13-3 (0.314 g, 0.88 mmol) was taken in 50 mL single necked flask, dissolved in THF (5 mL), followed by addition of TEA (0.88 g, 8.8 mmol) and diphosgene (0.35 g, 1.76 mmol) and stirred for half an hour at room temperature, followed by concentration under reduced pressure, the residue was dissolved in N,N-dimethylformamide (5 mL), then compound 1-3 (0.29 g, 1.05 mmol) was added and stirred for 1 hour at room temperature. After completion of reaction monitored by TLC, the reaction mixture was purified by column chromatography (acetonitrile / 0.1% aqueous formic acid) to get compound 13-4 (0.29 g, yield 60.12%).

[0402] LCMS (ESI) M / Z: 552.3 [M-BOC+H] + .

[0403] Step D: Compound 13-4 (0.37 g, 0.45 mmol) was taken in 50 mL single necked flask, dissolved in dichloromethane (9 mL), trifluoroacetic acid (3 mL) was added and stirred for 2 hours at room temperature. After completion of reaction monitored by LCMS, the reaction mixture was purified by column chromatography to get compound 13 (130 mg, yield 41.3%).

[0404] LCMS (ESI) M / Z: 552.3 [M+H] + .

[0405] 1H NMR (400 MHz, DMSO-d6) δ 10.98 (s, 1H), 8.88 (s, 1H), 8.16 (s, 1H), 7.68 (dd, J = 7.6, 4.9 Hz, 2H), 7.51 (s, 1H), 7.44 (d, J = 7.9 Hz, 1H), 7.26 - 7.12 (m, 2H), 6.92 (t, J = 5.9 Hz, 1H), 5.11 (dd, J = 13.3, 5.1 Hz, 1H), 4.49 - 4.29 (m, 4H), 3.59 - 3.52 (m, 6H), 3.30 (t, J = 2.2 Hz, 1H), 2.97 - 2.81 (m, 5H), 2.60 (dd, J = 15.5, 2.1 Hz, 1H), 2.38 (qd, J = 13.3, 4.4 Hz, 1H), 2.00 (dt, J = 5.0, 4.0 Hz, 1H).

[0406] Example 14:

[0407] Preparation of 4-(S)-2-(S)-2-(6-(2,5-dioxo-2,5-dihydro-1H-pyrrol-1-yl)hexanamido)-3- methylbutanamido)-5-uracilpentanamide) benzyl (2-(2-chloro-4-(3-(2-(2,6-dioxopyrimidin-3-yl)-1- oxoisoindolin-5-yl)methyl)uracil)phenethyl)oxy)ethyl) (prop-2-en-1-yl)carbamate.

[0408] Reaction Scheme:

[0409] Procedure:

[0410] Step A: Compound 13 (100 mg, 0.18 mmol) was added to a 25 mL single-neck flask, followed by the addition of N,N-dimethylformamide (4 mL), compound 1-10 (140 mg, 0.19 mmol), HOBT (13 mg, 0.10 mmol), and 2,4-dimethylpyridine (40 mg, 0.35 mmol). The reaction system was reacted at 40 °C for 12 hours under nitrogen protection. After the reaction was completed, LCMS monitoring, the reaction liquid was purified by high performance liquid chromatography to obtain compound 14 (30 mg, yield 14.4%).

[0411] LCMS (ESI) M / Z: 1150.6 [M+H] + .

[0412] 1H NMR (400 MHz, DMSO-d6) δ 10.99 (s, 1H), 10.01 (s, 1H), 8.87 (s, 1H), 8.10 (d, J = 7.3 Hz, 1H), 7.83 (d, J = 8.5 Hz, 1H), 7.72 - 7.64 (m, 2H), 7.60 (d, J = 8.4 Hz, 2H), 7.51 (s, 1H), 7.44 (d, J = 7.9 Hz, 1H), 7.30 (s, 2H), 7.15 (s, 2H), 6.96 (s, 2H), 6.91 (s, 1H), 5.11 (dd, J = 13.2, 5.0 Hz, 1H), 5.02 (s, 2H), 4.48 - 4.25 (m, 6H), 4.23 - 4.13 (m, 1H), 4.05 (s, 2H), 3.60 - 3.28 (m, 8H), 3.16 (s, 1H), 3.09 - 2.75 (m, 5H), 2.66 - 2.53 (m, 1H), 2.50 (s, 1H), 2.45 - 2.29 (m, 1H), 2.23 - 2.06 (m, 2H), 2.05 - 1.89 (m, 2H), 1.79 - 1.55 (m, 2H), 1.54 - 1.35 (m, 6H), 1.23 - 1.12 (m, 2H), 0.91 - 0.78 (m, 6H).

[0413] Example 15:

[0414] Preparation of 1-(3-chloro-4-(2-(2,2,2-trifluoroethyl)amino)ethoxy)ethyl)phenyl)-3-((2-(2,6-dioxopiperidin-3-yl)-1-oxoisoindolin-5-yl)methyl)urea.

[0415] Reaction Scheme:

[0416] Procedure:

[0417] Step A: Compound 15-1 (1.8 g, 7.04 mmol) was taken in 100 mL single necked flask, MeOH (40 mL), compound 15-2 (4 g, 29.5 mmol) was added sequentially, stirred at room temperature for 1 h under nitrogen protection, sodium cyanoborohydride (0.468 g, 7.44 mmol) was added and stirred at room temperature for 2 h.

[0418] LCMS monitoring reaction was complete, compound 15-3 (1.012 g, yield 44.00%) was obtained by flash purification (ACN / 0.1% FA in water).

[0419] LCMS (ESI) M / Z: 327.0 [M+H] + .

[0420] Step B: Compound 15-3 (1 g, 3.07 mmol) was taken in 100 mL single necked flask, after adding THF (15 mL), triethylamine (1 g, 9.9 mmol) and Boc20 (2 g, 9.17 mmol) were added, stirred at room temperature overnight.

[0421] After completion of reaction by LCMS, reaction mixture was concentrated, crude obtained was purified by silica gel column (PE / EA = 1:1) to get compound 15-4 (0.7 g, yield 53.57%).

[0422] LCMS (ESI): 371.1 [M-(t-Bu)+H] + .

[0423] Step C: Compound 15-4 (0.7 g, 1.6 mmol) was taken in 50 mL single necked flask, after adding dry DMF (10 mL), tetrahydroxyboron (584 mg, 6.56 mmol) and 4,4’-dipyridine (128 mg, 0.82 mmol) were added, reaction mixture was stirred at 0 degree Celsius for 5 minutes under nitrogen atmosphere.

[0424] After completion of reaction by LCMS, small amount of methanol was added, reaction mixture was purified by flash (ACN / 0.1% FA in water) to get compound 15-5 (470 mg, yield 72.23%).

[0425] LCMS (ESI): 341.0 [M-(t-Bu)+H] + .

[0426] Step D: Compound 15-5 (570 mg, 1.44 mmol) was taken in 25 mL single necked flask, after adding dry THF (6 mL), diphosgene (567 mg, 2.87 mmol) was added drop wise slowly, reaction mixture was stirred at room temperature for 1 hour under nitrogen atmosphere, solvent was evaporated at low temperature, residue was dissolved in dry DMF (3 mL) and added drop wise to compound 1-3 (548 mg, 1.77 mmol) and triethylamine (291 mg, 2.88 mmol) in DMF (5 mL) solution, reaction mixture was stirred at room temperature for 1 hour under nitrogen atmosphere.

[0427] After completion of reaction by LCMS, reaction mixture was purified by flash (ACN / 0.1% FA in water) to get compound 15-6 (800 mg, yield 79.84%).

[0428] LCMS (ESI): 596.2 [M-Boc+H] + .

[0429] Step E: Compound 15-6 (800 mg, 1.15 mmol) was taken in 50 mL single neck flask, DCM (20 mL) was added, TFA (10 mL) was added at 0 °C, the reaction mixture was stirred at room temperature for 1 h under nitrogen atmosphere.

[0430] After completion of reaction by LCMS, the reaction mixture was concentrated, the crude was purified by Prep-HPLC to get compound 15 (650 mg, yield 94.86 %).

[0431] LCMS (ESI): 596.3 [M+H] + .

[0432] 1 H NMR (400 MHz, DMSO-d6) δ 10.98 (s, 1H), 8.93 (s, 1H), 7.73 - 7.63 (m, 2H), 7.51 (s, 1H), 7.44 (d, J = 7.9 Hz, 1H), 7.26 - 7.14 (m, 2H), 6.98 (t, J = 5.9 Hz, 1H), 5.11 (dd, J = 13.3, 5.1 Hz, 1H), 4.50 - 4.25 (m, 4H), 3.95 (q, J = 9.6 Hz, 2H), 3.68 (t, J = 5.2 Hz, 2H), 3.62 (t, J = 7.0 Hz, 2H), 3.17 (t, J = 5.1 Hz, 2H), 2.97 - 2.86 (m, 3H), 2.66 - 2.55 (m, 1H), 2.44 - 2.32 (m, 1H), 2.06 - 1.94 (m, 1H).

[0433] Example 16:

[0434] Preparation of l-(3-chloro-4-(2-((cyanomethyl)amino)ethoxy)ethyl)phenyl)-3-((2-(2,6-dioxopiperidin-3-yl)-l-oxoisoindolin-5-yl)methyl)urea.

[0435] Reaction Scheme:

[0436] Procedure:

[0437] Step A: Compound 16-1 (900 mg, 3.52 mmol) was taken in 100 mL single neck flask, MeOH (10 mL) was added, compound 16-2 (2 g, 14.5 mmol) was added, stirred at room temperature for 1 h under nitrogen atmosphere, sodium cyanoborohydride (2348 mg, 3.72 mmol) was added and stirred at room temperature for 2 h.

[0438] After LCMS monitoring the reaction was completed, the reaction was concentrated, the obtained crude was purified by silica gel column (PE / EA = 1 : 1) to give compound 16-4 (350 mg, yield 57.82%).

[0439] LCMS (ESI): 284.0 [M+H] + .

[0440] Step B: Compound 16-3 (500 mg, 1.53 mmol) was added into a 100 mL single neck flask, after THF (5 mL) was added, triethylamine (500 mg, 4.45 mmol) and Boc20 (1 g, 4.52 mmol) were added, and stirred at room temperature overnight.

[0441] After LCMS monitoring the reaction was completed, the reaction was concentrated, the obtained crude was purified by silica gel column (PE / EA = 1 : 1) to give compound 16-4 (350 mg, yield 57.82%).

[0442] LCMS (ESI): 284.0 [M+H] + .

[0443] Step C: Compound 16-4 (350 mg, 0.8 mmol) was added into a 50 mL single neck flask, dry DMF (4 mL) was added, followed by tetrahydroxyboron (290 mg, 3.2 mmol) and 4,4'-dipyridine (64 mg, 0.4 mmol), and the reaction system was reacted at room temperature for 5 minutes under nitrogen protection.

[0444] After LCMS monitoring the reaction was completed, a small amount of methanol was added, and the system was purified by flash (ACN / 0.1% FA in water) to give compound 16-5 (270 mg, yield 70.32%).

[0445] LCMS (ESI): 284.0 [M+H] + .

[0446] Step D: Compound 16-5 (270 mg, 2.72 mmol) was added into a 25 mL single neck flask, after dry THF (3 mL) was added, diphosgene (280 mg, 1.43 mmol) was slowly added dropwise, and the reaction system was reacted at room temperature for 1 hour under nitrogen protection, and then the solvent was dried by low temperature rotation, and then dissolved in dry DMF (3 mL) and slowly added dropwise into a solution of compound 1-3 (274 mg, 1.77 mmol) and triethylamine (291 mg, 2.88 mmol) in DMF (5 mL), and the reaction system was reacted at room temperature for 1 hour under nitrogen protection.

[0447] LCMS monitoring reaction completion, compound 16-6 (400 mg, yield 82.46%) was obtained by Flash purification (ACN / 0.1% FA in water).

[0448] LCMS (ESI): 653.2 [M+H] + .

[0449] Step E: Compound 16-6 (300 mg, 0.81 mmol) was added to a 50 mL single-necked flask, DCM (5 mL) was added, TMSOTf (308 mg, 2.41 mmol) was added at 0 °C, the reaction system was protected by nitrogen, and the reaction was carried out at room temperature for 3 hours.

[0450] LCMS monitoring reaction completion, compound 16-6 (400 mg, yield 82.46%) was obtained by Flash purification (ACN / 0.1% FA in water).

[0451] LCMS (ESI): 553.2 [M+H] + .

[0452] 1 H NMR (400 MHz, DMSO-d6) δ 10.98 (s, 1H), 8.79 (s, 1H), 8.70 (s, 2H), 7.68 (dd, J = 8.6, 5.0 Hz, 2H), 7.51 (s, 1H), 7.44 (dd, J = 7.7, 5.0 Hz, 1H), 7.28 - 7.11 (m, 2H), 6.82 (t, J = 6.0 Hz, 1H), 5.19 - 5.01 (m, 1H), 4.51 - 4.25 (m, 4H), 3.74 (s, 2H), 3.62 - 3.44 (m, 4H), 2.98 - 2.83 (m, 3H), 2.80 (s, 2H), 2.70 - 2.57 (m, 1H), 2.43 - 2.31 (m, 1H), 2.05 - 1.95 (m, 1H).

[0453] Example 17:

[0454] Preparation of 2-amino-N-(2-(2-chloro-4-(3-(2-(2,6-dioxopiperidin-3-yl)-1- oxoisoindolin-5-yl)methyl)ureido)phenethoxy)ethyl)acetamide.

[0455] Reaction Scheme:

[0456] Procedure:

[0457] Step A:

[0458] Compound 16-6 (300 mg, 0.81 mmol) was taken in 50 mL single necked flask, DCM (5 mL) was added, TFA (2.5 mL) was added at 0 °C, the reaction mixture was stirred at room temperature for 3 h under nitrogen atmosphere.

[0459] After completion of reaction by LCMS, the reaction mixture was concentrated, the crude was purified by Prep-HPLC to get compound 17 (200 mg, yield 79.96 %).

[0460] LCMS (ESI): 571.3 [M+H] + .

[0461] 1 H NMR (400 MHz, DMSO-d6) δ 10.98 (s, 1H), 8.84 (s, 1H), 8.70 (s, 2H), 7.82 (s, 1H), 7.72 - 7.65 (m, 2H), 7.57 (s, 1H), 7.51 (s, 1H), 7.46 - 7.41 (m, 1H), 7.26 - 7.15 (m, 2H), 6.92 - 6.83 (m, 1H), 5.11 (dd, J = 13.3, 5.0 Hz, 1H), 4.51 - 4.25 (m, 4H), 3.72 - 3.64 (m, 4H), 3.61 - 3.57 (m, 2H), 3.16 - 3.10 (m, 2H), 2.97 - 2.86 (m, 3H), 2.63 - 2.57 (m, 1H), 2.44 - 2.33 (m, 1H), 2.06 - 1.93 (m, 1H).

[0462] Example 18:

[0463] Preparation of 1-(3-chloro-4-(2-(2-(cyclopropylamino)ethoxy)ethyl)phenyl)-3-((2-(2,6- dioxopiperidin-3-yl)-1-oxoisoindolin-5-yl)methyl)urea.

[0464] Reaction Scheme:

[0465] Procedure:

[0466] Step A: Compound 18-1 (1.5 g, 6.16 mmol) was taken in MeOH (20 mL), then cyclopropyl amine (1.406 g, 24.64 mmol) was added, after 0.5 h sodium cyano borohydride (1.161 g, 18.84 mmol) was added and the reaction mixture was stirred at room temperature for 1 h.

[0467] After completion of reaction, compound 18-2 (1.08 g, yield 62%) was obtained by Flash purification (ACN / 0.1% TFA in water).

[0468] LCMS (ESI): 285.0 [M+H] + .

[0469] Step B: Compound 18-2 (1 g, 3.51 mmol) was dissolved in THF (20 mL), TEA (3 mL, 21.64 mmol) and Boc20 (2298 mg, 10.53 mmol) were added at room temperature, then stirred at room temperature overnight.

[0470] After completion of reaction, the reaction was quenched by dropwise addition of methanol, the solvent was removed by distillation under reduced pressure, the crude product was purified by Flash purification (ACN / 0.1% FA in water) to obtain compound 18-3 (840 mg, yield 63%).

[0471] LCMS (ESI): 285.1 [M+H-Boc] + .

[0472] Step C: Compound 18-3 (840 mg, 2.18 mmol) was dissolved in DMF (10 mL), diboronic acid (782 mg, 8.72 mmol) was added with stirring, then 4-4’ bipyridine (374 mg, 2.40 mmol) was slowly added, and the reaction was carried out at room temperature until the blue color disappeared.

[0473] After completion of reaction, the reaction was purified by Flash purification (ACN / 0.1% NH4HCO3 in water) to obtain compound 18-4 (610 mg, yield 79%).

[0474] LCMS (ESI): 355.3 [M+H] + .

[0475] Step D: Compound 18-4 (560 mg, 1.58 mmol) was dissolved in THF (10 mL), then TEA (400 mg, 3.95 mmol) and double light gas (469 mg, 2.37 mmol) were added, and the reaction was carried out at room temperature for half an hour, then concentrated under reduced pressure, the residue was dissolved in DMF (10 mL), then compound 1-3 (475 mg, 1.74 mmol) was added, and the reaction was carried out at room temperature for 1 hour.

[0476] After completion of reaction, the reaction was purified by Flash purification (ACN / 0.1% FA in water) to obtain compound 18-5 (760 mg, yield 73%).

[0477] LCMS (ESI): 554.0 [M+H-Boc] + .

[0478] Step E: Compound 18-5 (570 mg, 0.87 mmol) was dissolved in DCM (10 mL), trifluoroacetic acid (5 mL) was added with stirring, and the reaction was allowed to proceed at room temperature for 1 hour.

[0479] After the reaction was monitored to be complete by LCMS, the reaction was purified by flash to give compound 18 (370 mg, yield 77%).

[0480] LCMS (ESI): 554.3 [M+H] + .

[0481] 1 H NMR (400 MHz, DMSO-d6) δ 10.98 (s, 1H), 9.15 - 8.97 (m, 1H), 8.78 (s, 2H), 7.75 - 7.61 (m, 2H), 7.51 (s, 1H), 7.44 (d, J = 7.9 Hz, 1H), 7.26 - 7.17 (m, 2H), 7.10 (t, J = 5.8 Hz, 1H), 5.11 (dd, J = 13.3, 5.1 Hz, 1H), 4.52 - 4.24 (m, 4H), 3.70 - 3.66 (m, 2H), 3.62 (t, J = 7.2 Hz, 2H), 3.26 - 3.12 (m, 2H), 2.93 - 2.83 (m, 3H), 2.74 - 2.55 (m, 2H), 2.44 - 2.30 (m, 1H), 2.04 - 1.94 (m, 1H), 0.84 - 0.66 (m, 4H).

[0482] Example 19:

[0483] Preparation of 1-(3-chloro-4-(2-(2-(methoxyamino)ethoxy)ethyl)phenyl)-3-((2-(2,6-dioxopiperidin-3-yl)-1-oxoisoindolin-5-yl)methyl)urea.

[0484] Reaction Scheme:

[0485] Procedure:

[0486] Step A: Compound 19-1 (8.3 g, 31.97 mmol) was dissolved in THF (200 mL), and borane dimethyl sulfide complex (4.71 mL, 47.1 mmol, 10 M) was added dropwise at 0 °C. The reaction was allowed to proceed at 70 °C under nitrogen for 3 hours.

[0487] After the reaction was completed, the reaction was quenched by slowly adding methanol under ice-bath, and then the solvent was removed by rotary evaporation. Compound 19-2 (6.5 g, yield 82.77%) was obtained by silica gel column purification (PE / EA=5:1).

[0488] TLC: Rf=0.2 (PE / EA=5:1) f

[0489] Step B: Compound 19-2 (6.5 g, 26.46 mmol) was dissolved in DCM (280 mL), and DMP (13.47 g, 31.75 mmol) was added. The reaction system was reacted at room temperature for 2 hours under nitrogen protection.

[0490] After the reaction was completed, the reaction was quenched by slowly adding methanol under ice-bath, and then the solvent was removed by rotary evaporation. Compound 19-2 (6.5 g, yield 82.77%) was obtained by silica gel column purification (PE / EA=5:1).

[0491] TLC: Rf=0.4 (PE / EA=5:1) f

[0492] Step C: Compound 19-3 (1.2 g, 4.93 mmol) was dissolved in methanol (500 mL), and compound 19-4 (1.65 g, 19.72 mmol) was added. The reaction system was reacted at room temperature for 1 hour under nitrogen protection, and then sodium cyanoborohydride (340 mg, 5.42 mmol) was added. The reaction was reacted at room temperature for 1 hour.

[0493] After the reaction was completed, the reaction was quenched by slowly adding methanol under ice-bath, and then the solvent was removed by rotary evaporation. Compound 19-2 (6.5 g, yield 82.77%) was obtained by silica gel column purification (PE / EA=5:1).

[0494] LCMS (ESI): 275.1 [M+H] + .

[0495] Step D: Compound 19-5 (860 mg, 3.13 mmol) was dissolved in THF (40 mL) and deionized water (20 mL), and Boc2O (1.37 g, 6.26 mmol) and sodium hydroxide (375.6 mg, 9.39 mmol) were added. The reaction system was reacted at room temperature for 12 hours under nitrogen protection.

[0496] After the reaction was completed, the reaction was quenched by slowly adding methanol under ice-bath, and then the solvent was removed by rotary evaporation. Compound 19-2 (6.5 g, yield 82.77%) was obtained by silica gel column purification (PE / EA=5:1). ​​

[0497] LCMS (ESI): 275.1 [M-Boc+H] + .

[0498] Step E: Dissolve tetrahydroxydiboron (602.5 mg, 6.72 mmol) in dry DMF (12 mL), then quickly add 4,4'-dipyridine (288.6 mg, 1.85 mmol) and compound 19-6 (630 mg, 1.68 mmol) in sequence. The reaction system is protected by nitrogen, and the reaction is carried out at room temperature for 5 minutes.

[0499] After the reaction is completed by LCMS monitoring, a small amount of methanol is added, filtered, and the filtrate is purified by Flash (ACN / 0.1% FA in water) to obtain compound 19-7 (430 mg, yield 74.19%).

[0500] LCMS (ESI): 345.2 [M+H] + .

[0501] Step F: Dissolve compound 19-7 (800 mg, 2.32 mmol) in dry THF (15 mL), then slowly drop bis-triphosgene (0.56 mL, 4.64 mmol). The reaction system is protected by nitrogen, and the reaction is carried out at room temperature for 1 hour. The solvent is dried by rotary evaporation at low temperature, and the residue is dissolved in dry DMF (5 mL), then slowly dropped into a solution of compound 1-3 (968.66 mg, 2.55 mmol) and triethylamine (3.22 mL, 23.2 mmol) in DMF (12 mL). The reaction system is protected by nitrogen, and the reaction is carried out at room temperature for 1 hour

[0502] After the reaction is completed by LCMS monitoring, the compound 19-8 (1.07 g, yield 71.60%) is obtained by Flash purification (ACN / 0.1% FA in water).

[0503] LCMS (ESI): 544.1 [M-Boc+H] + .

[0504] Step G: Dissolve compound 19-8 (1 g, 1.55 mmol) in DCM (30 mL), and add TFA (6 mL) at 0°C. The reaction system is protected by nitrogen, and the reaction is carried out at room temperature for 1 hour.

[0505] After the reaction is completed by LCMS monitoring, the reaction solution is concentrated, and the crude product is purified by Prep-HPLC to obtain compound 19 (780 mg, yield 92.36%).

[0506] 1H NMR (400 MHz, DMSO-d6) δ 10.98 (s, 1H), 8.80 (s, 1H), 7.68 (dd, J = 8.4, 5.0 Hz, 2H), 7.51 (s, 1H), 7.44 (d, J = 7.9 Hz, 1H), 7.22 (d, J = 8.4 Hz, 1H), 7.16 (dd, J = 8.4, 2.1 Hz, 1H), 6.82 (t, J = 6.0 Hz, 1H), 6.41 (s, 1H), 5.11 (dd, J = 13.3, 5.1 Hz, 1H), 4.43 (t, J = 12.3 Hz, 3H), 4.31 (d, J = 17.3 Hz, 1H), 3.54 (t, J = 7.1 Hz, 2H), 3.48 (t, J = 5.8 Hz, 2H), 3.37 (s, 3H), 2.97 - 2.81 (m, 5H), 2.60 (d, J = 17.4 Hz, 1H), 2.45 - 2.31 (m, 1H), 2.06 - 1.94 (m, 1H).

[0507] LCMS (ESI): 544.3 [M-Boc+H] + .

[0508] Example 20:

[0509] Preparation of 1-(3-chloro-4-(2-(2-guanidinoethoxy)ethyl)phenyl)-3-((2-(2,6-dioxopiperidin-3-yl)-1-oxoisoindolin-5-yl)methyl)urea.

[0510] Reaction Scheme:

[0511] Procedure:

[0512] Step A: Compound 20-1 (500 mg, 2.04 mmol) was dissolved in THF (10 mL), TEA (1.41 mL, 10.20 mmol) and Boc20 (1.41 mL, 6.12 mmol) were added at room temperature, followed by stirring at room temperature for 30 min.

[0513] After monitoring the completion of the reaction by LCMS, the reaction was quenched by dropwise addition of methanol, the solvent was removed under reduced pressure, and the crude product was purified by column chromatography (PE:EA = 10:1-8:1) to obtain compound 20-2 (500 mg, yield 71%).

[0514] LCMS (ESI): 367.2 [M+Na] + .

[0515] Step B: Compound 20-2 (400 mg, 1.16 mmol) was dissolved in DMF (10 mL), tetrahydroxydiboron (416 mg, 4.64 mmol) was added with stirring, then 4-4’ bipyridine (181 mg, 1.16 mmol) was added slowly, the reaction was allowed to proceed at room temperature until the blue color disappeared.

[0516] LCMS monitoring showed the reaction was completed, the reaction solution was purified by Flash (ACN / 0.1% NH4HCO3 in water) to give compound 20-3 (310 mg, yield 85%).

[0517] LCMS (ESI): 337.3 [M+Na] + .

[0518] Step C: Compound 20-3 (300 mg, 0.95 mmol) was dissolved in THF (5 mL), then TEA (240 mg, 2.38 mmol) and diphosgene (282 mg, 1.42 mmol) were added, the reaction was allowed to proceed at room temperature for half an hour, then concentrated under reduced pressure, the residue was dissolved in DMF (5 mL), then compound 1-3 (286 mg, 1.04 mmol) was added, the reaction was allowed to proceed at room temperature for 1 hour.

[0519] LCMS monitoring showed the reaction was completed, the reaction solution was purified by Flash (ACN / 0.1% FA in water) to give compound 20-4 (530 mg, yield 92%).

[0520] LCMS (ESI): 514.2 [M+H-Boc] + .

[0521] Step D: Compound 20-4 (500 mg, 0.81 mmol) was dissolved in DCM (10 mL), trifluoroacetic acid (5 mL) was added with stirring, the reaction was allowed to proceed at room temperature for 1 hour.

[0522] LCMS monitoring showed the reaction was completed, the reaction solution was purified by Flash (ACN / 0.1% FA in water) to give compound 20-5 (370 mg, yield 88%).

[0523] LCMS (ESI): 514.1 [M+H] + .

[0524] Step E: Compound 20-5 (130 mg, 0.25 mmol) was dissolved in DMF (5 mL), TEA (0.21 mL) and compound 20-6 (122 mg, 0.83 mmol) were added with stirring, the reaction was allowed to proceed at room temperature for 24 hours.

[0525] After LCMS monitoring the reaction was completed, the reaction solution was purified by Prep-Flash to obtain compound 20 (72 mg, yield 52%).

[0526] LCMS (ESI): 556.3 [M+H] + .

[0527] 1 H NMR (400 MHz, DMSO-d6) δ 10.98 (s, 1H), 9.73 - 9.51 (m, 1H), 8.46 (s, 1H), 8.18 (s, 1H), 7.76 - 7.54 (m, 6H), 7.51 (s, 1H), 7.43 (d, J = 7.8 Hz, 1H), 7.27 - 7.13 (m, 2H), 5.10 (dd, J = 13.2, 5.0 Hz, 1H), 4.51 - 4.26 (m, 4H), 3.58 (t, J = 7.1 Hz, 2H), 3.48 (t, J = 5.3 Hz, 2H), 3.25 (d, J = 3.7 Hz, 2H), 2.97 - 2.79 (m, 3H), 2.65 - 2.55 (m, 1H), 2.44 - 2.31 (m, 1H), 2.07 - 1.93 (m, 1H).

[0528] Example 21:

[0529] Preparation of 1-(3-chloro-4-(2-(2-(3-methylguanidino)ethoxy)ethyl)phenyl)-3-((2-(2,6-dioxopiperidin-3-yl)-1-oxoisoindolin-5-yl)methyl)urea.

[0530] Reaction Scheme:

[0531] Procedure:

[0532] Step A: Compound 20-5 (145 mg, 0.28 mmol) was dissolved in DMF (5 mL), TEA (141 mg, 1.40 mmol) and compound 21-1 (203 mg, 0.87 mmol) were added at room temperature, followed by stirring at 50 degrees Celsius for 48 hours.

[0533] After LCMS monitoring the reaction was completed, the reaction solution was purified by Prep-Flash to obtain compound 21 (60 mg, yield 38%).

[0534] LCMS (ESI): m / z [M+H] + calcd for C 27 H 33 ClN7O5: 570.22, mass found: 570.3

[0535] 1 H NMR (400 MHz, DMSO-d6) δ 10.97 (brs, 1H), 9.86 - 9.61 (m, 1H), 8.48 (brs, 2H), 7.94 - 7.74 (m, 1H), 7.73 - 7.53 (m, 4H), 7.51 (s, 1H), 7.43 (d, J = 7.9 Hz, 1H), 7.28 - 7.13 (m, 2H), 5.10 (dd, J = 13.3, 5.1 Hz, 1H), 4.57 - 4.19 (m, 4H), 3.58 (t, J = 7.0 Hz, 2H), 3.48 (t, J = 5.4 Hz, 2H), 3.26 (brs, 2H), 2.96 - 2.81 (m, 3H), 2.69 (s, 3H), 2.64 - 2.55 (m, 1H), 2.38 (ddd, J = 25.3, 12.7, 3.8 Hz, 1H), 2.06 - 1.94 (m, 1H).

[0536] Example 22:

[0537] 1-(3-chloro-4-(2-(2-(1-methylguanidino)ethoxy)ethyl)phenyl)-3-((2-(2,6-dioxopiperidin-3-yl)-1-oxoisoindolin-5-yl)methyl)urea preparation.

[0538] Reaction Scheme:

[0539] Operation Steps:

[0540] Step A: Compound 22-1 (4 g, 12.38 mmol) was added to a 250 mL single-neck flask, followed by the addition of N,N-dimethylformamide (60 mL), Pd(PPh3)4 (1.43 g, 1.24 mmol) and Zn(CN)2 (1.02 g, 8.67 mmol). The reaction system was reacted at 100 °C for 3 hours under nitrogen protection.

[0541] After the reaction was completed, the reaction liquid was poured into water, extracted with ethyl acetate, and the organic phase was washed with saturated sodium chloride solution, dried over anhydrous sodium sulfate, filtered, and the filtrate was concentrated. The obtained crude product was purified by silica gel column (dichloromethane / methanol = 10:1) to obtain compound 22-2.

[0542] LCMS (ESI) M / Z: 268.1 [M-H + ].

[0543] 1H NMR (400MHz, DMSO-d6) δ11.03(s,1H),8.18(d,J=10.5Hz,1H),7.98(dt,J=10.2,5.1Hz,1H),7.92(d,J=7.8Hz,1H),5.16(dd,J=13.3,5.1 Hz,1H),4.56(d,J=17.9Hz,1H),4.43(d,J=17.9Hz,1H),2.99–2.86(m,1H),2.61(d,J=17.4Hz,1H),2.49–2.36(m,1H),2.09–1.99(m,1H).

[0544] Step B: Compound 22-2 (1g, 3.71mmol) was added to a 100mL single-necked flask, followed by methanol (15mL), PtO2 (252.74mg, 1.11mmol) and concentrated hydrochloric acid (1.2mL). The reaction system was carried out under hydrogen protection at room temperature for 15 hours.

[0545] The reaction was monitored by LCMS until it was complete. The mixture was diluted with methanol (250 ml), filtered with diatomaceous earth, and the filtrate was evaporated to dryness. The residue was slurried with dichloromethane / methanol = 3:1 (20 ml) to obtain compound 22-3.

[0546] LCMS(ESI)M / Z:274.1[M+H + ].

[0547] 1 H NMR (400MHz, DMSO-d6) δ11.00(s,1H),8.64(s,3H),7.81–7.72(m,2H),7.65(d,J=7.8Hz,1H),5.13(dd,J=13.3,5.1Hz,1H),4.48(d,J=17. 5Hz,1H),4.34(d,J=17.5Hz,1H),4.14(d,J=5.5Hz,2H),2.99–2.86(m,1H),2.61(d,J=17.3Hz,1H),2.45–2.34(m,1H),2.07–1.96(m,1H).

[0548] Step C: NaH (2.29 g, 57.29 mmol) was added into a 500 mL flask, N,N- dimethylformamide (130 mL) was added, compound 22-5 (6.88 g, 52.08 mmol) was dissolved in N,N-dimethylformamide (8 mL), the reaction flask was added slowly drop by drop at 0 °C, and stirred for 30 min under nitrogen protection. Compound 22-4 (5 g, 26.04 mmol) was dissolved in N,N-dimethylformamide (8 mL), the reaction flask was added slowly drop by drop at 0 °C, and the reaction system was reacted at 70 °C for 16 h under nitrogen protection.

[0549] After the reaction was completed by LCMS monitoring, the reaction solution was poured into water, extracted with ethyl acetate, the organic phase was washed with saturated sodium chloride solution, dried over anhydrous sodium sulfate, filtered, and the filtrate was concentrated. The obtained crude product was slurried with anhydrous ethanol (50 mL) to obtain compound 22-6.

[0550] LCMS (ESI) M / Z: 286.1 [M-H + ].

[0551] 1 H NMR (400 MHz, DMSO-d6) δ 8.37 (d, J = 2.4 Hz, 1H), 8.24 (dd, J = 8.6, 2.4 Hz, 1H), 7.74 (d, J = 8.6 Hz, 1H), 5.45 (s, 1H), 3.75 (s, 6H).

[0552] Step D: Compound 22-6 (4.9 g, 17.03 mmol) was added into a 250 mL flask, methanol (15 mL) and aqueous NaOH solution (8.5 mL, 51 mmol, 6M) were added in turn, and the reaction system was reacted at 50 °C for 2 h under nitrogen protection. Then aqueous NaOH solution (1.4 mL, 8.4 mmol, 6M) was added, and the reaction was reacted at 50 °C for 1 h.

[0553] After the reaction was completed by LCMS monitoring, water (10 mL) was added to the reaction solution, the pH was adjusted to 5 with saturated aqueous citric acid solution under ice bath, and extracted with ethyl acetate. The organic phase was washed with saturated sodium chloride solution, dried over anhydrous sodium sulfate, filtered, and the filtrate was concentrated to obtain compound 22-7.

[0554] LCMS (ESI) M / Z: 214.1 [M-H + ].

[0555] Step E: Compound 22-7 (4.1 g, 19.02 mmol) was added to a 250 mL single-necked flask, THF (70 mL) was added, and then borane dimethyl sulfide (4.8 mL, 48 mmol, 10 M in THF) was added at 0 °C. The reaction system was carried out at 70 °C for 3 hours under nitrogen protection.

[0556] After the reaction was completed by TLC monitoring, methanol was added to quench the reaction, the reaction solution was concentrated, and the crude product was purified by silica gel column chromatography (PE / EA = 1:1) to obtain compound 22-8.

[0557] 1 H NMR (400MHz, DMSO-d6) δ8.25 (d, J=2.4Hz, 1H), 8.14 (dd, J=8.5, 2.4Hz, 1H), 7.66 (d, J= 8.5Hz, 1H), 4.83 (t, J = 5.3Hz, 1H), 3.68 (dd, J = 12.0, 6.6Hz, 2H), 2.97 (t, J = 6.6Hz, 2H).

[0558] Step F: Compound 22-8 (2.9 g, 14.38 mmol) was added to a 500 mL single-necked flask, followed by toluene (90 mL), compound 22-9 (22.44 g, 115.04 mmol), and Bu4NHSO4 (3.91 g, 11.5 mmol). Then, NaOH aqueous solution (290 mL, 5 M) was added dropwise at 0 °C. The reaction system was carried out under nitrogen protection at room temperature for 2 hours.

[0559] After the reaction was completed by TLC monitoring, the organic phase was extracted with ethyl acetate, washed with saturated sodium chloride solution, dried over anhydrous sodium sulfate, filtered, concentrated, and the crude product was purified by silica gel column chromatography (PE / EA = 10:1) to obtain compound 22-10.

[0560] 1 H NMR (400MHz, DMSO-d6) δ8.27(d,J=2.4Hz,1H),8.15(dd,J=8.5,2.4Hz,1H),7.74(d,J=8.5H z,1H),4.00(s,2H),3.76(t,J=6.5Hz,2H),3.08(t,J=6.5Hz,2H),1.99(s,1H),1.41(s,9H).

[0561] Step G: Compound 22-10 (4.2 g, 13.3 mmol) was added to a 100 mL single-necked flask, followed by dichloromethane (40 mL) and TFA (8 mL). The reaction system was carried out under nitrogen protection at room temperature for 1 hour.

[0562] After the reaction was completed by TLC monitoring, the reaction solution was concentrated, water (80 mL) was added and extracted with ethyl acetate, the organic phase was washed with saturated sodium chloride solution, dried over anhydrous sodium sulfate, filtered and the filtrate was concentrated to obtain compound 22-11.

[0563] 1 H NMR (400 MHz, DMSO) δ 12.47 (s, 1H), 8.26 (d, J = 2.1 Hz, 1H), 8.14 (dd, J = 8.5, 2.1 Hz, 1H), 7.75 (d, J = 8.5 Hz, 1H), 4.04 (s, 2H), 3.77 (t, J = 6.4 Hz, 2H), 3.08 (t, J = 6.4 Hz, 2H).

[0564] Step H: Compound 22-11 (2.5 g, 9.63 mmol) was added to a 100 mL single-necked flask, dry N,N-dimethylformamide (25 mL), methylamine hydrochloride (780.26 mg, 11.56 mmol), HATU (5.49 g, 14.45 mmol) and N,N-diisopropylethylamine (6.38 mL, 38.52 mmol) were added in turn, and the reaction system was reacted at room temperature for 2 hours under nitrogen protection.

[0565] After the reaction was completed by LCMS monitoring, saturated sodium chloride solution was added, extracted with ethyl acetate, and the organic phase was washed with saturated sodium chloride solution. The organic phase was dried over anhydrous sodium sulfate, filtered, and the filtrate was concentrated. The obtained crude product was purified by silica gel column (dichloromethane / methanol = 10:1) to obtain compound 22-12.

[0566] 1 H NMR (400 MHz, DMSO-d6) δ 8.27 (d, J = 2.3 Hz, 1H), 8.15 (dd, J = 8.5, 2.4 Hz, 1H), 7.75 (d, J = 8.5 Hz, 1H), 7.63 (s, 1H), 3.88 (s, 2H), 3.74 (t, J = 6.6 Hz, 2H), 3.12 (t, J = 6.5 Hz, 2H), 2.62 (d, J = 4.7 Hz, 3H).

[0567] Step I: Compound 22-12 (2.9 g, 10.64 mmol) was added to a 100 mL single-necked flask, THF (30 mL) was added, and then borane tetrahydrofuran (11.7 mL, 1M in THF) was added at room temperature. The reaction system was reacted at 70°C for 5 hours under nitrogen protection.

[0568] After the reaction was completed by LCMS monitoring, the reaction was quenched by adding methanol under ice bath. The pH was adjusted to 6 with 1M hydrochloric acid, extracted with ethyl acetate, the obtained aqueous phase was adjusted to pH = 8 with saturated sodium bicarbonate solution, and then extracted with ethyl acetate. All the organic phases were combined, dried over anhydrous sodium sulfate, filtered, and the filtrate was rotary evaporated to obtain compound 22-13 crude product.

[0569] LCMS (ESI) M / Z: 259.1 [M+H + ].

[0570] Step J: Compound 22-13 (2.5 g, 9.66 mmol) was added to a 100 mL single-necked flask, followed by THF (50 mL), Boc20 (2.53 g, 11.59 mmol) and TEA (2.01 mL, 14.49 mmol) successively. The reaction system was protected by nitrogen, and reacted at room temperature for 2 hours.

[0571] After the reaction was completed by LCMS monitoring, the solvent was rotary evaporated, and the crude product was purified by column chromatography (acetonitrile / 0.1% formic acid aqueous solution) to obtain compound 22-14.

[0572] LCMS (ESI) M / Z: 259.0 [M-Boc+H + ].

[0573] 1 H NMR (400 MHz, DMSO-d6) δ 8.26 (d, J = 2.3 Hz, 1H), 8.13 (dd, J = 8.5, 2.4 Hz, 1H), 7.67 (d, J = 8.5 Hz, 1H), 3.69 (t, J = 6.4 Hz, 2H), 3.49 (t, J = 5.5 Hz, 2H), 3.27 (t, J = 5.6 Hz, 2H), 3.06 (t, J = 6.3 Hz, 2H), 2.73 (s, 3H), 1.37 (s, 9H).

[0574] Step K: Compound 22-14 (900 mg, 2.51 mmol) was added to a 100 mL single-necked flask, followed by ethanol (45 mL), water (9 mL), ammonium chloride (402.78 mg, 7.53 mmol) and reduced iron powder (700.92 mg, 12.55 mmol) successively. The reaction system was protected by nitrogen, and reacted at 80°C for 5 hours.

[0575] After the reaction was completed by LCMS monitoring, the reaction was quenched by adding methanol under ice bath. The pH was adjusted to 6 with 1M hydrochloric acid, extracted with ethyl acetate, the obtained aqueous phase was adjusted to pH = 8 with saturated sodium bicarbonate solution, and then extracted with ethyl acetate. All the organic phases were combined, dried over anhydrous sodium sulfate, filtered, and the filtrate was rotary evaporated to obtain compound 22-13 crude product.

[0576] LCMS (ESI) M / Z: 259.1 [M+H + ].

[0577] 1 H NMR (400 MHz, DMSO-d6) δ 6.96 (d, J = 8.2 Hz, 1H), 6.59 (d, J = 2.3 Hz, 1H), 6.45 (dd, J = 8.2, 2.3 Hz, 1H), 5.20 (s, 2H), 3.48 (dd, J = 12.5, 6.3 Hz, 4H), 3.28 (t, J = 5.7 Hz, 2H), 2.74 (dd, J = 14.8, 7.5 Hz, 5H), 1.39 (s, 9H).

[0578] Step L: Compound 22-15 (500 mg, 1.52 mmol) was taken in 25 mL single necked flask, after adding dry THF (10 mL) compound 22-3 (563.74 mg, 1.82 mmol) and triethylamine (0.37 mL, 3.04 mmol) in dry N,N-dimethylformamide (15 mL) was added slowly drop wise, the reaction mixture was stirred at room temperature for 1 h under nitrogen atmosphere.

[0579] After completion of reaction by LCMS, the crude was purified by column chromatography (acetonitrile / 0.1% formic acid in water) to get compound 22-16.

[0580] LCMS (ESI) M / Z: 528.2 [M-Boc+H + ].

[0581] Step M: Compound 22-16 (600 mg, 0.96 mmol) was taken in 50 mL single necked flask, after adding dichloromethane (15 mL) TFA (2.5 mL) was added at 0°C, the reaction mixture was stirred at room temperature for 1 h under nitrogen atmosphere.

[0582] After completion of reaction by LCMS, the crude was purified by high pressure liquid chromatography (acetonitrile / 0.1% formic acid in water) to get compound 22-17.

[0583] LCMS (ESI) M / Z: 528.2 [M+H + ].

[0584] 1H NMR (400 MHz, DMSO-d6) δ 9.38 (s, 1H), 8.39 (s, 1H), 7.73 - 7.63 (m, 2H), 7.51 (s, 1H), 7.49 - 7.40 (m, 2H), 7.21 (s, 2H), 5.11 (dd, J = 13.3, 5.1 Hz, 1H), 4.50 - 4.24 (m, 4H), 3.59 (dd, J = 9.1, 5.7 Hz, 4H), 3.00 - 2.81 (m, 5H), 2.65 - 2.55 (m, 1H), 2.47 (s, 3H), 2.44 - 2.33 (m, 1H), 2.04 - 1.95 (m, 1H).

[0585] Step N: Compound 22-17 (50 mg, 0.095 mmol) was dissolved in DMF (2 mL), compound 22-18 (20.89 mg, 0.14 mmol) and DIEA (0.063 mL, 0.38 mmol) were added, the reaction system was protected by nitrogen, and reacted at room temperature for 12 hours.

[0586] After the completion of the reaction was monitored by LCMS, the crude product was purified by Prep-HPLC to obtain compound 22.

[0587] LCMS (ESI): 570.3 [M+H] + .

[0588] 1 H NMR (400 MHz, DMSO-d6) δ 10.98 (s, 1H), 8.95 (s, 1H), 7.75 - 7.63 (m, 2H), 7.51 (s, 1H), 7.44 (d, J = 7.9 Hz, 1H), 7.28 - 7.15 (m, 6H), 7.02 (t, J = 6.0 Hz, 1H), 5.11 (dd, J = 13.3, 5.1 Hz, 1H), 4.50 - 4.38 (m, 3H), 4.31 (d, J = 17.4 Hz, 1H), 3.61 - 3.53 (m, 4H), 3.47 (t, J = 4.9 Hz, 3H), 2.97 - 2.82 (m, 6H), 2.60 (d, J = 17.6 Hz, 1H), 2.45 - 2.31 (m, 1H), 2.00 (dd, J = 8.9, 3.6 Hz, 1H).

[0589] Example 23:

[0590] Preparation of 1-(3-chloro-4-(3-(methylamino)prop-1-yn-1-yl)phenyl)-3-((2-(2,6-dioxopiperidin-3-yl)-1-oxoisoindolin-5-yl)methyl)urea.

[0591] Reaction Scheme:

[0592] Procedure:

[0593] Step A: Compound 23-1 (900 mg, 5.32 mmol) was dissolved in dry THF (20 mL), then 1-bromo-2-chloro-4-nitrobenzene (1510 mg, 6.38 mmol), Pd(PPh3)4(615 mg, 0.53 mmol), CuI (101 mg, 0.53 mmol) and TEA (2.22 mL, 15.96 mmol) were added, the reaction system was heated to reflux at 95 degree Celsius under nitrogen protection for 1 hour.

[0594] After the reaction was completed by LCMS monitoring, the reaction solution was filtered, then the filtrate was obtained, and the crude product was purified by reverse phase Flash (ACN / 0.1% FA in water) to obtain compound 23-2 (1 g, yield 58%).

[0595] LCMS (ESI): 269.0 [M-(t-Bu)+H] + .

[0596] Step B: Tetrahydroxydiboron (553 mg, 6.22 mmol) was dissolved in dry DMF (20 mL), 4,4'-dipyridyl (242 mg, 1.56 mmol) and compound 23-2 (1000 mg, 3.08 mmol) were added quickly in turn, and the reaction system was reacted at room temperature under nitrogen protection for 5 minutes until the blue color faded.

[0597] After the reaction was completed by LCMS monitoring, a small amount of methanol was added, filtered, and the filtrate was purified by Flash (ACN / 0.1% FA in water) to obtain compound 23-3 (850 mg, yield 95%).

[0598] LCMS (ESI): 293.2 [M-H] - .

[0599] Step C: Compound 23-3 (210 mg, 0.71 mmol) was dissolved in dry THF (10 mL), and diphosgene (0.13 mL, 1.06 mmol) was added slowly dropwise, and the reaction system was reacted at room temperature under nitrogen protection for 1 hour, and the solvent was rotary evaporated at low temperature, and the obtained residue was dissolved in dry DMF (5 mL), then compound 1-3 (213 mg, 0.78 mmol) and triethylamine (0.25 mL, 1.77 mmol) in DMF (5 mL) were added slowly dropwise, and the reaction system was reacted at room temperature under nitrogen protection for 1.5 hours

[0600] LCMS monitoring reaction completion, compound 23-4 (270 mg, yield 64%) was obtained by flash purification (ACN / 0.1% FA in water).

[0601] LCMS (ESI): 592.3 [M-H] - .

[0602] Step D: Compound 23-4 (260 mg, 0.44 mmol) was dissolved in DCM (15 mL), then TFA (5 mL) was added, the reaction system was protected under nitrogen, and the reaction was carried out at room temperature for 1 hour.

[0603] LCMS monitoring reaction completion, compound 23-4 (270 mg, yield 64%) was obtained by flash purification (ACN / 0.1% FA in water).

[0604] LCMS (ESI): 494.3 [M+H] + .

[0605] 1 H NMR (400 MHz, DMSO-d6) δ 10.98 (s, 1H), 9.50 - 9.25 (m, 1H), 9.17 - 8.93 (m, 2H), 7.89 - 7.76 (m, 1H), 7.69 (d, J = 7.8 Hz, 1H), 7.51 (s, 1H), 7.49 - 7.41 (m, 2H), 7.35 - 7.10 (m, 2H), 5.11 (dd, J = 13.3, 5.0 Hz, 1H), 4.51 - 4.38 (m, 3H), 4.36 - 4.26 (m, 1H), 4.19 (s, 2H), 2.97 - 2.85 (m, 1H), 2.68 (s, 3H), 2.64 - 2.55 (m, 1H), 2.46 - 2.30 (m, 1H), 2.06 - 1.94 (m, 1H).

[0606] Example 24:

[0607] Preparation of 1-(3-chloro-4-(3-(3-methylguanidino)prop-1-yn-1-yl)phenyl)-3-((2-(2,6-dioxopiperidin-3-yl)-1-oxoisoindolin-5-yl)methyl)urea.

[0608] Reaction Scheme:

[0609] Procedure:

[0610] Step A: Dissolve tetrahydroxydiboron (1882 mg, 20.99 mmol) in dry DMF (20 mL), quickly add 4,4'-dipyridine (563 mg, 3.61 mmol) and compound 24-1 (1000 mg, 3.22 mmol) in sequence, the reaction system is protected by nitrogen, and the reaction is carried out at room temperature for 5 minutes until the blue color disappears.

[0611] After TLC monitoring of the completion of the reaction, a small amount of methanol is added, filtered, and the filtrate is purified by Flash (ACN / 0.1% FA in water) to obtain compound 24-2 (850 mg, yield 94%).

[0612] TLC: R f = 0.3 (PE:EA = 10:1)

[0613] Step B: Dissolve compound 24-2 (330 mg, 1.18 mmol) in dry THF (10 mL), slowly drop bis(trichloromethyl) carbonate (0.21 mL, 1.77 mmol), the reaction system is protected by nitrogen, and the reaction is carried out at room temperature for 1 hour, the solvent is rotary evaporated at low temperature, the obtained residue is dissolved in dry DMF (2 mL), and then slowly dropped into a solution of compound 1-3 (355 mg, 1.30 mmol) and triethylamine (0.41 mL, 2.95 mmol) in DMF (10 mL), the reaction system is protected by nitrogen, and the reaction is carried out at room temperature for 1 hour

[0614] After LCMS monitoring of the completion of the reaction, compound 24-3 (350 mg, yield 52%) is obtained by Flash purification (ACN / 0.1% FA in water).

[0615] LCMS (ESI): 578.3 [M-H] - .

[0616] Step C: Dissolve compound 24-3 (320 mg, 0.55 mmol) in DCM (10 mL), and then add TFA (5 mL), the reaction system is protected by nitrogen, and the reaction is carried out at room temperature for 1 hour.

[0617] After LCMS monitoring of the completion of the reaction, the reaction solution is concentrated, and the crude product is purified by Flash (ACN / 0.1% FA in water) to obtain compound 24-4 (200 mg, yield 76%).

[0618] LCMS (ESI): 480.3 [M+H] + .

[0619] Step D: Compound 24-4 (100 mg, 0.21 mmol) was dissolved in ACN (5 mL), DIEA (0.17 mL, 1.05 mmol) and compound 24-5 (97 mg, 0.42 mmol) were added at room temperature, followed by stirring at room temperature for 12 hours.

[0620] After the reaction was monitored by LCMS to be completed, the reaction solution was purified by Flash to obtain compound 24 (10 mg, yield 9%).

[0621] LCMS (ESI): 536.2 [M+H] + .

[0622] 1 H NMR (400 MHz, DMSO-d6) δ 10.98 (s, 1H), 9.25 (s, 1H), 7.92 (br s, 1H), 7.80 (s, 1H), 7.72 - 7.63 (m, 2H), 7.58 (s, 2H), 7.51 (s, 1H), 7.46 - 7.37 (m, 2H), 7.27 (dd, J = 8.5, 1.4 Hz, 1H), 7.18 - 7.10 (m, 1H), 5.10 (dd, J = 13.3, 5.0 Hz, 1H), 4.53 - 4.38 (m, 3H), 4.36 - 4.24 (m, 3H), 2.98 - 2.85 (m, 1H), 2.77 (d, J = 4.6 Hz, 3H), 2.65 - 2.56 (m, 1H), 2.38 (ddd, J = 26.3, 13.2, 4.1 Hz, 1H), 2.04 - 1.94 (m, 1H).

[0623] Example 25:

[0624] Preparation of 1-(3-chloro-4-(4-(3-methylguanidino)but-1-yn-1-yl)phenyl)-3-((2-(2,6-dioxopiperidin-3-yl)-1-oxoisoindolin-5-yl)methyl)urea.

[0625] Reaction Scheme:

[0626] Procedure:

[0627] Step A: Compound 25-1 (1.4 g, 7.1 mmol) was added into a 250 mL single-neck flask, followed by the addition of dry THF (140 mL), compound 25-2 (2.0 g, 8.52 mmol), Pd(PPh3)4(820 mg, 0.7 mmol), CuI (130 mg, 0.7 mmol) and TEA (2.96 mL, 21.3 mmol) successively. The reaction system was reacted at 90 °C for 5 hours under nitrogen protection.

[0628] After the reaction was completed by LCMS monitoring, the solvent was evaporated, and the obtained crude product was purified by silica gel column (PE / EA = 10: 1) to obtain compound 25-3 (2.1 g, yield 80.18%).

[0629] LCMS (ESI): 225.1 [M-Boc+H] + .

[0630] Step B: Tetrahydroxydiboron (1 g, 10.76 mmol) was added to a 50 mL vial, dry DMF (20 mL) was added, and then 4,4'-dipyridine (500.14 mg, 2.96 mmol) and compound 25-3 (1 g, 3.24 mmol) were added quickly in turn. The reaction system was protected by nitrogen, and the reaction was carried out at room temperature for 5 minutes.

[0631] After the reaction was completed by LCMS monitoring, a small amount of methanol was added, filtered, and the filtrate was purified by Flash (ACN / 0.1% FA in water) to obtain compound 25-4 (660 mg, yield 84.25%).

[0632] LCMS (ESI): 295.2 [M+H] + .

[0633] Step C: Compound 25-4 (600 mg, 1.81 mmol) was added to a 100 mL vial, dry THF (24 mL) was added, and then bis(trichloromethyl) carbonate (0.54 mL, 3.62 mmol) was added dropwise slowly. The reaction system was protected by nitrogen, and the reaction was carried out at room temperature for 1 hour. The solvent was evaporated at low temperature, and the obtained residue was dissolved with dry DMF (4 mL), and then added slowly dropwise to a solution of compound 1-3 (605.62 mg, 1.84 mmol) and triethylamine (2.43 mL, 18.1 mmol) in DMF (5 mL). The reaction system was protected by nitrogen, and the reaction was carried out at room temperature for 1 hour.

[0634] After the reaction was completed by LCMS monitoring, compound 25-5 (700 mg, yield 52.65%) was obtained by Flash purification (ACN / 0.1% FA in water).

[0635] LCMS (ESI): 594.1 [M+H] + .

[0636] Step D: Compound 25-5 (600 mg, 0.96 mmol) was added to a 25 mL vial, DCM (8 mL) was added, and then TFA (1.6 mL) was added at 0°C. The reaction system was protected by nitrogen, and the reaction was carried out at room temperature for 1 hour.

[0637] After the reaction was completed by LCMS monitoring, the reaction solution was concentrated, and the crude product was purified by Prep-HPLC (THF / 0.1% TFA in water) to obtain compound 25-6 (500 mg, yield 92.56%).

[0638] LCMS (ESI): 494.1 [M+H] + .

[0639] Step E: Compound 25-6 (100 mg, 0.15 mmol) was added to a 25 mL single-necked flask, followed by the addition of DMF (2 mL), compound 25-7 (240 mg, 1.50 mmol) and DIEA (300 mg, 2.31 mmol). The reaction system was reacted at 40 °C for 24 hours under nitrogen protection.

[0640] After the reaction was completed by LCMS monitoring, the reaction solution was purified by Prep-HPLC to obtain compound 25 (20 mg, yield 22.45%).

[0641] LCMS (ESI): 550.3 [M+H] + .

[0642] 1 H NMR (400 MHz, DMSO-d6) δ 10.97 (m, 1H), 10.33 - 10.12 (m, 1H), 8.47 (s, 2H), 8.25 - 8.06 (m, 1H), 7.93 - 7.55 (m, 4H), 7.50 (s, 1H), 7.43 (d, J = 7.9 Hz, 1H), 7.36 - 7.27 (m, 2H), 5.10 (dd, J = 13.2, 5.1 Hz, 1H), 4.50 - 4.25 (m, 4H), 2.98 - 2.85 (m, 1H), 2.71 (s, 3H), 2.68 - 2.57 (m, 3H), 2.38 (m, J = 13.1, 4.4 Hz, 1H), 2.05 - 1.94 (m, 1H).

[0643] Example 26:

[0644] Preparation of 1-(3-chloro-4-(2-(2-(1-cyclopropylguanidino)ethoxy)ethyl)phenyl)-3-((2-(2,6-dioxopiperidin-3-yl)-1-oxoisoindolin-5-yl)methyl)urea.

[0645] Reaction Scheme:

[0646] Procedure:

[0647] Step A: Compound 18 (30 mg, 0.054 mmol) was taken in 250 mL single neck flask, ACN (40 mL) was added followed by compound 26-1 (80 mg, 0.54 mmol) and DIEA (100 mg, 0.77 mmol) and the reaction mixture was allowed to react at 80 degree Celsius for 96 h under nitrogen atmosphere.

[0648] After completion of reaction by LCMS, the reaction mixture was concentrated and purified by Prep-HPLC to get compound 26 (5.02 mg, yield 13.05%).

[0649] LCMS (ESI): 596.1 [M+H] + .

[0650] 1 H NMR (400 MHz, DMSO-d6) δ 10.98 (s, 1H), 8.97 - 8.83 (m, 1H), 7.73 - 7.62 (m, 2H), 7.51 (s, 1H), 7.46 - 7.42 (m, 1H), 7.42 - 7.29 (m, 4H), 7.19 (s, 2H), 7.03 - 6.90 (m, 1H), 5.10 (dd, J = 13.3, 5.1 Hz, 1H), 4.48 - 4.41 (m, 2H), 4.41 - 4.28 (m, 2H), 3.60 - 3.55 (m, 4H), 3.52 - 3.49 (m, 2H), 2.97 - 2.88 (m, 1H), 2.87 - 2.83 (m, 2H), 2.65 - 2.57 (m, 2H), 2.43 - 2.31 (m, 1H), 2.03 - 1.95 (m, 1H), 0.90 - 0.86 (m, 2H), 0.75 - 0.70 (m, 2H).

[0651] Example 27:

[0652] Preparation of 1-(3-chloro-4-(2-(2-(1-methoxyguanidino)ethoxy)ethyl)phenyl)-3-((2-(2,6-dioxopiperidin-3-yl)-1-oxoisoindolin-5-yl)methyl)urea.

[0653] Scheme:

[0654] Procedure:

[0655] Step A: Compound 19 (50 mg, 0.092 mmol) was dissolved in dry DMF (6 mL), compound 27-1 (269.71 mg, 1.84 mmol) and DIEA (0.38 mL, 2.3 mmol) were added and the reaction mixture was allowed to react at 80 °C for 24 h under nitrogen atmosphere.

[0656] After the reaction was completed, the reaction solution was concentrated, and compound 27 (7 mg, yield 13.00%) was obtained by Prep-HPLC purification.

[0657] 1 H NMR (400 MHz, DMSO-d6) δ 10.98 (s, 1H), 9.19 (s, 1H), 7.83 (s, 3H), 7.72 - 7.63 (m, 2H), 7.51 (s, 1H), 7.44 (d, J = 7.8 Hz, 1H), 7.20 (s, 2H), 7.08 (t, J = 6.0 Hz, 1H), 5.11 (dd, J = 13.2, 5.0 Hz, 1H), 4.53 - 4.24 (m, 4H), 3.82 (t, J = 5.0 Hz, 2H), 3.63 (s, 3H), 3.61 - 3.52 (m, 4H), 3.00 - 2.77 (m, 3H), 2.68 - 2.55 (m, 1H), 2.41 - 2.31 (m, 1H), 2.04 - 1.97 (m, 1H).

[0658] LCMS (ESI): 586.4 [M+H] + .

[0659] Example 28:

[0660] Preparation of 1-(3-chloro-4-(2-(1-(prop-2-yn-1-yl)guanidino)ethoxy)ethyl)phenyl)-3-((2-(2,6-dioxopiperidin-3-yl)-1-oxoisoindolin-5-yl)methyl)urea.

[0661] Reaction Scheme:

[0662] Procedure:

[0663] Step A: Compound 13 (85 mg, 0.15 mmol) was added to a 250 mL single-necked flask, followed by ACN (40 mL), compound 28-1 (240 mg, 1.50 mmol) and DIEA (300 mg, 2.31 mmol). The reaction system was reacted at 80 degrees Celsius for 96 hours under nitrogen protection.

[0664] After the reaction was completed, the reaction solution was concentrated, and compound 28 (20 mg, yield 23.05%) was obtained by Prep-HPLC purification.

[0665] LCMS (ESI): 594.5 [M+H] + .

[0666] 1 H NMR (400 MHz, DMSO-d6) δ 10.98 (s, 1H), 9.65 (s, 1H), 8.56 - 7.88 (m, 4H), 7.83 - 7.65 (m, 3H), 7.51 (s, 1H), 7.44 (d, J = 7.8 Hz, 1H), 7.26 - 7.13 (m, 2H), 5.10 (dd, J = 13.3, 5.1 Hz, 1H), 4.47 - 4.28 (m, 4H), 4.17 (d, J = 2.0 Hz, 2H), 3.61 - 3.56 (m, 4H), 3.52 (d, J = 4.8 Hz, 2H), 3.42 (t, J = 2.1 Hz, 1H), 2.96 - 2.82 (m, 3H), 2.65 - 2.56 (m, 1H), 2.44 - 2.32 (m, 1H), 2.03 - 1.93 (m, 1H).

[0667] Example 29:

[0668] Preparation of 1-(3-chloro-4-(2-(2-(1-ethylguanidine)ethoxy)ethyl)phenyl)-3-((2-(2,6-dioxopiperidin-3-yl)-1-oxoisoindolin-5-yl)methyl)urea

[0669] Reaction Scheme:

[0670] Procedure:

[0671] Step A: Compound 29-1 (600 mg, 2.46 mmol) was taken in 100 mL single neck flask, MeOH (40 mL), compound 29-2 (2.4 g, 29.6 mmol) was added sequentially, stirred at room temperature for 1 h under nitrogen protection, sodium cyanoborohydride (156 mg, 2.48 mmol) was added and stirred at room temperature for 2 h.

[0672] After completion of reaction by LCMS, compound 29-3 (375 mg, yield 55.83%) was obtained by flash purification (ACN / 0.1% FA in water).

[0673] LCMS (ESI): 273.1 [M+H] + .

[0674] Step B: Compound 29-3 (375 mg, 1.38 mmol) was taken in 100 mL single neck flask, THF (15 mL), triethylamine (835 mg, 8.26 mmol) and Boc20 (645 mg, 2.96 mmol) were added and stirred at room temperature for 2 h.

[0675] After the reaction was completed by LCMS monitoring, the reaction solution was concentrated, and the obtained crude product was purified by silica gel column (PE / EA = 1:1) to obtain compound 29-4 (400 mg, yield 77.99%).

[0676] LCMS (ESI): 317.1 [M-(t-Bu)+H] + .

[0677] Step C: Compound 29-4 (400 mg, 1.075 mmol) was added to a 25 mL single-necked flask, dry DMF (5 mL) was added, and then tetrahydroxyboron (287 mg, 3.22 mmol) and 4,4'-dipyridine (62 mg, 0.397 mmol) were added in turn. The reaction system was protected by nitrogen, and the reaction was carried out at room temperature for 5 minutes.

[0678] After the reaction was completed by LCMS monitoring, a small amount of methanol was added, filtered, and the filtrate was purified by Flash (ACN / 0.1% FA in water) to obtain compound 29-5 (360 mg, yield 97.89%).

[0679] LCMS (ESI): 365.3 [M+Na] + .

[0680] Step D: Compound 29-5 (360 mg, 1.053 mmol) was added to a 25 mL single-necked flask, dry THF (6 mL) was added, and then diphosgene (452 mg, 2.286 mmol) was slowly added dropwise. The reaction system was protected by nitrogen, and the reaction was carried out at room temperature for 1 hour. The solvent was rotary evaporated at low temperature, and the residue was dissolved in dry DMF (3 mL), which was slowly added dropwise to a solution of compound 1-3 (339 mg, 1.097 mmol) and triethylamine (184 mg, 1.822 mmol) in DMF (5 mL). The reaction system was protected by nitrogen, and the reaction was carried out at room temperature for 1 hour.

[0681] After the reaction was completed by LCMS monitoring, the reaction solution was concentrated, and the obtained crude product was purified by silica gel column (PE / EA = 1:1) to obtain compound 29-4 (400 mg, yield 77.99%).

[0682] LCMS (ESI): 542.3 [M-Boc+H] + .

[0683] Step E: Compound 29-6 (310 mg, 0.484 mmol) was added to a 50 mL single-necked flask, DCM (20 mL) was added, and then TFA (10 mL) was added at 0°C. The reaction system was protected by nitrogen, and the reaction was carried out at room temperature for 1 hour.

[0684] After the reaction was completed by LCMS monitoring, the reaction solution was concentrated, and the crude product was purified by Prep-HPLC (ACN / 0.1% TFA in water) to obtain compound 29-7 (200 mg, yield 76.44%).

[0685] LCMS (ESI): 542.2 [M+H] + .

[0686] Step F: Compound 29-7 (30 mg, 0.054 mmol) was added to a 250 mL single-necked flask, followed by ACN (40 mL), compound 29-8 (80 mg, 0.54 mmol) and DIEA (100 mg, 0.77 mmol). The reaction system was reacted at 80 degrees Celsius for 96 hours under nitrogen protection.

[0687] After the reaction was completed by LCMS monitoring, the reaction solution was concentrated, and the crude product was purified by Prep-HPLC to obtain compound 29 (5.02 mg, yield 13.05%).

[0688] LCMS (ESI): 584.5 [M+H] + .

[0689] 1 H NMR (400 MHz, DMSO-d6) δ 10.98 (s, 1H), 8.98 - 8.84 (m, 1H), 8.51 (s, 2H), 7.74 - 7.62 (m, 2H), 7.51 (s, 1H), 7.44 (d, J = 7.7 Hz, 1H), 7.20 - 7.12 (m, 5H), 7.03 - 6.91 (m, 1H), 5.11 (dd, J = 13.2, 4.9 Hz, 1H), 4.48 - 4.24 (m, 4H), 3.63 - 3.56 (m, 2H), 3.55 - 3.50 (m, 2H), 3.47 - 3.43 (m, 2H), 3.31 - 3.28 (m, 2H), 2.98 - 2.88 (m, 1H), 2.88 - 2.81 (m, 2H), 2.65 - 2.55 (m, 1H), 2.42 - 2.32 (m, 1H), 2.07 - 1.92 (m, 1H), 1.05 (t, J = 6.9 Hz, 3H).

[0690] Example 30:

[0691] Preparation of N-(2-(2-chloro-4-(3-(2-(2,6-dioxopiperidin-3-yl)-1-oxoisoindolin-5- yl)methyl)ureido)phenethoxy)ethyl)-N-methylacetamide.

[0692] Reaction Scheme:

[0693] Procedure:

[0694] Step A: Compound 22-17 (50 mg, 0.095 mmol) was dissolved in THF (10 mL), compound 30-1 (176.1 mg, 1.43 mmol) and TEA (0.26 mL, 1.9 mmol) were added, the reaction system was protected under nitrogen, and the reaction was carried out at room temperature for 30 hours.

[0695] After the reaction was completed by LCMS monitoring, the crude product was purified by Prep-HPLC to obtain compound 30 (25 mg, yield 46.39%).

[0696] 1 H NMR (400 MHz, DMSO-d6) δ 10.98 (s, 1H), 9.08 (d, J = 7.4 Hz, 1H), 9.00 (d, J = 4.4 Hz, 1H), 8.40 (d, J = 6.2 Hz, 1H), 7.72 - 7.62 (m, 2H), 7.51 (s, 1H), 7.44 (d, J = 7.9 Hz, 1H), 7.22 - 7.14 (m, 2H), 7.11 - 7.04 (m, 1H), 5.11 (dd, J = 13.3, 5.1 Hz, 1H), 4.43 (dd, J = 15.0, 11.8 Hz, 3H), 4.31 (d, J = 17.4 Hz, 1H), 3.60 (t, J = 5.9 Hz, 6H), 3.08 (s, 1H), 2.99 (s, 2H), 2.96 - 2.81 (m, 3H), 2.60 (dd, J = 15.7, 1.4 Hz, 1H), 2.45 - 2.32 (m, 1H), 2.21 (s, 3H), 2.04 - 1.95 (m, 1H).

[0697] LCMS (ESI): 569.4 [M+H] + .

[0698] Example 31:

[0699] Preparation of N-(2-(2-chloro-4-(3-(2-(2,6-dioxopiperidin-3-yl)-1-oxoisoindolin-5- yl)methyl)ureido)phenethoxy)ethyl)-2-cyano-N-methylacetamide.

[0700] Reaction Scheme:

[0701] Procedure:

[0702] Step A: Compound 31-1 (10 g, 151.5 mmol), EtOH (6.97 g, 151.5 mmol) were dissolved in diethyl ether (120 mL), 2M hydrochloric acid diethyl ether solution (98.4 mL, 197 mmol) was added, and the reaction was stirred at room temperature for 16 hours. The reaction was filtered, the filter cake was collected and washed with diethyl ether (100 mL) to obtain compound 31-2 (8 g, yield 36%).

[0703] 1 H NMR (400 MHz, DMSO-d6) δ 4.17 (q, J = 7.1 Hz, 2H), 4.03 (s, 2H), 1.22 (t, J = 7.1 Hz, 3H)

[0704] Step B: Compound 22-17 (50 mg, 0.095 mmol) was dissolved in THF (10 mL), then compound 31-2 (424 mg, 2.86 mmol) and TEA (0.45 mL) were added, and the reaction was stirred at room temperature for 48 hours under nitrogen protection.

[0705] After the reaction was completed by LCMS monitoring, the crude product was purified by Prep-HPLC to obtain compound 31 (10 mg, yield 18%).

[0706] LCMS (ESI): 594.1 [M+H] + .

[0707] 1 H NMR (400 MHz, DMSO-d6) δ 10.98 (s, 1H), 8.86 (s, 1H), 8.16 (s, 1H), 7.79 - 7.64 (m, 2H), 7.51 (s, 1H), 7.44 (d, J = 7.9 Hz, 1H), 7.17 (dt, J = 8.4, 5.2 Hz, 2H), 6.88 (d, J = 5.5 Hz, 1H), 5.11 (dd, J = 13.3, 5.1 Hz, 1H), 4.54 - 4.27 (m, 4H), 3.56 (t, J = 6.9 Hz, 2H), 3.49 (t, J = 5.1 Hz, 2H), 3.37 - 3.33 (m, 2H), 2.99 - 2.72 (m, 7H), 2.69 - 2.54 (m, 2H), 2.46 - 2.31 (m, 1H), 2.07 - 1.93 (m, 1H).

[0708] Example 32:

[0709] Preparation of N-(2-(2-chloro-4-(3-((2-(2-6-dioxopiperidin-3-yl)-1-oxoisoindolin-5- yl)methyl)indolyl)phenoxy)ethyl)-N-cyclopropylacetamide.

[0710] Reaction Scheme:

[0711] Procedure:

[0712] Step A: Compound 18 (100 mg, 0.181 mmol) was taken in 5 mL sealed tube, THF (3 mL) and compound 32-1 (665 mg, 5.4 mmol) were added, the reaction mixture was stirred at 70 degree Celsius for 10 hours under nitrogen atmosphere.

[0713] After completion of reaction by LCMS, the reaction mixture was concentrated, the crude compound was purified by Prep-HPLC to get compound 32 (26.28 mg, yield 24.46%).

[0714] LCMS (ESI): 595.5 [M+H] + .

[0715] 1 H NMR (400 MHz, DMSO-d6) δ 10.12 - 9.82 (m, 1H), 8.52 (s, 1H), 8.23 - 7.96 (m, 1H), 7.73 - 7.61 (m, 2H), 7.51 (s, 1H), 7.43 (d, J = 7.9 Hz, 1H), 7.28 - 7.22 (m, 1H), 7.18 - 7.12 (m, 1H), 5.10 (dd, J = 13.3, 5.1 Hz, 1H), 4.51 - 4.24 (m, 4H), 3.65 - 3.55 (m, 6H), 2.99 - 2.87 (m, 1H), 2.87 - 2.80 (m, 2H), 2.74 - 2.56 (m, 2H), 2.44 - 2.32 (m, 1H), 2.24 (s, 3H), 2.05 - 1.95 (m, 1H), 1.01 - 0.89 (m, 2H), 0.82 (s, 2H).

[0716] Example 33:

[0717] Preparation of N-(2-(2-chloro-4-(3-(2-(2,6-dioxopiperidin-3-yl)-1-oxoisoindolin-5- yl)methyl)ureido)phenethoxy)ethyl)-N-methylcyclopropanecarboximidamide.

[0718] Reaction Scheme:

[0719] Procedure:

[0720] Step A: Compound 33-1 (12 g, 178.86 mmol) was dissolved in diethyl ether (40 mL), methanol (10.88 mL, 268.29 mmol) was added, hydrochloric acid-dioxane solution (70 mL, 4 M) was added slowly at 0 °C, the reaction system was protected by nitrogen, and the reaction was carried out at room temperature for 12 h.

[0721] After the reaction was completed by LCMS monitoring, the solvent was rotary evaporated, the residue was slurried with diethyl ether (170 mL) to obtain compound 33-2 (12.5 g, crude product, containing triethylamine hydrochloride).

[0722] LCMS (ESI): 100.1 [M+H] + .

[0723] Step B: Compound 22-17 (50 mg, 0.095 mmol) was dissolved in THF (10 mL), compound 33-2 (154.56 mg, 1.14 mmol) and TEA (0.21 mL, 1.52 mmol) were added, the reaction system was protected by nitrogen, and the reaction was carried out at room temperature for 30 h.

[0724] After the reaction was completed by LCMS monitoring, the crude product was purified by Prep-HPLC to obtain compound 33 (35 mg, yield 62.11%).

[0725] 1 H NMR (400 MHz, DMSO-d6) δ 10.98 (s, 1H), 9.02 (d, J = 4.4 Hz, 1H), 8.35 (s, 2H), 7.68 (dd, J = 12.1, 7.2 Hz, 2H), 7.51 (s, 1H), 7.44 (d, J = 7.9 Hz, 1H), 7.21 - 7.16 (m, 2H), 7.10 (t, J = 6.0 Hz, 1H), 5.11 (dd, J = 13.3, 5.1 Hz, 1H), 4.43 (dd, J = 14.7, 11.8 Hz, 3H), 4.31 (d, J = 17.4 Hz, 1H), 3.82 (t, J = 5.0 Hz, 1H), 3.70 - 3.55 (m, 5H), 3.23 (s, 1H), 3.02 (s, 2H), 2.97 - 2.82 (m, 3H), 2.60 (d, J = 16.7 Hz, 1H), 2.46 - 2.30 (m, 1H), 2.04 - 1.92 (m, 2H), 1.07 - 0.96 (m, 4H).

[0726] LCMS (ESI): 595.7 [M+H] + .

[0727] Example 34:

[0728] Preparation of 3-(2-chloro-4-(3-(2-(2,6-dioxopiperidin-3-yl)-1-oxoisoindolin-5- yl)methyl)ureido)phenyl)propionic acid piperidin-3-yl ester.

[0729] Reaction Scheme:

[0730] Procedure:

[0731] Step A: Compound 34-1 (10 g, 58.86 mmol) was dissolved in CCI4(70 mL), then NBS (10.48 g, 58.86 mmol) and dibenzoyl peroxide (0.12 mL, 0.59 mmol) were added. The reaction was stirred at 80 °C overnight under nitrogen.

[0732] After the reaction was completed by TLC, the solvent was evaporated, and the crude product was purified by reverse phase flash (ACN / 0.1% FA in water) to give compound 34-2 (6 g, yield 41%).

[0733] TLC: Rf = 0.2 (PE:EA = 10:1) f

[0734] Step B: NaH (1.05 g, 43.91 mmol) was added to DMSO (70 mL) in portions, then compound 34-3 (5.27 g, 39.92 mmol) was added, stirred for 0.5 h, then compound 34-2 (5 g, 19.61 mmol) was added. The reaction was stirred at 60 °C overnight under nitrogen.

[0735] After the reaction was completed by LCMS, it was diluted with ethyl acetate, then washed with water and saturated sodium chloride solution successively. The organic phases were combined and dried, and concentrated. The crude product 34-4 was used directly in the next reaction.

[0736] LCMS (ESI): 300.1 [M-H] -

[0737] Step C: The crude product 34-4 was dissolved in 6N hydrochloric acid solution (20 mL), and the reaction was stirred at 100 °C overnight under nitrogen.

[0738] After the reaction was completed by LCMS, the reaction was filtered, then washed with water and saturated sodium chloride solution successively. The organic phases were combined and dried, and concentrated. The crude product was purified by reverse phase flash (ACN / 0.1% FA in water) to give compound 34-5 (1 g, yield 22%).

[0739] ​LCMS (ESI): 228.0 [M-H] - .

[0740] Step D: Compound 34-5 (300 mg, 1.31 mmol) was dissolved in dry DCM (10 mL), then compound 34-6 (526 mg, 2.61 mmol), DMAP (1.6 mg, 0.0131 mmol) and EDCI (301 mg, 1.57 mmol) were added, the reaction system was protected by nitrogen, and reacted at room temperature for 1 hour.

[0741] After the reaction was completed by LCMS monitoring, the reaction solution was poured into water, extracted with ethyl acetate, and the organic phase was washed with saturated sodium chloride solution. The organic phase was dried over anhydrous sodium sulfate, filtered, and the filtrate was concentrated. The obtained crude product was purified by silica gel column (PE / EA = 5:1) to obtain compound 34-7 (520 mg, yield 96.34%).

[0742] LCMS (ESI): 435.3 [M+Na] + .

[0743] Step E: Tetrahydroxydiboron (870 mg, 9.70 mmol) was dissolved in dry DMF (10 mL), and 4,4'-dipyridyl (380 mg, 2.42 mmol) and compound 34-7 (500 mg, 1.21 mmol) were added quickly in turn. The reaction system was protected by nitrogen, and reacted at room temperature for 5 minutes until the blue color faded.

[0744] After the reaction was completed by LCMS monitoring, a small amount of methanol was added, filtered, and the filtrate was purified by Flash (ACN / 0.1% FA in water) to obtain compound 34-8 (450 mg, yield 97%).

[0745] LCMS (ESI): 283.0 [M-Boc+H] + .

[0746] Step F: Compound 34-8 (150 mg, 0.39 mmol) was dissolved in dry THF (4 mL), and diphosgene (117 mg, 0.60 mmol) was slowly added dropwise. The reaction system was protected by nitrogen, and reacted at room temperature for 1 hour. The solvent was rotary evaporated at low temperature, and the residue was dissolved in dry DMF (2 mL). Compound 1-3 (165 mg, 0.60 mmol) and triethylamine (0.17 mL) in DMF (5 mL) were slowly added dropwise. The reaction system was protected by nitrogen, and reacted at room temperature for 1 hour

[0747] LCMS monitoring reaction completion, compound 34-9 (220 mg, yield 82%) was obtained by Flash purification (ACN / 0.1% FA in water).

[0748] LCMS (ESI): 582.2 [M-Boc+H] + .

[0749] Step G: Compound 34-9 (220 mg, 0.32 mmol) was dissolved in DCM (8 mL), then TFA (4 mL) was added, the reaction system was protected by nitrogen, and the reaction was carried out at room temperature for 1 hour.

[0750] LCMS monitoring reaction completion, compound 34-9 (220 mg, yield 82%) was obtained by Flash purification (ACN / 0.1% FA in water).

[0751] 1 H NMR (400 MHz, DMSO-d6) δ 10.98 (s, 1H), 9.01 - 8.73 (m, 1H), 7.72 - 7.64 (m, 2H), 7.51 (s, 1H), 7.44 (d, J = 7.9 Hz, 1H), 7.18 (s, 2H), 6.93 (brs, 1H), 5.11 (dd, J = 13.3, 5.1 Hz, 1H), 4.90 (brs, 1H), 4.51 - 4.26 (m, 4H), 3.19 (dd, J = 12.7, 2.5 Hz, 1H), 3.07 - 2.85 (m, 6H), 2.67 - 2.56 (m, 3H), 2.38 (ddd, J = 16.8, 12.8, 3.5 Hz, 1H), 2.04 - 1.94 (m, 1H), 1.85 - 1.73 (m, 2H), 1.69 - 1.56 (m, 2H).

[0752] LCMS (ESI): 582.0 [M+H] + .

[0753] Example 35:

[0754] Preparation of tert-butyl 4-((3-(2-chloro-4-(3-((2-(2,6-dioxopiperidin-3-yl)-1- oxoisoindolin-5-yl)methyl)ureido)phenyl)propionyl)oxy)piperidine-1-carboxylate.

[0755] Reaction Scheme:

[0756] Procedure:

[0757] Step A: Compound 35-1 (300 mg, 1.31 mmol) was dissolved in dry DCM (10 mL), then compound 35-2 (526 mg, 2.61 mmol), DMAP (1.6 mg, 0.0131 mmol) and EDCI (301 mg, 1.57 mmol) were added, the reaction system was protected by nitrogen, and reacted at room temperature for 1 hour.

[0758] After the reaction was completed by LCMS monitoring, the reaction solution was poured into water, extracted with ethyl acetate, and the organic phase was washed with saturated sodium chloride solution. The organic phase was dried over anhydrous sodium sulfate, filtered, and the filtrate was concentrated. The obtained crude product was purified by silica gel column (PE / EA = 5:1) to obtain compound 35-3 (505 mg, yield 93.56%).

[0759] LCMS (ESI): 435.3 [M+Na] + .

[0760] Step B: Tetrahydroxydiboron (870 mg, 9.70 mmol) was dissolved in dry DMF (10 mL), and 4,4'-dipyridine (380 mg, 2.42 mmol) and compound 35-3 (500 mg, 1.21 mmol) were added quickly in turn. The reaction system was protected by nitrogen, and reacted at room temperature for 5 minutes until the blue color faded.

[0761] After the reaction was completed by LCMS monitoring, a small amount of methanol was added, filtered, and the filtrate was purified by Flash (ACN / 0.1% FA in water) to obtain compound 35-4 (450 mg, yield 97%).

[0762] LCMS (ESI): 283.0 [M-Boc+H] + .

[0763] Step C: Compound 35-4 (150 mg, 0.39 mmol) was dissolved in dry THF (4 mL), and diphosgene (117 mg, 0.60 mmol) was added slowly dropwise. The reaction system was protected by nitrogen, and reacted at room temperature for 1 hour. The solvent was rotary evaporated at low temperature, and the residue was dissolved in dry DMF (2 mL). Compound 1-3 (165 mg, 0.60 mmol) and triethylamine (0.17 mL) in DMF (5 mL) were added slowly dropwise. The reaction system was protected by nitrogen, and reacted at room temperature for 1 hour

[0764] After the reaction was completed by LCMS monitoring, compound 35-5 (110 mg, yield 42%) was obtained by Flash purification (ACN / 0.1% FA in water).

[0765] LCMS (ESI): 582.3 [M-Boc+H]+ .

[0766] Step D: Compound 35-5 (100 mg, 0.15 mmol) was dissolved in DCM (8 mL), then TFA (4 mL) was added, the reaction system was protected under nitrogen, and reacted at room temperature for 1 hour.

[0767] After the reaction was completed by LCMS monitoring, the reaction solution was concentrated, and the crude product was purified by Prep-HPLC to obtain compound 35 (50 mg, yield 59%).

[0768] 1 H NMR (400 MHz, DMSO-d6) δ 10.98 (brs, 1H), 9.04 (s, 1H), 7.79-7.64 (m, 2H), 7.51 (s, 1H), 7.44 (d, J = 7.9 Hz, 1H), 7.26-7.16 (m, 2H), 7.10 (s, 1H), 5.10 (dd, J = 133.3, 5.1 Hz, 1H), 4.93-4.73 (m, 1H), 4.53-4.22 (m, 4H), 3.15-3.01 (m, 2H), 2.97-2.76 (m, 5H), 2.65-2.56 (m, 3H), 2.38 (ddd, J = 26.0, 13.1, 4.3 Hz, 1H), 2.04-1.95 (m, 1H), 1.94–1.84 (m, 2H), 1.67–1.55 (m, 2H).

[0769] LCMS (ESI): 582.0 [M+H] + .

[0770] Example 36:

[0771] Preparation of (1S,4S)-4-(methylamino)cyclohexyl 3-(2-chloro-4-(3-((2-(2-2,6-dioxopiperidin-3-yl)-1-oxoisoindolin-5-yl)methyl)ureido)phenyl)propanoate.

[0772] Reaction Scheme:

[0773] Operation Steps:

[0774] Step A: Compound 36-1 (150 mg, 0.655 mmol) was added to a 100 mL three-necked flask, THF (5 mL), compound 36-2 (100 mg, 0.437 mmol), triphenylphosphine (171 mg, 0.653 mmol) were added in turn, and DEAD (132 mg, 0.653 mmol) was added dropwise at 0°C. The reaction system was protected under nitrogen, and reacted at room temperature for 3 hours.

[0775] After LCMS monitoring the reaction was completed, the reaction solution was poured into water, extracted with ethyl acetate, and the organic phase was washed with saturated sodium chloride solution, dried over anhydrous sodium sulfate, filtered, and the filtrate was concentrated. The obtained crude product was purified by silica gel column (PE / EA = 5: 1) to obtain compound 36-3 (80 mg, yield 41.63%).

[0776] 1 H NMR (400 MHz, CDC13) δ 8.25 (d, J = 2.2 Hz, 1H), 8.07 (dd, J = 8.4, 2.3 Hz, 1H), 7.48 (d, J = 8.5 Hz, 1H), 5.01 (s, 1H), 3.92 (s, 1H), 3.18 (t, J = 7.6 Hz, 2H), 2.79 - 2.66 (m, 5H), 1.92 (d, J = 9.0 Hz, 2H), 1.58 (s, 2H), 1.50 (d, J = 7.3 Hz, 2H), 1.46 (s, 9H), 1.34 - 1.19 (m, 2H).

[0777] Step B: Compound 36-3 (80 mg, 0.177 mmol) was added to a 50 mL vial, dry DMF (5 mL) was added, and then tetrahydroxyboron (63 mg, 0.708 mmol) and 4,4'-dipyridyl (14 mg, 0.090 mmol) were added quickly in turn. The reaction system was protected by nitrogen, and reacted at room temperature for 5 minutes.

[0778] After LCMS monitoring the reaction was completed, a small amount of methanol was added, filtered, and the filtrate was purified by Flash (ACN / 0.1% FA in water) to obtain compound 36-4 (74 mg, yield 99.27%).

[0779] LCMS (ESI): 411.4 [M+H] + .

[0780] Step C: Compound 36-4 (74 mg, 0.18 mmol) was added to a 25 mL vial, dry THF (6 mL) was added, and then diphosgene (71 mg, 0.359 mmol) was added dropwise slowly. The reaction system was protected by nitrogen, and reacted at room temperature for 1 hour. The solvent was dried by rotary evaporation under low temperature and nitrogen. After being dissolved in dry DMF (3 mL), it was added dropwise slowly to a solution of compound 1-3 (220 mg, 0.712 mmol) and triethylamine (101 mg, 1.0 mmol) in DMF (5 mL). The reaction system was protected by nitrogen, and reacted at room temperature for 1 hour

[0781] After completion of reaction, compound 36-5 (110 mg, yield 85.93%) was obtained by flash purification (ACN / 0.1% TFA in water).

[0782] LCMS (ESI): 610.2 [M-Boc+H] + .

[0783] Step D: Compound 36-5 (110 mg, 0.155 mmol) was added to a 25 mL single neck flask, DCM (5 mL) was added, TFA (5 mL) was added at 0 °C, the reaction system was protected under nitrogen, and the reaction was carried out at room temperature for 1 hour.

[0784] After completion of reaction, the reaction solution was concentrated, and the crude product was purified by Prep-HPLC to obtain compound 36 (48.97 mg, yield 38.24%).

[0785] 1 H NMR (400 MHz, DMSO-d6) δ 10.98 (s, 1H), 8.89 (s, 1H), 8.45 (s, 2H), 7.73 - 7.61 (m, 2H), 7.51 (s, 1H), 7.44 (d, J = 7.9 Hz, 1H), 7.17 (s, 2H), 6.94 (t, J = 6.0 Hz, 1H), 5.11 (dd, J = 13.3, 5.1 Hz, 1H), 4.64 - 4.52 (m, 1H), 4.47 - 4.26 (m, 4H), 2.99 - 2.82 (m, 4H), 2.62 - 2.53 (m, 6H), 2.44 - 2.32 (m, 1H), 2.06 - 1.90 (m, 5H), 1.42 - 1.32 (m, 4H).

[0786] LCMS (ESI): 610.5 [M+H] + .

[0787] Example 37:

[0788] Preparation of (1R,4R)-4-(methylamino)cyclohexyl 3-(2-chloro-4-(3-((2-(2-2,6-dioxopiperidin-3-yl)-1-oxoisoindolin-5-yl)methyl)ureido)phenyl)propanoate.

[0789] Reaction Scheme:

[0790] Procedure:

[0791] Step A: Compound 37-1 (300 mg, 1.31 mmol) was added to a 100 mL flask, followed by the addition of DCM (10 mL), compound 37-2 (600 mg, 2.62 mmol), DMAP (1.6 mg, 0.013 mmol), EDCI (300 mg, 1.57 mmol). The reaction system was protected by nitrogen, and the reaction was carried out at room temperature for 3 hours.

[0792] After the reaction was completed by LCMS monitoring, the reaction solution was poured into water, extracted with ethyl acetate, and the organic phase was washed with saturated sodium chloride solution. The organic phase was dried over anhydrous sodium sulfate, filtered, and the filtrate was concentrated. The obtained crude product was purified by silica gel column (PE / EA = 5:1) to obtain compound 37-3 (527 mg, yield 91.60%).

[0793] 1 H NMR (400 MHz, CDCl3) δ 8.24 (d, J = 2.3 Hz, 1H), 8.06 (dd, J = 8.5, 2.3 Hz, 1H), 7.46 (d, J = 8.5 Hz, 1H), 4.69-4.58 (m, 1H), 3.91 (s, 1H), 3.14 (t, J = 7.5 Hz, 2H), 2.70 (s, 3H), 2.67 (t, J = 7.5 Hz, 2H), 2.02-1.95 (m, 2H), 1.77-1.68 (m, 2H), 1.66-1.59 (m, 2H), 1.56-1.49 (m, 2H), 1.46 (s, 9H).

[0794] Step B: Compound 37-3 (264 mg, 0.60 mmol) was added to a 50 mL flask, followed by the addition of dry DMF (5 mL), and then tetrahydroxyboron (213 mg, 2.39 mmol) and 4,4'-dipyridine (46 mg, 0.29 mmol) were quickly added. The reaction system was protected by nitrogen, and the reaction was carried out at room temperature for 5 minutes.

[0795] After the reaction was completed by LCMS monitoring, a small amount of methanol was added, filtered, and the filtrate was purified by Flash (ACN / 0.1% FA in water) to obtain compound 37-4 (230 mg, yield 93.50%).

[0796] LCMS (ESI): 411.4 [M+H] + .

[0797] Step D: Compound 37-4 (230 mg, 0.561 mmol) was taken in 25 mL vial, after addition of dry THF (6 mL), dropwise added with bis(trichloromethyl) carbonate (216 mg, 1.092 mmol), the reaction was stirred at room temperature for 1 h under nitrogen atmosphere, the solvent was evaporated at low temperature, the residue was dissolved in dry DMF (3 mL) and added dropwise to a solution of compound 1-3 (220 mg, 0.711 mmol) and triethylamine (101 mg, 1.0 mmol) in DMF (5 mL), the reaction was stirred at room temperature for 1 h under nitrogen atmosphere.

[0798] After completion of reaction as monitored by LCMS, compound 37-5 (272 mg, yield 68.54%) was obtained by flash purification (ACN / 0.1% TFA in water).

[0799] LCMS (ESI): 610.2 [M-Boc+H] + .

[0800] Step E: Compound 37-5 (200 mg, 0.282 mmol) was taken in 25 mL vial, added with DCM (5 mL), added with TFA (5 mL) at 0 °C, the reaction was stirred at room temperature for 1 h under nitrogen atmosphere.

[0801] After completion of reaction as monitored by LCMS, the reaction was concentrated, the crude was purified by Prep-HPLC to obtain compound 37 (69.13 mg, yield 40.24%).

[0802] 1 H NMR (400 MHz, DMSO-d6) δ 10.98 (s, 1H), 8.89 (s, 1H), 8.45 (s, 2H), 7.73 - 7.61 (m, 2H), 7.51 (s, 1H), 7.44 (d, J = 7.9 Hz, 1H), 7.17 (s, 2H), 6.94 (t, J = 6.0 Hz, 1H), 5.11 (dd, J = 13.3, 5.1 Hz, 1H), 4.64 - 4.52 (m, 1H), 4.47 - 4.26 (m, 4H), 2.99 - 2.82 (m, 4H), 2.62 - 2.53 (m, 6H), 2.44 - 2.32 (m, 1H), 2.06 - 1.90 (m, 5H), 1.42 - 1.32 (m, 4H).

[0803] LCMS (ESI): 610.5 [M+H] + .

[0804] Example 38:

[0805] Preparation of (R) 1-(2-chloro-4-(3-(2-(2,6-dioxopiperidin-3-yl)-1-oxoisoindolin-5- yl)methyl)ureido)phenyl)cyclopropane-1-carboxylic acid piperidin-3-yl ester.

[0806] Reaction Scheme:

[0807] Operation Steps:

[0808] Step A: Compound 38-1 (400 mg, 1.86 mmol) was dissolved in dry DCM (25 mL), compound 38-2 (748.69 mg, 3.72 mmol), EDCI (427.87 mg, 2.23 mmol) and DMAP (22.72 mg, 0.19 mmol) were added, and the reaction system was reacted at room temperature for 2 hours under nitrogen protection.

[0809] After the reaction was completed by LCMS monitoring, the solvent was rotary evaporated, and the obtained crude product was purified by silica gel column (PE / EA = 3:1) to obtain compound 38-3 (500 mg, yield 67.57%).

[0810] LCMS (ESI): 397.1 [M-H] - .

[0811] Step B: Compound 38-3 (450 mg, 1.13 mmol) was dissolved in DMSO (16 mL), compound 38-4 (614.25 mg, 1.69 mmol) was added, and after stirring for 5 min, DBU (0.51 mL, 3.39 mmol) was added, and the reaction system was reacted at room temperature for 3 hours under nitrogen protection.

[0812] After the reaction was completed by LCMS monitoring, the reaction liquid was poured into water, extracted with ethyl acetate, and the organic phase was washed with saturated sodium chloride solution. After drying the organic phase with anhydrous sodium sulfate, filtering, concentrating the filtrate, the obtained crude product was purified by silica gel column (PE / EA = 6:1) to obtain compound 38-5 (440 mg, yield 91.97%).

[0813] LCMS (ESI): 325.1 [M-Boc+H] + .

[0814] Step C: Tetrahydroxyboron (377.08 mg, 3.76 mmol) was dissolved in dry DMF (12 mL), and then 4,4'-dipyridine (146.81 mg, 0.94 mmol) and compound 38-5 (400 mg, 0.94 mmol) were added quickly in sequence, and the reaction system was reacted at room temperature for 5 minutes under nitrogen protection.

[0815] After LCMS monitoring the completion of the reaction, a small amount of methanol was added, filtered, and the filtrate was purified by Flash (ACN / 0.1% FA in water) to give compound 38-6 (350 mg, yield 94.15%).

[0816] LCMS (ESI): 295.1 [M-Boc+H] + .

[0817] Step D: After compound 38-6 (100 mg, 0.25 mmol) was dissolved in dry THF (4 mL), diphosgene (0.06 mL, 0.5 mmol) was added dropwise slowly. The reaction system was protected by nitrogen, and reacted at room temperature for 1 hour. The solvent was dried and spun down at low temperature. The residue was dissolved in dry DMF (3 mL), and a solution of compound 1-3 (118.30 mg, 0.28 mmol) and triethylamine (0.35 mL, 2.5 mmol) in DMF (3 mL) was added dropwise slowly. The reaction system was protected by nitrogen, and reacted at room temperature for 1 hour.

[0818] After LCMS monitoring the completion of the reaction, the compound 38-7 (110 mg, yield 62.58%) was obtained by Flash purification (ACN / 0.1% FA in water).

[0819] LCMS (ESI): 594.2 [M-Boc+H] + .

[0820] Step E: Compound 38-7 (90 mg, 0.13 mmol) was dissolved in (10 mL), and TFA (2 mL) was added at 0°C. The reaction system was protected by nitrogen, and reacted at room temperature for 1 hour.

[0821] After LCMS monitoring the completion of the reaction, the reaction solution was concentrated, and the crude product was purified by Prep-HPLC to give compound 38 (65 mg, yield 70.80%).

[0822] 1H NMR (400 MHz, DMSO-d6) δ 10.98 (s, 1H), 9.07 (s, 1H), 8.71 (d, J = 48.8 Hz, 2H), 7.70 (t, J = 5.0 Hz, 2H), 7.51 (s, 1H), 7.44 (d, J = 7.9 Hz, 1H), 7.25 - 7.18 (m, 2H), 7.09 (t, J = 6.0 Hz, 1H), 5.11 (dd, J = 13.3, 5.1 Hz, 1H), 4.94 - 4.84 (m, 1H), 4.43 (t, J = 12.4 Hz, 3H), 4.31 (d, J = 17.3 Hz, 1H), 3.22 (d, J = 10.3 Hz, 1H), 3.05 - 2.86 (m, 4H), 2.60 (d, J = 17.3 Hz, 1H), 2.44 - 2.32 (m, 1H), 2.04 - 1.95 (m, 1H), 1.85 - 1.77 (m, 1H), 1.77 - 1.68 (m, 2H), 1.68 - 1.60 (m, 2H), 1.58 - 1.50 (m, 1H), 1.22 (d, J = 12.6 Hz, 2H).

[0823] LCMS (ESI): 594.4 [M+H] + .

[0824] Example 39:

[0825] Preparation of (S) 1-(2-chloro-4-(3-(2-(2,6-dioxopiperidin-3-yl)-1-oxoisoindolin-5- yl)methyl)ureido)phenyl)cyclopropane-1-carboxylic acid piperidin-3-yl ester.

[0826] Reaction Scheme:

[0827] Procedure:

[0828] Step A: Compound 39-1 (400 mg, 1.86 mmol) was dissolved in dry DCM (25 mL), compound 39-2 (748.69 mg, 3.72 mmol), EDCI (427.87 mg, 2.23 mmol) and DMAP (22.72 mg, 0.19 mmol) were added, the reaction system was protected by nitrogen, and reacted at room temperature for 2 hours.

[0829] After the reaction was completed by LCMS monitoring, the solvent was rotary evaporated, and the obtained crude product was purified by silica gel column (PE / EA = 3: 1) to obtain compound 39-3 (400 mg, yield 54.05%).

[0830] LCMS (ESI): 396.9 [M-H] - .

[0831] Step B: Compound 39-3 (350 mg, 0.88 mmol) was dissolved in DMSO (10 mL), compound 39-4 (478.35 mg, 1.32 mmol) was added, and after stirring for 5 min, DBU (401.91 mg, 2.64 mmol) was added. The reaction system was protected by nitrogen, and the reaction was carried out at room temperature for 3 h.

[0832] After the reaction was completed by LCMS monitoring, the reaction solution was poured into water, extracted with ethyl acetate, and the organic phase was washed with saturated sodium chloride solution. After drying the organic phase over anhydrous sodium sulfate, filtration, concentration of the filtrate, and purification of the obtained crude product by silica gel column (PE / EA = 5:1), compound 39-5 (350 mg, yield 93.87%) was obtained.

[0833] LCMS (ESI): 447.3 [M+Na] + .

[0834] Step C: Tetrahydroxydiboron (294.05 mg, 3.28 mmol) was dissolved in dry DMF (12 mL), and then 4,4'-dipyridine (128.07 mg, 0.82 mmol) and compound 39-5 (350 mg, 0.82 mmol) were added quickly in sequence. The reaction system was protected by nitrogen, and the reaction was carried out at room temperature for 5 min.

[0835] After the reaction was completed by LCMS monitoring, a small amount of methanol was added, filtered, and the filtrate was purified by Flash (ACN / 0.1% FA in water) to obtain compound 39-6 (270 mg, yield 83%).

[0836] LCMS (ESI): 295.0 [M-Boc+H] + .

[0837] Step D: Compound 39-6 (100 mg, 0.25 mmol) was dissolved in dry THF (4 mL), and then bis-triphosgene (0.06 mL, 0.5 mmol) was added dropwise slowly. The reaction system was protected by nitrogen, and the reaction was carried out at room temperature for 1 h. After the solvent was rotary evaporated at low temperature, the residue was dissolved in dry DMF (3 mL), and then added dropwise slowly to a solution of compound 1-3 (118.30 mg, 0.28 mmol) and triethylamine (0.35 mL, 2.5 mmol) in DMF (3 mL). The reaction system was protected by nitrogen, and the reaction was carried out at room temperature for 1 h

[0838] After the reaction was completed by LCMS monitoring, purification by Flash (ACN / 0.1% FA in water) gave compound 39-7 (140 mg, yield 79.64%).

[0839] LCMS (ESI): 594.3 [M-Boc+H] + .

[0840] Step E: Compound 39-7 (70 mg, 0.10 mmol) was dissolved in DCM (8 mL), TFA (1.6 mL) was added at 0 °C, the reaction system was protected under nitrogen, and the reaction was carried out at room temperature for 1 hour.

[0841] After the reaction was completed by LCMS monitoring, the reaction solution was concentrated, and the crude product was purified by Prep-HPLC to obtain compound 39 (58 mg, yield 96.82%).

[0842] 1 H NMR (400 MHz, DMSO-d6) δ 10.98 (s, 1H), 9.07 (s, 1H), 8.87 - 8.56 (m, 2H), 7.69 (d, J = 8.1 Hz, 2H), 7.51 (s, 1H), 7.44 (d, J = 7.9 Hz, 1H), 7.21 (dt, J = 8.6, 5.1 Hz, 2H), 7.09 (t, J = 6.0 Hz, 1H), 5.11 (dd, J = 13.3, 5.1 Hz, 1H), 4.94 - 4.85 (m, 1H), 4.43 (t, J = 12.4 Hz, 3H), 4.31 (d, J = 17.3 Hz, 1H), 3.27 - 3.18 (m, 1H), 3.07 - 2.86 (m, 4H), 2.60 (d, J = 16.7 Hz, 1H), 2.46 - 2.30 (m, 1H), 2.06 - 1.95 (m, 1H), 1.87 - 1.77 (m, 1H), 1.77 - 1.68 (m, 2H), 1.68 - 1.59 (m, 2H), 1.58 - 1.50 (m, 1H), 1.28 - 1.14 (m, 2H).

[0843] LCMS (ESI): 594.3 [M+H] + .

[0844] Example 40:

[0845] Preparation of (R)-1-(methylamino)propan-2-yl 1-(2-chloro-4-(3-((2-(2,6-dioxopiperidin-3-yl)-1-oxoisoindolin-5-yl)methyl)ureido)phenyl)cyclopropane-1-carboxylate.

[0846] Reaction Scheme:

[0847] Procedure:

[0848] Step A: Compound 40-2 (189 mg, 0.879 mmol) was added to a 100 mL three-necked flask, followed by the addition of DCM (10 mL), compound 40-1 (250 mg, 1.322 mmol), DMAP (1.6 mg, 0.013 mmol), EDCI (200 mg, 1.047 mmol). The reaction system was protected by nitrogen, and reacted at room temperature for 3 hours.

[0849] After the reaction was completed by LCMS monitoring, the reaction solution was poured into water, extracted with ethyl acetate, and the organic phase was washed with saturated sodium chloride solution. The organic phase was dried over anhydrous sodium sulfate, filtered, and the filtrate was concentrated. The obtained crude product was purified by silica gel column (PE / EA = 5:1) to obtain compound 40-3 (141 mg, yield 41.55%).

[0850] LCMS (ESI): 287.0 [M+H] + .

[0851] Step B: Compound 40-3 (140 mg, 0.365 mmol) was added to a 50 mL single-necked flask, followed by the addition of dry DMSO (10 mL), DBU (165 mg, 1.086 mmol) and compound 40-4. The reaction system was protected by nitrogen, and reacted at room temperature overnight.

[0852] After the reaction was completed by LCMS monitoring, 30 mL of ethyl acetate was added, washed with saturated brine three times, and the organic phase was concentrated. The residue was purified by silica gel column (PE / EA = 8:1) to obtain compound 40-5 (118 mg, yield 85.17%).

[0853] LCMS (ESI): 313.2 [M-Boc+H] + .

[0854] Step C: Compound 40-5 (118 mg, 0.286 mmol) was added to a 50 mL single-necked flask, followed by the addition of dry DMF (5 mL), tetrahydroxyboron (103 mg, 1.157 mmol) and 4,4'-dipyridine (23 mg, 0.147 mmol) were added quickly. The reaction system was protected by nitrogen, and reacted at room temperature for 5 minutes.

[0855] After LCMS monitoring the reaction was complete, a small amount of methanol was added, filtered, and the filtrate was purified by Flash (ACN / 0.1% FA in water) to give compound 40-6 (102 mg, yield 93%), which was separated by SFC to give 40-6-P1 (41 mg) and 40-6-P2 (40 mg). Separation conditions Mobile phase: supercritical carbon dioxide and 0.1% ammonia in methanol; flow rate: 150 mL / min; gradient: hold 0.1% ammonia in isopropanol at 15%; detection wavelength: 214 nm; cycle time: 4.0 min.

[0856] Compound 40-6-P1 retention time Rt = 6.0 min; compound 40-6-P2 retention time Rt = 7.2 min.

[0857] LCMS (ESI): 283.1 [M-Boc+H] + .

[0858] Step D: Compound 40-6-P1 (40 mg, 0.105 mmol) was added to a 25 mL vial, after adding dry THF (6 mL), bis(trichloromethyl) carbonate (216 mg, 0.202 mmol) was added dropwise slowly, the reaction system was protected by nitrogen, and reacted at room temperature for 1 hour. The solvent was rotary evaporated at low temperature, and the residue was dissolved in dry DMF (3 mL), then added dropwise to a solution of compound 1-3 (42 mg, 0.136 mmol) and triethylamine (21 mg, 0.209 mmol) in DMF (5 mL). The reaction system was protected by nitrogen, and reacted at room temperature for 1 hour.

[0859] After LCMS monitoring the reaction was complete, the reaction solution was concentrated, and the crude product was purified by Prep-HPLC to give compound 40 (12.78 mg, yield 49.93%).

[0860] LCMS (ESI): 582.0 [M-Boc+H] + .

[0861] Step E: Compound 40-7 (30 mg, 0.044 mmol) was added to a 25 mL vial, DCM (5 mL) was added, and TFA (5 mL) was added at 0°C. The reaction system was protected by nitrogen, and reacted at room temperature for 1 hour.

[0862] After LCMS monitoring the reaction was complete, the reaction solution was concentrated, and the crude product was purified by Prep-HPLC to give compound 40 (12.78 mg, yield 49.93%).

[0863] LCMS (ESI): 582.5 [M+H] + .

[0864] 1 H NMR (400 MHz, DMSO-d6) δ 10.98 (s, 1H), 9.12 - 8.90 (m, 1H), 8.63 - 8.35 (m, 2H), 7.74 - 7.64 (m, 2H), 7.51 (s, 1H), 7.44 (d, J = 7.9 Hz, 1H), 7.26 - 7.18 (m, 2H), 7.09 - 6.96 (m, 1H), 5.11 (dd, J = 13.2, 5.1 Hz, 1H), 5.05 - 4.96 (m, 1H), 4.49 - 4.26 (m, 4H), 3.09 - 2.85 (m, 3H), 2.64 - 2.56 (m, 1H), 2.49 (s, 3H), 2.47 - 2.31 (m, 1H), 2.04 - 1.95 (m, 1H), 1.79 - 1.71 (m, 1H), 1.60 - 1.52 (m, 1H), 1.27 - 1.22 (m, 1H), 1.16 - 1.06 (m, 4H).

[0865] LCMS (ESI): 582.5 [M+H] + .

[0866] Example 41:

[0867] Preparation of (S)-1-(methylamino)propan-2-yl 1-(2-chloro-4-(3-((2-(2,6-dioxopiperidin-3-yl)-1-oxoisoindolin-5-yl)methyl)ureido)phenyl)cyclopropane-1-carboxylate.

[0868] Reaction Scheme:

[0869] Procedure:

[0870] Step A: Compound 40-6-P2 (40 mg, 0.105 mmol) was added to a 25 mL vial, after adding dry THF (6 mL), bis(trichloromethyl) carbonate (216 mg, 0.202 mmol) was added slowly dropwise, the reaction system was protected by nitrogen, and reacted at room temperature for 1 hour. The solvent was rotary evaporated at low temperature, the residue was dissolved in dry DMF (3 mL), and then slowly added to a solution of compound 1-3 (42 mg, 0.136 mmol) and triethylamine (21 mg, 0.209 mmol) in DMF (5 mL). The reaction system was protected by nitrogen, and reacted at room temperature for 1 hour

[0871] After the reaction was completed by LCMS monitoring, compound 41-2 (48 mg, yield 89.67%) was obtained by Flash purification (ACN / 0.1% TFA in water).

[0872] LCMS (ESI): 582.2 [M-Boc+H] + .

[0873] Step B: Compound 41-2 (48 mg, 0.070 mmol) was taken in 25 mL single necked flask, DCM (5 mL) was added, TFA (5 mL) was added at 0 °C, the reaction was stirred at room temperature for 1 h under nitrogen atmosphere.

[0874] After completion of reaction by LCMS, the reaction mixture was concentrated, the crude was purified by Prep-HPLC to get compound 41 (31.61 mg, yield 77.18%).

[0875] 1 H NMR (400 MHz, DMSO-d6) δ 10.98 (s, 1H), 9.00 (s, 1H), 8.47 (s, 2H), 7.73 - 7.67 (m, 2H), 7.51 (s, 1H), 7.44 (d, J = 7.9 Hz, 1H), 7.26 - 7.17 (m, 2H), 7.02 (t, J = 6.0 Hz, 1H), 5.20 - 4.93 (m, 2H), 4.49 - 4.25 (m, 4H), 3.12 - 2.84 (m, 3H), 2.69 - 2.53 (m, 1H), 2.48 (s, 3H), 2.45 - 2.30 (m, 1H), 2.03 - 1.94 (m, 1H), 1.80 - 1.71 (m, 1H), 1.61 - 1.52 (m, 1H), 1.31 - 1.21 (m, 1H), 1.17 - 1.10 (m, 4H).

[0876] LCMS (ESI): 582.5 [M+H] + .

[0877] Example 42:

[0878] Preparation of 1-(3-chloro-4-(2-(2-(methylamino)ethyl)sulfonyl)ethyl)phenyl)-3-(2-(2,6-dioxopiperidin-3-yl)-1-oxoisoindolin-5-yl)methyl)urea.

[0879] Reaction Scheme:

[0880] Procedure:

[0881] Step A: Compound 42-1 (1 g, 5.71 mmol) was dissolved in THF (20 mL), PPh3 (2.40 g, 9.14 mmol) and CBr4 (2.27 g, 6.85 mmol) were added successively at 0 °C, the reaction system was protected by nitrogen, and the reaction was carried out at room temperature for 2 hours.

[0882] After TLC monitoring of the completion of the reaction, the reaction solution was concentrated, and the crude product was purified by silica gel column (PE / EA = 20:1) to obtain compound 42-2 (1.01 g, yield 74.32%).

[0883] 1 H NMR (400 MHz, DMSO-d6) δ 3.58-3.48 (m, 4H), 2.82 (d, J = 11.5 Hz, 3H), 1.40 (s, 9H).

[0884] Step B: Compound 42-3 (2 g, 9.92 mmol) was dissolved in DCM (100 mL), PPh3 (4.16 g, 15.87 mmol) and CBr4 (3.95 g, 11.90 mmol) were added successively at 0 °C, the reaction system was protected by nitrogen, and the reaction was carried out at room temperature for 2 hours.

[0885] After TLC monitoring of the completion of the reaction, the reaction solution was concentrated, and the crude product was purified by silica gel column (PE / EA = 10:1) to obtain compound 42-4 (2.4 g, yield 91.47%).

[0886] 1 H NMR (400 MHz, DMSO-d6) δ 8.29 (d, J = 2.3 Hz, 1H), 8.19 (dd, J = 8.5, 2.3 Hz, 1H), 7.75 (d, J = 8.5 Hz, 1H), 3.80 (t, J = 7.0 Hz, 2H), 3.40 (t, J = 7.0 Hz, 2H).

[0887] Step C: Compound 42-4 (2.4 g, 9.07 mmol) was dissolved in dry DMF (30 mL), compound 42-5 (1.24 g, 10.88 mmol) was added, the reaction system was protected by nitrogen, and the reaction was carried out at room temperature for 2 hours.

[0888] After TLC monitoring of the completion of the reaction, the reaction solution was poured into water, extracted with ethyl acetate, and the organic phase was washed with saturated sodium chloride solution. After drying the organic phase with anhydrous sodium sulfate, filtering, and concentrating the filtrate, the obtained crude product was purified by silica gel column (PE / EA = 10:1) to obtain compound 42-6 (2 g, yield 84.87%).

[0889] 1H NMR (400 MHz, DMSO-d6) δ 8.27 (d, J = 2.3 Hz, 1H), 8.16 (dd, J = 8.5, 2.3 Hz, 1H), 7.68 (d, J = 8.5 Hz, 1H), 3.16 (t, J = 6.9 Hz, 2H), 3.07 (t, J = 6.9 Hz, 2H), 2.32 (s, 3H).

[0890] Step D: Compound 42-6 (2 g, 7.7 mmol) was dissolved in methanol (30 mL), sodium methoxide (2.08 g, 38.5 mmol) was added at 0 °C, the reaction system was protected by nitrogen, and reacted at 0 °C for 2 hours.

[0891] After the completion of the reaction was monitored by TLC, the pH value of the reaction solution was adjusted to 4 by adding acetic acid under ice bath, the reaction solution was concentrated, and the crude product was purified by silica gel column (PE / EA = 10:1) to obtain compound 42-7 (1.6 g, yield 95.45%).

[0892] 1 H NMR (400 MHz, DMSO-d6) δ 8.27 (d, J = 2.3 Hz, 1H), 8.16 (dd, J = 8.5, 2.3 Hz, 1H), 7.68 (d, J = 8.5 Hz, 1H), 3.16 (t, J = 6.9 Hz, 2H), 3.07 (t, J = 6.9 Hz, 2H), 2.32 (s, 3H).

[0893] Step E: Compound 42-7 (400 mg, 1.84 mmol) was dissolved in methanol (30 mL), compound 42-2 (481.95 mg, 2.02 mmol) and KOH (227.13 mg, 4.05 mmol) were added, the reaction system was protected by nitrogen, and reacted at 50 °C for 4 hours.

[0894] After the completion of the reaction was monitored by TLC, the reaction solution was concentrated, and the crude product was purified by silica gel column (PE / EA = 10:1) to obtain compound 42-8 (180 mg, yield 26.13%).

[0895] LCMS (ESI): 274.8 [M-Boc+H] + .

[0896] Step F: Compound 42-8 (160 mg, 0.43 mmol) was dissolved in dry DCM (20 mL), m-CPBA (222.62 mg, 1.29 mmol) was added, the reaction system was protected by nitrogen, and reacted at room temperature for 2 hours.

[0897] After the reaction was completed by LCMS monitoring, the reaction solution was concentrated, and the crude product was purified by Flash (ACN / 0.1% FA in water) to obtain compound 42-9 (140 mg, yield 80.62%).

[0898] LCMS (ESI): 351.0 [M-(t-Bu)+H] + .

[0899] Step G: Tetrahydroxydiboron (121.92 mg, 1.36 mmol) was dissolved in dry DMF (8 mL), and then 4,4'-dipyridine (53.10 mg, 0.34 mmol) and compound 42-9 (140 mg, 0.34 mmol) were added quickly in turn. The reaction system was protected by nitrogen, and the reaction was carried out at room temperature for 5 minutes.

[0900] After the reaction was completed by LCMS monitoring, a small amount of methanol was added, filtered, and the filtrate was purified by Flash (ACN / 0.1% FA in water) to obtain compound 42-10 (110 mg, yield 84.82%).

[0901] LCMS (ESI): 399.3 [M+Na] + .

[0902] Step H: Compound 42-10 (100 mg, 0.27 mmol) was dissolved in dry THF (8 mL), and then diphosgene (106.83 mg, 0.54 mmol) was added dropwise slowly. The reaction system was protected by nitrogen, and the reaction was carried out at room temperature for 1 hour. After the reaction solution was concentrated, the residue was dissolved in dry DMF (4 mL), and then a solution of compound 1-3 (81.17 mg, 0.3 mmol) and triethylamine (0.38 mL, 2.7 mmol) in DMF (4 mL) was added dropwise slowly. The reaction system was protected by nitrogen, and the reaction was carried out at room temperature for 1 hour.

[0903] After the reaction was completed by LCMS monitoring, the crude product was purified by Flash (ACN / 0.1% FA in water) to obtain compound 42-11 (140 mg, yield 78.04%).

[0904] LCMS (ESI): 576.1 [M-Boc+H] + .

[0905] Step I: Compound 42-11 (80 mg, 0.12 mmol) was dissolved in DCM (8 mL), and then TFA (1.2 mL) was added at 0°C. The reaction system was protected by nitrogen, and the reaction was carried out at room temperature for 1 hour.

[0906] After the reaction was completed by LCMS monitoring, the reaction solution was concentrated, and the crude product was purified by Prep-HPLC to obtain compound 42 (45 mg, yield 66.03%).

[0907] 1 H NMR (400 MHz, DMSO-d6) δ 10.98 (s, 1H), 8.99 (s, 1H), 8.60 (s, 2H), 7.70 (dd, J = 8.2, 5.0 Hz, 2H), 7.51 (s, 1H), 7.44 (d, J = 8.0 Hz, 1H), 7.31 - 7.19 (m, 2H), 7.01 (t, J = 6.0 Hz, 1H), 5.11 (dd, J = 13.3, 5.1 Hz, 1H), 4.43 (dd, J = 14.4, 11.8 Hz, 3H), 4.31 (d, J = 17.4 Hz, 1H), 3.59 - 3.53 (m, 2H), 3.52 - 3.45 (m, 2H), 3.41 - 3.36 (m, 2H), 3.09 - 3.01 (m, 2H), 2.97 - 2.86 (m, 1H), 2.64 - 2.56 (m, 4H), 2.44 - 2.32 (m, 1H), 2.04 - 1.95 (m, 1H).

[0908] LCMS (ESI): 576.3 [M+H] + .

[0909] Example 43:

[0910] Preparation of 1-(3-chloro-4-(2-(piperidin-4-ylsulfonyl)ethyl)phenyl)-3-((2-(2-(2,6-dioxopiperidin-3-yl)-1-oxoisoindolin-5-yl)methyl)urea.

[0911] Reaction Scheme:

[0912] Procedure:

[0913] Step A: Compound 43-1 (200 mg, 0.922 mmol) was dissolved in DMF (4 mL), compound 43-2 (429 mg, 1.378 mmol) and potassium carbonate (381 mg, 2.76 mmol) were added, and the reaction was carried out at 50 degrees Celsius for 3 hours.

[0914] After the reaction was completed by LCMS monitoring, the reaction solution was poured into water, extracted with ethyl acetate, and the organic phase was washed with saturated sodium chloride solution. After drying the organic phase with anhydrous sodium sulfate, filtering, concentrating the filtrate, the obtained crude product was purified by silica gel column (PE / EA = 10:1) to obtain compound 43-3 (133 mg, yield 36.00%).

[0915] LCMS (ESI): 345.0 [M-(t-Bu)+H] + .

[0916] Step B: Compound 43-3 (133 mg, 0.332 mmol) was dissolved in dry DCM (5 mL), m-CPBA (158 mg, 0.831 mmol, 85% purity) was added, the reaction was stirred at 0 °C for 1 h under nitrogen.

[0917] After the reaction was completed by LCMS monitoring, the reaction was concentrated, the crude product was purified by Flash (ACN / 0.1% FA in water) to give compound 43-4 (130 mg, yield 90.52%).

[0918] LCMS (ESI): 333.2 [M-Boc+H] + .

[0919] Step C: Compound 43-4 (130 mg, 0.301 mmol) was dissolved in dry DMF (10 mL), tetrahydroxyboron (107 mg, 1.202 mmol) and 4,4'-dipyridine (23 mg, 0.147 mmol) were added quickly, the reaction was stirred at room temperature for 5 min under nitrogen.

[0920] After the reaction was completed by LCMS monitoring, a small amount of methanol was added, filtered, the filtrate was purified by Flash (ACN / 0.1% FA in water) to give compound 43-5 (110 mg, yield 90.91%).

[0921] LCMS (ESI): 303.0 [M-Boc+H] + .

[0922] Step D: Compound 43-5 (110 mg, 0.274 mmol) was dissolved in dry THF (6 mL), diphosgene (110 mg, 0.556 mmol) was added slowly dropwise, the reaction was stirred at room temperature for 1 h under nitrogen, the solvent was rotary evaporated at low temperature, the residue was dissolved in dry DMF (3 mL), then added slowly dropwise to a solution of compound 1-3 (130 mg, 0.421 mmol) and triethylamine (65 mg, 0.644 mmol) in DMF (5 mL), the reaction was stirred at room temperature for 1 h under nitrogen.

[0923] After the reaction was completed by LCMS monitoring, compound 43-6 (110 mg, yield 57.38%) was obtained by Flash purification (ACN / 0.1% TFA in water).

[0924] LCMS (ESI): 602.2 [M-Boc+H] + .

[0925] Step E: Compound 43-6 (90 mg, 0.128 mmol) was dissolved in DCM (5 mL), TFA (5 mL) was added at 0 °C, the reaction system was protected under nitrogen, and the reaction was carried out at room temperature for 1 hour.

[0926] After the reaction was completed by LCMS monitoring, the reaction solution was concentrated, and the crude product was purified by Prep-HPLC to obtain compound 43 (30.82 mg, yield 39.94%).

[0927] 1 H NMR (400 MHz, DMSO-d6) δ 10.98 (s, 1H), 9.08 (s, 1H), 8.29 (s, 1H), 7.74 - 7.62 (m, 2H), 7.51 (s, 1H), 7.44 (d, J = 8.1 Hz, 1H), 7.32 - 7.27 (m, 1H), 7.24 - 7.19 (m, 1H), 7.10 (t, J = 5.9 Hz, 1H), 5.10 (dd, J = 13.3, 5.1 Hz, 1H), 4.49 - 4.26 (m, 4H), 3.44 - 3.36 (m, 1H), 3.35 - 3.19 (m, 5H), 3.06 - 3.00 (m, 2H), 2.96 - 2.86 (m, 1H), 2.81 - 2.68 (m, 2H), 2.65 - 2.54 (m, 1H), 2.44 - 2.32 (m, 1H), 2.14 - 2.04 (m, 2H), 2.04 - 1.95 (m, 1H), 1.76 - 1.61 (m, 2H).

[0928] LCMS (ESI): 602.6 [M+H] + .

[0929] Example 44:

[0930] Preparation of (4S)-4-amino-5-((2-(2-chloro-4-(3-(2-(2,6-dioxopiperidin-3-yl)-1- oxoisoindolin-5-yl)methyl)ureido)phenethoxy)ethyl)(prop-2-yn-1-yl)amino)-5- oxopentanoic acid.

[0931] Reaction Scheme:

[0932] Procedure:

[0933] Step A: Compound 44-1 (35 mg, 0.064 mmol) was dissolved in dry DMF (3 mL), HATU (27 mg, 0.070 mmol), compound 44-2 (23 mg, 0.076 mmol) and DIEA (18 mg, 0.134 mmol) were added at room temperature, the reaction was stirred at room temperature for 1 hour.

[0934] After the reaction was completed by LCMS monitoring, the reaction solution was purified by reverse phase Flash (ACN / 0.1% FA in water) to give compound 44-3 (45 mg, yield 85%).

[0935] LCMS (ESI): 837.4 [M+H] + .

[0936] Step B: Compound 44-3 (45 mg, 0.054 mmol) was dissolved in DCM (4 mL), then TFA (4 mL) was added, the reaction system was protected by nitrogen, and reacted at room temperature for 3 hours.

[0937] After the reaction was completed by LCMS monitoring, the reaction solution was concentrated, and the crude product was purified by Prep-HPLC to give compound 44 (36 mg, yield 99%).

[0938] 1 H NMR (400 MHz, DMSO-d6) δ 10.98 (s, 1H), 9.00-8.80 (m, 1H), 8.17 (br s, 2H), 7.74-7.60 (m, 2H), 7.51 (s, 1H), 7.44 (d, J = 8.0 Hz, 1H), 7.26-7.13 (m, 2H), 7.07-6.82 (m, 1H), 5.11 (dd, J = 13.3, 5.1 Hz, 1H), 4.47-4.10 (m, 7H), 3.78-3.52 (m, 6H), 3.50 - 3.40 (m, 1H), 3.24 (s, 1H), 2.98 - 2.81 (m, 3H), 2.69-2.58 (m, 1H), 2.45 - 2.27 (m, 3H), 2.05 - 1.94 (m, 2H), 1.91 - 1.82 (m, 1H).

[0939] LCMS (ESI): 681.0 [M+H] + .

[0940] Example 45:

[0941] 4-((S)-2-(S)-2-(6-(2,5-dioxo-2,5-dihydro-1H-pyrrol-1-yl)hexanamine)-3- methylbutanamine)-5-ureidopentanamine)benzyl(2-(2-(2,6-dioxo-3-yl)-1- oxoisoindol-5-yl)methyl)ureido)phenethyloxy)(2,2,2-trifluoroethyl)carbamate.

[0942] Reaction Scheme:

[0943] Procedure:

[0944] Step A: Compound 15 (50 mg, 0.083 mmol) was added to a 25 mL single neck flask, followed by DMF (2 mL), compound 1-10 (70 mg, 0.095 mmol), HOPO (5 mg, 0.045 mmol), the reaction system was protected by nitrogen, and reacted at room temperature for 16 hours.

[0945] After the reaction was completed by LCMS monitoring, a small amount of methanol was added, filtered, and the filtrate was purified by Flash to obtain compound 45 (20.33 mg, yield 20.31%).

[0946] LCMS (ESI): 1194.6 [M+H] + .

[0947] 1H NMR (400 MHz, DMSO-d6) δ 10.98 (s, 1H), 10.00 (s, 1H), 8.80 (s, 1H), 8.08 (d, J = 7.4 Hz, 1H), 7.80 (d, J = 8.6 Hz, 1H), 7.71 - 7.64 (m, 2H), 7.62 - 7.57 (m, 2H), 7.51 (s, 1H), 7.44 (d, J = 7.8 Hz, 1H), 7.33 - 7.25 (m, 2H), 7.23 - 7.09 (m, 2H), 6.99 (s, 2H), 6.83 (t, J = 5.8 Hz, 1H), 6.01 (dr, 1H), 5.11 (dd, J = 13.3, 5.1 Hz, 1H), 5.04 (s, 2H), 4.44 - 4.30 (m, 5H), 4.20 - 4.17 (m, 1H), 4.06 - 4.01 (m, 2H), 3.58 - 3.47 (m, 4H), 3.46 - 3.41 (m, 2H), 3.36 (t, J = 7.0 Hz, 2H), 3.08 - 2.87 (m, 3H), 2.86 - 2.77 (m, 2H), 2.66 - 2.55 (m, 1H), 2.45 - 2.32 (m, 1H), 2.23 - 2.06 (m, 2H), 2.04 - 1.92 (m, 2H), 1.76 - 1.55 (m, 2H), 1.51 - 1.34 (m, 6H), 1.21 - 1.13 (m, 2H), 0.83 (dd, J = 12.4, 6.7 Hz, 6H).

[0948] Example 46:

[0949] Preparation of 4-((S)-2-(S)-2-(6-(2,5-dioxo-2,5-dihydro-1H-pyrrol-1-yl)hexanamido)-3- methylbutanamido)-5-ureidopentanamido)benzyl(2-(2-chloro-4-(3-(2-(2,6-dioxopiperidin- 3-yl)-1-oxoisoindolin-5-yl)methyl)ureido)phenethyl)ethyl)(cyanomethyl)carbamate.

[0950] Reaction Scheme:

[0951] Procedure:

[0952] Step A: Compound 16 (50 mg, 0.083 mmol) was added to a 25 mL vial followed by DMF (2 mL), compound 1-10 (70 mg, 0.095 mmol), HOBt (22 mg, 0.166 mmol), Lutidine (0.02 mL). The reaction was allowed to stir at room temperature for 16 hours under a nitrogen atmosphere.

[0953] After LCMS monitoring the completion of the reaction, a small amount of methanol was added, filtered, and the filtrate was purified by Flash to obtain compound 46 (25 mg, yield 22.56%).

[0954] LCMS (ESI): 1149.6 [M-H] - .

[0955] 1 H NMR (400 MHz, DMSO-d6) δ 10.97 (s, 1H), 10.01 (s, 1H), 8.93 (s, 1H), 8.09 (d, J = 7.5 Hz, 1H), 7.80 (d, J = 8.7 Hz, 1H), 7.69 (d, J = 8.2 Hz, 2H), 7.60 (d, J = 8.5 Hz, 2H), 7.51 (s, 1H), 7.44 (d, J = 7.9 Hz, 1H), 7.32 (s, 2H), 7.16 (s, 2H), 7.04 - 6.92 (m, 3H), 5.98 (t, J = 5.6 Hz, 1H), 5.41 (s, 2H), 5.19 - 4.99 (m, 3H), 4.43 - 4.28 (m, 6H), 4.23 - 4.15 (m, 1H), 3.58 - 3.49 (m, 4H), 3.49 - 3.43 (m, 2H), 3.39 - 3.34 (m, 2H), 3.05 - 2.81 (m, 5H), 2.66 - 2.54 (m, 1H), 2.44 - 2.32 (m, 1H), 2.22 - 2.08 (m, 2H), 2.02 - 1.90 (m, 2H), 1.70 - 1.34 (m, 8H), 1.28 - 1.07 (m, 3H), 0.88 - 0.78 (m, 6H).

[0956] Example 47:

[0957] Preparation of 4-((S)-2-(S)-2-(6-(2,5-dioxo-2,5-dihydro-1H-pyrrol-1-yl)hexanamido)-3- methylbutanamido)-5-ureidopentanamido)benzyl(2-amino-2-oxoethyl)(2-(2-(2,6- dioxopiperidin-3-yl)-1-oxoisoindolin-5-yl)methyl)ureido)phenethyl)oxycarbamate.

[0958] Reaction Scheme:

[0959] Procedure:

[0960] Step A: Compound 17 (50 mg, 0.083 mmol) was added to a 25 mL vial, followed by DMF (2 mL), compound 1-10 (70 mg, 0.095 mmol), HOBt (22 mg, 0.166 mmol), Lutidine (0.2 mL). The reaction was stirred at room temperature for 16 hours under nitrogen.

[0961] After LCMS monitoring the reaction was complete, a small amount of methanol was added, filtered, and the filtrate was purified by Flash (ACN / 0.1% FA in water) to give compound 47 (30 mg, yield 33.72%).

[0962] LCMS (ESI): 1169.7 [M+H] + .

[0963] 1 H NMR (400 MHz, DMSO-d6) δ 10.98 (s, 1H), 9.98 (d, J = 4.7 Hz, 1H), 8.79 (s, 1H), 8.15 - 8.04 (m, 1H), 7.80 (d, J = 8.6 Hz, 1H), 7.73 - 7.64 (m, 2H), 7.61 - 7.54 (m, 2H), 7.51 (s, 1H), 7.44 (d, J = 7.8 Hz, 1H), 7.33 - 7.12 (m, 5H), 7.03 - 6.92 (m, 3H), 6.82 (t, J = 4.6 Hz, 1H), 5.98 (s, 1H), 5.41 (s, 1H), 5.13 - 5.05 (m, 1H), 4.98 (d, J = 9.6 Hz, 2H), 4.49 - 4.28 (m, 5H), 4.23 - 4.14 (m, 1H), 3.81 (d, J = 4.5 Hz, 2H), 3.55 - 3.51 (d, J = 6.9 Hz, 2H), 3.49 - 3.46 (m, 2H), 3.40 - 3.34 (m, 4H), 3.02 - 2.77 (m, 5H), 2.66 - 2.56 (m, 1H), 2.43 - 2.32 (m, 1H), 2.21 - 2.07 (m, 2H), 2.03 - 1.92 (m, 2H), 1.77 - 1.38 (m, 8H), 1.33 - 1.11 (m, 3H), 0.87 - 0.78 (m, 6H).

[0964] Example 48:

[0965] 4-((S)-2-(S)-2-(6-(2,5-dioxo-2,5-dihydro-1H-pyrrol-1-yl)hexanoylamido)-3- methylbutanamido)-5-ureidopentanamido)benzyl(2-(2-chloro-4-(3-(2-(2,6- dioxopiperidin-3-yl)-1-oxoisoindolin-5-yl)methyl)ureido)phenethyl)ethyl)(cyclopropyl)carbamic acid

[0966] Reaction Scheme:

[0967] Procedure:

[0968] Step A: Compound 18 (100 mg, 0.18 mmol) was dissolved in DMF (4 mL), then 1-10 (135 mg, 0.18 mmol), HOBt (12 mg, 0.09 mmol) and 2,4-dimethylpyridine (19 mg, 0.18 mmol) were added. The reaction system was reacted at 40 °C for 12 hours under nitrogen protection.

[0969] After the reaction was completed by LCMS monitoring, compound 48 (75 mg, yield 36%) was obtained by Prep-HPLC purification.

[0970] LCMS (ESI): 1152.7 [M+H] + .

[0971] 1H NMR (400 MHz, DMSO-d6) δ 10.98 (s, 1H), 9.98 (s, 1H), 8.80 (s, 1H), 8.08 (d, J = 7.5 Hz, 1H), 7.80 (d, J = 7.7 Hz, 1H), 7.71 - 7.64 (m, 2H), 7.59 (d, J = 8.4 Hz, 2H), 7.51 (s, 1H), 7.44 (d, J = 8.3 Hz, 1H), 7.29 (d, J = 8.7 Hz, 2H), 7.21 - 7.11 (m, J = 9.0 Hz, 2H), 7.00 (s, 2H), 6.83 (s, 1H), 5.97 (s, 1H), 5.41 (s, 2H), 5.10 (dd, J = 13.1, 4.8 Hz, 1H), 5.00 (s, 2H), 4.50 - 4.28 (m, 5H), 4.18 (t, J = 7.7 Hz, 1H), 3.58 - 3.47 (m, 5H), 3.39 - 3.36 (m, 3H), 3.07 - 2.85 (m, 3H), 2.82 (t, J = 6.7 Hz, 2H), 2.69 - 2.58 (m, 4H), 2.45 - 2.31 (m, 1H), 2.20 - 2.06 (m, 2H), 2.03 - 1.93 (m, 4H), 1.51 - 1.42 (m, 4H), 1.22 - 1.12 (m, 2H), 0.85 - 0.79 (m, 6H), 0.66 (d, J = 5.1 Hz, 2H), 0.57 (s, 2H).

[0972] Example 49:

[0973] Preparation of 4-((S)-2-((S)-2-(6-(2,5-dioxo-2,5-dihydro-1H-pyrrol-1- yl)hexanamido)-3-methylbutanamido)-5-ureidopentanamido)benzyl (2-(2-chloro-4- (3-((2-(2,6-dioxopiperidin-3-yl)-1-oxoisoindolin-5-yl)methyl)ureido)phenethyl)ethyl) (methoxy)carbamate.

[0974] Reaction Scheme:

[0975] Procedure:

[0976] Step A: Compound 19 (80 mg, 0.15 mmol) was dissolved in DMF (6 mL), followed by the addition of compound 1-10 (119.52 mg, 0.16 mmol), HOBt (10.13 mg, 0.075 mmol) and 2,4-dimethylpyridine (32.15 mg, 0.30 mmol). The reaction system was protected by nitrogen and reacted at 40 °C for 12 hours.

[0977] LCMS monitoring of the reaction completion, compound 49 was obtained by Prep-HPLC purification (55 mg, yield 29.46%).

[0978] 1 H NMR (400 MHz, DMSO-d6) δ 10.98 (s, 1H), 10.00 (s, 1H), 8.80 (s, 1H), 8.08 (d, J = 7.5 Hz, 1H), 7.80 (d, J = 8.7 Hz, 1H), 7.73 - 7.55 (m, 2H), 7.60 (d, J = 8.5 Hz, 2H), 7.51 (s, 1H), 7.44 (d, J = 8.2 Hz, 1H), 7.31 (d, J = 8.6 Hz, 2H), 7.15 (dt, J = 8.4, 5.2 Hz, 2H), 7.00 (s, 2H), 6.83 (t, J = 5.9 Hz, 1H), 6.02 (s, 1H), 5.60 (brs, 2H), 5.14 - 5.02 (m, 3H), 4.50 - 4.27 (m, 5H), 4.23 - 4.15 (m, 1H), 3.63 - 3.48 (m, 9H), 3.36 (t, J = 7.0 Hz, 2H), 3.08 - 2.85 (m, 3H), 2.81 (t, J = 7.0 Hz, 2H), 2.69 - 2.55 (m, 1H), 2.44 - 2.32 (m, 1H), 2.22 - 2.06 (m, 2H), 2.03 - 1.91 (m, 2H), 1.76 - 1.64 (m, 1H), 1.64 - 1.54 (m, 1H), 1.52 - 1.32 (m, 6H), 1.22 - 1.13 (m, 2H), 0.87 - 0.75 (m, 6H).

[0979] LCMS (ESI): 1142.6 [M+H] + .

[0980] Example 50:

[0981] Preparation of 4-((S)-2-((S)-2-(6-(2,5-dioxo-2,5-dihydro-1H-pyrrol-1- yl)hexanamido)-3-methylbutanamido)-5-ureidopentanamido)benzyl (4-(2-chloro-4- (3-((2-(2,6-dioxopiperidin-3-yl)-1-oxoisoindolin-5-yl)methyl)ureido)phenethyl oxy)-N- (N-(2-hydroxyethyl)amidinyl))carbamate.

[0982] Reaction Scheme:

[0983] Procedure:

[0984] Step A: Compound 20 (90 mg, 0.16 mmol) was dissolved in THF (7 mL), then 1-10 (160 mg, 0.21 mmol) and TEA (49 mg, 0.48 mmol) were added, the reaction system was protected by nitrogen, and it was reacted at 40 °C for 12 hours.

[0985] After the reaction was completed by LCMS monitoring, compound 50 (35 mg, yield 19%) was obtained by Prep-HPLC purification.

[0986] LCMS (ESI): 1154.5 [M+H] + .

[0987] 1 H NMR (400 MHz, DMSO-d6) δ 10.97 (s, 1H), 9.96 (s, 1H), 8.92 - 8.77 (m, 1H), 8.07 (d, J = 7.3 Hz, 1H), 7.80 (d, J = 8.6 Hz, 1H), 7.72 - 7.61 (m, 2H), 7.59 - 7.53 (m, 2H), 7.51 (s, 1H), 7.44 (d, J = 7.7 Hz, 1H), 7.27 - 7.19 (m, 3H), 7.17 - 7.13 (m, 1H), 7.00 (s, 2H), 6.92 - 6.83 (m, 1H), 5.97 (t, J = 5.6 Hz, 1H), 5.40 (s, 2H), 5.17 - 5.04 (m, 1H), 4.89 (s, 2H), 4.48 - 4.34 (m, 4H), 4.34 - 4.26 (m, 1H), 4.22 - 4.13 (m, 1H), 3.55 (t, J = 7.0 Hz, 2H), 3.49 - 3.41 (m, 3H), 3.07 - 2.77 (m, 6H), 2.70 - 2.54 (m, 2H), 2.45 - 2.31 (m, 2H), 2.19 - 2.08 (m, 2H), 2.02 - 1.94 (m, 2H), 1.74 - 1.65 (m, 1H), 1.63 - 1.54 (m, 1H), 1.53 - 1.30 (m, 7H), 1.27 - 1.12 (m, 4H), 0.87 - 0.79 (m, 6H).

[0988] Example 51:

[0989] 4-((S)-2-((S)-2-(6-(2,5-dioxo-2,5-dihydro-1H-pyrrol-1-yl)hexanamido)-3- methylbutanamido)-5-ureidopentanamido)benzyl (4-(2-chloro-4-(3-((2-(2,6- dioxopiperidin-3-yl)-1-oxoisoindolin-5-yl)methyl)ureido)phenethylthio)-N-(N-(2- hydroxyethyl)-N-methyl)amidinyl)carbamate

[0990] Reaction Scheme:

[0991] Procedure:

[0992] Step A: Compound 22 (100 mg, 0.18 mmol) was dissolved in THF (20 mL), compound 1-10 (146.08 mg, 0.20 mmol) and TEA (0.075 mL, 0.54 mmol) were added, and the reaction system was reacted at 40 °C for 12 hours under nitrogen protection.

[0993] After the reaction was completed by LCMS monitoring, compound 51 (50 mg, yield 24.39%) was obtained by Prep-HPLC purification.

[0994] LCMS (ESI): 1168.6 [M+H] + .

[0995] 1H NMR (400 MHz, DMSO) δ 10.98 (s, 1H), 10.03 (s, 1H), 8.88 - 8.79 (m, 3H), 8.09 (d, J = 7.4 Hz, 1H), 7.80 (d, J = 8.5 Hz, 1H), 7.74 - 7.57 (m, 4H), 7.51 (s, 1H), 7.44 (d, J = 7.8 Hz, 1H), 7.36 (d, J = 8.5 Hz, 2H), 7.25 - 7.07 (m, 2H), 7.07 - 6.90 (m, 3H), 6.01 (s, 1H), 5.23 - 5.02 (m, 3H), 4.47 - 4.27 (m, 7H), 4.21 - 4.16 (m, 1H), 3.59 (m, 6H), 3.37 (t, J = 7.0 Hz, 2H), 3.08 - 2.86 (m, 6H), 2.82 (t, J = 6.7 Hz, 2H), 2.63 - 2.59 (m, 1H), 2.44 - 2.31 (m, 1H), 2.24 - 2.06 (m, 2H), 2.04 - 1.90 (m, 2H), 1.79 - 1.53 (m, 3H), 1.52 - 1.31 (m, 6H), 1.23 - 1.13 (m, 2H), 0.88 - 0.79 (m, 6H).

[0996] Example 52:

[0997] Preparation of (2S,3S,4S,5R,6S)-6-(4-(((2-(2-chloro-4-(3-((2-(2-(2-6-dioxopiperidin-3-yl)-1- oxoisoindolin-5-yl)methyl)ureido)phenoxy)ethyl)(prop-2-yn-1-yl)carbamoyl)oxy)methyl)-2-(4-(6-(2,5- dioxo-1H-pyrrol-1-yl)hexan-1-ylidene)-but-1-yn-1-yl)phenoxy)-3,4,5-trihydroxytetrahydro-2H-pyran-2- carboxylic acid.

[0998] Reaction Scheme:

[0999] Procedure:

[1000] Step A: Compound 52-1 (4.30 g, 17.34 mmol) and 52-2 (13.73 g, 34.68 mmol) were dissolved in acetonitrile (100 mL), silver oxide (20.20 g, 86.70 mmol) was added, and the reaction system was reacted at 30 degrees Celsius for 16 hours under nitrogen protection.

[1001] After the completion of the reaction, the reaction mixture was filtered, the filtrate was evaporated, and the crude product was purified by silica gel column to obtain compound 52-3 (8.70 g, yellow oily substance, 88.93% yield).

[1002] 1 H NMR (400 MHz, DMSO-d6) δ 9.93 - 9.82 (m, 1H), 8.33 (d, J = 2.0 Hz, 1H), 7.95 (dd, J = 8.5, 2.0 Hz, 1H), 7.30 (d, J = 8.6 Hz, 1H), 5.81 (d, J = 7.8 Hz, 1H), 5.49 (dd, J = 19.0, 9.4 Hz, 1H), 5.23 (dd, J = 9.6, 7.8 Hz, 1H), 5.12 (t, J = 9.7 Hz, 1H), 4.85 - 4.74 (m, 1H), 3.63 (d, J = 14.2 Hz, 3H), 2.06 - 2.00 (m, 9H).

[1003] Step B: Compound 52-3 (1.5 g, 2.66 mmol) and 52-4 (900 mg, 5.32 mmol) were dissolved in dry tetrahydrofuran (30 mL), triethylamine (13.47 g, 53.2 mmol), cuprous iodide (253 mg, 1.33 mmol) and Pd(PPh3)2Cl2 (187 mg, 0.266 mmol) were added, and the reaction system was reacted at 40 degrees Celsius for 2 hours under nitrogen protection.

[1004] After the completion of the reaction, the reaction mixture was filtered, the filtrate was evaporated, and the crude product was purified by silica gel column to obtain compound 52-3 (8.70 g, yellow oily substance, 88.93% yield).

[1005] LCMS (ESI): m / z [M+H] + 606.1.

[1006] Step C: Compound 52-5 (1.2 g, 1.98 mmol) was dissolved in methanol (30 mL), and sodium borohydride (83 mg, 2.18 mmol) was added under ice water bath, and the reaction system was reacted at 0 degrees Celsius for 1 hour.

[1007] After the completion of the reaction, the reaction mixture was filtered, the filtrate was evaporated, and the crude product was purified by silica gel column to obtain compound 52-3 (8.70 g, yellow oily substance, 88.93% yield).

[1008] LCMS (ESI): m / z [M+H] + 608.2.

[1009] Step D: Compound 52-6 (640 mg, 1.05 mmol) was dissolved in N,N- dimethylformamide (15 mL), N,N-diisopropylethylamine (203 mg, 1.58 mmol) and compound 52-7 (641 mg, 2.11 mmol) were added, the reaction system was reacted at room temperature for 16 hours.

[1010] After the reaction was completed by LCMS monitoring, the reaction solution was poured into saturated ammonium chloride solution, extracted with ethyl acetate, and the organic phase was washed with saturated ammonium chloride solution. The organic phase was dried over anhydrous sodium sulfate, filtered, and the filtrate was concentrated. The obtained crude product was purified by silica gel column to obtain compound 52-8 (770 mg, 94.60% yield).

[1011] LCMS (ESI): m / z [M+H] + 773.2

[1012] Step E: Compound 52-8 (210 mg, 0.28 mmol) was dissolved in dry N,N- dimethylformamide (12 mL), and then compound 13 (140 mg, 0.25 mmol) and N,N- diisopropylethylamine (97 mg, 0.75 mmol) were added quickly in turn. The reaction system was reacted at 40 degrees Celsius for 16 hours under nitrogen protection.

[1013] After the reaction was completed by LCMS monitoring, the reaction solution was poured into saturated ammonium chloride solution, extracted with ethyl acetate, and the organic phase was washed with saturated ammonium chloride solution. The organic phase was dried over anhydrous sodium sulfate, filtered, and the filtrate was concentrated. The obtained crude product was purified by silica gel column to obtain compound 52-9 (190 mg, yield 63.19%).

[1014] LCMS (ESI): m / z [M+H] + 1185.2.

[1015] Step F: Compound 52-9 (200 mg, 0.17 mmol) was dissolved in tetrahydrofuran (4 mL), hydrochloric acid solution (4 mL, 6 mol / L) was added, and the reaction system was reacted at 45 degrees Celsius for 5 hours.

[1016] After the reaction was completed by LCMS monitoring, the reaction solution was concentrated, and the crude product was purified by Prep-HPLC (ACN / 0.1% FA in water) to obtain compound 52-10 (90 mg, yield 56.08%).

[1017] LCMS (ESI): m / z [M+H] + 945.2.

[1018] Step G: Compound 52-10 (120 mg, 0.13 mmol) was dissolved in N,N- dimethylformamide (3 mL), compound 11 (59 mg, 0.19 mmol) and N,N- diisopropylethylamine (50 mg, 0.39 mmol) were added, and the reaction system was reacted at room temperature for 2 hours.

[1019] After the reaction was completed by LCMS monitoring, the reaction solution was concentrated, and the crude product was purified by Prep-HPLC to obtain compound 52 (51.11 mg, yield 34.58%).

[1020] 1 H NMR (400 MHz, DMSO-d6) δ 12.77 (s, 1H), 10.98 (s, 1H), 8.78 (s, 1H), 7.96 (t, J = 5.6 Hz, 1H), 7.68 (dd, J = 9.3, 4.9 Hz, 2H), 7.51 (s, 1H), 7.44 (d, J = 8.0 Hz, 1H), 7.31 (d, J = 31.9 Hz, 2H), 7.20 - 7.08 (m, 3H), 6.99 (s, 2H), 6.81 (t, J = 5.9 Hz, 1H), 5.64 - 5.05 (m, 4H), 4.97 (s, 2H), 4.50 - 4.26 (m, 4H), 4.05 (s, 2H), 3.90 (d, J = 9.6 Hz, 1H), 3.53 (d, J = 5.3 Hz, 5H), 3.45 - 3.39 (m, 7H), 3.31 - 3.15 (m, 5H), 2.97 - 2.76 (m, 3H), 2.60 (d, J = 17.6 Hz, 1H), 2.38 (dt, J = 13.3, 10.9 Hz, 1H), 2.03 (dt, J = 10.4, 6.2 Hz, 3H), 1.45 (td, J = 14.9, 7.5 Hz, 4H), 1.25 - 1.09 (m, 2H).

[1021] LCMS (ESI): m / z [M+H] + 1138.1.

[1022] Example 53:

[1023] Preparation of (2S,3S,4S,5R,6S)-6-(4-(2-(2-chloro-4-(3-((2-(2-2-dioxopiperidin-3-yl)-1- oxoisoindolin-5-yl)methyl)ureido)phenoxy)(prop-2-yn-1-yl)aminocarbonyl)methyl)-2-(3-(6-(2,5- dioxo-2,5-dihydro-1H-pyrrol-1-yl)hexanoylamino)prop-1-yn-1-yl)phenoxy)-3,4,5- trihydroxytetrahydro-2H-pyran-2-carboxylic acid.

[1024] Reaction Scheme:

[1025] Procedure:

[1026] Step A: Compound 53-1 (4.30 g, 17.34 mmol) and 53-2 (13.73 g, 34.68 mmol) were dissolved in acetonitrile (100 mL), silver oxide (20.20 g, 86.70 mmol) was added, and the reaction system was reacted at 30 degrees Celsius for 16 hours under nitrogen protection in the dark.

[1027] After the reaction was completed by TLC monitoring, the reaction solution was filtered, the filtrate was rotary evaporated, and the crude product was purified by silica gel column to obtain compound 53-3 (8.70 g, yellow oil, 88.93% yield).

[1028] 1 H NMR (400 MHz, DMSO-d6) δ 9.93 - 9.82 (m, 1H), 8.33 (d, J = 2.0 Hz, 1H), 7.95 (dd, J = 8.5, 2.0 Hz, 1H), 7.30 (d, J = 8.6 Hz, 1H), 5.81 (d, J = 7.8 Hz, 1H), 5.49 (dd, J = 19.0, 9.4 Hz, 1H), 5.23 (dd, J = 9.6, 7.8 Hz, 1H), 5.12 (t, J = 9.7 Hz, 1H), 4.85 - 4.74 (m, 1H), 3.63 (d, J = 14.2 Hz, 3H), 2.06 - 2.00 (m, 9H).

[1029] Step B: Compound 53-3 (1.5 g, 2.66 mmol) and 53-4 (824 mg, 5.32 mmol) were dissolved in dry tetrahydrofuran (30 mL), triethylamine (13.47 g, 53.2 mmol), cuprous iodide (253 mg, 1.33 mmol) and Pd(PPh3)2Cl2 (187 mg, 0.266 mmol) were added, and the reaction system was reacted at 40 degrees Celsius for 2 hours under nitrogen protection.

[1030] After the reaction was completed by TLC monitoring, the reaction solution was filtered with diatomite, the filtrate was concentrated, and the obtained crude product was purified by silica gel column to obtain compound 53-5 (1.4 g, yellow oil, 89.03% yield).

[1031] LCMS (ESI): m / z [M-Boc+H] + 492.1.

[1032] Step C: Compound 53-5 (1.46 g, 2.47 mmol) was dissolved in methanol (30 mL), sodium borohydride (103 mg, 2.71 mmol) was added under ice water bath, the reaction system was reacted at 0 degree Celsius for 1 hour.

[1033] After the reaction was completed by LCMS monitoring, the reaction solution was poured into saturated ammonium chloride solution for quenching, extracted with ethyl acetate, the organic phase was washed with saturated sodium chloride solution, dried over anhydrous sodium sulfate, filtered, and the filtrate was concentrated. The obtained crude product was purified by silica gel column to obtain compound 53-6 (840 mg, 57.34% yield).

[1034] LCMS (ESI): m / z [M-Boc+H] + 494.1.

[1035] Step D: Compound 53-6 (940 mg, 1.58 mmol) was dissolved in N,N- dimethylformamide (15 mL), N,N-diisopropylethylamine (310 mg, 2.37 mmol) and compound 53-7 (960 mg, 3.16 mmol) were added, and the reaction system was reacted at 25 degree Celsius for 16 hours.

[1036] After the reaction was completed by LCMS monitoring, the reaction solution was poured into saturated ammonium chloride solution for quenching, extracted with ethyl acetate, the organic phase was washed with saturated sodium chloride solution, dried over anhydrous sodium sulfate, filtered, and the filtrate was concentrated. The obtained crude product was purified by silica gel column to obtain compound 53-6 (840 mg, 57.34% yield).

[1037] LCMS (ESI): m / z [M-Boc+H] + 659.1.

[1038] Step E: Compound 53-8 (210 mg, 0.28 mmol) was dissolved in dry N,N- dimethylformamide (12 mL), compound 13 (140 mg, 0.25 mmol) and N,N- diisopropylethylamine (97 mg, 0.75 mmol) were added quickly in turn, and the reaction system was reacted at 40 degree Celsius for 16 hours under nitrogen protection.

[1039] After the reaction was completed by LCMS monitoring, the reaction solution was poured into saturated ammonium chloride solution for quenching, extracted with ethyl acetate, the organic phase was washed with saturated sodium chloride solution, dried over anhydrous sodium sulfate, filtered, and the filtrate was concentrated. The obtained crude product was purified by silica gel column to obtain compound 53-6 (840 mg, 57.34% yield).

[1040] LCMS (ESI): m / z [M+H] + 1171.2.

[1041] Step F: Compound 53-9 (210 mg, 0.18 mmol) was dissolved in tetrahydrofuran (4 mL), hydrochloric acid solution (4 mL, 6 mol / L) was added, and the reaction system was reacted at 45 degrees Celsius for 5 hours.

[1042] After the reaction was completed by LCMS monitoring, the reaction solution was concentrated, and the crude product was purified by Prep-HPLC (ACN / 0.1% FA in water) to obtain compound 53-10 (90 mg, 53.91% yield).

[1043] LCMS (ESI): m / z [M+H] + 931.1.

[1044] Step G: Compound 53-10 (90 mg, 0.097 mmol) was dissolved in N,N- dimethylformamide (3 mL), compound 53-11 (60 mg, 0.19 mmol) and N,N- diisopropylethylamine (38 mg, 0.29 mmol) were added, and the reaction system was reacted at room temperature for 2 hours.

[1045] After the reaction was completed by LCMS monitoring, the reaction solution was concentrated, and the crude product was purified by Prep-HPLC (ACN / 0.1% TFA in water) to obtain compound 53 (25.63 mg, 23.51% yield).

[1046] LCMS (ESI): m / z [M+H] + 1124.1.

[1047] 1H NMR (400 MHz, DMSO-d6) δ 12.79 (s, 1H), 10.97 (s, 1H), 8.77 (s, 1H), 8.28 (d, J = 5.0 Hz, 1H), 7.70 - 7.66 (m, 2H), 7.51 (s, 1H), 7.44 (d, J = 7.8 Hz, 1H), 7.38 (d, J = 10.7 Hz, 1H), 7.32 (d, J = 8.8 Hz, 1H), 7.15 - 7.12 (m, 3H), 6.98 (s, 2H), 6.80 (t, J = 6.0 Hz, 1H), 5.34 (s, 2H), 5.17 (d, J = 7.2 Hz, 1H), 5.10 (dd, J = 13.3, 5.1 Hz, 1H), 4.99 (s, 2H), 4.43 (t, J = 12.4 Hz, 3H), 4.31 (d, J = 17.3 Hz, 1H), 4.15 - 4.02 (m, 4H), 3.91 (d, J = 9.6 Hz, 1H), 3.53 - 3.52 (m, 5H), 3.42 - 3.41 (m, 4H), 3.33 (d, J = 3.4 Hz, 3H), 3.18 (t, J = 2.3 Hz, 1H), 2.95 - 2.82 (m, 3H), 2.60 (d, J = 16.8 Hz, 1H), 2.38 (dt, J = 16.4, 10.3 Hz, 1H), 2.09 (t, J = 7.4 Hz, 2H), 2.04 - 1.92 (m, 1H), 1.56 - 1.40 (m, 4H), 1.25 - 1.11 (m, 2H).

[1048] Example 54:

[1049] Preparation of (2S,3S,4S,5R,6S)-6-(4-((((2-(2-chloro-4-(3-((2-(2,6-dioxopiperidin-3-yl)-1- oxoisoindolin-5-yl)methyl)ureido)phenethoxy)ethyl)(prop-2-yn-1-yl)carbamoyl)oxy)methyl)- 2-((Z)-N'-((2-(6-(2,5-dioxo-2,5-dihydro-1H-pyrrol-1-yl)hexanamido)ethoxy)carbonyl)amino- carbamimidoyl)phenoxy)-3,4,5-trihydroxytetrahydro-2H-pyran-2-carboxylic acid

[1050] Reaction Scheme:

[1051] Procedure:

[1052] Step A: Compound 54-1 (5 g, 31.02 mmol) was dissolved in dichloromethane (200 mL), triethylamine (7.85 g, 77.55 mmol) was added, p-nitrophenyl chloroformate (9.44 g, 31.02 mmol) was added under ice bath, and stirred for 1 h.

[1053] After completion of the reaction as monitored by TLC, the reaction mixture was evaporated under reduced pressure and the residue was purified by silica gel column chromatography to afford compound 54-2 (7.2 g, yield 71 %)

[1054] LCMS (ESI): m / z [M+H-(t-Bu)] + 270.9.

[1055] Step B: Compound 54-3 (1.3 g, 8.84 mmol) was dissolved in ethanol (50 mL), acetyl chloride (50 mL) was added drop wise at -78 °C and stirred at room temperature for 72 h.

[1056] After completion of the reaction as monitored by TLC, to the reaction mixture was added diethyl ether (500 mL) and filtered to get compound 54-4 (1.6 g, yield 93 %).

[1057] LCMS (ESI): m / z [M+H] + 194.1.

[1058] Step C: Compound 54-4 (1.6 g, 8.28 mmol) was dissolved in ammonia in methanol solution (50 mL, 100 mmol, 2 M) and stirred at room temperature for 5 h.

[1059] After completion of the reaction as monitored by TLC, the reaction mixture was evaporated under reduced pressure, to which diethyl ether (50 mL) was added, stirred for 10 min and filtered to get compound 54-5 (1.2 g, yield 88 %).

[1060] LCMS (ESI): m / z [M+H] + 165.1.

[1061] Step D: Compound 54-5 (1.2 g, 7.31 mmol) was dissolved in DMF (10 mL), compound 54-2 (3.6 g, 10.96 mmol), triethylamine (2.8 g, 21.93 mmol) was added and stirred at room temperature for 16 h.

[1062] After completion of the reaction as monitored by TLC, it was diluted with water and extracted with ethyl acetate three times, combined organic layer was dried over anhydrous sodium sulfate and concentrated under reduced pressure, the residue was purified by Flash (0.1 % FA in water / ACN) to afford compound 54-6 (0.76 g, yield 30 %).

[1063] LCMS (ESI): m / z [M+H] + 352.1.

[1064] Step E: Compound 54-6 (0.76 g, 2.16 mmol) was dissolved in acetonitrile (10 mL), 1-bromo-1-deoxy-2,3,4-tri-O-acetyl-BETA-D-glucuronic acid methyl ester (1.72 g, 4.32 mmol), silver oxide (2.5 g, 10.8 mmol) were added, stirred at 30 °C for 16 hours.

[1065] After the completion of the reaction was monitored by TLC, the reaction was filtered, the filtrate was concentrated under reduced pressure, the residue was purified by silica gel column to get compound 54-7 (0.92 g, yield 64%).

[1066] LCMS (ESI): m / z [M+H] + 668.5.

[1067] Step F: Compound 54-7 (0.82 g, 1.23 mmol) was dissolved in methanol (10 mL), sodium borohydride (0.051 g, 1.35 mmol) was added under ice bath, stirred for 10 minutes.

[1068] After the completion of the reaction was monitored by TLC, the reaction was diluted with water, extracted with ethyl acetate three times, the organic phase was combined, dried over anhydrous sodium sulfate, concentrated under reduced pressure, the residue was purified by silica gel column to get compound 54-8 (0.54 g, yield 66%).

[1069] LCMS (ESI): m / z [M+H] + 670.4.

[1070] Step G: Compound 54-8 (0.54 g, 0.80 mmol) was dissolved in DMF (6 mL), N,N- diisopropylethylamine (155 mg, 1.2 mmol), 4-nitrophenyl carbonate (487 mg, 1.60 mmol) were added, stirred at room temperature for 1 hour.

[1071] After the completion of the reaction was monitored by TLC, the reaction was diluted with water, extracted with ethyl acetate three times, the organic phase was combined, dried over anhydrous sodium sulfate, concentrated under reduced pressure, the residue was purified by silica gel column to get compound 54-9 (0.31 g, yield 66%).

[1072] LCMS (ESI): m / z [M+H] + 835.3.

[1073] Step H: Compound 54-9 (0.25 g, 0.30 mmol) was dissolved in DMF (3 mL), compound 13 (138 mg, 0.25 mmol), N,N-diisopropylethylamine (97 mg, 0.75 mmol) were added, stirred at 45 °C for 16 h.

[1074] After completion of the reaction as monitored by TLC, the reaction mixture was diluted with water and extracted with ethylacetate (3 x 50 mL). The combined organic extracts were washed with water (2 x 50 mL), brine (1 x 50 mL), dried over Na2S04and evaporated under reduced pressure. The crude product was purified by silica gel column to afford compound 54-10 (0.21 g, 67% yield).

[1075] LCMS (ESI): m / z [M+H] + 1247.4.

[1076] Step I: Compound 54-10 (0.21 g, 0.17 mmol) was dissolved in tetrahydrofuran (3 mL), hydrochloric acid (3 mL, 6 M) was added, stirred at 45 °C for 5 h.

[1077] After completion of the reaction as monitored by TLC, the reaction mixture was evaporated under reduced pressure. The residue was purified by Flash (0.1% FA in water / ACN) to afford compound 54-11 (96 mg, 56% yield).

[1078] LCMS (ESI): m / z [M+H] + 1007.3.

[1079] Step J: Compound 54-11 (96 mg, 0.095 mmol) was dissolved in DMF (2 mL), 6-(maleimido)hexanoic acid succinimidyl ester (59 mg, 0.19 mmol), N,N- diisopropylethylamine (37 mg, 0.29 mmol) were added, stirred at room temperature for 1 h.

[1080] After completion of the reaction as monitored by TLC, the reaction mixture was filtered and the residue was purified by prep-HPLC to afford compound 54 (21 mg, 18% yield).

[1081] LCMS (ESI): m / z [M+H] + 1200.1.

[1082] 1H NMR (400 MHz, DMSO) δ 10.97 (s, 1H), 9.43 (brs, 1H), 8.83 (brs, 2H), 7.97 (t, J = 5.5 Hz, 1H), 7.90 (d, J = 2.0 Hz, 1H), 7.68 (d, J = 7.7 Hz, 2H), 7.57 - 7.50 (m, 2H), 7.44 (d, J = 8.6 Hz, 1H), 7.31 (d, J = 8.5 Hz, 1H), 7.11 (brs, 2H), 6.98 (s, 2H), 5.75 (s, 1H), 5.32 (d, J = 3.7 Hz, 1H), 5.21 - 5.04 (m, 4H), 4.48 - 4.28 (m, 4H), 4.08 - 3.89 (m, 5H), 3.58 - 3.51 (m, 4H), 3.43 - 3.36 (m, 6H), 3.28 - 3.26 (m, 1H), 3.18 (s, 1H), 2.96 - 2.86 (m, 1H), 2.80 (brs, 2H), 2.63 - 2.55 (m, 1H), 2.43 - 2.35 (m, 1H), 2.09 - 1.97 (m, 3H), 1.51 - 1.43 (m, 4H), 1.20 - 1.11 (m, 2H).

[1083] Example 55:

[1084] Preparation of compound 55

[1085] Reaction Scheme:

[1086] Procedure:

[1087] Step A: Compound 55-1 (200 mg, 0.673 mmol) was dissolved in THF (15 mL), compound 55-2 (99 mg, 0.804 mmol), EEDQ (250 mg, 1.011 mmol) were added, the reaction system was protected under nitrogen, and reacted at room temperature for 24 hours.

[1088] After the reaction was monitored by LCMS, the reaction solution was concentrated, and the crude product was purified by Flash (ACN / 0.1% FA in water) to obtain compound 55-3 (80 mg, 36.94% yield).

[1089] LCMS (ESI): m / z [M+H] + 403.1.

[1090] Step B: Compound 55-3 (100 mg, 0.249 mmol) was dissolved in DMF (10 mL), added compound 55-4 (378 mg, 1.243 mmol), Lutidine (186 mg, 1.736 mmol), the reaction system was protected by nitrogen, and reacted at room temperature for 16 hours.

[1091] After the reaction was completed by LCMS monitoring, the reaction solution was concentrated, and the crude product was purified by flash (ACN / 0.1% FA in water) to obtain compound 55-4 (45 mg, 31.91% yield).

[1092] LCMS (ESI): m / z [M+Na] + 590.28.

[1093] Step C: Compound 22 (20 mg, 0.04 mmol) was dissolved in DMF (2 mL), added compound 55-4 (20 mg, 0.04 mmol), DIEA (18 mg, 0.14 mmol), the reaction system was protected by nitrogen, and reacted at 40 degrees Celsius for 16 hours.

[1094] After the reaction was completed by LCMS monitoring, the reaction solution was purified by Prep-HPLC to obtain compound 55 (20.11 mg, 57.41% yield).

[1095] LCMS (ESI): m / z [M+H] + 998.5.

[1096] 1H NMR (400 MHz, DMSO-d6) δ 10.98 (s, 1H), 10.69 (s, 1H), 9.92 (s, 1H), 9.00 - 8.52 (m, 3H), 8.43 (d, J = 7.3 Hz, 1H), 8.19 (d, J = 7.2 Hz, 1H), 7.73 - 7.58 (m, 4H), 7.51 (s, 1H), 7.44 (d, J = 8.2 Hz, 1H), 7.35 (d, J = 8.4 Hz, 2H), 7.21 - 7.14 (m, 2H), 7.09 (s, 2H), 6.97 - 6.84 (m, 1H), 5.26 - 5.01 (m, 3H), 4.49 - 4.26 (m, 6H), 4.18 - 4.00 (m, 2H), 3.61 - 3.55 (m, 6H), 3.02 (s, 3H), 2.97 - 2.86 (m, 1H), 2.85 - 2.78 (m, 2H), 2.64 - 2.56 (m, 1H), 2.43 - 2.31 (m, 1H), 2.05 - 1.93 (m, 1H), 1.31 (d, J = 7.1 Hz, 3H), 1.21 (d, J = 7.1 Hz, 3H).

[1097] Example 56:

[1098] Preparation of compound 56.

[1099] Reaction Scheme:

[1100] Procedure:

[1101] Step A: Compound 27 (40 mg, 0.068 mmol) was added to a 25 mL single neck flask, followed by THF (8 mL), compound 1-10 (60.2 mg, 0.082 mmol) and triethylamine (0.028 mL, 0.2 mmol). The reaction system was reacted at 40 °C for 12 hours under nitrogen protection.

[1102] After the reaction was completed by LCMS monitoring, compound 56 (15 mg, 18.55% yield) was obtained by Prep-HPLC purification.

[1103] LCMS (ESI): m / z [M+H] + 1184.5.

[1104] 1H NMR (400 MHz, DMSO) δ 10.98 (s, 1H), 10.02 (s, 1H), 9.16 (s, 1H), 8.87 (s, 1H), 8.09 (d, J = 7.4 Hz, 1H), 7.80 (d, J = 8.6 Hz, 1H), 7.69 (d, J = 7.8 Hz, 1H), 7.67 - 7.59 (m, 3H), 7.51 (s, 1H), 7.44 (d, J = 8.0 Hz, 1H), 7.35 (d, J = 8.5 Hz, 2H), 7.20 - 7.10 (m, 2H), 7.00 (s, 2H), 6.92 (t, J = 5.8 Hz, 1H), 6.02 (s, 1H), 5.23 - 5.05 (m, 3H), 4.47 - 4.27 (m, 5H), 4.21 - 4.14 (m, 1H), 3.92 (d, J = 4.5 Hz, 2H), 3.66 (s, 3H), 3.57 (d, J = 5.0 Hz, 4H), 3.52 (d, J = 7.1 Hz, 3H), 3.36 (t, J = 7.0 Hz, 2H), 3.06 - 2.86 (m, 3H), 2.76 (t, J = 6.9 Hz, 2H), 2.63 (t, J = 18.3 Hz, 1H), 2.44 - 2.30 (m, 1H), 2.24 - 2.06 (m, 2H), 2.05 - 1.90 (m, 2H), 1.75 - 1.65 (m, 1H), 1.65 - 1.55 (m, 1H), 1.54 - 1.32 (m, 6H), 1.25 - 1.14 (m, 2H), 0.89 - 0.79 (m, 6H).

[1105] Example 57:

[1106] Preparation of (2S)-2-amino-N1-(2-(2-chloro-4-(3-((2-(2-2-dioxopiperidin-3-yl)-1- oxoisoindolin-5-yl)methyl)ureido)phenethyl)-N1-(propyl-2-yne-1-yl)pentanediamide.

[1107] Reaction Scheme:

[1108] Procedure:

[1109] Step A: Compound 13 (50 mg, 0.091 mmol) was dissolved in dry DMF (3 mL), HATU (51.90 mg, 0.14 mmol), compound 57-1 (26.79 mg, 0.11 mmol) and DIEA (47.04 mg, 0.36 mmol) were added at room temperature and the reaction was stirred at room temperature for 1 h.

[1110] After LCMS monitoring the reaction completion, the reaction solution was purified by reverse phase flash to obtain compound 57-2 (40 mg 62.34% yield).

[1111] LCMS (ESI): m / z [M+H] + 780.2.

[1112] Step B: Compound 57-2 (30 mg, 0.051 mmol) was dissolved in dichloromethane (5 mL), then trifluoroacetic acid (1 mL) was added, the reaction system was protected by nitrogen, and the reaction was carried out at room temperature for 1 hour.

[1113] After LCMS monitoring the reaction completion, the reaction solution was concentrated, and the crude product was purified by Prep-HPLC (to obtain compound 57 (9 mg, 30.21% yield).

[1114] LCMS (ESI): m / z [M+H] + 680.0.

[1115] 1 H NMR (400 MHz, DMSO-d6) δ 10.98 (s, 1H), 9.06 - 8.87 (m, 1H), 8.21 (s, 1H), 7.78 - 7.63 (m, 2H), 7.51 (s, 1H), 7.44 (d, J = 7.8 Hz, 1H), 7.40 - 7.30 (m, 1H), 7.26 - 7.14 (m, 2H), 7.08 - 6.94 (m, 1H), 6.90 - 6.77 (m, 1H), 5.10 (dd, J = 13.3, 5.1 Hz, 1H), 4.55 - 4.03 (m, 6H), 4.01 - 3.87 (m, 1H), 3.80 - 3.70 (m, 1H), 3.66 - 3.50 (m, 5H), 3.48 - 3.32 (m, 1H), 3.17 (t, J = 2.3 Hz, 1H), 3.00 - 2.76 (m, 3H), 2.70 - 2.57 (m, 1H), 2.45 - 2.09 (m, 3H), 2.05 - 1.94 (m, 1H), 1.92 - 1.74 (m, 1H), 1.65 - 1.52 (m, 1H).

[1116] Example 58:

[1117] Preparation of 4-((S)-2-((S)-2-(6-(2,5-dioxo-2,5-dihydro-1H-pyrrol-1-yl)hexanoyl-amido)-3-methylbutanoylamido)-5-ureidopentanoyl)benzyl ((2S)-5-amino-1-((2-(2-chloro-4-(3-((2-(2-dioxopiperidin-3-yl)-1-oxoisoindolin-5-yl)methyl)ureido)phenoxy)ethyl)(prop-2-yn-1-yl)amino)-1,5-dioxopentan-2-yl)carbamate.

[1118] Reaction Scheme:

[1119] Operation Steps:

[1120] Step A: Compound 57 (80 mg, 0.118 mmol) was added to a 25 mL single-neck flask, followed by the addition of DMF (2 mL), compound 1-10 (130 mg, 0.176 mmol), DIEA (45 mg, 0.348) and HOPO (12 mg, 0.176 mmol). The reaction system was reacted at room temperature for 1 hour under nitrogen protection.

[1121] After the reaction was completed by LCMS monitoring, the reaction solution was purified by Prep-HPLC to obtain compound 58 (20.49 mg, 13.62% yield).

[1122] LCMS (ESI): m / z [M+H] + calcd for C 62 H 77 ClN 13 O 15 : 1278.53, mass found: 1279.1

[1123] 1H NMR (400 MHz, DMSO-d6) δ 10.98 (s, 1H), 9.98 (s, 1H), 8.79 (s, 1H), 8.08 (d, J = 7.3 Hz, 1H), 7.80 (d, J = 8.6 Hz, 1H), 7.72 - 7.63 (m, 2H), 7.58 (d, J = 8.1 Hz, 2H), 7.53 - 7.40 (m, 3H), 7.29-7.27 (m, 3H), 7.24 - 7.18 (m, 1H), 7.17 - 7.09 (m, 1H), 7.00 (s, 2H), 6.88 - 6.70 (m, 2H), 5.99 (s, 1H), 5.10 (dd, J = 13.3, 5.0 Hz, 1H), 4.98 - 4.87 (m, 2H), 4.45 - 4.26 (m, 7H), 4.25 - 4.13 (m, 2H), 4.00 (d, J = 18.0 Hz, 1H), 3.69 (s, 2H), 3.64 - 3.47 (m, 5H), 3.37 (t, J = 7.0 Hz, 2H), 3.13 (s, 1H), 3.08 - 2.71 (m, 6H), 2.64 - 2.56 (m, 1H), 2.43 - 2.30 (m, 1H), 2.23 - 2.07 (m, 4H), 2.03 - 1.92 (m, 2H), 1.87 - 1.75 (m, 1H), 1.72 - 1.56 (m, 3H), 1.53 - 1.32 (m, 6H), 1.24 - 1.14 (m, 2H), 0.86 - 0.81 (m, 6H).

[1124] Example 59:

[1125] Preparation of ((R)-1-(methylamino)propan-2-yl 2-(2-chloro-4-(3-((2-(2,6-dioxopiperidin-3-yl)-1- oxoisoindolin-5-yl)methyl)ureido)phenyl)acetate

[1126] Reaction Scheme:

[1127] Procedure:

[1128] Step A: Compound 59-1 (1 g, 11.24 mmol) was dissolved in dry THF (20 mL), then Boc2O (4.90 g, 22.47 mmol) and TEA (3.40 g, 33.71 mmol) were added. The reaction was stirred at room temperature under nitrogen overnight.

[1129] After completion of the reaction, the reaction mixture was diluted with water (100 mL) and extracted with ethyl acetate (2 x 100 mL). The organic layer was washed with water (2 x 100 mL), dried over anhydrous sodium sulfate, filtered and concentrated under reduced pressure to obtain the crude product. The crude product was purified by column chromatography (PE:EA = 20:1-1:1) to obtain compound 59-2 (1.9 g, colorless oily liquid, 90% yield).

[1130] TLC: Rf = 0.3 (PE:EA = 1:1)

[1131] Step B: To a stirred solution of compound 59-2 (1 g, 5.29 mmol) in dry dichloromethane (15 mL), compound 59-3 (0.95 g, 4.41 mmol), EDCI (1.11 g, 5.81 mmol) and DMAP (7 mg, 0.06 mmol) were added and the reaction mixture was stirred at room temperature for 2 h under nitrogen atmosphere.

[1132] After completion of the reaction, the reaction mixture was diluted with water (100 mL) and extracted with ethyl acetate (2 x 100 mL). The organic layer was washed with water (2 x 100 mL), dried over anhydrous sodium sulfate, filtered and concentrated under reduced pressure to obtain the crude product. The crude product was purified by column chromatography (PE:EA = 20:1-1:1) to obtain compound 59-2 (1.9 g, colorless oily liquid, 90% yield).

[1133] LCMS (ESI): m / z [M-H] - 385.5.

[1134] Step C: To a stirred solution of compound 59-4 (1.3 g, 3.37 mmol) in dry DMF (20 mL), 4,4'-dipyridyl (0.52 g, 3.37 mmol) was added followed by compound 59-5 (1.19 g, 3.34 mmol) and the reaction mixture was stirred at room temperature for 5 min under nitrogen atmosphere till blue color fades.

[1135] After completion of the reaction, the reaction mixture was diluted with water (100 mL) and extracted with ethyl acetate (2 x 100 mL). The organic layer was washed with water (2 x 100 mL), dried over anhydrous sodium sulfate, filtered and concentrated under reduced pressure to obtain the crude product. The crude product was purified by column chromatography (PE:EA = 20:1-1:1) to obtain compound 59-2 (1.9 g, colorless oily liquid, 90% yield).

[1136] LCMS (ESI): m / z [M+H] + 357.2.

[1137] Step D: Compound 59-5 (1.19 g, 3.34 mmol) was resolved by chiral resolution (column: mobile phase: MEOH (+0.1% 7.0 mol / l Ammonia in MEOH), flow rate: 140 mL / min) to obtain compound compound 59-5-P1 (0.47 g, retention time 2.832 min) and compound 59-5-P2 (0.45 g, retention time 2.302 min).

[1138] Step E: Compound 59-5-P1 (200 mg, 0.56 mmol) was dissolved in dry THF (5 mL), and diphosgene (0.16 mL, 1.12 mmol) was added dropwise slowly. The reaction was stirred at room temperature for 1 h under nitrogen. Then, 3-(5-(aminomethyl)-l-oxoisoindolin-2-yl)piperidine-2,6-dione (191 mg, 0.62 mmol) and triethylamine (0.39 mL, 2.8 mmol) in DMF (5 mL) were added dropwise slowly. The reaction was stirred at room temperature for 1 h under nitrogen.

[1139] After the reaction was completed by LCMS monitoring, compound 59-6-P1 (220 mg, 60% yield) was obtained by flash purification.

[1140] LCMS (ESI): m / z [M-Boc+H] + 556.2.

[1141] Step F: Compound 59-6-P1 (280 mg, 0.43 mmol) was dissolved in DCM (8 mL), and then TFA (4 mL) was added. The reaction was stirred at room temperature for 1 h under nitrogen.

[1142] After the reaction was completed by LCMS monitoring, the reaction was concentrated, and the crude product was purified by Prep-HPLC to give compound 59 (230 mg, 97% yield).

[1143] LCMS (ESI): m / z [M+H] + 556.0.

[1144] 1 H NMR (400 MHz, DMSO-d6) δ 10.97 (br s, 1H), 9.07 (d, J = 8.0 Hz, 1H), 8.25 (d, J = 3.6 Hz, 1H), 7.75 - 7.61 (m, 2H), 7.51 (s, 1H), 7.44 (d, J = 7.9 Hz, 1H), 7.29 - 7.18 (m, 2H), 7.15 - 7.02 (m, 1H), 5.11 (dd, J = 13.3, 5.1 Hz, 1H), 5.05 - 4.93 (m, 1H), 4.47 - 4.29 (m, 4H), 3.75 - 3.67 (m, 2H), 2.97 - 2.86 (m, 3H), 2.64 - 2.56 (m, 1H), 2.45 (s, 3H), 2.43 - 2.31 (m, 1H), 2.04 - 1.95 (m, 1H), 1.19 (d, J = 6.4 Hz, 3H).

[1145] Example 60:

[1146] (R)-1-(((4-(S)-2-((S)-2-(6-(2,5-dioxo-2,5-dihydro-1H-pyrrol-1-yl)hexanoylamido)-3- methylbutanamido)-5-ureidopentanamido)benzyl)oxy)carbonyl)(methyl)amino)propan-2-yl- 2-(2-chloro-4-(3-(2-(2,6-dioxopiperidin-3-yl)-1-oxoisoindolin-5-yl)methyl)ureido)phenyl) acetate

[1147] Reaction Scheme:

[1148] Procedure:

[1149] Step A: Compound 59 (50 mg, 0.09 mmol) was dissolved in THF (4 mL), then compound 1-10 (70 mg, 0.10 mmol), HOBt (6 mg, 0.05 mmol) and 2,6-Lutidine (19 mg, 0.18 mmol) were added, the reaction system was protected by nitrogen, and reacted at 40 °C overnight.

[1150] After the reaction was completed by LCMS monitoring, the reaction solution was purified by Prep-HPLC to obtain compound 60 (60 mg, 58% yield).

[1151] LCMS (ESI): m / z [M+H] + 1154.1.

[1152] 1H NMR (400 MHz, DMSO-d6) δ 10.98 (s, 1H), 9.99 (s, 1H), 8.90 (s, 1H), 8.08 (d, J = 7.4 Hz, 1H), 7.80 (d, J = 8.7 Hz, 1H), 7.74 - 7.64 (m, 2H), 7.59 (d, J = 7.9 Hz, 2H), 7.51 (s, 1H), 7.44 (d, J = 7.9 Hz, 1H), 7.28 (d, J = 8.5 Hz, 2H), 7.19 (s, 2H), 7.00 (s, 2H), 6.90 (t, J = 5.6 Hz, 1H), 5.97 (t, J = 5.6 Hz, 1H), 5.41 (s, 2H), 5.10 (dd, J = 13.3, 5.0 Hz, 1H), 5.06 - 4.95 (m, 3H), 4.48 - 4.26 (m, 5H), 4.22 - 4.15 (m, 1H), 3.69 - 3.57 (m, 2H), 3.39 - 3.34 (m, 3H), 3.07 - 2.85 (m, 3H), 2.79 (d, J = 10.9 Hz, 3H), 2.68 - 2.57 (m, 1H), 2.44 - 2.31 (m, 1H), 2.23 - 2.06 (m, 2H), 2.03 - 1.93 (m, 2H), 1.76 - 1.64 (m, 1H), 1.63 - 1.54 (m, 1H), 1.52 - 1.32 (m, 6H), 1.27 - 1.08 (m, 6H), 0.88 - 0.79 (m, 6H).

[1153] Example 61:

[1154] Preparation of (S)-1-(methylamino)propan-2-yl 2-(2-chloro-4-(3-((2-(2,6-dioxopiperidin-3-yl)-1- oxoisoindolin-5-yl)methyl)ureido)phenyl)acetate.

[1155] Reaction Scheme:

[1156] Procedure:

[1157] Step A: Compound 61-1 (100 mg, 1.12 mmol) was dissolved in dry THF (5 mL), then Boc20 (490 mg, 2.25 mmol) and TEA (340 mg, 3.37 mmol) were added. The reaction was stirred at room temperature under nitrogen overnight.

[1158] After the reaction was completed by LCMS monitoring, the solvent was removed by rotary evaporation under reduced pressure, and the resulting crude product was purified by normal phase column chromatography (PE:EA = 20:1-1:1) to obtain compound 61-2 (200 mg, colorless oily liquid, 94% yield).

[1159] TLC: Rf = 0.3 (PE:EA = 1:1)

[1160] Step B: Compound 61-2 (200 mg, 1.06 mmol) was dissolved in dry DCM (5 mL), then compound 59-3 (0.19 g, 0.88 mmol), EDCI (0.22 g, 1.16 mmol) and DMAP (2 mg, 0.02 mmol) were added, the reaction system was stirred at room temperature for 2 hours under nitrogen protection.

[1161] After the reaction was completed by LCMS monitoring, the solvent was rotary evaporated under reduced pressure, and the obtained crude product was purified by reverse phase Flash (ACN / 0.1% FA in water) to obtain compound 61-3 (200 mg, brown oily liquid, 49% yield).

[1162] LCMS (ESI): m / z [M-H] - 385.1.

[1163] Step C: Tetrahydroxydiboron (47 mg, 0.52 mmol) was dissolved in dry DMF (2 mL), 4,4'-dipyridine (20 mg, 0.13 mmol) and compound 61-3 (50 mg, 0.13 mmol) were added quickly in turn, the reaction system was reacted at room temperature for 5 minutes under nitrogen protection until the blue color faded.

[1164] After the reaction was completed by LCMS monitoring, a small amount of methanol was added, filtered, and the filtrate was purified by Flash (ACN / 0.1% FA in water) to obtain compound 59-5-P2 (30 mg, brown oily liquid, 65% yield).

[1165] LCMS (ESI): m / z [M+H] + 357.1.

[1166] Step D: Compound 59-5-P2 (200 mg, 0.56 mmol) was dissolved in dry THF (5 mL), and diphosgene (0.14 mL, 1.12 mmol) was slowly added dropwise, the reaction system was reacted at room temperature for 1 hour under nitrogen protection, then the solvent was rotary evaporated under reduced pressure at low temperature, and then dissolved in dry DMF (2 mL), and then a solution of compound 3-(5-(aminomethyl)-1-oxoisoindolin-2-yl)piperidine-2,6-dione (191 mg, 0.62 mmol) and triethylamine (0.39 mL, 2.8 mmol) in DMF (5 mL) was slowly added dropwise, the reaction system was reacted at room temperature for 1 hour under nitrogen protection, and then LCMS monitoring was completed, and then compound 61-4 (240 mg, 65% yield) was obtained by Flash purification.

[1167] LCMS (ESI): m / z [M-Boc+H] + 556.2

[1168] Step E: Compound 61-4 (300 mg, 0.46 mmol) was dissolved in DCM (8 mL), then TFA (4 mL) was added, the reaction system was protected under nitrogen, and reacted at room temperature for 1 hour.

[1169] After the reaction was monitored by LCMS, the reaction solution was concentrated, and the crude product was purified by Prep-HPLC (THF / 0.1% TFA in water) to obtain compound 61 (250 mg, 98% yield).

[1170] LCMS (ESI): m / z [M+H] + 556.0.

[1171] 1 H NMR (400 MHz, DMSO-d6) δ 10.98 (brs, 1H), 9.18 - 8.91 (m, 1H), 8.35 - 8.14 (m, 1H), 7.78 - 7.60 (m, 2H), 7.52 (s, 1H), 7.44 (d, J = 7.9 Hz, 1H), 7.26 - 7.18 (m, 2H), 7.16 - 6.96 (m, 1H), 5.11 (dd, J = 13.3, 5.1 Hz, 1H), 5.03 - 4.89 (m, 1H), 4.39 (dt, J = 46.8, 17.7 Hz, 4H), 3.76 - 3.64 (m, 2H), 2.97 - 2.78 (m, 3H), 2.69 - 2.56 (m, 1H), 2.43 - 2.32 (m, 4H), 2.04 - 1.95 (m, 1H), 1.18 (d, J = 6.3 Hz, 3H).

[1172] Example 62:

[1173] (S)-1-(((4-(S)-2-((S)-2-(6-(2,5-dioxo-2,5-dihydro-1H-pyrrol-1-yl)hexanamido)-3- methylbutanamido)-5-ureidopentanamido)benzyl)oxy)carbonyl)(methyl)amino)propan- 2-yl 2-(2-chloro-4-(3-(2-(2,6-dioxopiperidin-3-yl)-1-oxoisoindolin-5-yl)methyl)ureido) phenyl)acetate

[1174] Reaction Scheme:

[1175] Procedure:

[1176] Step A: Compound 61 (50 mg, 0.09 mmol) was dissolved in THF (4 mL), then 1-10 (70 mg, 0.10 mmol), HOBt (6 mg, 0.05 mmol) and 2,6-Lutidine (19 mg, 0.18 mmol) were added, the reaction system was reacted at 40 °C overnight under nitrogen protection.

[1177] After the reaction was completed by LCMS monitoring, the reaction solution was purified by Prep-HPLC to obtain compound 62 (60 mg, 58% yield).

[1178] LCMS (ESI): m / z [M+H] + 1154.1.

[1179] 1 H NMR (400 MHz, DMSO-d6) δ 10.98 (s, 1H), 9.99 (s, 1H), 8.99 (s, 1H), 8.09 (d, J = 7.4 Hz, 1H), 7.80 (d, J = 8.5 Hz, 1H), 7.69 (d, J = 7.4 Hz, 2H), 7.59 (d, J = 7.8 Hz, 2H), 7.51 (s, 1H), 7.44 (d, J = 7.9 Hz, 1H), 7.28 (d, J = 8.5 Hz, 2H), 7.19 (s, 2H), 7.00 (s, 3H), 5.98 (t, J = 5.7 Hz, 1H), 5.42 (s, 2H), 5.10 (dd, J = 13.4, 5.1 Hz, 1H), 5.05 - 4.93 (m, 3H), 4.51 - 4.26 (m, 5H), 4.19 (dd, J = 8.4, 7.0 Hz, 1H), 3.73 - 3.52 (m, 2H), 3.37 (d, J = 7.0 Hz, 3H), 3.08 - 2.85 (m, 3H), 2.79 (d, J = 10.7 Hz, 3H), 2.67 - 2.55 (m, 2H), 2.43 - 2.32 (m, 1H), 2.22 - 2.06 (m, 2H), 2.04 - 1.90 (m, 2H), 1.74 - 1.64 (m, 1H), 1.62 - 1.54 (m, 1H), 1.52 - 1.33 (m, 6H), 1.27 - 1.08 (m, 6H), 0.88 - 0.79 (m, 6H).

[1180] Example 63:

[1181] Preparation of (2S, 3S, 4S, 5R, 6S)-6-(4-(2-(2-chloro-4-(3-((2-(2-dioxopiperidin-3-yl)-1- oxoisoindol-5-yl)methyl)phenoxy)ethyl)(prop-2-yn-1-yl)aminocarbonyl)methyl)-2-((N-(2-(6-(2-(2-5- dioxo-1H-pyrrol-1-yl)hexanoyl)ethyl)sulfonyl)aminocarbonyl)phenoxy)-3, 4, 5-trihydroxytetrahydro-2H- pyran-2-carboxylic acid.

[1182] Reaction Scheme:

[1183] Procedure:

[1184] Step A: Compound 63-1 (6.0 g, 37.5 mmol) was dissolved in 1,4-dioxane (30 mL), then sulfonamide (7.2 g, 75.0 mmol) was added, and the reaction system was reacted at 90 degrees Celsius for 5 hours.

[1185] After the reaction was completed by LCMS monitoring, 1M hydrochloric acid was added to the reaction solution to adjust Ph to 3, and ethyl acetate was extracted twice. The combined organic phase was dried over anhydrous sodium sulfate, filtered, and the filtrate was concentrated under reduced pressure to obtain the crude compound 63-2 (3.7 g, 41.27% yield).

[1186] LCMS (ESI): m / z [M+Na] + 262.0.

[1187] Step B: Compound 63-3 (5.0 g, 30.12 mmol) was dissolved in tert-butyl alcohol (50 mL), then DCC (9.3 g, 45.18 mmol) and DMAP (367 mg, 3.01 mmol) were added, and the reaction system was reacted at 85 degrees Celsius for 16 hours.

[1188] After the reaction was completed by LCMS monitoring, the reaction solution was poured into ice water to quench, extracted with ethyl acetate, and the organic phase was washed with saturated sodium chloride solution. After the organic phase was dried over anhydrous sodium sulfate, it was filtered and the filtrate was concentrated. The obtained crude product was purified by silica gel column to obtain compound 63-4 (4.62 g, 69.09% yield).

[1189] LCMS (ESI): m / z [M+H] + 223.1.

[1190] Step C: Compound 63-4 (3 g, 13.51 mmol) and (2S,3R,4S,5S,6S)-2-bromo-6- (methoxycarbonyl)tetrahydro-2H-pyran-3,4,5-triyl triacetate (10.73 g, 27.03 mmol) were dissolved in acetonitrile (75 mL), silver oxide (15.65 g, 67.55 mmol) was added and the reaction mixture was stirred at 30 °C for 16 h under nitrogen.

[1191] After completion of the reaction as monitored by TLC, the reaction mixture was filtered, the filtrate was evaporated and the crude compound was purified by silica gel column to afford compound 63-5 (6.98 g, 96.04 % yield).

[1192] LCMS (ESI): m / z [M+NH4] + 556.2.

[1193] Step D: Compound 63-5 (6.98 g, 12.97 mmol) was dissolved in dichloromethane (70 mL), trifluoroacetic acid (35 mL) was added and the reaction mixture was stirred at 25 °C for 2 h.

[1194] After completion of the reaction as monitored by LCMS, the reaction mixture was concentrated under reduced pressure, most of the trifluoroacetic acid was removed and the residue was added to ice cold water. The organic layer was extracted with ethyl acetate (3x), the organic layers were combined and dried over anhydrous sodium sulfate, filtered and the filtrate was concentrated to obtain the crude compound 63-6 (5 g, 79.98 % yield).

[1195] LCMS (ESI): m / z [M-H] - 481.0.

[1196] Step E: Compound 63-6 (5.0 g, 10.37 mmol) was dissolved in dichloromethane (50 mL), EDCI (20.74 g, 45.18 mmol), DMAP (2.53 g, 20.74 mmol) and compound 63-2 (3.72 g, 15.56 mmol) were added and the reaction mixture was stirred at 25 °C for 16 h.

[1197] After completion of the reaction as monitored by LCMS, the reaction mixture was poured into ice cold water and quenched, extracted with dichloromethane and the organic layer was washed with saturated sodium chloride solution. The organic layer was dried over anhydrous sodium sulfate, filtered and the filtrate was concentrated. The crude compound was purified by silica gel column to afford compound 63-7 (1 g, 13.71 % yield).

[1198] LCMS (ESI): m / z [M-H] - 702.3.

[1199] Step F: Compound 63-7 (1.4 g, 1.99 mmol) was dissolved in methanol (30 mL), sodium borohydride (90 mg, 2.39 mmol) was added under ice water bath, the reaction system was reacted at 0 degree Celsius for 1 hour.

[1200] After the reaction was completed by LCMS monitoring, the reaction solution was poured into saturated ammonium chloride solution to quench, extracted with ethyl acetate, the organic phase was washed with saturated sodium chloride solution, dried over anhydrous sodium sulfate, filtered, and the filtrate was concentrated. The obtained crude product was purified by silica gel column to obtain compound 63-8 (1.0 g, 71.23% yield).

[1201] LCMS (ESI): m / z [M-Boc+H] + 606.1.

[1202] Step G: Compound 63-8 (1.0 g, 1.42 mmol) was dissolved in N,N-dimethylformamide (15 mL), N,N-diisopropylethylamine (274 mg, 2.13 mmol) and compound bis(p-nitrophenyl) carbonate (863 mg, 2.84 mmol) were added, and the reaction system was reacted at 25 degrees Celsius for 16 hours.

[1203] After the reaction was completed by LCMS monitoring, the reaction solution was poured into saturated ammonium chloride solution to quench, extracted with ethyl acetate, the organic phase was washed with saturated sodium chloride solution, dried over anhydrous sodium sulfate, filtered, and the filtrate was concentrated. The obtained crude product was purified by silica gel column to obtain compound 63-8 (1.0 g, 71.23% yield).

[1204] LCMS (ESI): m / z [M-Boc+H] + 771.0.

[1205] Step H: Compound 63-9 (340 mg, 0.40 mmol) was dissolved in dry N,N-dimethylformamide (8 mL), and then compound 13 (200 mg, 0.36 mmol) and N,N-diisopropylethylamine (140 mg, 1.08 mmol) were added quickly in turn. The reaction system was reacted at 40 degrees Celsius for 16 hours under nitrogen protection.

[1206] After the reaction was completed by LCMS monitoring, the reaction solution was poured into saturated ammonium chloride solution to quench, extracted with ethyl acetate, the organic phase was washed with saturated sodium chloride solution, dried over anhydrous sodium sulfate, filtered, and the filtrate was concentrated. The obtained crude product was purified by silica gel column to obtain compound 63-8 (1.0 g, 71.23% yield).

[1207] LCMS (ESI): m / z [M-Boc+H] + 1183.1.

[1208] Step I: Compound 63-10 (120 mg, 0.094 mmol) was dissolved in tetrahydrofuran (4 mL), hydrochloric acid solution (4 mL, 6 mol / L) was added, and the reaction system was reacted at 45 degrees Celsius for 5 hours.

[1209] After the reaction was completed by LCMS monitoring, the reaction solution was concentrated, and the crude product was purified by Prep-HPLC (ACN / 0.1% FA in water) to obtain compound 63-11 (90 mg, 91.89% yield).

[1210] LCMS (ESI): m / z [M+H] + 1043.0.

[1211] Step J: Compound 63-11 (90 mg, 0.086 mmol) was dissolved in N,N-dimethylformamide (3 mL), compound 2,5-dioxopyrrolidin-1-yl 6-(2,5-dioxo-2,5-dihydro-1H-pyrrol-1- yl)hexanoate (32 mg, 0.10 mmol) and N,N-diisopropyl ethylamine (33 mg, 0.26 mmol) were added, and the reaction system was reacted at room temperature for 2 hours.

[1212] After the reaction was completed by LCMS monitoring, the reaction solution was concentrated, and the crude product was purified by Prep-HPLC to obtain compound 63 (52.63 mg, 23.51% yield).

[1213] LCMS (ESI): m / z [M+2H] 2+ :619.0.

[1214] 1H NMR (400 MHz, DMSO-d6) δ 12.87 (s, 1H), 11.03 (s, 1H), 10.98 (s, 1H), 8.77 (s, 1H), 7.86 (t, J = 5.7 Hz, 1H), 7.77 (t, J = 5.6 Hz, 1H), 7.73 - 7.64 (m, 3H), 7.56 (d, J = 7.3 Hz, 1H), 7.51 (s, 1H), 7.44 (d, J = 7.8 Hz, 1H), 7.29 (d, J = 8.7 Hz, 1H), 7.21 - 7.10 (m, 2H), 6.98 (s, 2H), 6.80 (t, J = 6.0 Hz, 1H), 5.41 (s, 2H), 5.26 - 5.19 (m, 1H), 5.10 (dd, J = 13.4, 5.1 Hz, 3H), 4.52 - 4.23 (m, 3H), 4.31 (d, J = 17.4 Hz, 1H), 4.06 (s, 2H), 3.98 (d, J = 9.5 Hz, 1H), 3.58 - 3.48 (m, 5H), 3.47 - 3.38 (m, 7H), 3.17 (dd, J = 12.0, 5.7 Hz, 3H), 3.05 - 2.97 (m, 2H), 2.97 - 2.86 (m, 1H), 2.83 (t, J = 6.2 Hz, 2H), 2.65 - 2.55 (m, 1H), 2.45 - 2.30 (m, 1H), 2.07 - 1.96 (m, 3H), 1.53 - 1.39 (m, 4H), 1.22 - 1.10 (m, 2H).

[1215] Example 64:

[1216] Preparation of ((2S)-2-amino-N-(2-(2-chloro-4-(3-(2-(2,6-dioxopiperidin-3-yl)-1- oxoisoindolin-5-yl)methyl)ureido)phenethoxy)ethyl)-3-hydroxy-N-(prop-2-yn-1-yl)propanamide

[1217] Reaction Scheme:

[1218] Procedure:

[1219] Step A: Compound 13 (60 mg, 0.11 mmol) was dissolved in dry DMF (2 mL) and HATU (46 mg, 0.12 mmol), (tert-butoxycarbonyl)-L-serine (23 mg, 0.11 mmol) and DIEA (29 mg, 0.22 mmol) were added at room temperature. The reaction was stirred at room temperature for 1 hour.

[1220] After the reaction was completed by LCMS monitoring, the reaction solution was purified by reverse phase flash (ACN / 0.1% FA and water) to give compound 64-1.

[1221] LCMS (ESI): m / z [M-Boc+H] + 639.5.

[1222] Step B: Compound 64-1 (50 mg, 0.07 mmol) was dissolved in DCM (2 mL), then TFA (2 mL) was added, the reaction system was protected by nitrogen, and reacted at room temperature for 1 hour.

[1223] After the reaction was completed by LCMS monitoring, the reaction solution was concentrated, and the crude product was purified by Prep-HPLC to give compound 64.

[1224] LCMS (ESI): m / z [M+H] + 639.1.

[1225] 1 H NMR (400 MHz, DMSO) δ 10.98 (s, 1H), 8.86 (brs, 1H), 8.17 (s, 3H), 7.75-7.62 (m, 2H), 7.51 (s, 1H), 7.44 (d, J = 8.2 Hz, 1H), 7.26-7.13 (m, 2H), 6.91 (brs, 1H), 5.47 (s, 1H), 5.11 (dd, J = 13.2, 5.1 Hz, 1H), 4.49-4.24 (m, 6H), 4.20 (d, J = 1.6 Hz, 1H), 3.82-3.70 (m, 1H), 3.68-3.52 (m, 7H), 3.24 (t, J = 2.3 Hz, 1H), 2.98-2.81 (m, 3H), 2.68-2.56 (m, 1H), 2.04-1.95 (m, 1H), 2.04-1.95 (m, 1H).

[1226] Example 65:

[1227] Preparation of 4-((S)-2-(S)-2-(6-(2,5-dioxo-2,5-dihydro-1H-pyrrol-1-yl)hexanamido)-3- methylbutanamido)-5-ureidopentanamido)-benzyl-((2S)-1-(2-(2-chloro-4-(3-(2-(2,6- dioxopiperidin-3-yl)-1-oxoisoindolin-5-yl)methyl)ureido)phenethyl oxy)ethyl)(prop-2-yn-1- yl)amino)-3-hydroxy-1-oxoprop-2-yl)carbamate

[1228] Reaction Scheme:

[1229] Procedure:

[1230] Step A: Compound 64 (25 mg, 0.04 mmol) was dissolved in DMF (3 mL), then compound 1-10 (40 mg, 0.05 mmol) and 2,4-Lutidine (13 mg, 0.12 mmol) were added, the reaction system was stirred at room temperature overnight under nitrogen protection.

[1231] After the reaction was completed by LCMS monitoring, compound 65 was obtained by Prep-HPLC purification.

[1232] LCMS (ESI): m / z [M+2H] 2+ 619.6.

[1233] 1 H NMR (400 MHz, DMSO-d6) δ 10.98 (s, 1H), 9.98 (s, 1H), 8.79 (s, 1H), 8.16 - 8.03 (m, 1H), 7.80 (d, J = 8.6 Hz, 1H), 7.74 - 7.65 (m, 2H), 7.58 (d, J = 8.3 Hz, 2H), 7.51 (s, 1H), 7.46 - 7.37 (m, 2H), 7.35 - 7.24 (m, 2H), 7.21 (d, J = 8.5 Hz, 1H), 7.14 (d, J = 8.3 Hz, 1H), 7.00 (s, 2H), 6.82 (t, J = 5.8 Hz, 1H), 6.00 (br s, 1H), 5.19 - 5.05 (m, 1H), 5.04 - 4.85 (m, 2H), 4.59 - 4.52 (m, 1H), 4.46 - 4.35 (m, 5H), 4.33 - 4.26 (m, 2H), 4.23 - 4.05 (m, 3H), 3.77 - 3.63 (m, 2H), 3.59 - 3.43 (m, 5H), 3.40 - 3.30 (m, 2H), 3.15 - 2.80 (m, 5H), 2.67 - 2.56 (m, 1H), 2.43 - 2.31 (m, 1H), 2.24 - 2.06 (m, 2H), 2.04 - 1.89 (m, 2H), 1.72 - 1.33 (m, 8H), 1.27 - 1.15 (m, 3H), 0.89 - 0.79 (m, 6H).

[1234] Example 92:

[1235] (2S,3S,4S,5R,6S)-6-(4-(11-(2-chloro-4-(3-(2-(2,6-dioxopiperidin-3-yl)-1- oxoisoindolin-5-yl)methyl)ureido)phenyl)-5-imino-6-methyl-3-oxo-2,9-dioxa-4,6- diazadodecyl)-2-(4-(6-(2,5-dioxo-2,5-dihydro-1H-pyrrol-1-yl)hexanamido)but-1-yn-1- yl)phenoxy)-3,4,5-trihydroxytetrahydro-2H-pyran-2-carboxylic acid

[1236] Reaction Scheme:

[1237] Procedure:

[1238] Step A: Compound 52-8 (351.58 mg, 0.45 mmol) was dissolved in dry THF (12 mL), then compound 22 (200 mg, 0.35 mmol) and TEA (106.25 mg, 1.05 mmol) were added successively. The reaction system was reacted at 40 degrees Celsius for 20 hours under nitrogen protection. After the reaction was completed by LCMS monitoring, the reaction solution was purified by reverse phase flash (THF / 0.1% FA in water) to obtain compound 92-1 (170 mg, 40.26% yield). LCMS (ESI): 1203.3 [M+H] + .

[1239] Step B: Compound 92-1 (200 mg, 0.17 mmol) was dissolved in tetrahydrofuran (8 mL), and hydrochloric acid solution (4 mL, 6 mol / L) was added. The reaction system was reacted at 45 degrees Celsius for 8 hours. After the reaction was completed by LCMS monitoring, the reaction solution was concentrated, and the crude product was purified by Prep-HPLC (ACN / 0.1% FA in water) to obtain compound 92-2 (130 mg, 81.21% yield). LCMS (ESI): 963.3 [M+H] + Step C: Compound 92-2 (120 mg, 0.12 mmol) was dissolved in dry DMF (3 mL), and compound 3 (55.49 mg, 0.18 mmol) and DIEA (46.53 mg, 0.36 mmol) were added. The reaction system was reacted at room temperature for 1 hour. After the reaction was completed by LCMS monitoring, the reaction solution was concentrated, and the crude product was purified by Prep-HPLC (ACN / 0.1% TFA in water) to obtain compound 92 (50 mg, 34.71% yield).

[1240] 1 H NMR (400 MHz, DMSO-d6) δ 12.75 (br s, 1H), 10.98 (s, 1H), 10.66 (br s, 1H), 8.89 (s, 1H), 8.60 (br s, 1H), 7.97 (t, J = 5.7 Hz, 1H), 7.73 - 7.61 (m, 2H), 7.51 (s, 1H), 7.47 - 7.36 (m, 2H), 7.31 (d, J = 8.3 Hz, 1H), 7.18 (s, 2H), 7.12 (d, J = 8.7 Hz, 1H), 7.03 - 6.88 (m, 3H), 5.54 - 5.28 (m, 2H), 5.21 - 5.14 (m, 1H), 5.13 - 4.96 (m, 3H), 4.52 - 4.24 (m, 4H), 3.91 (d, J = 9.5 Hz, 1H), 3.64 - 3.53 (m, 6H), 3.34 - 3.18 (m, 8H), 3.09 - 2.76 (m, 7H), 2.68 - 2.53 (m, 2H), 2.44 - 2.31 (m, 1H), 2.10 - 1.94 (m, 3H), 1.51 - 1.42 (m, 4H), 1.22 - 1.15 (m, 2H).

[1241] LCMS (ESI): 1156.7 [M+H] + .

[1242] Example 93:

[1243] ((2S,3S,4S,5R,6S)-6-(4-((5S)-12-(2-chloro-4-(3-(2-(2,6-dioxopiperidin-3-yl)-1- oxoisoindolin-5-yl)methyl)ureido)phenyl)-5-(hydroxymethyl)-3,6-dioxo-7-(prop-2-yn-1-yl)- 2,10-dioxa-4,7-diazadodecyl)-2-(4-(6-(2,5-dioxo-2,5-dihydro-1H-pyrrol-1-yl)hexanamido)but-1- ynyl)phenoxy)-3,4,5-trihydroxytetrahydro-2H-pyran-2-carboxylic acid

[1244] Reaction Scheme:

[1245] Procedure:

[1246] Step A: Compound 64 (190 mg, 0.30 mmol) was dissolved in dry N,N- dimethylformamide (6 mL), followed by the addition of 52-8 (255 mg, 0.33 mmol) and N,N-diisopropyl ethylamine (91 mg, 0.90 mmol). The reaction was stirred at room temperature overnight under nitrogen. After the reaction was completed by LCMS, the reaction was purified by reverse phase flash (THF / 0.1% FA in water) to give compound 93-1 (360 mg, 95.15% yield).

[1247] LCMS (ESI): 1272.5 [M+H] + .

[1248] Step B: Compound 93-1 (220 mg, 0.17 mmol) was dissolved in tetrahydrofuran (5 mL), followed by the addition of hydrochloric acid solution (5 mL, 6 mol / L). The reaction was stirred at 40 °C for 7 hours. After the reaction was completed by LCMS, the reaction was concentrated, and the crude product was purified by Prep-HPLC (THF / 0.1% FA in water) to give compound 93-2 (100 mg, 56.03% yield).

[1249] LCMS (ESI): 1032.3 [M+H] + .

[1250] Step C: Compound 93-2 (30 mg, 0.029 mmol) was dissolved in N,N- dimethylformamide (2 mL), followed by the addition of compound 6-(maleimido) hexanoic acid succinimidyl ester (12 mg, 0.038 mmol) and N,N- diisopropyl ethylamine (11 mg, 0.087 mmol). The reaction was stirred at room temperature for 1 hour. After the reaction was completed by LCMS, the reaction was concentrated, and the crude product was purified by Prep-HPLC (ACN / 0.1% TFA in water) to give compound compound 93 (10 mg, 28.08% yield).

[1251] 1H NMR (400 MHz, DMSO-d6) δ 10.97 (s, 1H), 9.00 - 8.67 (m, 1H), 8.16 - 7.86 (m, 1H), 7.77 - 7.61 (m, 2H), 7.54 - 7.48 (m, 1H), 7.46 - 7.42 (m, 1H), 7.37 - 7.31 (m, 1H), 7.27 - 7.08 (m, 5H), 6.99 (s, 2H), 6.73 - 6.55 (m, 1H), 5.50 - 5.28 (m, 3H), 5.25 - 5.06 (m, 3H), 5.00 - 4.82 (m, 3H), 4.64 - 4.23 (m, 7H), 4.20 - 4.04 (m, 2H), 3.94 - 3.86 (m, 1H), 3.74 - 3.65 (m, 3H), 3.58 - 3.52 (m, 4H), 3.14 - 3.08 (m, 2H), 2.94 - 2.83 (m, 4H), 2.68 - 2.66 (m, 1H), 2.34 - 2.32 (m, 1H), 2.08 - 1.93 (m, 8H), 1.53 - 1.41 (m, 6H).

[1252] LCMS (ESI): 1225.7 [M+H] + .

[1253] Example 94:

[1254] 4-((S)-2-((S)-2-(2-(2,5-dioxo-lH-pyrrol-l-yl)acetamido)propanamido)propanamido)propanamido)benzyl ((2-(2-chloro-4-(3-((2-(2-(2-dioxopiperidin-3-yl)-l-oxoisoindolin-5-yl)methyl)phenoxy)ethyl)(propan-2-yl-l-yl)amino)-3-hydroxy-l-oxopropan-2-yl)carbamate

[1255] Reaction Scheme:

[1256] Procedure:

[1257] Step A: Compound 64 (100 mg, 0.156 mmol) was dissolved in DMF (5 mL), compound 55-5 (97 mg, 0.172 mmol), DIEA (60 mg, 0.465 mmol) was added, the reaction was stirred at 40 °C under nitrogen atmosphere for 48 h. After completion of the reaction as monitored by LCMS, the reaction mixture was purified by Prep-HPLC (ACN / 0.1% TFA in water) to get compound 94 (51.15 mg, 30.62% yield).

[1258] 1 H NMR (400MHz, DMSO-d6) δ10.98(s,1H),9.87(s,1H),8.79(s,1H),8.43(d,J=7.2Hz,1H),8.19(d,J=7.2Hz,1H),7.74–7.63(m,2H),7.57 (d,J=8.3Hz,2H),7.51(s,1H),7.48–7.30(m,2H),7.27(d,J=8.3Hz,2H),7.23–7.11(m,2H),7.09(s,2H),6.81(t,J=5.9Hz,1H),5.15– 5.05(m,1H),5.03–4.86(m,2H),4.62–4.50(m,1H),4.50–4.22(m,7H),4.15–4.02(m,3H),3.72–3.64(m,3H),3.59–3.41(m,6H),3.32– 3.10(m,1H),3.02–2.72(m,3H),2.65–2.56(m,1H),2.45–2.29(m,1H),2.05–1.95(m,1H),1.30(d,J=7.1Hz,3H),1.22(d,J=7.0Hz,3H).

[1259] LCMS(ESI): 1067.6 [M+H] + .

[1260] Gemtuzumab antibody was purchased from Sanyou Biopharmaceutical (Shanghai) Co., Ltd. (Catalog No.: JQX20230805).

[1261] Example 95: Synthesis of ADC-1

[1262] Add 10 mM TCEP solution (2.6 equivalents) to Gemtuzumab antibody solution (10 mg / mL, 5 mL), adjust pH to 6.5, and then reduce at 37°C for 21 hours. Then add 10 mM...

[1263] A solution of N-[2-(2-[2-chloro-4-[([[2(2,6-dioxopiperidin-3-yl)-1-oxo-3H- isoindol-5-yl]methyl]aminoformyl)amino]phenyl]ethoxy)ethyl]-N- methylcarbamic acid [4-[(2S)-5-(carbamoylamino)-2-[(2S)-2-[6(2,5- dioxopyrrol-1-yl)hexanoylamino]-3-methylbutanoylamino]pentanoyl]phenyl] methyl ester (obtained by the method of synthesis of compound (la) in the patent WO2021198965A1) in dimethyl sulfoxide (6.5 equivalents), DMSO accounted for 10%, was mixed at 8°C for 2 hours. After the completion of the coupling reaction, according to the quenching ratio of 20:1 (quenching agent: antibody), 100 mM N-acetyl-cysteine solution was added, and the coupling reaction was terminated at room temperature for 30 min. After the reaction solution was concentrated by tangential flow ultrafiltration, it was washed and filtered 15 times with 10 mM His-HCl (pH 5.5), and then filtered with a 0.22 μm filter to obtain ADC-1 (9.50 mg / mL, 4.2 mL), which was stored at -20°C. The average value of the amount of toxin connected to each antibody molecule (DAR) was 4.0 by RP-HPLC, and the SEC purity was 98.7%.

[1264] Example 96: Synthesis of ADC-2

[1265] To the Gemtuzumab antibody solution (10 mg / mL, 5 mL), 10 mM TCEP solution (2.6 eq) was added, and the pH was adjusted to 6.5 before reduction at 37 °C for 21 h. Subsequently, 10 mM (2S,3S,4S,5R,6S) 6-{4-[({[2-(2-{2-chloro-4[({[2-(2,6-dioxopiperidin-3-yl)-1-oxo-3H-indol-5-yl]methyl}aminoformyl)amino]phenyl}ethoxy)ethyl](methyl)aminoformyl}oxy)methyl]-2({2[6-(2,5-dioxopyrrolidin-1-yl)hexanoylamino]ethyl}aminoformyl)phenoxy}-3,4,5-trihydroxyoxan-2-carboxylic acid (obtained by the method of synthesis of Compound I(a) in the patent WO2022254376A1) dimethyl sulfoxide solution (6.5 eq) was added, and the mixture was stirred at 8 °C for 2 h. After the completion of the coupling reaction, 100 mM N-acetyl-cysteine solution was added at a quenching ratio of 20:1 (quencher: antibody), and the quenching was performed at room temperature for 30 min to terminate the coupling reaction. The reaction solution was concentrated by tangential flow ultrafiltration, washed with 10 mM His-HCl (pH 5.5) for 15 times, and filtered with a 0.22 μm filter to obtain ADC-2 (9.54 mg / mL, 4.1 mL), which was stored at -20 °C. The average value of the amount of toxin (DAR) connected to each antibody molecule was 4.1 by RP-HPLC, and the SEC purity was 99.2%.

[1266] Example 97: Synthesis of ADC-3

[1267] To the Gemtuzumab antibody solution (10 mg / mL, 5 mL), 10 mM TCEP solution (2.45 eq) was added, and the pH was adjusted to 6.5 before reduction at 37 °C for 21 h. Subsequently, 10 mM Compound 51 dimethyl sulfoxide solution (6.5 eq) was added, and the mixture was stirred at 8 °C for 2 h. After the completion of the coupling reaction, 100 mM N-acetyl-cysteine solution was added at a quenching ratio of 20:1 (quencher: antibody), and the quenching was performed at room temperature for 30 min to terminate the coupling reaction. The reaction solution was concentrated by tangential flow ultrafiltration, washed with 10 mM His-HCl (pH 5.5) for 15 times, and filtered with a 0.22 μm filter to obtain ADC-3 (5.78 mg / mL, 7.8 mL), which was stored at -20 °C. The average value of the amount of toxin (DAR) connected to each antibody molecule was 4.2 by RP-HPLC, and the SEC purity was 99.0%.

[1268] Example 98: Synthesis of ADC-4

[1269] To the Gemtuzumab antibody solution (10 mg / mL, 5 mL), 10 mM TCEP solution (2.45 eq) was added, and the pH was adjusted to 6.5 before reduction at 37 °C for 21 h. Subsequently, 10 mM dimethyl sulfoxide solution of compound 55 (6.5 eq) was added, and the mixture was stirred at 8 °C for 2 h. After the completion of the conjugation reaction, 100 mM N-acetyl-cysteine solution was added at a quenching ratio of 20:1 (quencher: antibody), and the conjugation reaction was terminated by quenching at room temperature for 30 min. The reaction solution was concentrated by tangential flow ultrafiltration, washed with 10 mM His-HCl (pH 5.5) for 15 times, and filtered with a 0.22 μm filter to obtain ADC-4 (5.37 mg / mL, 7.9 mL), which was stored at -20 °C. The average amount of toxin linked to each antibody molecule (DAR) was 4.3 by RP-HPLC, and the SEC purity was 99.1%.

[1270] Example 99: Synthesis of ADC-5

[1271] Gemtuzumab antibody (10 mg / mL, 5 mL) in 20 mM PB (pH 7.0) buffer was reduced with 10 mM TCEP solution (2.6 eq) at 37 °C for 21 h after adjusting the pH to 6.5. Subsequently, 10 mM dimethyl sulfoxide solution of compound 65 (6.5 eq) was added, and the mixture was stirred at 8 °C for 2 h. After the completion of the conjugation reaction, 100 mM N-acetyl-cysteine solution was added at a quenching ratio of 20:1 (quencher: antibody), and the conjugation reaction was terminated by quenching at room temperature for 30 min. The reaction solution was concentrated by tangential flow ultrafiltration, washed with 10 mM His-HCl (pH 5.5) for 15 times, and filtered with a 0.22 μm filter to obtain ADC-5 (6.05 mg / mL, 7.2 mL), which was stored at -20 °C. The average amount of toxin linked to each antibody molecule (DAR) was 4.2 by RP-HPLC, and the SEC purity was 99.1%.

[1272] The following ADCs can be obtained according to the above-described synthesis method for antibody conjugation:

[1273] Biological Test Example 1: Cytotoxicity Test of Compounds

[1274] Purpose of Experiment: To test the in vitro cytotoxic activity of the compounds of the present application on SK-BR-3 (human breast cancer cells).

[1275] Method 1

[1276] Experimental Equipment:

[1277] 96-well transparent flat-bottom white plates (Corning, #3610)

[1278] RPMI1640 medium (Gibco, #A10491-01)

[1279] DMEM medium (Gibco, #11995065)

[1280] Fetal Bovine Serum (Sigma-Aldrich, #F8687-500ML)

[1281] Luminescent Cell Viability Assay (Promega, #G7572)

[1282] Experimental method:

[1283] Human breast cancer cells SK-BR-3 (Cell Resource Center of Shanghai Life Sciences, Chinese Academy of Sciences, Item No.: TCHu225) were washed with PBS, and the cells were trypsinized with 0.25% Trypsin-EDTA for about 3-10 minutes. The digestion was terminated with complete culture medium, and the cells were centrifuged at 1000 rpm for 3 minutes. The supernatant was discarded, and the cells were resuspended with complete cell culture medium. The cells were counted with a cell counter, and the cells were adjusted to the desired density. 90.0 μL of cell suspension (3000 cells / well) was added to each well, and the cell plate was incubated in a 37°C, 5% CO2 cell incubator overnight.

[1284] In the experiment, DMSO control group, positive control group and test sample group were set up. The cell plate was taken out of the incubator to observe the cell adhesion state. After the cells adhered, 10.00 μL of sample was added to each well (with a final concentration of 10 μM, 5-fold dilution, 9 concentrations), and gently shaken and placed in a 37°C, 5% CO2 incubator for incubation.

[1285] After 3 days of incubation, 100.0 μL / well of CellTiter-Glo TM (Promega, Item No.: G7572) working solution was added, and the cells were lysed by constant temperature shaker. After 10 minutes, the plate was read on a microplate reader.

[1286] The cell proliferation inhibition rate calculation formula is: cell proliferation inhibition rate % = (1-RLU 供试品孔 / RLU DMSO对照孔 ) x 100%.

[1287] Data analysis: plot the Log value of sample concentration as the horizontal coordinate and Inhibition% as the vertical coordinate, and perform Nonlinear regression (curve fit) analysis on the data to obtain the IC 50 value of each test product.

[1288] The positive control structure is as follows:

[1289] Experimental results: the SKBR-3 cell inhibition activity results of the compounds of the present disclosure are shown in Table 1.

[1290] Table 1. Inhibition activity of some compounds of the present disclosure on cell proliferation

[1291] Method two,

[1292] Experimental equipment:

[1293] 384-well cell culture plate (Corning, #707003)

[1294] McCoy's 5A medium (Gibco, #16600-082)

[1295] Fetal Bovine Serum (Avantor, #76294-180)

[1296] Cell Counting-Lite 2.0 Luminescent Cell Viability Assay (Vazyme, #DD1101-03)

[1297] Experimental method:

[1298] Human breast cancer cells SK-BR-3 (manufacturer ATCC, item number HTB-30) were washed with PBS, and then the cells were trypsinized for about 5-6 minutes, the digestion was terminated with complete culture medium, the cells were centrifuged at 1000 rpm for 3 minutes, the supernatant was discarded, the cells were resuspended with complete cell culture medium, and the cells were counted with a cell counter. The cells were adjusted to the required density, 30.0 μL of cell suspension (3000 cells / well) was added to each well, and the cell plate was incubated in a 37°C, 5% CO2 cell incubator overnight.

[1299] DMSO control group, positive control group and test product group were set in the experiment. The cell plate was taken out of the incubator to observe the cell adhesion state, and after the cells adhered, 10.00 μL of sample was added to each well (with a final concentration of 10 μM, 5-fold dilution, 9 concentrations), gently shaken and then placed in a 37°C, 5% CO2 incubator for incubation.

[1300] After 3 days of incubation, CellCounting-Lite 2.0 Luminescent Cell Viability Assay was added, and the constant temperature shaker was shaken to lyse the cells. After 20 min of incubation, the plate was read on the microplate detector.

[1301] Cell proliferation inhibition rate calculation formula: Cell proliferation inhibition rate % = (Ave_RLU DMSO对照孔 -RLU 供试品孔 ) / (Ave_RLU DMSO对 照孔 -Ave_RLU 仅培养基对照孔 )×100%.

[1302] Data analysis: take the Log value of the sample concentration as the abscissa, and the Inhibition% as the ordinate to plot, and analyze the data by Nonlinear regression (curve fit) to obtain the IC 50 value of each test product:

[1303] Experimental results:

[1304] The results of the inhibitory activity of the compounds of the present disclosure on human SK-BR-3 cells are shown in Table 2.

[1305] Table 2. Inhibitory activity of some compounds of the present disclosure on cell proliferation

[1306] Conclusion: The compounds of the present disclosure have good inhibitory activity on SK-BR-3 cells.

[1307] Biological test example 2: GSPT1 protein degradation ability test of some compounds of the present disclosure

[1308] Purpose of the experiment: To detect the GSPT1 protein degradation activity of the compounds in the present disclosure.

[1309] Experimental apparatus:

[1310] 384-well cell culture plate (Corning, #3764)

[1311] DMEM medium (HyClone, #SH30022.01)

[1312] Fetal Bovine Serum (Avantor, #76294-180)

[1313] HiBiT Lytic Detection System (Promega, #N3040)

[1314] Experimental method:

[1315] HiBiT assay in HEK293-GSPT1-HiBiT-KI Cells (Manufacturer: Elysia, Catalog: None), after washing with PBS, the cells were digested with 0.25% Trypsin-EDTA for about 2-4 minutes, and the digestion was terminated with complete culture medium. The cells were centrifuged at 1000 rpm for 3 minutes, the supernatant was discarded, and the cells were resuspended with complete cell culture medium. The cells were counted with a cell counter, and the cells were adjusted to the required density. 40.0 μL of cell suspension (5000 cells / well) was added to each well. In the experiment, DMSO control group, positive control group and test sample group were set up respectively. Through the ECHO acoustic pipetting workstation (Manufacturer: Bechman, Catalog: ECHO 655), 40 nL of sample was added to each well (with a final concentration of 10 μM, 3-fold dilution, 9 concentrations, and a final DMSO concentration of 0.1%). After centrifugation, the cells were incubated in a 37°C, 5% CO2 incubator. After incubation for 24 h, the cell culture plate was equilibrated at room temperature for 10 minutes. The LgBiT Protein was diluted at a ratio of 1:100, and the HiBiT Protein was diluted at a ratio of 1:50 HiBiT Lytic Substrate, mix well, add 20 μL of Nano-Glo HiBiT Lytic reagent to each well, and shake at 300 rpm in the dark for 3 minutes. After incubation of the cell culture plate at 25°C for 15 min, detection was performed in a microplate detector.

[1316] Cell degradation activity calculation formula: H = Ave (DMSO), L = Ave (Medium) Degradation% = (Ave_H-Sample) / Ave_H*100

[1317] Data analysis: plot the compound concentration as the horizontal coordinate and the degradation rate as the vertical coordinate, and perform Nonlinear regression (curve fit) analysis on the data to obtain the DC 50 value of each test sample.

[1318] Experimental results: The results of the GSPT1 protein degradation activity of the compounds of the present disclosure are shown in Table 3.

[1319] Table 3. GSPT1 protein degradation activity of compounds of the present disclosure

[1320] Conclusion: The compounds of the present disclosure have good GSPT1 protein degradation ability.

[1321] Biological test example 3: liver microsomal metabolic stability test

[1322] Purpose of the experiment: to detect the metabolic stability of the compounds in the present application in human / rat liver microsomes.

[1323] Experimental materials:

[1324] Human Liver Microsomes (Hui Zhi and Source, 0121A1.03)

[1325] SD Rat Liver Microsomes (Hui Zhi and Source, 0121D1.01)

[1326] Experimental method: each incubation system contains phosphate buffer (PBS, pH 7.4), liver microsomal protein, sample to be tested (acetonitrile solution) and NADPH, incubation is carried out in a water bath at 37°C, and 3 times the volume of ice-cold acetonitrile is added to terminate the reaction (containing internal standard and 0.2% formic acid) at 0, 5, 15, 30, 45 and 60 min of reaction, respectively. The sample solution is centrifuged, 100 μl of supernatant is taken, 100 μl of pure water containing 0.2% formic acid is added, and the remaining amount of substrate original form is detected by LC-MS / MS method. The negative control is incubated with the corresponding species of liver microsomal NCF group (without NADPH). Compound 1 is used as a positive control.

[1327] Data analysis:

[1328] The T 1 / 2 , CL int(mic) and CL int(liver) are calculated by using the first-order kinetic equation:

[1329] When

[1330] The in vitro liver microsomal intrinsic clearance (CL int(mic) ) and the hepatic intrinsic clearance (CL int(liver) ) are calculated by k e :

[1331] CL int(mic) = 0.693 / T 1 / 2 / microsomal protein content (microsomal concentration mg / mL during incubation)

[1332] CL int(liver) =CL int(mic) × Liver microsomal protein content (mg / g) × Liver weight to body weight ratio

[1333] The parameters used in the formula are as follows:

[1334] Table 4. Liver microsomal stability of the compounds disclosed herein

[1335] Conclusion: Compared with Comparative Compound 1, the compounds of Examples 13 and 64 of this disclosure have a faster metabolic rate and a shorter metabolic half-life in terms of stability in human liver microsomes, which can reduce the risk of toxic side effects caused by ADC load shedding. Therefore, the compounds of this disclosure have better metabolic properties as ADC loads.

[1336] Biological Test Example 4: Cytotoxicity Assay of ADC

[1337] Experimental objective: To detect the in vitro cytotoxic activity of the compounds in this invention against SK-BR-3 (human breast cancer cells).

[1338] Experimental equipment:

[1339] 96-hole transparent flat-bottom whiteboard (Corning, #3610)

[1340] RPMI 1640 medium (Gibco, #A10491-01)

[1341] DMEM medium (Gibco, #11995065)

[1342] Fetal Bovine Serum (Sigma-Aldrich, #F8687-500ML)

[1343] Luminescent Cell Viability Assay (Promega, #G7572)

[1344] Cytotoxicity assay methods:

[1345] Human myeloid monocytic leukemia cells MV-4-11 (Kyoho Cell Bank) and human promyelocytic leukemia cells HL60 (Kyoho Cell Bank) were counted using a cell counter. The cells were adjusted to the required density, and 180 μL was added to each well (2000 cells / well). The cell plate was placed in a 37°C, 5% CO2 cell incubator.

[1346] The control group, positive group and test product group were set in the experiment, and ADC-1 and ADC-2 were used as positive control samples. The cell plate was taken out of the incubator to observe the cell state, and after the cells adhered, 20 μL of 10X sample (starting at 100 μg / mL, 5-fold dilution, 9 concentrations) was added to each well, gently shaken and then placed in the incubator for incubation. After 6 days of incubation, 100 μL / well of Luminescent Cell Viability Detection Kit (Vkeybio, Catalog No: A2010004N) working solution was added, and the cells were lysed by gently shaking for 30 min. The plate was read on a multifunctional enzyme labeler.

[1347] Cell survival rate calculation formula: cell survival rate = (sample well / control well) x 100%. Data analysis: plot the Log value of the sample concentration as the abscissa and the % of surviving cells as the ordinate, and perform Nonlinear regression (curve fit) analysis on the data to obtain the IC 50 value of each test product.

[1348] Experimental results: The results of the proliferation inhibition activity of the ADC of the present disclosure on human myeloblast leukemia cells HL60 are shown in Table 5.

[1349] Table 5. Inhibition activity of the compound of the present disclosure on cell proliferation

[1350] Conclusion: The ADC of the present disclosure has good inhibitory effect on the proliferation of human myeloblast leukemia cells HL60.

[1351] Biological test example 5: in vivo efficacy of ADC in MV-4-11 xenograft model

[1352] Test purpose: to evaluate the in vivo anti-tumor activity of the ADC of the present disclosure in a CD33-positive human leukemia MV-4-11 xenograft model.

[1353] Test method: BALB / c Nude mice (purchased from Shanghai Jihui Experimental Animal Breeding Co., Ltd.) were used as test animals to evaluate the efficacy of anti-CD33-ADC in a human leukemia MV-4-11 xenograft model after intravenous administration. 1 x 10 7 MV-4-11 cells were resuspended in 100 μL PBS and mixed with an equal volume of Matrigel, and inoculated subcutaneously in the right axillary of the mouse back, with a volume of about 200 μL. Before inoculation, the mice were anesthetized with 3-4% isoflurane. When the tumors grew to an average of about 150 mm 3When the tumor volume and body weight were measured, the mice were randomly divided into groups according to the tumor volume and body weight, with 6 mice in each group. The day of administration was defined as day 0.

[1354] The tail vein injection was used for administration once. The tumor volume and body weight were measured twice a week, and the data were recorded. The tumor inhibition rate TGI (%) = [1-(T28-T0) / (V28-V0)]x100, T28 and T0 were the tumor volumes of the experimental group on day 28 after administration and the day of administration, respectively, and V28 and V0 were the tumor volumes of the blank control group (Vehicle, PBS) on day 28 after administration and the day of administration, respectively. The specific experimental results at the end of the experiment on day 28 after the start of administration are shown in Table 6.

[1355] Table 6 In vivo efficacy evaluation of the ADC compounds of the present disclosure

[1356] The experimental results show that in the CD33-positive MV-4-11 transplanted tumor model, the in vivo efficacy of the ADC of the present disclosure is better than that of ADC-1 and ADC-2.

[1357] Although the specific embodiments of the present application are described above, those skilled in the art should understand that these are only illustrative, and various changes or modifications can be made to these embodiments without departing from the principles and essence of the present application. Therefore, the protection scope of the present application is defined by the appended claims.

Claims

a compound represented by formula (I), a stereoisomer thereof, or a pharmaceutically acceptable salt thereof: wherein Ring A is selected from R 1 selected from H or halogen; R 2 -(CH2) m -O-(CH2) n -NR 2a R 2b -(CH2) m -O-C 3-6 cycloalkyl-NR 2a R 2b -(CH2) m -O-3 to 6 membered heterocyclyl, -C(O)NR 2a -(CH2) n -NR 2a R 2b -C 2-6 alkynyl-NR 2a R 2b -O-C 2-6 alkynyl-NR 2a R 2b -C 2-6 alkenyl-NR 2a R 2b -O-C 2-6 alkenyl-NR 2a R 2b -NR 2a C(O)NR 2a (CH2) n -NR 2a R 2b -3 to 6 membered heterocyclyl-(CH2) n -NR 2a R 2b , -(CH2) m -SO2-(CH2) n -NR 2a R 2b -(CH2) m -SO2-(CH2) n -3 to 8 membered heterocyclyl, -(CH2) m -COO-3 to 8 membered heterocyclyl, -(CH2) m -COO-C 3-6 cycloalkyl-NR 2a R 2b -(CH2) m -O-(CH2) n -NR 2a CO-C 1-6 alkyl, and said C 1-6 alkyl is optionally further substituted by one or more substituents selected from the group consisting of OH, NH2, -CONH2, -COOH, -NHC(=NH)NH2; m, n are each independently selected from 0, 1, 2, 3, 4 or 5; R 2a , R 2b are each independently selected from H, C 1-4 alkyl, C 2-4 alkenyl, C 2-4 alkynyl, C 1-4 alkoxy, -C(=NH)-NH2, -C(=NH)-NHCH3, -C(=NH)-R 2h , -CO-R y , C 3-6 cycloalkyl, -CH2C(O)NH2or 5- to 12-membered heterocyclyl, said C 1-4 alkyl groups can be further substituted by one or more substituents selected from halogen, CN or OH; R y selected from C 1-6 alkyl or 5-12 membered heterocyclyl, and said C 1-6 alkyl or 5-12 membered heterocyclyl can be further substituted by one or more substituents selected from OH, NH2, -CONH2, -S-CH3, phenyl, 4-10 membered heterocyclyl or 7-10 membered heteroaryl, said phenyl can be further substituted by OH, NH2, CN or halogen; or R 2a , R 2b and the nitrogen atom to which they are attached form a 3- to 8-membered heterocyclyl; R 2c , R 2d are each independently selected from H or C 1-4 alkyl; or R 2c , R 2d together with the carbon atom to which they are attached form a C 3-6 cycloalkyl; R 2e , R 2f are each independently selected from H or C 1-4 alkyl; or R 2e , R 2a together with the atom to which they are attached form a 3- to 8-membered heterocyclyl; or R 2e , R 2b together with the atom to which they are attached form a 3- to 8-membered heterocyclyl; R 2h selected from H, C 1-4 alkyl, C 2-4 alkynyl or C 3-6 cycloalkyl, said C 1-4 alkyl or C 3-6 cycloalkyl can be further substituted by one or more substituents selected from halogen, CN or OH; R 3 , R 4 , and R 5 are each independently selected from H or halogen; with the proviso that the compound of formula (I) is not The compound of Formula (I), stereoisomer, or pharmaceutically acceptable salt thereof according to claim 1: wherein, Ring A is selected from R 1 selected from H or halogen; R 2 -(CH2) m -O-(CH2) n -NR 2a R 2b -(CH2) m -O-C 3-6 alkyl-NR 2a R 2b -(CH2) m -O-3 to 6 membered heterocyclyl, -C(O)NR 2a -(CH2) n -NR 2a R 2b -C 2-6 alkynyl-NR 2a R 2b -O-C 2-6 alkynyl-NR 2a R 2b -C 2-6 alkenyl-NR 2a R 2b -O-C 2-6 alkenyl-NR 2a R 2b -NR 2a C(O)NR 2a (CH2) n -NR 2a R 2b -3 to 6 membered heterocyclyl-(CH2) n -NR 2a R 2b , -(CH2) m -SO2-(CH2) n -NR 2a R 2b -(CH2) m -SO2-(CH2) n -3 to 8 membered heterocyclyl, -(CH2) m -COO-3 to 8 membered heterocyclyl, -(CH2) m -COO-C 3-6 cycloalkyl-NR 2a R 2b -(CH2) m -O-(CH2) n -NR 2a CO-C 1-6 alkyl, and said C 1-6 alkyl is optionally further substituted by one or more substituents selected from the group consisting of OH, NH2, -CONH2, -COOH, -NHC(=NH)NH2; m, n are each independently selected from 0, 1, 2, 3, 4 or 5; R 2a , R 2b are each independently selected from H, C 1-4 alkyl, C 2-4 alkenyl, C 2-4 alkynyl, C 1-4 alkoxy, -C(=NH)-NH2, -C(=NH)-NHCH3, -C(=NH)-R 2h , -CO-R y , C 3-6 cycloalkyl or 5-12 membered heterocyclyl, said C 1-4 alkyl can be further substituted by one or more substituents selected from halogen, CN or OH; R y selected from C 1-6 alkyl, and said C 1-6 alkyl can be further substituted with one or more substituents selected from OH, NH2, -CONH2, -S-CH3, phenyl, 4-10 membered heterocyclyl, or 7-10 membered heteroaryl, said phenyl can be further substituted with OH, NH2, CN, or halogen; or R 2a , R 2b and the nitrogen atom to which they are attached form a 3- to 8-membered heterocyclyl; R 2c , R 2d are each independently selected from H or C 1-4 alkyl; or R 2c , R 2d together with the carbon atom to which they are attached form a C 3-6 cycloalkyl; R 2e , R 2f are each independently selected from H or C 1-4 alkyl; or R 2e , R 2a together with the atom to which they are attached form a 3- to 8-membered heterocyclyl; or R 2e , R 2b together with the atom to which they are attached form a 3- to 8-membered heterocyclyl; R 2h selected from H, C 1-4 alkyl, C 2-4 alkynyl or C 3-6 cycloalkyl, said C 1-4 alkyl or C 3-6 cycloalkyl can be further substituted by one or more substituents selected from halogen, CN or OH; R 3 , R 4 , and R 5 are each independently selected from H or halogen; with the proviso that the compound of formula (I) is not The compound of Formula (I), a stereoisomer thereof, or a pharmaceutically acceptable salt thereof according to claim 1 or 2: wherein Ring A is selected from R 1 selected from H or halogen; R 2 -(CH2) m -O-(CH2) n -NR 2a R 2b -(CH2) m -O-C 3-6 cycloalkyl-NR 2a R 2b -(CH2) m -O-3 to 6 membered heterocyclyl, -C(O)NR 2a -(CH2) n -NR 2a R 2b -C 2-6 alkynyl-NR 2a R 2b -O-C 2-6 alkynyl-NR 2a R 2b -C 2-6 alkenyl-NR 2a R 2b -O-C 2-6 alkenyl-NR 2a R 2b -NR 2a C(O)NR 2a (CH2) n -NR 2a R 2b -3 to 6 membered heterocyclyl-(CH2) n -NR 2a R 2b , -(CH2) m -SO2-(CH2) n -NR 2a R 2b -(CH2) m -SO2-(CH2) n -3 to 8 membered heterocyclyl, -(CH2) m -COO-3 to 8 membered heterocyclyl, -(CH2) m -COO-C 3-6 cycloalkyl-NR 2a R 2b -(CH2) m -O-(CH2) n -NR 2a CO-C 1-6 alkyl, and said C 1-6 alkyl is optionally further substituted by one or more substituents selected from the group consisting of OH, NH2, -CONH2, -COOH, -NHC(=NH)NH2; m, n are each independently selected from 0, 1, 2, 3, 4 or 5; R 2a , R 2b each independently is selected from H, C 1-4 alkyl, C 2-4 alkenyl, C 2-4 alkynyl, C 1-4 alkoxy, -C(=NH)-NH2, -C(=NH)-NHCH3, -C(=NH)-R 2h , C 3-6 cycloalkyl or 5-12 membered heterocyclyl, said C 1-4 alkyl can be further substituted by one or more substituents selected from halogen, CN or OH; or R 2a , R 2b together with the nitrogen atom to which they are attached form a 3- to 8-membered heterocyclyl; R 2c , R 2d are each independently selected from H or C 1-4 alkyl; or R 2c , R 2d together with the carbon atom to which they are attached form a C 3-6 cycloalkyl; R 2e , R 2f are each independently selected from H or C 1-4 alkyl; or R 2e , R 2a together with the atom to which they are attached form a 3- to 8-membered heterocyclyl; or R 2e , R 2b together with the atom to which they are attached form a 3- to 8-membered heterocyclyl; R 2h selected from H, C 1-4 alkyl, C 2-4 alkynyl or C 3-6 cycloalkyl, said C 1-4 alkyl or C 3-6 cycloalkyl can be further substituted by one or more substituents selected from halogen, CN or OH; R 3 , R 4 , and R 5 are each independently selected from H or halogen; with the proviso that the compound of formula (I) is not The compound represented by Formula (I), a stereoisomer thereof, or a pharmaceutically acceptable salt thereof according to any one of Items 1-3: wherein ring A is selected from R 1 selected from H or halogen; R 2 -(CH2) m -O-(CH2) n -NR 2a R 2b -(CH2) m -O-C 3-6 cycloalkyl-NR 2a R 2b -(CH2) m -O-3 to 6 membered heterocyclyl, -C(O)NR 2a -(CH2) n -NR 2a R 2b -C 2-6 alkynyl-NR 2a R 2b -O-C 2-6 alkynyl-NR 2a R 2b -C 2-6 alkenyl-NR 2a R 2b -O-C 2-6 alkenyl-NR 2a R 2b -NR 2a C(O)NR 2a (CH2) n -NR 2a R 2b -3 to 6 membered heterocyclyl-(CH2) n -NR 2a R 2b or m, n are each independently selected from 0, 1, 2, 3, 4 or 5; R 2a , R 2b are each independently selected from H, C 1-4 alkyl, C 2-4 alkenyl, C 2-4 alkynyl, C 1-4 alkoxy, -C(=NH)-NH2, -C(=NH)-NHCH3, C 3-6 cycloalkyl or 5-12 membered heterocyclyl, said C 1-4 alkyl can be further substituted by one or more substituents selected from halogen, CN or OH; R 2c , R 2d are each independently selected from H or C 1-4 alkyl; or R 2c , R 2d together with the carbon atom to which they are attached form a C 3-6 cycloalkyl; R 2e , R 2f are each independently selected from H or C 1-4 alkyl; R 3 , R 4 , and R 5 are each independently selected from H or halogen; with the proviso that the compound of formula (I) is not The compound represented by Formula (I), a stereoisomer thereof, or a pharmaceutically acceptable salt thereof according to any one of claims 1-4, wherein R 1 is halogen; preferably, R 1 is F or Cl, more preferably Cl. The compound represented by Formula (I), a stereoisomer thereof, or a pharmaceutically acceptable salt thereof according to any one of claims 1-5, wherein the R 2 -(CH2) m -O-(CH2) n -NR 2a R 2b -(CH2) m -O-cyclobutyl-NR 2a R 2b -(CH2) m -O-4- to 6-membered heterocyclyl, -C(O)NR 2a -O-(CH2) n -NR 2a R 2b -C 3-5 alkynyl-NR 2a R 2b -O-C 3-5 alkynyl-NR 2a R 2b -4-membered heterocyclyl-(CH2) n -NR 2a R 2b , -(CH2) m -SO2-(CH2) n -NR 2a R 2b -(CH2) m -SO2-(CH2) n -4 to 6 membered heterocyclyl, -(CH2) m -COO-4 to 6 membered heterocyclyl, -(CH2) m -COO-C 3-6 cycloalkyl-NR 2a R 2b -(CH2) m -O-(CH2) n -NR 2a CO-C 1-6 alkyl, and said C 1-6 alkyl is optionally further substituted by one or more substituents selected from the group consisting of OH, NH2, -CONH2, -COOH, -NHC(=NH)NH2; m and n are each independently selected from 0, 1 or 2; R 2a , R 2b are each independently selected from the group consisting of H, methyl, ethyl, propenyl, ethynyl, propynyl, -CH2CF3, -CH2CN, -CH2C(O)NH2, methoxy, cyclopropyl, -CO-R y , -C(=NH)-NH2, -C(=NH)-NHCH3, -C(=NH)-R 2h or 5-membered heterocyclyl; R y selected from C 1-6 alkyl or and the C 1-6 alkyl can be further substituted by one or more substituents selected from the group consisting of OH, NH2, -CONH2, -S-CH3, phenyl or said phenyl group is further substituted by OH, NH2, CN or halogen; or R 2a or R 2b with the nitrogen atom to which it is attached forms R 2c , R 2d are each independently selected from H or methyl; or R 2c , R 2d together with the carbon atom to which they are attached form a cyclopropyl group; R 2e , R 2f are each independently selected from H or methyl; or R 2e or R 2a together with the atom to which it is attached form or R 2e or R 2b together with the atom to which it is attached form R 2h is selected from H, methyl, -CH2CN or cyclopropyl. The compound represented by Formula (I), a stereoisomer thereof, or a pharmaceutically acceptable salt thereof according to any one of claims 1-6, wherein the R 2 selected from Preferably, R 2 selected from The compound represented by Formula (I), a stereoisomer thereof, or a pharmaceutically acceptable salt thereof according to any one of claims 1-7, wherein R 3 and R 5 each independently is H. The compound represented by Formula (I), a stereoisomer thereof, or a pharmaceutically acceptable salt thereof according to any one of claims 1-8, wherein R 4 is selected from H or F; and / or, R 2 selected from -(CH2) m -O-(CH2) n -NR 2a R 2b , -C 2-6 alkynyl-NR 2a R 2b or -(CH2) m -(CH2) n -NR 2a R 2b ; m and n are 2; R 2a , R 2b are each independently selected from the group consisting of H, C 1-4 alkyl, C 2-4 alkenyl, C 2-4 alkynyl, C 1-4 alkoxy, -C(=NH)-NH2, -C(=NH)-NHCH3, -CH2C(O)NH2, -CO-R y , -C(=NH)-R 2h or C 3-6 cycloalkyl, said C 1-4 alkyl groups can be further substituted by one or more substituents selected from the group consisting of halogen or CN; R y selected from C 1-6 alkyl or 5-12 membered heterocyclyl, and said C 1-6 alkyl or 5-12 membered heterocyclyl can be further substituted by one or more substituents selected from OH, NH2, -CONH2, -S-CH3, phenyl or 7-10 membered heteroaryl, said phenyl can be further substituted by OH or NH2; R 2c , R 2d are each independently selected from H or C 1-4 alkyl; or R 2c , R 2d together with the carbon atom to which they are attached form a C 3-6 cycloalkyl; R 2e , R 2f are each independently selected from H or C 1-4 alkyl; R 2h selected from C 1-4 alkyl or C 3-6 cycloalkyl, said C 1-4 alkyl can be further substituted by CN. The compound, stereoisomer, or pharmaceutically acceptable salt thereof of any one of claims 1-9, which is selected from: wherein R 1 , R 2 and R 4 are as defined in any one of claims 1 to 9. A compound represented by Formula (I) according to claim 1, a stereoisomer thereof, or a pharmaceutically acceptable salt thereof: wherein, Ring A is R 1 is halogen; R 2 selected from -(CH2) m -O-(CH2) n -NR 2a R 2b , -C 2-6 alkynyl-NR 2a R 2b or preferably -(CH2) m -O-(CH2) n -NR 2a R 2b ; m and n are 2; R 2a , R 2b are each independently selected from the group consisting of H, C 1-4 alkyl, C 2-4 alkenyl, C 2-4 alkynyl, C 1-4 alkoxy, -C(=NH)-NH2, -C(=NH)-NHCH3, -CH2C(O)NH2, -CO-R y , -C(=NH)-R 2h or C 3-6 cycloalkyl, said C 1-4 alkyl groups can be further substituted by one or more substituents selected from the group consisting of halogen or CN; R y selected from C 1-6 alkyl or 5-12 membered heterocyclyl, and said C 1-6 alkyl or 5-12 membered heterocyclyl can be further substituted by one or more substituents selected from OH, NH2, -CONH2, -S-CH3, phenyl or 7-10 membered heteroaryl, said phenyl can be further substituted by OH or NH2; R 2c , R 2d are each independently selected from H or C 1-4 alkyl; or R 2c , R 2d together with the carbon atom to which they are attached form a C 3-6 cycloalkyl; R 2e , R 2f are each independently selected from H or C 1-4 alkyl; R 2h selected from C 1-4 alkyl or C 3-6 cycloalkyl, said C 1-4 alkyl can be further substituted by CN; R 3 , R 4 , and R 5 are each independently selected from H; with the proviso that the compound of formula (I) is not The compound of Formula (I), a stereoisomer thereof, or a pharmaceutically acceptable salt thereof according to any one of claims 1-11, is selected from: wherein R 1 is halogen; preferably, R 1 is CI; R 2 selected from Preferably, R 2 selected from The compound of Formula (I), a stereoisomer thereof, or a pharmaceutically acceptable salt thereof according to Claim 1, wherein the compound of Formula (I) is selected from any one of: The protein degrader antibody conjugate, stereoisomer thereof, or pharmaceutically acceptable salt thereof is selected from the structures shown in Formula (A) or Formula (A-1): wherein, Ab is an antibody or an antigen binding fragment; preferably, Ab is an anti-CD33 antibody; further preferably, Ab is Gemtuzumab; L is a linker group; s, t are each independently selected from a number from 1 to 10; preferably from a number from 2 to 8; for example from a number from 3 to 5; R 2x R 2 R 2 R R 1 , R 2 , ring A is as defined in any one of claims 1 to 13. The protein degrader antibody conjugate, stereoisomer, or pharmaceutically acceptable salt thereof of claim 14, wherein L is selected from Preferably, L is selected from The protein degrader antibody conjugate, stereoisomer, or pharmaceutically acceptable salt thereof according to claim 14 or 15, wherein R 2x selected from wherein R 2x the N-containing end is attached to L; preferably, R 2x is selected from The protein degrader antibody conjugate, stereoisomer, or pharmaceutically acceptable salt thereof according to any one of claims 14-16 is selected from the structures shown in Formula (A-2): wherein, Ab is an antibody or an antigen binding fragment; preferably, Ab is an anti-CD33 antibody; further preferably, Ab is Gemtuzumab; L is selected from t is selected from a number from 2 to 8; preferably from a number from 3 to 5; R 2x selected from R 2x The N-containing end is attached to L. the protein degradation agent linker, stereoisomer thereof, or pharmaceutically acceptable salt thereof is selected from the structure represented by Formula (B): wherein, L a selected from R 2x R 2 the structure with H removed; preferably, R 2 the divalent structure formed by removing H from -NH- or -NH2; for example, R 2x as defined in claim 16; R 1 , R 2 , ring A is as defined in any one of claims 1-13. The protein degrader linker of claim 18, a stereoisomer thereof, or a pharmaceutically acceptable salt thereof, is selected from the structure represented by Formula (B-1): L a selected from R 2x selected from R 2x The upper N-containing end is connected with L a . The protein degrader linker of claim 18, a stereoisomer thereof, or a pharmaceutically acceptable salt thereof, is selected from: The protein degrader antibody conjugate, stereoisomer, or pharmaceutically acceptable salt thereof of any one of claims 14-17, which is selected from: Ab is selected from an antibody or an antigen fragment; preferably, Ab is an anti-CD33 antibody; further preferably, Ab is Gemtuzumab; t is selected from a number from 1 to 10; preferably from a number from 2 to 8; for example from a number from 3 to 5; Alternatively, the proteasome inhibitor antibody conjugate is selected from any one of the following: A pharmaceutical composition comprising a compound of Formula (I), a stereoisomer thereof, or a pharmaceutically acceptable salt thereof, as defined in any one of claims 1-13, a proteolysis targeting chimera, a stereoisomer thereof, or a pharmaceutically acceptable salt thereof, as defined in any one of claims 14-17, 21, a proteolysis targeting chimera linker, a stereoisomer thereof, or a pharmaceutically acceptable salt thereof, as defined in any one of claims 18-20, and a pharmaceutically acceptable carrier. Use of a compound of Formula (I), a stereoisomer thereof, or a pharmaceutically acceptable salt thereof, as defined in any one of claims 1-13, a proteolysis targeting chimera, a stereoisomer thereof, or a pharmaceutically acceptable salt thereof, as defined in any one of claims 14-17, 21, a proteolysis targeting chimera linker, a stereoisomer thereof, or a pharmaceutically acceptable salt thereof, as defined in any one of claims 18-20, or a pharmaceutical composition as defined in claim 22, for the manufacture of a medicament for the treatment of cancer. The use according to claim 23, wherein the cancer is selected from the group consisting of oesophageal cancer, brain cancer, lung cancer, squamous cell cancer, bladder cancer, gastric cancer, ovarian cancer, peritoneal cancer, pancreatic cancer, breast cancer, head and neck cancer, cervical cancer, endometrial cancer, colorectal cancer, liver cancer, renal cancer, urothelial cancer, non-Hodgkin lymphoma, central nervous system tumour, prostate cancer, thyroid cancer, acute myeloid leukaemia or myelodysplastic syndrome. The use according to claim 23, wherein the cancer is selected from the group consisting of oesophageal cancer, brain cancer, lung cancer, squamous cell cancer, bladder cancer, gastric cancer, ovarian cancer, peritoneal cancer, pancreatic cancer, breast cancer, head and neck cancer, cervical cancer, endometrial cancer, colorectal cancer, liver cancer, renal cancer, urothelial cancer, non-Hodgkin lymphoma, central nervous system tumour, prostate cancer, thyroid cancer, acute myeloid leukaemia or myelodysplastic syndrome. A method of treating or preventing cancer, wherein, The use of a therapeutically effective amount of a compound represented by Formula (I), a stereoisomer thereof, or a pharmaceutically acceptable salt thereof of any one of claims 1-13, a proteolysis targeting chimera antibody conjugate, a stereoisomer thereof, or a pharmaceutically acceptable salt thereof of any one of claims 14-17, 21, a proteolysis targeting chimera linker, a stereoisomer thereof, or a pharmaceutically acceptable salt thereof of any one of claims 18-20, or a pharmaceutical composition of claim 22 to a subject in need thereof; preferably, the cancer is selected from the group consisting of esophageal cancer, brain cancer, lung cancer, squamous cell carcinoma, bladder cancer, gastric cancer, ovarian cancer, peritoneal cancer, pancreatic cancer, breast cancer, head and neck cancer, cervical cancer, endometrial cancer, colorectal cancer, liver cancer, kidney cancer, urothelial cancer, non-Hodgkin's lymphoma, central nervous system tumor, prostate cancer, thyroid cancer, acute myeloid leukemia, or myelodysplastic syndrome.

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