Patent Application
20250129088
The present invention relates to a cereblon E3 ubiquitin ligase inhibitor, a Proteolysis Targeting Chimeras (PROTAC) compound comprising the inhibitor, and a use thereof in medicine.
Cereblon (CRBN) is a brain-associated protein with ionic protease activity, which can interact with DNA damage binding protein-1 (DDB1), cadherin 4 (Cullin4, Cul4A or Cul4B), and the regulatory factor Cullins 1 (RoC1) to form a functional E3 ubiquitin ligase complex (CRBN-CRL4), which binds to the substrate and undergoes proteolysis through the ubiquitin-proteasome pathway. Immunomodulators (IMiDs), such as thalidomide, lenalidomide, and pomalidomide, etc., bind to CRBN to recruit new substrates that bind them to CRBN-CRL4, resulting in its ubiquitination and increased proteasome-dependent degradation to treat cancer and other diseases.
On this basis, CRBN ligands have been widely used in protein degradation, and a series of CRBN ligand-based protein degradation targeting chimeras (PROTACs) molecules have been developed. Due to the influence of the CRBN ligand itself on the target, it may additionally degrade the domain protein. So the design and synthesis of new CRBN ligands are of great significance for the further development of PROTACs molecules.
The present invention will disclose a series of novel CRBN ligands and can further synthesize corresponding PROTACs molecules.
The first aspect of the present invention provides a compound represented by formula I, or an isomer, an isotopic derivative, a polymorph, a prodrug thereof, or a pharmaceutically acceptable salt or a solvate thereof:
In one embodiment, W1 and W2 are identical or different, and are each independently CH2 or C(═O), and at least one of W1 and W2 is C(═O);
In another embodiment, G and Z are O.
In another embodiment, R3b and R3c together with the carbon atoms to which they are attached form
and R3a and R3d are each independently selected from H, deuterium atom, halogen, C1-C6 alkyl, heteroalkyl, C2-C6 alkenyl, C2-C6 alkynyl, C1-C6 alkoxy, C1-C6 haloalkyl, C1-C6 haloalkoxy, hydroxyl, C1-C6 hydroxyalkyl, nitro, cyano, amino, C3-C8 cycloalkyl, C4-C10 heterocyclyl, aryl and heteroaryl, wherein the C1-C6 alkyl, C1-C6 heteroalkyl, C2-C6 alkenyl, C2-C6 alkynyl, C1-C6 alkoxy, C3-C8 cycloalkyl, C4-C10 heterocyclyl, aryl and heteroaryl are each independently substituted with one or more substituents selected from halogen, C1-C6 alkyl, C1-C6 heteroalkyl, C1-C6 alkoxy, C1-C6 haloalkyl, hydroxyl, C1-C6 hydroxyalkyl, cyano, amino, nitro, C3-C8 cycloalkyl, C4-C10 heterocyclyl, aryl and heteroaryl, preferably R3a and R3d are each independently selected from H, deuterium atom, F, Cl, Br, I, C1-C6 alkyl, C1-C6 heteroalkyl, C2-C6 alkenyl, C2-C6 alkynyl, C1-C6 alkoxy, hydroxyl, nitro, cyano, amino, C3-C8 cycloalkyl, C4-C10 heterocyclyl, C6-C10 aryl and C5-C10 heteroaryl, wherein the C1-C6 alkyl, C1-C6 heteroalkyl, C2-C6 alkenyl, C2-C6 alkynyl, C1-C6 alkoxy, C3-C8 cycloalkyl, C4-C10 heterocyclyl, C6-C10 aryl and C5-C10 heteroaryl each independently optionally substituted with one or more substituents selected from F, Cl, Br, I, C1-C6 alkyl, C1-C6 heteroalkyl, C1-C6 alkoxy, C1-C6 haloalkyl, hydroxyl, C1-C6 hydroxyalkyl, cyano, amino, nitro, C3-C8 cycloalkyl, C4-C10 heterocyclyl, C6-C10 aryl and C5-C10 heteroaryl, preferably R3a and R3d are each independently selected from H, deuterium atom, halogen, C1-C3 alkyl, and C1-C3 alkoxy;
In another embodiment, R3c and R3d together with the carbon atoms to which they are attached
and R3a and R3b are each independently selected from H, deuterium atom, halogen C1-C6 alkyl, heteroalkyl, C2-C6 alkenyl, C2-C6 alkynyl, C1-C6 alkoxy, C1-C6 haloalkyl, C1-C6 haloalkoxy, hydroxyl, C1-C6 hydroxyalkyl, nitro, cyano, amino, C3-C8 cycloalkyl, C4-C10 heterocyclyl, C6-C10 aryl, and C5-C10 heteroaryl, wherein the C1-C6 alkyl, C1-C6 heteroalkyl, C2-C6 alkenyl, C2-C6 alkynyl, C1-C6 alkoxy, C3-C8 cycloalkyl, C4-C10 heterocyclyl, C6-C10 aryl, C5-C10 heteroaryl are each independently optionally substituted with one or more substituents selected from the groups of halogen, C1-C6 alkyl, C1-C6 heteroalkyl, C1-C6 alkoxy, C1-C6 haloalkyl, hydroxyl, C1-C6 hydroxyalkyl, cyano, amino, nitro, C3-C8 cycloalkyl, C4-C10 heterocyclyl, C6-C10 aryl and C5-C10 heteroaryl, preferably R3a and R3b are each independently selected from H, deuterium atom, F, Cl, Br, I, C1-C6 alkyl, C1-C6 heteroalkyl, C2-C6 alkenyl, C2-C6 alkynyl, C1-C6 alkoxy, hydroxyl, nitro, cyano, amino, C3-C8 cycloalkyl, C4-C10 heterocyclyl, C6-C10 aryl and C5-C10 heteroaryl, wherein the C1-C6 alkyl, C1-C6 heteroalkyl, C2-C6 alkenyl, C2-C6 alkynyl, C1-C6 alkoxy, C3-C8 cycloalkyl, C4-C10 heterocyclyl, C6-C10 aryl and C5-C10 heteroaryl are each independently optionally substituted with one or more substituents selected from F, Cl, Br, I, C1-C6 alkyl, C1-C6 heteroalkyl, C1-C6 alkoxy, C1-C6 haloalkyl, hydroxyl, C1-C6 hydroxyalkyl, cyano, amino, nitro, C3-C8 cycloalkyl, C4-C10 heterocyclyl, C6-C10 aryl, and C5-C10 heteroaryl, preferably R3a and R3b are each independently selected from H, deuterium atom, halogen, C1-C3 alkyl, and C1-C3 alkoxy;
In another embodiment, R3a and R3b together with the carbon atoms to which they are attached form
and R3c and R3d are each independently selected from H, deuterium, halogen C1-C6 alkyl, heteroalkyl, C2-C6 alkenyl, C2-C6 alkynyl, C1-C6 alkoxy, C1-C6 haloalkoxy, C1-C6 haloalkoxy, hydroxyl, C1-C6 hydroxyalkyl, nitro, cyano, amino, C3-C8 cycloalkyl, C4-C10 heterocyclyl, C6-C10 aryl, and C5-C10 heteroaryl, wherein the C1-C6 alkyl, C1-C6 heteroalkyl, C2-C6 alkenyl, C2-C6 alkynyl, C1-C6 alkoxy, C3-C8 cycloalkyl, C4-C10 heterocyclyl, C6-C10 aryl, C5-C10 heteroaryl are each independently optionally substituted with one or more substituents selected from the group of halogen, C1-C6 alkyl, C1-C6 heteroalkyl, C1-C6 alkoxy, C1-C6 haloalkyl, hydroxyl, C1-C6 hydroxyalkyl, cyano, amino, nitro, C3-C8 cycloalkyl, C4-C10 heterocyclyl, C6-C10 aryl and C5-C10 heteroaryl, preferably R3c and R3d each independently selected from H, deuterium atom, F, Cl, Br, I, C1-C6 alkyl, C1-C6 heteroalkyl, C2-C6 alkenyl, C2-C6 alkynyl, C1-C6 alkoxy, hydroxyl, nitro, cyano, amino, C3-C8 cycloalkyl, C4-C10 heterocyclyl, C6-C10 aryl and C5-C10 heteroaryl, wherein the C1-C6 alkyl, C1-C6 heteroalkyl, C2-C6 alkenyl, C2-C6 alkynyl, C1-C6 alkoxy, C3-C8 cycloalkyl, C4-C10 heterocyclyl, C6-C10 aryl and C5-C10 heteroaryl are each independently optionally substituted with one or more substituents selected from F, Cl, Br, I, C1-C6 alkyl, C1-C6 heteroalkyl, C1-C6 alkoxy, C1-C6 haloalkyl, hydroxyl, C1-C6 hydroxyalkyl, cyano, amino, nitro, C3-C8 cycloalkyl, C4-C10 heterocyclyl, C6-C10 aryl, and C5-C10 heteroaryl, preferably R3c and R3d each independently selected from H, deuterium atom, halogen, C1-C3 alkyl, and C1-C3 alkoxy.
In another embodiment, at each occurrence, Rd, Re, Rf, Rg, RD, RE, RF, and RG at each occurrence are independently C(Rm)2 or O.
In another embodiment, W3 and W4 are CH.
In another embodiment, R1h, R2h, and R3h are each independently selected from H, deuterium atom, halogen, C1-C6 alkyl, C1-C6 deuterated alkyl, C1-C6 heteroalkyl, C2-C6 alkenyl, C2-C6 alkynyl, C1-C6 alkoxy, C1-C6 haloalkyl, C1-C6 haloalkoxy, hydroxyl, C1-C6 hydroxyalkyl, nitro, cyano, amino, C3-C8 cycloalkyl, C4-C10 heterocyclyl, C6-C10 aryl, and C5-C10 heteroaryl, wherein the C1-C6 alkyl, C1-C6 heteroalkyl, C2-C6 alkenyl, C2-C6 alkynyl, C1-C6 alkoxy, C3-C8 cycloalkyl, C4-C10 heterocyclyl, C6-C10 aryl, and C5-C10 heteroaryl are independently and optionally substituted with one or more substituents selected from the group consisting of halogen, C1-C6 alkyl, C1-C6 heteroalkyl, C1-C6 alkoxy, C1-C6 haloalkyl, hydroxyl, C1-C6 hydroxyalkyl, cyano, amino, nitro, C3-C8 cycloalkyl, C4-C10 heterocyclyl, C6-C10 aryl, and C5-C10 heteroaryl.
In another embodiment, m1 and m2 at each occurrence are each independently an integer of 0, 1, 2, or 3, and m1+m2≤3; preferably m1+m2=1 or m1+m2=2.
In another embodiment, m3 at each occurrence is independently an integer of 0, 1, 2, 3, or 4, and m4 is an integer of 1, 2, 3, 4, or 5, and m3+m4≤5; preferably m3+m4=2, m3+m4=3 or m3+m4=4.
In another embodiment, m5 and m6 at each occurrence are each independently an integer of 0, 1, 2, 3 or 4, and m5+m6≤4, preferably m5+m6=2 or m5+m6=3.
In another embodiment, m7 and m8 at each occurrence are independently an integer of 0, 1, 2, 3 or 4, and m7+m8≤4, preferably m7+m8=2 or m7+m8=3.
In another embodiment, Rm at each occurrence is independently selected from H, deuterium atom, halogen, C1-C6 alkyl, C1-C6 heteroalkyl, C2-C6 alkenyl, C2-C6 alkynyl, C1-C6 alkoxy, C1-C6 haloalkyl, C1-C6 haloalkoxy, hydroxyl, C1-C6 hydroxyalkyl, nitro, cyano, amino, C3-C8 cycloalkyl, C4-C10 heterocyclyl, aryl and heteroaryl, wherein the C1-C6 alkyl, C1-C6 heteroalkyl, C2-C6 alkenyl, C2-C6 alkynyl, C1-C6 alkoxy, C3-C8 cycloalkyl, C4-C10 heterocyclyl, C6-C10 aryl, and C5-C10 heteroaryl are each independently optionally substituted with one or more substituents selected from the group consisting of halogen, C1-C6 alkyl, C1—C heteroalkyl, C1-C6 alkoxy, C1-C6 haloalkyl, hydroxyl, C1-C6 hydroxyalkyl, cyano, amino, nitro, C3-C8 cycloalkyl, C4-C10 heterocyclyl, C6-C10 aryl and C5-C10 heteroaryl.
In another embodiment, R1 is selected from H, halogen, C1-C3 alkyl, and hydroxyl; preferably, R1 is selected from H, F, Cl, Br, I, C1-C3 alkyl, and hydroxyl.
In another embodiment, R2 is selected from H, and C1-C3 alkyl.
In another embodiment, n is 0 or 1.
In another embodiment, the compound is selected from the following structures:
In another embodiment, the compound is selected from the following structures:
In another embodiment, the compound is selected from the following structures:
In another embodiment the compound is selected from the following structures:
In another embodiment, the compound is selected from the following structures:
A second aspect of the present invention discloses a compound, wherein the compound has the following chemical structure:
CLM-L-PTM,
In one embodiment, W1 and W2 are identical or different, and are each independently CH2 or C(═O), and at least one of W1 and W2 is C(═O).
In another embodiment, R3a, R3b, R3c, and R3a are each independently selected from H, deuterium atom, halogen, C1-C6 alkyl, heteroalkyl, C2-C6 alkenyl, C2-C6 alkynyl, C1-C6 alkoxy, C1-C6 haloalkyl, C-C6 haloalkoxy, hydroxyl, C1-C6 hydroxyalkyl, nitro, cyano, amino, C3-C8 cycloalkyl, C4-C10 heterocyclyl, C6-C10 aryl, and C5-C10 heteroaryl, wherein the C1-C6 alkyl, C1-C6 heteroalkyl, C2-C6 alkenyl, C2-C6 alkynyl, C1-C6 alkoxy, C3-C8 cycloalkyl, C4-C10 heterocyclyl, C6-C10 aryl and C5-C10 heteroaryl are each independently optionally substituted with one or more substituents selected from the group consisting of halogen, C1-C6 alkyl, C1-C6 heteroalkyl, C1-C6 alkoxy, C1-C6 haloalkyl, hydroxyl, C1-C6 hydroxyalkyl, cyano, amino, nitro, C3-C8 cycloalkyl, C4-C10 heterocyclyl, C6-C10 aryl and C5-C10 heteroaryl, preferably R3a, R3b, R3c, and R3d are each independently selected from H, deuterium atom, F, Cl, Br, I, C1-C6 alkyl, C1-C6 heteroalkyl, C2-C6 alkenyl, C2-C6 alkynyl, C1-C6 alkyl, hydroxyl, nitro, cyano, amino, C3-C8 cycloalkyl, C4-C10 heterocyclyl, C6-C10 aryl, and C5-C10 heteroaryl, wherein the C1-C6 alkyl, C1-C6 heteroalkyl, C2-C6 alkenyl, C2-C6 alkynyl, C1-C6 alkoxy, C3-C8 cycloalkyl, C4-C10 heterocyclyl, C6-C10 aryl and C5-C10 heteroaryl are each independently substituted with one or more of the substituents selected from F, Cl, Br, I, C1-C6 alkyl, C1-C6 heteroalkyl, C1-C6 alkoxy, C1-C6 haloalkyl, hydroxyl, C1-C6 hydroxyalkyl, cyano, amino, nitro, C3-C8 cycloalkyl, C4-C10 heterocyclyl, C6-C10 aryl and C5-C10 heteroaryl; preferably R3a and R3b are each independently selected from H, deuterium atom, halogen, C1-C3 alkyl, and C1-C3 alkoxy, preferably R3c and R3d are each independently selected from H, deuterium atom, F, Cl, Br, I, C1-C3 alkyl, and C1-C3 alkoxy.
In another embodiment, at each occurrence, Rd, Re, Rf and Rg at each occurrence are each independently C(Rm)2 or O.
In another embodiment, at each occurrence, RD, RE, RF and RG at each occurrence are each independently C(Rm)2 or O.
In another embodiment, W3 and W4 are CH.
In another embodiment, R2h is selected from H, deuterium atom, halogen, C1-C6 alkyl, C1-C6 deuterated alkyl, C1-C6 heteroalkyl, C2-C6 alkenyl, C2-C6 alkynyl, C1-C6 alkoxy, C1-C6 haloalkyl, C1-C6 haloalkoxy, hydroxyl, C1-C6 hydroxyalkyl, nitro, cyano, amino, C3-C8 cycloalkyl, C4-C10 heterocyclyl, C6-C10 aryl, and C5-C10 heteroaryl, wherein the C1-C6 alkyl, C1-C6 heteroalkyl, C2-C6 alkenyl, C2-C6 alkynyl, C1-C6 alkoxy, C3-C8 cycloalkyl, C4-C10 heterocyclyl, C6-C10 aryl and C5-C10 heteroaryl are each independently optionally substituted with one or more substituents selected from halogen, C1-C6 alkyl, C1-C6 heteroalkyl, C1-C6 alkoxy, C1-C6 haloalkyl, hydroxyl, C1-C6 hydroxyalkyl, cyano, amino, nitro, C3-C8 cycloalkyl, C4-C10 heterocyclyl, C6-C10 aryl and C5-C10 heteroaryl; preferably R2h is selected from H, deuterium atom, F, Cl, Br, I, C1-C6 alkyl, C1-C6 deuterated alkyl, C1-C6 heteroalkyl, C2-C6 alkenyl, C2-C6 alkynyl, C1-C6 alkoxy, hydroxyl, nitro, cyano, amino, C3-C8 cycloalkyl, C4-C10 heterocyclyl, C6-C10 aryl, and C5-C10 heteroaryl, wherein the C1-C6 alkyl, C1-C6 heteroalkyl, C2-C6 alkenyl, C2-C6 alkynyl, C1-C6 alkoxy, C3-C8 cycloalkyl, C4-C10 heterocyclyl, C6-C10 aryl and C5-C10 heteroaryl are each independently substituted with one or more substituents selected from F, Cl, Br, I, C1-C6 alkyl, C1-C6 heteroalkyl, C1-C6 alkoxy, C1-C6 haloalkyl, hydroxyl, C1-C6 hydroxyalkyl, cyano, amino, nitro, C3-C8 cycloalkyl, C4-C10 heterocyclyl, C6-C10 aryl and C5-C10 heteroaryl; preferably R2h is selected from H, deuterium atom, halogen, C1-C3 alkyl, and C1-C3 alkoxy; preferably R2h is selected from H, deuterium atom, F, Cl, Br, I, C1-C3 alkyl, C1-C3 haloalkyl, C1-C3 alkoxy, and C1-C3 haloalkoxy.
In another embodiment, m1 and m2 at each occurrence are each independently an integer of 0, 1, 2, or 3, and m1+m2≤3, preferably m1+m2=1 or m1+m2=2.
In another embodiment, m3 at each occurrence is independently an integer of 0, 1, 2, 3, or 4, and m4 is an integer of 1, 2, 3, 4, or 5, and m3+m4≤5, preferably m3+m4=2, m3+m4=3 or m3+m4=4.
In another embodiment, at each occurrence, m5 and m6 at each occurrence are each independently an integer of 0, 1, 2, 3 or 4, and m5+m6≤4, preferably m5+m6=2 or m5+m6=3.
In another embodiment, m7 and m8 at each occurrence are each independently an integer of 0, 1, 2, 3 or 4, and m7+m8≤4, preferably m7+m8=2, or m7+m8=3.
In another embodiment, Rm at each occurrence is each independently selected from H, deuterium atom, halogen, C1-C6 alkyl, C1-C6 heteroalkyl, C2-C6 alkenyl, C2-C6 alkynyl, C1-C6 alkoxy, C1-C6 haloalkyl, C1-C6 haloalkoxy, hydroxyl, C1-C6 hydroxyalkyl, nitro, cyano, amino, C3-C8 cycloalkyl, C4-C10 heterocyclyl, aryl and heteroaryl, wherein the C1-C6 alkyl, C1-C6 heteroalkyl, C2-C6 alkenyl, C2-C6 alkynyl, C1-C6 alkoxy, C3-C5 cycloalkyl, C4-C10 heterocyclyl, C6-C10 aryl and C5-C10 heteroaryl are each independently optionally substituted with one or more substituents selected from halogen, C1-C6 alkyl, C1-C6 heteroalkyl, C1-C6 alkoxy, C1-C6 haloalkyl, hydroxyl, C1-C6 hydroxyalkyl, cyano, amino, nitro, C3-C8 cycloalkyl, C4-C10 heterocyclyl, C6-C10 aryl and C5-C10 heteroaryl; preferably, Rm at each occurrence is independently selected from H, deuterium atom, F, Cl, Br, I, C1-C6 alkyl, C1-C6 heteroalkyl, C2-C6 alkenyl, C2-C6 alkynyl, C1-C6 alkoxy, hydroxyl, nitro, cyano, amino, C3-C8 cycloalkyl, C4-C10 heterocyclyl, C6-C10 aryl and C5-C10 heteroaryl, wherein the C1-C6 alkyl, C1-C6 heteroalkyl, C2-C6 alkenyl, C2-C6 alkynyl, C1-C6 alkoxy, C3-C8 cycloalkyl, C4-C10 heterocyclyl, C6-C10 aryl and C5-C10 heteroaryl are each independently optionally substituted with one or more substituents selected from F, Cl, Br, I, C1-C6 alkyl, C1-C6 heteroalkyl, C1-C6 alkoxy, C1-C6 haloalkyl, hydroxyl, C1-C6 hydroxyalkyl, cyano, amino, nitro, C3-C8 cycloalkyl, C4-C10 heterocyclyl, C6-C10 aryl and C5-C10 heteroaryl; preferably Rm at each occurrence is independently selected from H, deuterium atom, halogen, C1-C3 alkyl, and C1-C3 alkoxy.
In another embodiment, R1 is selected from H, halogen, C1-C3 alkyl, and hydroxyl; preferably, R1 is selected from H, F, Cl, Br, I, C1-C3 alkyl, and hydroxyl.
In another embodiment, R2 is selected from H, and C1-C3 alkyl.
In another embodiment, n is 0 or 1.
In another embodiment, said CLM is selected from the following structures:
In another embodiment, the CLM is selected from the following structures:
In another embodiment, the CLM is selected from the following structures:
In another embodiment, the CLM is selected from the following structures:
In another embodiment, the CLM is selected from the following structures:
In another embodiment, L is a bond or —(B)Lq-;
In another embodiment BL is selected from one or more of the following structures; —O—, —S—, —SO—, —SO2—, —CH2—, —CO—, —NH—,
is the point of attachment.
In another embodiment, L is selected from the following structures:
is the point of attachment with CLM or PTM;
Preferably L is selected from covalent bonds, —CH2—, —(CH2)2—, —(CH2)3—, —(CH2)4—, —(CH2)5—, —(CH2)6—, —(CH2)7—, —(CH2)8—, —NH—CH2—, —NH—(CH2)2—, —NH—(CH2)3—, —NH—(CH2)4—, —NH—(CH2)5—, —NH—(CH2)6—, —NH—(CH2)7—, —NH—(CH2)8—, —CO—NH—CH2—, —CO—NH—(CH2)2—, —CO—NH—(CH2)3—, —CO—NH—(CH2)4—, —CO—NH—(CH2)5—, —CO—NH—(CH2)6—, —CO—NH—(CH2)7—, —CO—NH—(CH2)8—, —CH2—NH—, —(CH2)2—NH—, —(CH2)3—NH—, —(CH2)4—NH—, —(CH2)5—NH—, —(CH2)6—NH—, —(CH2)7—NH—, —(CH2)8—NH—, —NH—CH2—NH—, —NH—(CH2)2—NH—, —NH—(CH2)3—NH—, —NH—(CH2)4—NH—, —NH—(CH2)5—NH—, —NH—(CH2)6—NH—, —NH—(CH2)7—NH—, —NH—(CH2)8—NH—, —(CH2—CH2—O)—CH2—CH2—, —(CH2—CH2—O)2—CH2—CH2—, —(CH2—CH2—O)3—CH2—CH2—, —NH—(CH2—CH2—O)—CH2—CH2—, —NH—(CH2—CH2—O)2—CH2—CH2—, —NH—(CH2—CH2—O)3—CH2—CH2—, —CO—NH—(CH2—CH2—O)—CH2—CH2—, —CO—NH—(CH2—CH2—O)2—CH2—CH2—, —CO—NH—(CH2—CH2—O)3—CH2—CH2—, —(CH2—CH2—O)—CH2—CH2—NH—, —(CH2—CH2—O)2—CH2—CH2—NH—, —(CH2—CH2—O)3—CH2—CH2—NH—, —NH—(CH2—CH2—O)—CH2—CH2—NH—, —NH—(CH2—CH2—O)2—CH2—CH2—NH—, —NH—(CH2—CH2—O)3—CH2—CH2—NH—, —CO—NH—(CH2—CH2—O)—CH2—CH2—NH—, —CO—NH—(CH2—CH2—O)2—CH2—CH2—NH—, —CO—NH—(CH2—CH2—O)3—CH2—CH2—NH—, —CH2—CH2—(O—CH2—CH2)—, —CH2—CH2—(O—CH2—CH2)2—, —CH2—CH2—(O—CH2—CH2)3—, —NH—CH2—CH2—(O—CH2—CH2)—, —NH—CH2—CH2—(O—CH2—CH2)2—, —NH—CH2—CH2—(O—CH2—CH2)3—, —CO—NH—CH2—CH2—(O—CH2—CH2)—, —CO—NH—CH2—CH2—(O—CH2—CH2)2—, —CO—NH—CH2—CH2—(O—CH2—CH2)3—, —CH2—CH2—(O—CH2—CH2)—NH—, —CH2—CH2—(O—CH2—CH2)2—NH—, —CH2—CH2—(O—CH2—CH2)3—NH—, —NH—CH2—CH2—(O—CH2—CH2)—NH—, —NH—CH2—CH2—(O—CH2—CH2)2—NH—, —NH—CH2—CH2—(O—CH2—CH2)3—NH—, —NH—CH2—CH2—O—CH2—CH2—CO—, —CO—CH2—CH2—O—CH2—CH2—NH—, —NH—(CH2)4—CO—, —NH—(CH2)5—CO—, —NH—(CH2)6—CO—, —CO—(CH2)4—NH—, —CO—(CH2)5—NH—, —CO—(CH2)6—NH—, —NH—(CH2—CH2—O)—(CH2)3—, —NH—(CH2—CH2—O)—(CH2)4—, —NH—(CH2—CH2—O)—(CH2)5—, —NH—(CH2—CH2—O)—(CH2)6—, —(CH2)3—(O—CH2—CH2)—NH—, —(CH2)4—(O—CH2—CH2)—NH—, —(CH2)5—(O—CH2—CH2)—NH—, —(CH2)6—(O—CH2—CH2)—NH—, —CH2—CH2—O—(CH2)2—CO—, —CH2—CH2—O—(CH2)3—CO—, —CH2—CH2—O—(CH2)4—CO—, —CO—(CH2)2O—CH2—CH2—, —CO—(CH2)3O—CH2—CH2—, —CO—(CH2)4O—CH2—CH2—, —CO—(CH2)2—, —CO—(CH2)3—, —CO—(CH2)4—, —CO—(CH2)5—, —CO—(CH2)6—, —(CH2)2—CO—, —(CH2)3—CO—, —(CH2)4—CO—, —(CH2)5—CO—, —(CH2)6—CO—, —CO—(CH2)2—CO—, —CO—(CH2)3—CO—, —CO—(CH2)4—CO—, —CO—(CH2)5—CO—, —CO—(CH2)6—CO—, —CH2—CO—CH2—, —CH2—CO—(CH2)2—, —CH2—CO—(CH2)3—, —CH2—CO—(CH2)4—, —(CH2)2—CO—CH2—, —(CH2)2—CO—(CH2)2—, —(CH2)2—CO—(CH2)3—, —(CH2)2—CO—(CH2)4—, —(CH2)3—CO—CH2—, —(CH2)3—CO—(CH2)2—, —(CH2)3—CO—(CH2)3—, —(CH2)3—CO—(CH2)4—, —(CH2)4—CO—CH2—, —(CH2)4—CO—(CH2)2—, —(CH2)4—CO—(CH2)3—, —(CH2)4—CO—(CH2)4—, —CH2—O—CH2—, —CH2—O—(CH2)2—, —CH2—O—(CH2)3—, —CH2—O—(CH2)4—, —(CH2)2—O—CH2—, —(CH2)2—O—(CH2)2—, —(CH2)2—O—(CH2)3—, —(CH2)2—O—(CH2)4—, —(CH2)3—O—CH2—, —(CH2)3—O—(CH2)2—, —(CH2)3—O—(CH2)3—, —(CH2)3—O—(CH2)4—, —(CH2)4—O—CH2—, —(CH2)4—O—(CH2)2—, —(CH2)4—O—(CH2)3—, —(CH2)4—O—(CH2)4—,
In another embodiment, the PTM is a moiety that binds to a target protein or polypeptide, wherein the target protein is selected from the following proteins: structural proteins, receptors, enzymes, cell surface proteins; proteins pertinent to the integrated function of a cell, including proteins involved in catalytic activity, aromatase activity, motor activity, helicase activity, metabolic process, antioxidant activity, proteolysis, biosynthesis; proteins with kinase activity, oxidoreductase activity, transferase activity, hydrolase activity, lyase activity, isomerase activity, ligase activity, enzyme regulator activity, signal transducer activity, structural molecule activity, binding activity, receptor activity, cell motility, membrane fusion, cell communication, regulation of biological process, development, cell differentiation, response to stimulus; behavioral proteins; cell adhesion proteins; proteins involved in cell death; and proteins involved in transportation, including proteins of protein transporter activity, nuclear transporter activity, ion transporter activity, channel transporter activity, carrier activity, permease activity, secretion activity, electron transporter activity, pathogenesis, chaperone regulator activity, nucleic acid binding activity, transcription regulator activity, extracellular tissues and biogenesis activity, and translation regulator activity.
In another embodiment, PTM is a moiety binding to a target protein or polypeptide, wherein the target protein is selected from the group consisting of B7.1 and B7, TNFR2, NADPH oxidase, BclIBax, and other conjugates in the apotosis pathway, C5a receptor, HMG-CoA reductase, PDE V phosphodiesterase type, PDEIV phosphodiesterase type 4, PDEI I, PDEI II, PDE III, squalene epoxidation enzyme, CXCR1, CXCR2, nitric oxide (NO) synthase, cyclo-oxygenase 1, cyclo-oxygenase 2, 5HT receptors, dopamine receptors, G proteins (ie Gq), histamine receptors, 5-lipoxygenase, tryptase seine protease, thymidylate synthase, purine nucleoside phosphorylase, GAPDH trypanosomal, glycogen phosphorylase, carbonic anhydrase, chemokine receptor, JAW STAT, RXR and its analog, HIV1 protease, HIV1 integrase, influenza neuraminidase, hepatitis B reverse transcriptase, sodium channel, multi drug resistant bacteria, protein P-glycoprotein, tyrosine kinases, CD23, CD124, tyrosine kinase p561ck, CD4, CD5, IL-2 receptor, IL-1 receptor, TNF-αR, ICAM1, Ca2+ channels, VCAM, VLA-4 integrin, selectins, CD40/CD40L, inosine monophosphate dehydrogenase, p38 MAP kinase, RaslRaflMEWERK pathway, interleukin-1 converting enzyme, caspase, HCV, NS3 protease, HCV NS3, RNA helicase, glycinamide ribonucleotide formyl transferase, rhinovirus, 3C protease, herpes simplex virus-I (HSV-I), protease, cytomegalovirus (CMV) protease, poly(ADP-ribose) polymerase, cyclindependent kinase 4/6, vascular endothelial growth factor, oxytocin receptor, microsomal transfer protein inhibitor, bile acid transport inhibitor, 5a reductase inhibitors, angiotensin 11, glycine receptor, noradrenaline reuptake receptor, endothelin receptor, neuropeptide Y and receptor, adenosine receptors, adenosine kinase and AMP deaminase, purinergic receptors (P2Y1, P2Y2, P2Y4, P2Y6, P2X1-7), farnesyltransferases, geranylgeranyl transferase, TrkA a receptor for NGF, β-amyloid, tyrosine kinase Flk-II KDR, vitronectin receptor, integrin receptor, Her-21 neu, telomerase inhibition, cytosolic phospholipase A2, EGF receptor tyrosine kinase, ecdysone 20-monooxygenase, ion channel of GABA-gated chloride channel, acetylcholinesterase, voltage-sensitive sodium channel protein, calcium release channel and chloride ion channel, acetyl-CoA carboxylase, adenylosuccinate synthetase, protoporphyrinogen oxidase, enolpyruvylshikimate phosphate synthase, MYC protein, androgen receptor, estrogen receptor, aurora kinase A, Bcr-Abl protein, epidermal growth factor receptor, anaplastic lymphoma kinase, Bruton's tyrosine kinase, bromodomain protein 4.
In another embodiment, the PTM is an Hsp90 inhibitor, a kinase inhibitor, a phosphatase inhibitor, a MDM2 inhibitor, a compound that binds to a protein comprising the human BET bromodomain, a HDAC inhibitor, a human lysine methyltransferase inhibitor, a compound that binds to RAF receptor, a compound that binds to FKBP, an angiogenesis inhibitor, an immunosuppressive compound, an compound that binds to aryl hydrocarbon receptor, a compound that binds to androgen receptor, a compound that binds to estrogen receptors, a compound that binds to thyroid hormone receptor, a compound that binds to HIV protease, a compound that binds to HIV integrase, a compound that binds to HCV protease, a compound that binds to acyl protein thioesterase 1 and/or 2, a compound that binds to c-MYC protein, a compound that binds to laser kinase A, a compound that binds to Bcr-Abl protein, a compound that binds to bromodomain protein 4 (BRD4), a compound that binds to anaplastic lymphoma kinase (ALK), a compound that binds to Bruton's tyrosine kinase (BTK), a compound that binds to cyclin-dependent kinase 4/6 (CDK4/6), or a compound that binds to epidermal growth factor receptor.
In another embodiment, the PTM is a moiety binding to the androgen receptor.
In another embodiment, the PTM is selected from the following structures:
In another embodiment, the compound is selected from the following structures:
In another embodiment, the PTM is a moiety binding to aurora kinase A (AURORA-A).
In another embodiment, the PTM has the following structure:
In another embodiment, the compound has the following structures:
In another embodiment, the PTM is a moiety binding to Bcr-Abl protein.
In another embodiment, the PTM has the following structure:
In another embodiment, the compound has the following structures:
In another embodiment, the PTM is a moiety that binds to epidermal growth factor receptor (EGFR).
In another embodiment, the PTM has the following structure:
In another embodiment, the compound has the following structures:
In another embodiment, the PTM is a moiety binding to anaplastic lymphoma kinase (ALK).
In another embodiment, the compound has the following structures:
In another embodiment, the compound has the following structures:
In another embodiment, the PTM is a moiety that binds to Bruton's tyrosine kinase (BTK).
In another embodiment, the PTM has the following structure:
In another embodiment, the compound has the following structures:
In another embodiment, the PTM is a moiety binding to cyclin-dependent kinase 4/6 (CDK4/6).
In another embodiment, the PTM has the following structures:
In another embodiment, the compound has the following structures:
In another embodiment, the PTM is a moiety binding to bromodomain protein 4 (BRD4).
In another embodiment, the PTM has the following structure:
In another embodiment, the compound has the following structures:
In another embodiment, the PTM is a moiety binding to estrogen receptor (ER).
In another embodiment, the PTM has the following structure:
In another embodiment, the compound has the following structures:
A third aspect of the present invention provides a pharmaceutical composition, the pharmaceutical composition comprising a compound provided in the first or second aspect of the present invention.
In one embodiment, use of a compound described in the first or second aspect of the present invention, or a pharmaceutical composition described in the third aspect of the present invention, in the preparation of a medicament for the treatment or prevention of diseases treated by degrading target protein bound to a ligand of the target protein, the target protein is preferably cyclin-dependent kinase 4/6, androgen receptor, estrogen receptor, aurora kinase A, Bcr-Abl protein, epidermal growth factor receptor, anaplastic lymphoma kinase, Bruton's tyrosine kinase, or bromodomain protein 4.
In another embodiment, the use of a compound described in the first or second aspect of the present invention, or a pharmaceutical composition described in the third aspect of the present invention, in the preparation of a medicament for the treatment or prevention of diseases treated by binding to cereblon in vivo.
In another embodiment, the use of a compound described in the first or second aspect of the present invention, or a pharmaceutical composition described in the third aspect of the present invention, in the treatment or prevention of a disease associated with the accumulation and/or aggregation of a target protein, the target protein is preferably cyclin-dependent kinase 4/6, androgen receptor, estrogen receptor, aurora kinase A, Bcr-Abl protein, epidermal growth factor receptor, mesenchymal lymphoma kinase, Bruton's tyrosine kinase, or bromodomain protein 4.
In another embodiment, wherein the disease is tumor or cancer, preferably the tumor or cancer is breast cancer, ductal carcinoma of the breast, prostate cancer, condylomatous lymphoma, chronic myeloid leukemia, acute myeloid leukemia, myelomonocytic leukemia, non-small cell lung cancer, lung adenocarcinoma, and/or cervical cancer.
In another embodiment, a compound as described in the first or second aspect of the present invention, or a pharmaceutical composition as described in the third aspect of the present invention, for use in the treatment or prevention of disease treated by degradation of a target protein bound to the target protein ligand, the target protein preferably is cyclin-dependent kinase 4/6, androgen receptor, estrogen receptor, aurora kinase A, Bcr-Abl protein, epidermal growth factor receptor, mesenchymal lymphoma kinase, Bruton's tyrosine kinase, or bromodomain protein 4.
In another embodiment, a compound as described in the first or second aspect of the present invention, or a pharmaceutical composition as described in the third aspect of the present invention, for use in the treatment or prevention of disease that is treated by binding to a cereblon protein in vivo.
In another embodiment, a compound as described in the first or second aspect of the present invention, or a pharmaceutical composition as described in the third aspect of the present invention, for use in the treatment or prevention of a disease associated with the accumulation and/or aggregation of a target protein, the target protein is preferably cyclin-dependent kinase 4/6, androgen receptor, estrogen receptor, aurora kinase A, Bcr-Abl protein, epidermal growth factor receptor, mesenchymal lymphoma kinase, Bruton's tyrosine kinase, or bromodomain protein 4.
In another embodiment, the disease is a tumor or cancer, preferably the tumor or cancer is breast cancer, ductal carcinoma of the breast, prostate cancer, condylomatous lymphoma, chronic myeloid leukemia, acute myeloid leukemia, myelomonocytic leukemia, non-small cell lung cancer, lung adenocarcinoma, and/or cervical cancer.
In another embodiment, a method for treating or preventing a disease treated by degrading a target protein bound to a target protein ligand, comprising administering to a subject in need thereof a therapeutically effective amount of a compound described in the first or second aspect of the present invention, or a pharmaceutical composition described in the third aspect of the present invention, the target protein preferably is cyclin-dependent kinase 4/6, androgen receptor, estrogen receptor, aurora kinase A, Bcr-Abl protein, epidermal growth factor receptor, mesenchymal lymphoma kinase, Bruton's tyrosine kinase, or bromodomain protein 4.
In another embodiment, a method for treating or preventing a disease treated by binding to a cereblon protein, comprising administering a therapeutically effective amount of the compound described in the first or second aspect of the present invention, or a pharmaceutical composition described in the third aspect of the present invention.
In another embodiment, a method for treating or preventing a disease associated with the accumulation and/or aggregation of a target protein, comprising administering to a subject in need thereof a therapeutically effective amount of the compound described in the first or second aspect of the present invention, or a pharmaceutical composition described in the third aspect of the present invention, the target protein is preferably cyclin-dependent kinase 4/6, androgen receptor, estrogen receptor, aurora kinase A, Bcr-Abl protein, epidermal growth factor receptor, anaplastic lymphoma kinase, Bruton's tyrosine kinase, or bromodomain protein 4.
In another embodiment, the disease is a tumor or cancer, preferably the tumor or cancer is a breast cancer, breast ductal carcinoma, prostate cancer, mantle cell lymphoma, chronic myelogenous leukemia, acute myeloid leukemia, myelomonocytic leukemia, non-small cell lung cancer, lung adenocarcinoma, and/or cervical cancer.
A fourth aspect of the present invention discloses a method for inducing degradation of a target protein in cells, the method comprising administering an effective amount of the compound as described in the first or second aspect of the present invention, or a pharmaceutical composition as described in the third aspect of the present invention, the target protein is preferably cyclin-dependent kinase 4/6, androgen receptor, estrogen receptor, aurora kinase A, Bcr-Abl protein, epidermal growth factor receptor, anaplastic lymphoma kinase, Bruton's tyrosine Kinase, or bromodomain protein 4.
The term “alkyl” as considered herein refers to a saturated aliphatic hydrocarbon group which is a straight or branched chain group containing 1 to 20 carbon atoms, preferably containing 1 to 12 (e.g. 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 11 and 12) carbon atoms, more preferably an alkyl group containing 1 to 6 carbon atoms. Non-limiting examples include methyl, ethyl, n-propyl, isopropyl, n-butyl, isobutyl, t-butyl, sec-butyl, n-pentyl, 1,1-dimethylpropyl, 1,2-dimethylpropyl, 2,2-dimethylpropyl, 1-ethylpropyl, 2-methylbutyl, 3-methylbutyl, n-hexyl, 1-ethyl-2-methylpropyl, 1,1,2-trimethylpropyl, 1,1-dimethylbutyl, 1,2-dimethylbutyl, 2,2-dimethylbutyl, 1,3-dimethylbutyl, 2-ethylbutyl, 2-methylpentyl, 3-methylpentyl, 4-methylpentyl, 2,3-dimethylbutyl, n-heptyl, 2-methylhexyl, 3-methylhexyl, 4-methylhexyl, 5-methylhexyl, 2,3-dimethylpentyl, 2,4-dimethylpentyl, 2,2-dimethylpentyl, 3,3-dimethylpentyl, 2-ethylpentyl, 3-ethylpentyl, n-octyl, 2,3-dimethylhexyl, 2,4-dimethylhexyl, 2,5-dimethylhexyl, 2,2-dimethylhexyl, 3,3-dimethylhexyl, 4,4-dimethylhexyl, 2-ethylhexyl, 3-ethylhexyl, 4-ethylhexylylhexyl, 2-methyl-2-ethylpentyl, 2-methyl-3-ethylpentyl, n-nonyl, 2-methyl-2-ethylhexyl, 2-methyl-3-ethyl 2,2-diethylhexyl, 2,2-diethylpentyl, n-decyl, 3,3-diethylhexyl, 2,2-diethylhexyl, and various branched chain isomers, etc.
More preferably are lower alkyl groups containing 1 to 6 carbon atoms, non-limiting examples include methyl, ethyl, n-propyl, isopropyl, n-butyl, isobutyl, tert-butyl, sec-butyl, n-pentyl, 1,1-dimethylpropyl, 1,2-dimethylpropyl, 2,2-dimethylpropyl, 1-ethylpropyl, 2-methylbutyl, 3-methylbutyl, n-hexyl, 1-ethyl-2-methylpropyl, 1,1,2-trimethylpropyl, 1,1-dimethylbutyl, 1,2-dimethyl dimethylbutyl, 2,2-dimethylbutyl, 1,3-dimethylbutyl, 2-ethylbutyl, 2-methylpentyl, 3-methylpentyl, 4-methylpentyl, 2,3-dimethylbutyl, etc.
Alkyl groups may be substituted or unsubstituted, and when substituted, the substituents could substitute at any available point of attachment, preferably the substituents are independently and optionally selected from one or more of the groups consisting of H atom, D atom, halogen, alkyl, alkoxy, haloalkyl, hydroxyl, hydroxyalkyl, cyano, amino, nitro, cycloalkyl, heterocyclyl, aryl, heteroaryl.
The term “heteroalkyl” means that one or more —CH2— in the alkyl group is replaced by a heteroatom selected from NH, O and S or one or more —CH— in the alkyl group is replaced by an N atom; The alkyl group is as defined above; the heteroalkyl group may be substituted or unsubstituted, and when substituted, the substituent could substitute at any available point of attachment, preferably the substituents are independently and optionally selected from one or more of the groups consisting of H atom, D atom, halogen, alkyl, alkoxy, haloalkyl, hydroxyl, hydroxyalkyl, cyano, amino, nitro, cycloalkyl, heterocyclyl, aryl, heteroaryl.
The term “alkoxy” refers to —O-(alkyl) and —O-(unsubstituted cycloalkyl), wherein alkyl or cycloalkyl is as defined herein. Non-limiting examples of alkoxy include: methoxy, ethoxy, propoxy, butoxy, cyclopropoxy, cyclobutoxy, cyclopentyloxy, cyclohexyloxy. Alkoxy may be optionally substituted or unsubstituted, and when substituted, the substituents are preferably one or more of the following groups, independently selected from one or more of the groups consisting of H atom, D atom, halogen, alkyl, alkoxy, Haloalkyl, hydroxyl, hydroxyalkyl, cyano, amino, nitro, cycloalkyl, heterocyclyl, aryl, heteroaryl.
The term “alkenyl” refers to an alkyl compound containing a carbon-carbon double bond in the molecule, wherein the definition of alkyl is as above. Alkenyl groups may be substituted or unsubstituted, and when substituted, the substituents are preferably one or more of the following groups, independently selected from one or more of the groups consisting of hydrogen atom, alkyl, alkoxy, halogen, haloalkyl, hydroxyl, hydroxyalkyl, cyano, amino, nitro, cycloalkyl, heterocyclyl, aryl and heteroaryl.
The term “alkynyl” refers to an alkyl compound containing a carbon-carbon triple bond in the molecule, wherein alkyl is as defined above. The alkynyl group may be substituted or unsubstituted, and when substituted, the substituents are preferably one or more of the following groups, independently selected from one or more of the groups consisting of hydrogen atom, alkyl, alkoxy, halogen, haloalkyl, hydroxyl, hydroxyalkyl, cyano, amino, nitro, cycloalkyl, heterocyclyl, aryl and heteroaryl.
The term “cycloalkyl” refers to a saturated or partially unsaturated monocyclic cyclic hydrocarbon substituent, the cycloalkyl ring contains 3 to 20 carbon atoms, preferably 3 to 12 carbon atoms, more preferably 3 to 8 (e.g. 3, 4, 5, 6, 7 and 8) carbon atoms, more preferably comprising 4 to 7 carbon atoms.
Non-limiting examples of monocyclic cycloalkyls include cyclopropyl, cyclobutyl, cyclopentyl, cyclopentenyl, cyclohexyl, cyclohexenyl, cyclohexadienyl, cycloheptyl, cycloheptatrienyl, cyclooctyl, etc.
Cycloalkyl groups may be substituted or unsubstituted, and when substituted, the substituents could substitute at any available point of attachment, the substituents are preferably independently and optionally selected from one or more of the group consisting of hydrogen atom, halogen, alkyl, alkoxy, haloalkyl, hydroxyl, hydroxyalkyl, cyano, amino, nitro, cycloalkyl, heterocyclyl, aryl, heteroaryl.
The term “heterocyclyl” refers to a saturated or partially unsaturated monocyclic cyclic hydrocarbon substituent comprising 3 to 20 ring atoms, one or more of which are heteroatoms selected from nitrogen, oxygen or S(O)m (where m is an integer from 0 to 2), excluding ring portions of —O—O—, —O—S— or —S—S—, the remaining ring atoms being carbon. Preferably comprising 3 to 12 (e.g. 3, 4, 5, 6, 7, 8, 9, 10, 11 and 12) ring atoms, of which 1 to 4 (e.g. 1, 2, 3 and 4) are hetero atoms; more preferably containing 3 to 8 ring atoms, of which 1-3 are heteroatoms; more preferably containing 3 to 6 ring atoms, of which 1-3 are heteroatoms; most preferably containing 5 or 6 ring atoms, of which 1-3 are heteroatoms. Non-limiting examples of monocyclic heterocyclyl groups include pyrrolidinyl, tetrahydropyranyl, 1,2.3.6-tetrahydropyridyl, piperidinyl, piperazinyl, morpholinyl, thiomorpholinyl, homopiperazinyl, etc.
The heterocyclyl group may be substituted or unsubstituted, and when substituted, the substituents may be substituted at any available attachment point, the substituents are preferably independently and optionally selected from one or more of the group consisting of hydrogen atoms, halogens, alkyls, alkoxy, haloalkyl, hydroxyl, hydroxyalkyl, cyano, amino, nitro, cycloalkyl, heterocyclyl, aryl, heteroaryl. The term “aryl” refers to a 6 to 14 membered all-carbon monocyclic or fused polycyclic (a fused polycyclic is a ring sharing adjacent pairs of carbon atoms) group having a conjugated π-electron system, preferably 6 to 10 membered, such as phenyl and naphthyl. The aryl ring includes an aryl ring as described above fused to a heteroaryl, heterocyclyl or cycloalkyl ring, wherein the ring bonded to the parent structure is an aryl ring. Aryl groups may be substituted or unsubstituted, and when substituted, substituents may be substituted at any available point of attachment, the substituents are preferably independently and optionally selected from one or more of the group consisting of hydrogen atoms, halogen, alkyl, alkane oxy, haloalkyl, hydroxyl, hydroxyalkyl, cyano, amino, nitro, cycloalkyl, heterocyclyl, aryl, heteroaryl.
The term “heteroaryl” refers to a heteroaromatic system comprising 1 to 4 (e.g. 1, 2, 3 and 4) heteroatoms, 5 to 14 ring atoms, wherein the heteroatoms are selected from oxygen, sulfur and nitrogen. Heteroaryl is preferably 5 to 10 membered (e.g. 5, 6, 7, 8, 9 or 10 membered), more preferably 5 or 6 membered, e.g. furyl, thienyl, pyridyl, pyrrolyl, N-alkyl Pyrrolyl, pyrimidinyl, pyrazinyl, pyridazinyl, imidazolyl, pyrazolyl, triazolyl, tetrazolyl, etc. The heteroaryl ring includes a heteroaryl as described above fused to an aryl, heterocyclyl or cycloalkyl ring, wherein the ring attached to the parent structure is a heteroaryl ring. Heteroaryl groups may be substituted or unsubstituted, and when substituted, the substituents could substitute at any available point of attachment, preferably independently and optionally selected from one or more of the groups consisting of hydrogen atom, halogens, alkyl, alkoxy, haloalkyl, hydroxyl, hydroxyalkyl, cyano, amino, nitro, cycloalkyl, heterocyclyl, aryl, heteroaryl. The term “haloalkyl” refers to an alkyl group substituted with one or more halogens, wherein alkyl is as defined above.
The term “hydroxyalkyl” refers to an alkyl group substituted with one or more hydroxy groups, wherein alkyl is as defined above.
The term “halogen” refers to fluorine, chlorine, bromine or iodine.
The term “amino” refers to —NH2.
The term “cyano” refers to —CN.
The term “nitro” refers to —NO2.
The term “ubiquitin ligase” refers to a family of proteins that facilitate the transfer of ubiquitin to specific substrate proteins and target substrate proteins for degradation. For example, Cerebellin is an E3 ubiquitin ligase protein that alone or in combination with E2 ubiquitin conjugating enzymes results in the attachment of ubiquitin to lysines on target proteins and subsequent targeting of specific protein substrates for degradation by the proteasome. Thus, E3 ubiquitin ligases alone or in complex with E2 ubiquitin conjugating enzymes is the cause of the transfer of ubiquitin to target proteins. In general, ubiquitin ligases are involved in polyubiquitination so that the second ubiquitin is attached to the first ubiquitin, the third ubiquitin is attached to the second ubiquitin, and so on. Polyubiquitination marks proteins for degradation by the proteasome. However, there are some ubiquitination events that are restricted to monoubiquitination, where only a single ubiquitin is added to a substrate molecule by an ubiquitin ligase. Monoubiquitinated proteins are not targeted to the proteasome for degradation, but may instead be altered in their cellular location or function, for example via binding to other proteins with domains capable of binding ubiquitin. To complicate matters, different lysines on ubiquitin can be targeted by E3 to make chains. The most common lysine is Lys48 on the ubiquitin chain. This is the lysine used to make polyubiquitin, which is recognized by the proteasome.
The term “target protein” refers to proteins and peptides that have any biological function or activity (including structural, regulatory, hormonal, enzymatic, genetic, immune, contractile, storage, transport, and signal transduction). In some embodiments, target proteins include structural proteins, receptors, enzymes, cell surface proteins, proteins associated with the integrated function of the cell, including proteins involved in each of the following: catalytic activity, aromatase activity, motor activity, helical Enzyme activity, metabolic processes (anabolism and catabolism), antioxidant activity, proteolysis, biosynthesis, proteins with kinase activity, oxidoreductase activity, transferase activity, hydrolase activity, lyase activity, isomerase activity, ligase activity, enzyme regulator activity, signal transduction factor activity, structural molecule activity, binding activity (protein, lipid carbohydrate), receptor activity, cell motility, membrane fusion, cell communication, regulation of biological processes, development, cell differentiation, stimulus response, behavioral proteins, cell adhesion proteins, white matter involved in cell death, proteins involved in transport (including protein transport activity, nuclear transport, ion transport activity, channel transport activity, carrier activity), permease activity, secretion activity, electron transport activity, pathogenicity, chaperone regulator activity, nucleic acid binding activity, transcription regulator activity, extracellular conformation and biogenesis activity, translation regulator activity. The proteins include proteins from eukaryotes and prokaryotes, the eukaryotes and prokaryotes including microorganisms, viruses, fungi, and parasites, and numerous others, including humans, microorganisms, viruses, fungi, and parasites as targets for drug therapies. Other animals including domestic animals, microorganisms for determining the targets of antibiotics and other antimicrobials and plants and even viruses and numerous others.
The term “isomer” means that the compounds of the present invention may have asymmetric centers and racemates, racemic mixtures and individual diastereomers, all of which, including stereoisomers, geometric isomers are included in the present invention. In the present invention, when a compound or a salt thereof exists in a stereoisomeric form (for example, it contains one or more asymmetric carbon atoms), individual stereoisomers (enantiomers and diastereoisomers) and mixtures thereof are included within the scope of the present invention. The invention also includes individual isomers of the compounds or salts, as well as mixtures of isomers in which one or more chiral centers are inverted. The scope of the present invention includes mixtures of stereoisomers, as well as purified enantiomers or enantiomerically/diastereomerically enriched mixtures. The present invention includes mixtures of stereoisomers of all enantiomers and diastereoisomers in all possible different combinations. The present invention includes all combinations and subsets of stereoisomers of all specific groups defined above. The present invention also includes geometric isomers of the compounds or salts thereof, including cis-trans isomers.
The term “isotopically derivative” refers to compounds that differ in structure only by the presence of one or more isotopically enriched atoms. For example, having the structure of the present disclosure except replacing hydrogen with “deuterium” or “tritium”, or replacing fluorine with 18F-fluorine label (18F isotope), or replacing carbon with 11C-, 13C- or 14C-enriched carbon (11C-, 13C-, or 14C-carbon labels; 11C-, 13C-, or 14C-isotopes) are within the scope of the present disclosure. Such compounds are useful, for example, as analytical tools or probes in biological assays, or as tracers for in vivo diagnostic imaging of disease, or as tracers for pharmacodynamic, pharmacokinetic or receptor studies.
“Optional” or “optionally” means that the subsequently described event or circumstance can but need not occur, and the description includes occasions where the event or circumstance occurs or does not occur. For example, “optionally substituted cyclopropyl” means that cyclopropyl group may but need not be substituted, and the description includes cases where the cyclopropyl group is substituted and cases where the cyclopropyl group is unsubstituted.
“Substituted” means that one or more hydrogen atoms in a group, preferably up to 5, more preferably 1 to 3 hydrogen atoms are independently substituted by the corresponding number of substituents. It goes without saying that substituents are only in their possible chemical positions and that a person skilled in the art can determine possible or impossible substitutions without undue effort (by experiment or theory).
“Medicinal salts” or “pharmaceutically acceptable salts” refers to salts of the disclosed compounds, which are safe and effective when used in mammals, and have the desired biological activity.
The following examples are offered by way of illustration and not limitation.
Abbreviations used in this article are as follows:
The following examples are directed to intermediate compounds and final products identified in the specification and synthetic schemes. The preparation of the compounds of the invention is described in detail using the following examples, but the chemical reactions described are disclosed in terms of their general applicability to the preparation of the compounds of the invention. Occasionally, the reaction may not be applicable to every compound within the scope of the invention as described. Compounds in which this occurs are readily identified by those skilled in the art. In these cases, the reactions can be carried out successfully by routine modifications known to those skilled in the art. In all preparative methods, all starting materials are known or can be readily prepared using known starting materials.
The starting materials, chemical reagents, and solvents used in this disclosure are all commercially available and purchased from Energy Chemical, Bide Pharmatech, Beijing Inochem, Jiangsu Aikon, Sinopharm Group, Beijing J&K Scientific, and Yunnan Xinlan Jing and other companies.
Compound structures are determined by nuclear magnetic resonance (NMR) and/or mass spectroscopy (MS). The determination of nuclear magnetic resonance (NMR) is to use Bruke AVANCE-400/600 nuclear magnetic instrument, and the deuterated solvent used is deuterated dimethyl sulfoxide (DMSO-d6), deuterated chloroform (CDCl3), deuterated methanol (CD3OD), the internal standard is tetramethylsilane (TMS). Mass spectrometry (MS) is determined using Waters Acquity UPLC® Plus equipment. Waters 2489 equipment is used for HPLC preparation. Medium pressure flash preparative chromatograph uses COMBIFLASH NEXTGEN 300+ equipment. The thin-layer chromatography silica gel plate uses Silica gel 60 thin-layer chromatography silica gel plate (aluminum plate, containing fluorescence). Silica gel (100-200 mesh, 200-300 mesh) used in silica gel thin-layer chromatography was purchased from Inochem.
The reaction process detection in the embodiment adopts thin-layer chromatography (TLC), and the system of the developing agent used for monitoring the reaction and the eluent used for purifying the compound by column chromatography includes: petroleum ether/ethyl acetate system, dichloromethane/methanol system.
To a solution of compound 1a (30 g, 164.7 mmol) in methanol (300 mL) was slowly added concentrated sulfuric acid (45 mL). The reaction solution was stirred at 65° C. for 5 hours. The resulting mixture was poured into ice water, filtered and washed with water, and dried in vacuo to give white solid compound 1b (30.0 g, 87%).
1H NMR (600 MHz, DMSO-d6) δ 10.66 (s, 1H), 7.70 (d, J=8.5 Hz, 1H), 6.96 (dd, J=8.5, 2.5 Hz, 1H), 6.93 (d, J=2.5 Hz, 1H), 3.79 (s, 3H), 3.76 (s, 3H).
LC-MS (ESI): [M−H]+=209.22
To a solution of compound 1b (20.0 g, 95.2 mmol) in trifluoroacetic acid (60 mL), N-iodosuccinimide (23.6 g, 104.7 mmol) was added slowly. The reaction was stirred overnight at room temperature. The resulting mixture was concentrated under vacuum and purified by C18 reverse phase column to give compound 1c (16.3 g, 51%) as a white solid.
1H NMR (600 MHZ, DMSO-d6) δ 11.59 (s, 1H), 8.11 (s, 1H), 7.02 (s, 1H), 3.79 (s, 3H), 3.77 (s, 3H).
LC-MS (ESI): [M−H]+=335.06
Under the condition of nitrogen protection, compound 1c (15.0 g, 44.6 mmol), tributyl (1-ethoxyethylene) tin (32.2 g, 89.3 mmol) and bis(triphenylphosphine) palladium dichloride (II) (3.1 g, 4.5 mmol) was dissolved in tetrahydrofuran (75 mL), heated to 65° C. and stirred for 15 h. After the reaction was completed, 1M dilute hydrochloric acid solution was added to the system and stirred for 0.5 h, and then potassium fluoride was added. The mixture was filtered and the filtrate was concentrated to give crude product. And purified by column chromatography (PE:EA=0-40%) to obtain compound 1d (9.8 g, 87%) as a yellow oil.
1H NMR (400 MHZ, CDCl3) δ 12.65 (s, 1H), 8.37 (s, 1H), 7.12 (s, 1H), 3.96 (s, 3H), 3.92 (s, 3H), 2.73 (s, 3H).
LC-MS (ESI): [M+H]+=253.15
Compound 1d (5.0 g, 19.8 mmol), N-(tert-butoxycarbonyl)-4-piperidone (1.4 g, 19.8 mmol) and tetrahydropyrrole (4.0 g, 19.8 mmol) were dissolved in methanol (50 mL), the mixture was stirred overnight at 70° C. After spin-drying, column chromatography (PE:EA=0-35%) purified to obtain compound 1e (6.9 g, 80%) as a yellow oil.
1H NMR (600 MHZ, DMSO-d6) δ 8.17 (s, 1H), 7.36 (s, 1H), 3.83 (s, 3H), 3.82 (s, 3H), 3.73 (s, 2H), 3.13 (d, J=40.7 Hz, 2H), 2.96 (s, 2H), 1.95-1.86 (m, 2H), 1.66 (dq, J=13.2, 5.0 Hz, 2H), 1.40 (s, 9H).
LC-MS (ESI): [M−H]+=432.22
To a solution of compound 1e (5.0 g, 11.5 mmol) in methanol (50 mL) was added sodium borohydride (0.7 g, 17.3 mmol) under ice-bath conditions. The reaction was stirred overnight at 70° C. The resulting mixture was concentrated in vacuo and purified by column chromatography (PE:EA=0-50%) to give compound 1f (3.3 g, 66%) as a white solid.
1H NMR (600 MHz, DMSO-d6) δ 7.92 (s, 1H), 7.02 (s, 1H), 5.73 (d, J=6.2 Hz, 1H), 4.74 (dt, J=9.3, 6.1 Hz, 1H), 3.79 (s, 6H), 3.74-3.64 (m, 2H), 3.06 (s, 2H), 2.18 (dd, J=13.6, 6.1 Hz, 1H), 1.78 (ddt, J=23.0, 13.5, 6.3 Hz, 2H), 1.72-1.64 (m, 2H), 1.57 (ddd, J=13.7, 11.3, 4.7 Hz, 1H), 1.40 (s, 9H).
LC-MS(ESI): [M−H]+=464.39
Compound 1f (3.0 g, 6.9 mmol) and triethylsilane (3.2 g, 27.6 mmol) were dissolved in trifluoroacetic acid (30 mL), and the mixture was stirred overnight at 80° C. The resulting mixture was concentrated in vacuo and purified by C18 reverse phase column to give Compound 1g (1.8 g, 83%) as a colorless oil.
1H NMR (400 MHz, CDCl3) δ 7.53 (s, 1H), 7.11 (s, 1H), 6.51 (d, J=9.9 Hz, 1H), 5.72 (d, J=9.8 Hz, 1H), 4.66 (s, 2H), 3.93 (s, 3H), 3.89 (s, 3H), 3.42-3.31 (m, 4H), 2.23 (d, J=14.6 Hz, 2H), 2.12-2.02 (m, 2H).
LC-MS(ESI): [M+H]+=318.25
Compound 1g (1.6 g, 5.0 mmol) was dissolved in methanol (20 mL), and palladium on carbon (0.2 g) was added. The reaction was stirred overnight at room temperature under hydrogen. The mixture was filtered and spin-dried to give compound 1h (1.3 g, 81%) as a yellow oil.
1H NMR (400 MHz, CDCl3) δ 7.61 (s, 1H), 7.14 (s, 1H), 3.93 (s, 3H), 3.89 (s, 3H), 3.35 (d, J=6.2 Hz, 4H), 3.30 (s, 2H), 2.87 (t, J=6.8 Hz, 2H), 2.05-2.00 (m, 2H), 1.95 (t, J=6.7 Hz, 2H).
LC-MS(ESI): [M+H]+=320.28
Compound 1h (1.5 g, 4.7 mmol) was dissolved in a suspension of methanol and water, and lithium hydroxide (1.7 g, 70.5 mmol) was added. The reaction was stirred overnight at room temperature. The reaction mixture was concentrated in vacuo. The crude product was used directly in the next step without further purification.
LC-MS(ESI): [M+H]+=292.21
Compound 1i (1.3 g, 7.7 mmol) was dissolved in acetic acid (15 mL) solution, 3-aminopiperidine-2,6-dione hydrochloride (1.52 g, 9.2 mmol) and sodium acetate (2.1 g, 15.5 mmol), the reaction was stirred for 4 hours at 110° C. After the reaction was completed, the mixture was purified by reverse phase column to obtain white solid compound 1 (1.6 g, 81%).
1H NMR (400 MHz, DMSO-d6) δ 11.10 (s, 1H), 7.74 (s, 1H), 7.38 (s, 1H), 5.10 (dd, J=12.9, 5.4 Hz, 1H), 3.28-3.14 (m, 4H), 2.95 (t, J=6.8 Hz, 2H), 2.91-2.83 (m, 1H), 2.65-2.53 (m, 2H), 2.07-1.99 (m, 1H), 1.93 (q, J=6.2 Hz, 4H), 1.88-1.78 (m, 2H).
LC-MS(ESI): [M+H]+=384.32.
Compound 1d was prepared with reference to Step 1 to Step 3 of Example 1. The obtained compound 1d (5.0 g, 19.8 mmol), 1-tert-butoxycarbonyl-3-pyrrolidone (1.4 g, 19.8 mmol) and tetrahydropyrrole (4.0 g, 19.8 mmol) were dissolved in methanol (50 mL), and the mixture was stirred overnight at 70° C. After spin-drying, purified by column chromatography (PE:EA=0-35%) to obtain compound 2a (5.0 g, 60%) as a yellow oil.
1H NMR (400 MHz, CDCl3) δ 8.44 (s, 1H), 7.18 (s, 1H), 3.95 (s, 3H), 3.91 (s, 3H), 3.89-3.83 (m, 1H), 3.76-3.64 (m, 1H), 3.62-3.50 (m, 1H), 3.40 (dd, J=17.4, 12.4 Hz, 1H), 3.06-2.86 (m, 2H), 2.36-2.25 (m, 1H), 1.97 (ddd, J=13.5, 10.4, 9.0 Hz, 1H), 1.47 (d, 9H).
LC-MS(ESI): [M−H]+=418.40
Compound 2b was prepared referring to Step 5 of Example 1.
LC-MS(ESI): [M+Na]+=444.33
Compound 2c was prepared referring to Step 6 of Example 1.
1H NMR (600 MHz, CDCl3) δ 7.53 (s, 1H), 7.09 (s, 1H), 6.61 (d, J=9.8 Hz, 1H), 5.79 (d, J=9.9 Hz, 1H), 4.53 (br s, 2H), 3.91 (s, 3H), 3.89 (s, 3H), 3.70-3.55 (m, 3H), 3.27 (d, J=12.5 Hz, 1H), 2.54 (dd, J=14.2, 6.4 Hz, 1H), 2.14 (ddd, J=13.8, 11.0, 6.5 Hz, 1H).
LC-MS(ESI): [M+H]+=304.27
Compound 2d was prepared referring to Step 7 of Example 1.
1H NMR (600 MHz, CDCl3) δ 7.61 (s, 1H), 7.09 (s, 1H), 3.90 (s, 3H), 3.88 (s, 3H), 3.52 (dtd, J=17.8, 11.4, 7.8 Hz, 2H), 3.44 (dd, J=12.7, 1.6 Hz, 1H), 3.25 (d, J=12.6 Hz, 1H), 2.90 (dtd, J=17.3, 10.7, 6.9 Hz, 2H), 2.28-2.23 (m, 1H), 2.14-2.10 (m, 2H), 2.08-2.02 (m, 1H).
LC-MS(ESI): [M+H]+=306.22
Compound 2e was prepared referring to Step 8 of Example 1.
LC-MS(ESI): [M+H]+=278.22
Compound 2 was prepared referring to Step 9 of Example 1.
1H NMR (400 MHz, DMSO-d6) δ 11.11 (s, 1H), 7.78 (s, 1H), 7.18 (s, 1H), 5.10 (dd, J=12.9, 5.4 Hz, 1H), 3.50-3.21 (m, 4H), 3.03-2.84 (m, 4H), 2.66-2.53 (m, 2H), 2.24-2.00 (m, 4H).
LC-MS(ESI): [M+H]+=370.34
7′-(2,6-dioxopiperidin-3-yl)-3′,4′-dihydro-6′H-spiro[azetidine-3,2′-pyrano[2,3-f]isoindole]-6′,8′(7′H)-dione Prepare 7′-(2,6-dioxopiperidin-3-yl)-3′,4′-dihydro-6′H-spiro[azetidine-3,2′-pyrano[2,3-f]isoindole]-6′,8′(7′H)-dione by using the similar step from step 4 to step 9 in example 1.
1H NMR (600 MHz, DMSO-d6) δ 11.12 (s, 1H), 7.76 (s, 1H), 7.30 (s, 1H), 5.10 (dd, J=12.9, 5.4 Hz, 1H), 4.16 (t, J=8.7 Hz, 4H), 2.97 (t, J=6.5 Hz, 2H), 2.88 (ddd, J=17.0, 13.9, 5.5 Hz, 1H), 2.59 (dt, J=17.1, 3.1 Hz, 1H), 2.54 (dd, J=13.1, 4.5 Hz, 1H), 2.22 (t, J=6.5 Hz, 2H), 2.06-2.00 (m, 1H).
LC-MS(ESI): [M+H]+=356.26
Compound 4a (10 g, 55.56 mmol) was dissolved in 100 mL of methanol, then 15 mL of concentrated sulfuric acid was added, and reacted overnight at room temperature. The reaction solution was poured into ice water, extracted with ethyl acetate and concentrated to obtain compound 4b (10.6 g, 91%) as a colorless oil, which was directly used in the next step without purification.
1H NMR (400 MHz, CDCl3) δ 7.66 (d, J=7.9 Hz, 1H), 7.46 (d, J=1.7 Hz, 1H), 7.32 (dd, J=7.9, 1.8 Hz, 1H), 3.89 (s, 3H), 3.88 (s, 3H), 2.40 (s, 3H).
Compound 4b (10 g, 48 mmol) was dissolved in 100 mL of concentrated sulfuric acid, then concentrated nitric acid (25 mL, 68%) was slowly added, and reacted overnight at room temperature. After the reaction, the reaction solution was poured into ice water, extracted with ethyl acetate and concentrated. The crude product was purified by column chromatography to obtain compound 4c (5.5 g, 45%) as a white solid.
1H NMR (400 MHz, CDCl3) δ 8.43 (s, 1H), 7.64 (s, 1H), 3.97 (s, 3H), 3.96 (s, 3H), 2.69 (s, 3H).
Compound 4c (5.1 g, 20 mmol) was dissolved in 100 mL of methanol, then 0.51 g of palladium on carbon was added, and the reaction system was replaced with nitrogen and hydrogen, and reacted overnight at room temperature. After the reaction was completed, it was filtered with celite, and the filtrate was collected and concentrated to obtain compound 4d (4 g, 91%) as a yellow oil, which was directly used in the next step without purification.
1H NMR (400 MHz, DMSO-d6) δ 7.46 (s, 1H), 6.68 (s, 1H), 5.91 (s, 2H), 3.75 (s, 3H), 3.71 (s, 3H).
LC-MS(ESI): [M+Na]+=246.18
Compound 4d (4 g, 17.92 mmol) was added into 10% fluoboric acid (32 ml), and the solution was suspended. After stirring for 0.5 h, cool to 0-5° C., and slowly add 4 mL NaNO2 (1.36 g, 19.71 mmol) aqueous solution for diazotization reaction, and stir in ice bath for 0.5 h. The tetrafluoroborate was filtered and the cake was collected and the solid was dried under vacuum. Then, the toluene solution of tetrafluoroborate was placed in an oil bath at 110° C. and stirred. After the reaction was completed, it was extracted with ethyl acetate, and the organic phase was washed with water and saturated brine, and dried with anhydrous sodium sulfate. The crude product was concentrated and purified by column chromatography to give compound 4e (2.31 g, 57%) as a light yellow oil.
1H NMR (400 MHz, CDCl3) δ 7.61 (dd, J=7.2, 0.9 Hz, 1H), 7.38 (d, J=9.4 Hz, 1H), 3.92 (s, 3H), 3.91 (s, 3H), 2.35 (d, J=2.0 Hz, 3H).
Compound 4e (2.26 g, 10 mmol) was dissolved in carbon tetrachloride, then N-bromosuccinimide (2.14 g, 12 mmol) and azobisisobutyronitrile (0.16 g, 1 mmol) were added, and heated to reflux overnight.
After concentration, purification was performed by reverse column to obtain white solid compound 4f (2.14 g, 70%).
1H NMR (400 MHz, CDCl3) δ 7.87 (d, J=7.1 Hz, 1H), 7.40 (d, J=9.4 Hz, 1H), 4.51 (d, J=1.0 Hz, 2H), 3.94 (s, 3H), 3.93 (s, 3H).
Compound 4f (2.14 g, 7 mmol) was dissolved in anhydrous tetrahydrofuran (30 mL), and potassium tert-butoxide (1.18 g, 10.5 mmol) was slowly added at −30° C. After reacting for 0.5 h, a solution of 1-tert-butoxycarbonylpiperidine-4-carbaldehyde (1.79 g, 8.4 mmol) in tetrahydrofuran (5 mL) was added dropwise at the same temperature. The reaction was moved to room temperature and stirred overnight. The reaction system was quenched with saturated ammonium chloride, and extracted with ethyl acetate. The organic phase was washed with saturated brine, dried over anhydrous sodium sulfate, concentrated and purified by flash silica gel column chromatography to obtain compound 4g (1.68 g, 55%) as a colorless oil.
1H NMR (400 MHz, CDCl3) δ 9.58 (d, J=2.3 Hz, 1H), 7.50 (d, J=7.1 Hz, 1H), 7.34 (d, J=9.5 Hz, 1H), 3.96-3.84 (m, 8H), 2.91-2.74 (m, 4H), 1.96 (d, J=13.0 Hz, 2H), 1.59-1.46 (m, 2H), 1.44 (s, 9H).
LC-MS(ESI): [M-Boc+H]+=338.29
Compound 4g (1.66 g, 3.8 mmol) was dissolved in methanol (15 mL), and sodium borohydride (0.215 g, 5.7 mmol) was added under ice-cooling conditions, and stirred at room temperature for 3 h. The mixture was quenched with saturated ammonium chloride, and extracted with ethyl acetate, the organic phase was washed with saturated brine, dried over anhydrous sodium sulfate, concentrated and purified by column chromatography to obtain colorless oil compound 4h (1.47 g, 88%).
1H NMR (400 MHz, DMSO-d6) δ 7.74 (d, J=7.0 Hz, 1H), 7.54 (d, J=9.7 Hz, 1H), 4.81 (t, J=5.0 Hz, 1H), 3.82 (s, 3H), 3.82 (s, 3H), 3.48-3.39 (m, 2H), 3.21 (d, J=5.0 Hz, 4H), 2.74 (s, 2H), 1.42-1.32 (m, 11H), 1.27-1.14 (m, 2H).
LC-MS(ESI): [M-Boc+H]+=340.41
Compound 4h (1.45 g, 3.3 mmol) was dissolved in dry N,N-dimethylformamide (6 mL), sodium hydride (0.4 g, 16.5 mmol) was added, and the reaction mixture was carried out at 110° C. for 2 h. After the reaction was completed, it was quenched with acetic acid and purified by a reverse column to obtain white solid compound 4i (0.65 g, 51%).
1H NMR (400 MHz, DMSO-d6) δ 12.93 (s, 2H), 7.50 (s, 1H), 6.92 (s, 1H), 3.99 (s, 2H), 3.53-3.42 (m, 2H), 3.33-3.23 (m, 2H), 2.75 (s, 2H), 1.40 (s, 9H), 1.39-1.28 (m, 4H).
LC-MS(ESI): [M-Boc+H]+=292.23
Compound 4i (0.63 g, 1.6 mmol) was dissolved in acetic acid (4 mL), and 3-amino-2,6-piperidinedione hydrochloride (0.33 g, 2.0 mmol) and sodium acetate (0.39 g, 4.8 mmol) were added, the reaction mixture was carried out overnight at 110° C. After the reaction was completed, it was purified by reverse column to obtain white solid compound 4 (0.5 g, 82%).
1H NMR (400 MHz, DMSO-d6) δ 11.12 (s, 1H), 8.51 (s, 2H), 7.69 (s, 1H), 7.29 (s, 1H), 5.10 (dd, J=12.7, 5.4 Hz, 1H), 4.16 (s, 2H), 3.25-3.05 (m, 4H), 2.92 (s, 2H), 2.90-2.82 (m, 1H), 2.64-2.52 (m, 2H), 2.05-1.99 (m, 1H), 1.67-1.51 (m, 4H).
LC-MS(ESI): [M+H]+=384.36
To a solution of compound 5a (5.5 g, 30.0 mmol) in methanol (100 mL) was slowly added concentrated sulfuric acid (15 mL) dropwise. The reaction was stirred overnight at room temperature. The reaction solution was poured into ice water, extracted with ethyl acetate, the organic phases were combined, washed with saturated brine, and dried with anhydrous sodium sulfate. After filtration and concentration under reduced pressure, Compound 5b (5.7 g, 90%) was obtained as a colorless oily liquid, and the crude product was used in the next reaction without further purification.
1H NMR (400 MHz, CDCl3) δ 7.82 (dd, J=8.6, 5.3 Hz, 1H), 7.38 (dd, J=8.6, 2.6 Hz, 1H), 7.82 (ddd, J=8.6, 5.3, 2.6 Hz, 1H), 3.96 (s, 3H), 3.92 (s, 3H).
Pinacol diboronate (4.5 g, 17.5 mmol), 4,4′-di-tert-butyl-2,2′-dipyridine (0.13 g, 0.47 mmol) and methoxy(cyclooctadiene) Iridium dimer (0.16 g, 0.23 mmol) were added in the methyl tert-butyl ether solution of 10 ml. The methyl tert-butyl ether (10 mL) solution of compound 5b (2.5 g, 11.8 mmol) was added, and replaced with nitrogen. The reaction mixture was stirred at 100° C. for 4 h. After the reaction was completed, it was filtered through diatomaceous earth, and the filtrate was concentrated to obtain crude product (3.2 g), which was used in the next reaction without further purification.
To a solution of compound 5c (3.38 g, 10 mmol) in acetonitrile (50 mL) was added potassium peroxymonosulfonate (6.15 g), and the reaction was stirred at room temperature overnight. The reaction solution was filtered and the filtrate was concentrated, and the crude product was subjected to silica gel column chromatography to obtain compound 5d (1.6 g, 70%) as a white solid.
1H NMR (400 MHz, CDCl3) δ 8.39 (s, 1H), 7.50 (d, J=10.8 Hz, 1H), 7.19 (d, J=8.1 Hz, 1H), 3.88 (s, 3H), 3.86 (s, 3H).
LC-MS(ESI): [M+H]+=229.18
Compound 5d (23 mg, 0.1 mmol) was dissolved in 1 ml of ultra-dry N,N-dimethylformamide, followed by the addition of benzyl 1-oxa-6-azaspiro[2.5]octane-6-carboxylate (25 mg, 0.1 mmol) and sodium hydride (6 mg, 0.25 mmol), the reaction was heated to 110° C. and stirred for 2 days. The reaction solution was prepared by reverse preparation to obtain white solid compound 5e (13 mg, 31%).
1H NMR (600 MHz, CD3OD) δ 7.41-7.35 (m, 4H), 7.33 (dd, J=5.9, 2.9 Hz, 1H), 7.29 (s, 1H), 7.27 (s, 1H), 5.15 (s, 2H), 4.07 (s, 2H), 4.01 (dt, J=13.6, 4.0 Hz, 2H), 3.42-3.32 (m, 2H), 1.82 (d, J=13.9 Hz, 2H), 1.76-1.67 (m, 2H).
LC-MS(ESI): [M+H]+=428.36
Add 3-aminopiperidine-2,6-dione hydrochloride (6 mg, 0.036 mmol) and sodium acetate (7 mg, 0.09 mmol) to a solution of compound 5e (13 mg, 0.03 mmol) in acetic acid (1 mL), and the reaction mixture was heated to 110° C. and stirred for 4 hours. After the reaction was completed, the reaction solution was subjected to reverse purification to obtain white solid compound 5f (12 mg, 77%).
1H NMR (600 MHz, CD3OD) δ 7.42 (s, 1H), 7.40-7.31 (m, 6H), 5.15 (s, 2H), 5.08 (dd, J=12.9, 5.5 Hz, 1H), 4.14 (s, 2H), 4.01 (dt, J=13.8, 4.0 Hz, 2H), 3.43-3.32 (m, 2H), 2.91-2.83 (m, 1H), 2.79-2.69 (m, 2H), 2.14-2.09 (m, 1H), 1.83 (d, J=13.9 Hz, 2H), 1.74 (td, J=14.4, 12.9, 4.8 Hz, 2H).
LC-MS(ESI): [M+H]+=520.29
Palladium carbon (5 mg) was added to a methanol solution of compound 5f (11 mg, 0.02 mmol), replaced by hydrogen, and the reaction solution was stirred under hydrogen for 6 hours. The reaction solution was filtered through celite, and the filtrate was concentrated to obtain compound 5 (7 mg, 90%) as a white solid.
1H NMR (400 MHz, DMSO-d6) δ 11.11 (s, 1H), 7.50 (s, 1H), 7.48 (s, 1H), 5.09 (dd, J=12.8, 5.3 Hz, 1H), 4.27 (s, 2H), 3.25 (d, J=12.7 Hz, 2H), 3.18-3.11 (m, 2H), 2.89 (ddd, J=16.7, 13.7, 5.3 Hz, 1H), 2.64-2.53 (m, 2H), 2.03 (ddd, J=13.3, 5.7, 3.4 Hz, 1H), 1.92-1.86 (m, 4H).
LC-MS(ESI): [M+H]+=386.32
Under the condition of nitrogen protection, compound 6a (5 g, 25.1 mmol) and triphenylphosphine (26.3 g, 100.4 mmol) were dissolved in anhydrous acetonitrile (50 mL), and tetrabromo carbonization (16.7 g, 50.2 mmol) was added to the reaction system in batches at 0° C., after reacting for 30 min, the reaction system was raised to room temperature, and reacted overnight. After the reaction was completed, the reaction system was filtered, and the obtained filtrate was concentrated under reduced pressure to obtain crude product, which was purified by column chromatography to obtain a white solid compound 6b (5.5 g, 62%).
1H NMR (600 MHz, DMSO-d6) δ 3.37 (s, 4H), 2.41 (dd, J=7.2, 4.7 Hz, 4H), 1.41 (d, J=0.9 Hz, 9H).
Under the condition of nitrogen protection, compound 6b (5.2 g, 18.9 mmol) was dissolved in tetrahydrofuran (34 mL) and methanol (17 mL), ammonium chloride (4.04 g, 75.6 mmol) was added to the reaction system at 0° C., After reacting at 0° C. for 30 min, zinc powder (4.9 g, 75.6 mmol) was added in batches, and reacted overnight at room temperature. After the reaction was completed, the reaction system was filtered, the filter cake was washed with methanol, the obtained filtrate was spin-dried under reduced pressure, and the concentrated crude product was purified by column chromatography to obtain white oily compound 6c (2.6 g, 50%).
1H NMR (600 MHz, DMSO-d6) δ 6.26 (t, J=1.3 Hz, 1H), 3.34 (dt, J=16.4, 6.3 Hz, 4H), 2.32-2.27 (m, 2H), 2.23 (ddd, J=7.2, 4.4, 1.2 Hz, 2H), 1.41 (s, 9H).
Under the condition of nitrogen protection, compound 6c (2 g, 7.3 mmol), compound 5c (2.64 g, 7.8 mmol) prepared by step 1 to step 2 of Example 5, palladium acetate (163 mg, 0.73 mmol), triphenylphosphine (382 mg, 1.46 mmol), cesium carbonate (7.14 g, 21.9 mmol) were added to the reaction flask in turn, 1,4-dioxane (20 mL) and water (2 mL) were added, and after nitrogen replacement, the reaction was carried out at 110° C. for 4 hours. After the reaction was completed, the reaction system was cooled to room temperature, and after adding water, it was extracted with ethyl acetate. The organic phase was washed with saturated brine and dried over anhydrous sodium sulfate. The concentrated crude product was purified by column chromatography to obtain light yellow oily compound 6d (1.4 g, 47%).
1H NMR (600 MHz, DMSO-d6) δ 7.65 (d, J=7.0 Hz, 1H), 7.60 (d, J=9.7 Hz, 1H), 6.32 (s, 1H), 3.83 (s, 3H), 3.82 (s, 3H), 3.43 (t, J=5.8 Hz, 2H), 3.35 (s, 1H), 3.33 (s, 1H), 2.37-2.32 (m, 2H), 2.25 (t, J=5.9 Hz, 2H), 1.42 (s, 9H).
LC-MS(ESI): [M-Boc+H]+=308.28.
Under the condition of nitrogen protection, compound 6d (7 g, 17.2 mmol) was dissolved in tetrahydrofuran (140 mL) and water (140 mL), N-bromosuccinimide (6.12 g, 34.4 mmol) was added, and the reaction mixture was carried out overnight at room temperature. After the reaction was completed, excess tetrahydrofuran was removed by concentration under reduced pressure, and after adding water, it was extracted with ethyl acetate. The organic phase was washed with saturated brine and dried over anhydrous sodium sulfate. The concentrated crude product was purified by column chromatography to obtain light yellow oily compound 6e (4.9 g, 56%).
LC-MS(ESI): [M-Boc+H]+=404.30/406.30.
Under the condition of nitrogen protection, compound 6e (10.9 g, 21.61 mmol) was dissolved in toluene (80 ml), and tris(trimethylsilyl)silane (8.06 g, 32.42 mmol) and azobisisobutyronitrile (357 mg, 2.16 mmol) were added, the reaction mixture was carried out overnight at 90° C. After the reaction was completed, excess toluene was removed by concentration under reduced pressure, and the concentrated crude product was purified by column chromatography to obtain light yellow oily compound 6f (1.5 g, 16%).
1H NMR (600 MHz, DMSO-d6) δ 7.77 (d, J=6.9 Hz, 1H), 7.52 (d, J=9.5 Hz, 1H), 4.62 (s, 1H), 3.82 (d, J=2.1 Hz, 6H), 3.66 (s, 2H), 3.34 (s, 4H), 2.79 (s, 2H), 1.38 (s, 9H).
LC-MS(ESI): [M-Boc+H]+=326.37.
Under nitrogen protection, compound 6f (1.6 g, 3.76 mmol) was dissolved in N,N-dimethylformamide (2 mL), sodium hydride (300 mg, 7.52 mmol) was added, and the reaction mixture was carried out overnight at 110° C. After the reaction was completed, water was added, followed by extraction with ethyl acetate. The organic phase was washed with saturated brine, and dried over anhydrous sodium sulfate. The concentrated crude product was purified by reverse purification to obtain 6 g (660 mg, 43%) of a light yellow solid compound.
1H NMR (600 MHz, CDCl3) δ 7.58 (s, 1H), 7.15 (s, 1H), 3.93 (s, 3H), 3.89 (s, 3H), 3.56 (t, J=5.7 Hz, 2H), 3.42 (t, J=5.8 Hz, 2H), 2.42 (t, J=5.9 Hz, 2H), 2.24 (t, J=5.9 Hz, 2H), 1.61 (s, 2H), 1.50 (s, 9H).
LC-MS(ESI): [M-Boc+H]+=306.23.
Under the condition of nitrogen protection, compound 6g (660 mg, 1.63 mmol) was dissolved in methanol (9 ml) and water (1 ml), lithium hydroxide (389 mg, 16.28 mmol) was added, and the reaction mixture was carried out overnight at room temperature. The concentrated crude product was purified by reverse purification to obtain light yellow solid compound 6h (600 mg, 97%).
1H NMR (400 MHz, DMSO-d6) δ 7.39 (s, 1H), 6.37 (s, 1H), 3.17 (s, 2H), 2.29 (d, J=51.9 Hz, 4H), 1.98 (dd, J=13.7, 6.9 Hz, 2H), 1.64 (s, 2H), 1.41 (s, 9H).
LC-MS(ESI): [M-Boc+H]+=278.26.
Compound 6h (400 mg, 1.06 mmol) was dissolved in acetic acid (5 mL) solution, 3-aminopiperidine-2,6-dione hydrochloride (169 mg, 1.32 mmol) and sodium acetate (260 mg, 3.18 mmol) were added, the reaction was stirred at 110° C. for 4 hours. After the reaction was completed, the mixture was purified by reverse phase column to obtain compound 6 (170 mg, 43%) as a white solid.
1H NMR (400 MHz, DMSO-d6) δ 7.46 (s, 1H), 7.11 (s, 1H), 5.04 (dd, J=12.8, 5.4 Hz, 1H), 2.99-2.79 (m, 5H), 2.62-2.54 (m, 1H), 2.53 (s, 1H), 2.38 (q, J=6.3 Hz, 4H), 2.04-1.96 (m, 1H), 1.90 (s, 2H).
LC-MS(ESI): [M+H]+=370.34.
Compound 7a (5 g, 33 mmol) was dissolved in anhydrous acetonitrile (50 mL). Tetrabutylamonium bromide (1.23 g, 3.3 mmol) and anhydrous potassium carbonate (14 g, 99 mmol) were added to the solution. Benzyl chloride (5.5 g, 43 mmol) was slowly added to the system and the reaction was carried out overnight at 30° C. The reaction was quenched with water and extracted with ethyl acetate. The organic phase was washed with saturated brine, dried over anhydrous sodium sulfate, and concentrated to obtain compound 7b (7.5 g, 94) as a white solid.
1H NMR (400 MHz, CDCl3) δ 7.37-7.20 (m, 5H), 5.95-5.83 (m, 2H), 4.63 (s, 2H), 3.20-3.01 (m, 2H), 2.61 (m, 2H), 2.31-2.10 (m, 2H).
LC-MS(ESI): [M+H]+=242.22
At 0° C., lithium aluminum tetrahydrogen (3.2 g, 82 mmol) was added to 40 mL of anhydrous tetrahydrofuran, then compound 7b (5 g, 20.7 mmol) was added, and after 5 minute s of ice bathing. the reaction system was moved to a 70C oil bath, reacted for 3 h. After the reaction was completed, water was slowly added dropwise to the reaction solution under ice bath conditions until no bubbles were generated. The filtrate was collected by filtration, extracted with ethyl acetate, and the organic phase was washed with saturated brine and dried over anhydrous sodium sulfate. The compound 7c (3.2 g, 73%) as a yellow oil was obtained by concentration.
1H NMR (400 MHz, CDCl3) δ 7.43-7.24 (m, 5H), 5.87 (t, J=2.7 Hz, 2H), 3.67 (s, 2H), 3.05-2.90 (m, 2H), 2.44 (m, 2H), 2.29, 2.16 (m, 4H), 1.92 (m, 2H).
Compound 7c (5 g, 23.44 mmol) was dissolved in 20 mL of tetrahydrofuran at 0° C., and 25 mL of 2M borane-tetrahydrofuran complex was added under ice-cooling conditions. After 12 hours of reaction, 13 mL of anhydrous methanol, 10.5 mL 3M NaOH solution and 10.5 mL hydrogen peroxide were added to the reaction solution. React at 60° C. for 6 h. After the reaction was completed, it was cooled to room temperature and extracted with ethyl acetate. The organic phase was saturated with saline and dried over anhydrous sodium sulfate. The organic phase was concentrated to obtain a crude product, which was purified by column chromatography to obtain compound 7d (1.2 g, 22.20).
1H NMR (400 MHz, CDCl3) δ 7.43-7.18 (m, 5H), 3.84 (m, 1H), 3.78 (s, 2H), 2.94 (dd, J=9.8, 6.7 Hz, 1H), 2.82 (dd, J=9.5, 7.7 Hz, 1H), 2.64 (dd, J=9.5, 8.4 Hz, 1H), 2.60-2.44 (m, 2H), 2.12 (m, 1H), 1.89-1.72 (m, 3H), 1.54 (m, 1H), 1.47-1.35 (m, 1H), 1.35-1.22 (in, 2H).
LC-MS(ESI): [M+H]+=232.31
Compound 7d (1.2 g, 12.97 mmol) was dissolved in anhydrous dichloromethane, and Dess Martin oxidant (11 g, 25.95 mmol) was added under ice-cooling conditions, and the reaction was reacted for 12 h. Quench the reaction with saturated sodium bicarbonate and saturated sodium thiosulfate 1:1, collect the filtrate, after filtration, the filtrate was extracted with dichloromethane. Wash with saturated brine, combine the organic phases and dry over anhydrous sodium sulfate. The crude product (1 g) was obtained by concentration and was directly used in the next reaction.
N,N-Dimethylformamide (1.01 mL, 13.08 mmol) was dissolved in 2 mL of dichloromethane at 0° C. Phosphorus tribromide (1.13 mL, 11.77 mmol) was slowly added dropwise, and stirred at 0° C. for 1 hour. The dichloro solution of compound 7e (1 g, 2.62 mmol) was added dropwise to the above system, and the temperature was raised to room temperature for 10 h. After the reaction, the reaction solution was placed at 0° C. and saturated sodium bicarbonate was added until no bubbles were generated. Extract with dichloromethane. The organic phase was washed with saturated brine and dried with anhydrous sodium sulfate. The rude product was obtained by concentration and purified by column chromatography to obtain the target compound 7f (270 mg, 20%).
1H NMR (400 MHz, CDCl3) δ 10.04 (s, 1H), 7.32 (d, J=5.8 Hz, 5H), 3.60 (s, 2H), 3.21-3.10 (m, 2H), 2.85-2.72 (m, 3H), 2.50-2.43 (m, 1H), 2.33 (dd, J=9.2, 5.0 Hz, 1H), 2.24-2.18 (m, 1H), 1.83-1.74 (m, 2H)
LC-MS(ESI): [M+H]+=320.16
Compound 7f (270 mg, 0.84 mmol), dimethyl itaconate (133.4 mg, 0.84 mmol), palladium acetate (9.5 mg, 0.042 mmol), triphenylphosphine (22.11 mg, 0.084 mmol), sodium acetate (207.5 mg, 2.53 mmol) were sequentially added into a pressure vessel, dissolved with tetrahydrofuran, replaced with nitrogen, and heated to 120° C. for overnight. After the reaction system was cooled to room temperature, THF was spin-dried, and then the solution was separated and purified by preparative liquid chromatography to obtain compound 7g (71 mg, 22%).
1H NMR (400 MHz, CDCl3) δ 7.52-7.39 (m, 7H), 4.38-4.14 (m, 3H), 3.91 (d, J=2.8 Hz, 7H), 3.27 (s, 3H), 2.94 (d, J=12.0 Hz, 1H), 2.85-2.72 (m, 3H), 1.55 (d, J=6.6 Hz, 1H)
LC-MS(ESI): [M+H]+=380.36
Compound 7g (71 mg, 0.19 mmol) was dissolved in methanol and water, lithium hydroxide (90 mg, 3.74 mmol) was added, and reacted at room temperature for 20 h. After the reaction, the water and methanol were spin-dried to obtain the crude product (62 mg, 95%). The crude product was used directly in the next step.
LC-MS (ESI): [M+H]+=352.34
Dissolve compound 7h (62 mg, 0.18 mmol) in glacial acetic acid, then add sodium acetate (43.2 mg, 0.53 mmol) and 3-amino-2,6-piperidinedione hydrochloride (44 mg, 0.26 mmol), and the reaction mixture was reacted in a 110° C. oil bath for overnight. Then methanol and water were spin-dried and the residual product was purified by preparative liquid chromatography to obtain compound 7i (39 mg, 49%).
1H NMR (400 MHz, DMSO-d6) δ 11.14 (s, 1H), 7.98-7.72 (m, 2H), 7.64-7.41 (m, 5H), 5.14 (dd, J=12.9, 5.4 Hz, 1H), 4.53-4.24 (m, 2H), 4.01 (dd, J=13.9, 7.6 Hz, 1H), 3.76 (s, 3H), 3.12 (d, J=9.7 Hz, 1H), 3.06-2.91 (m, 2H), 2.92-2.76 (m, 3H), 2.67-2.54 (m, 2H), 2.04 (dd, J=12.5, 6.2 Hz, 1H), 1.90-1.59 (m, 2H)
LC-MS(ESI): [M+H]+=444.41
Compound 7i (39 mg, 0.088 mmol) was dissolved in 2 mL of methanol, 4 mg of palladium on carbon was added, the reaction was replaced by hydrogen, and reacted overnight at room temperature. Palladium carbon was removed by filtration and methanol was spin-dried, and compound 7 (12 mg, 38%) was obtained by purification by preparative liquid chromatography.
1H NMR (400 MHz, DMSO-d6) δ 11.13 (s, 1H), 9.07-8.95 (m, 1H), 7.95-7.68 (m, 2H), 5.14 (dd, J=13.0, 5.3 Hz, 1H), 3.96-3.87 (m, 2H), 3.51 (m, 2H), 3.12 (m, 1H), 3.04-2.82 (m, 4H), 2.72-2.55 (m, 2H), 2.06 (m, 1H), 1.86 (m, 1H), 1.65-1.46 (m, 1H) LC-MS(ESI): [M+H]+=354.41
Compound 8a (10.0 g, 64.0 mmol) was dissolved in 200 mL of dichloromethane, followed by addition of 1,3-propanedithiol (7.0 g, 64.0 mmol). 32 mmol of boron trifluoride ether solution was added dropwise thereto at −18° C. After the addition was complete, the mixture was stirred at −18° C. for 4 h. After the reaction was complete, the solvent was removed under reduced pressure, 500 mL of water was added, and the solid was filtered by suction and purified by C18 column chromatography to obtain compound 8b (2 g, 12.7%).
1H NMR (400 MHz, DMSO-d6) δ 3.85 (s, 4H), 2.84-2.78 (m, 4H), 2.06-1.98 (m, 4H), 1.93-1.80 (m, 2H), 1.70-1.59 (m, 4H)
LC-MS(ESI): [M+H]+=247.07
Compound 8b (2.0 g, 8.0 mmol) was dissolved in 200 mL of dichloromethane, followed by the addition of 100 mL of trifluoroacetic acid at room temperature. After the addition was complete, the mixture was stirred at room temperature for 4 h. After the reaction was complete, sodium bicarbonate (aq) solution was added to the system to adjust the pH to 7. After extraction, the organic phase was dried, concentrated and purified by column chromatography to obtain compound 8c (1.5 g, 91.0%).
1H NMR (400 MHz, DMSO-d6) δ 2.93-2.84 (m, 4H), 2.41-2.33 (m, 4H), 2.31-2.25 (m, 4H), 1.94-1.88 (m, 2H)
LC-MS(ESI): [M+H]+=203.05
Compound 8c (2.0 g, 9.9 mmol) was dissolved in 200 mL of tetrahydrofuran, then compound 4 (2.0 g, 9.8 mmol), (2.0 g, 28.0 mmol) tetrahydropyrrole were added at room temperature. After the addition was complete, the mixture was stirred at 70° C. for 4 h. After the reaction was completed, the solvent was removed from the system under reduced pressure, and the residue was separated and purified by column chromatography to obtain compound 8d (2.0 g, 46.0%).
1H NMR (400 MHz, DMSO-d6) δ 8.18 (s, 1H), 7.35 (s, 1H), 3.83 (s, 3H), 3.81 (s, 3H), 2.92 (s, 2H), 2.90-2.85 (m, 2H), 2.81-2.74 (m, 2H), 2.14-2.10 (m, 2H), 2.02-1.77 (m, 8H).
LC-MS(ESI): [M+H]+=437.10
Compound 8d (2.0 g, 4.5 mmol) was dissolved in 50 mL methanol and 50 mL tetrahydrofuran, then sodium borohydride (350.0 mg, 9.0 mmol) was added at room temperature. After the addition was completed, the mixture was stirred at 70° C. for 4 h. After the reaction was completed, the solvent was removed from the system under reduced pressure, and the residue was separated and purified by column chromatography to obtain compound 8e (1.5 g, 75.0%).
1H NMR (600 MHz, DMSO-d6) δ 7.90 (s, 1H), 7.00 (s, 1H), 5.68 (d, J=6.2 Hz, 1H), 4.77-4.71 (m, 1H), 3.79 (s, 6H), 2.96-2.72 (m, 4H), 2.16-2.01 (m, 4H), 1.93-1.79 (m, 5H), 1.77-1.70 (m, 3H)
LC-MS(ESI): [M+H]+=439.12
Compound 8e (700 mg, 1.5 mmol) was dissolved in 200 mL of dichloromethane, followed by addition of triethylamine (480.0 mg, 4.5 mmol) and 4-dimethylaminopyridine (100.0 mg, 0.8 mmol) at room temperature. Sulfonyl chloride (720.0 mg, 6.0 mmol) was added dropwise thereto at 0° C. After the addition was completed, the mixture was stirred at 0° C. for 4 h, and the reaction was monitored by TLC. After the reaction was completed, the solvent was removed under reduced pressure. After the addition was completed, the residue was dissolved in 100 mL of toluene, followed by the addition of 1,8-diazabicycloundec-7-ene (1.6 g, 10.0 mmol) at room temperature. The mixture was stirred at 110° C. for 16 h. After the reaction was completed, the solvent was removed from the system under reduced pressure, and the residue was separated and purified by column chromatography to obtain compound 8f (400 mg, 59.0%).
1H NMR (400 MHz, DMSO-d6) δ 7.58 (s, 1H), 7.05 (s, 1H), 6.59 (d, J=9.9 Hz, 1H), 5.95 (d, J=11.4 Hz, 1H), 3.78 (s, 3H), 3.77 (s, 3H), 2.93-2.85 (m, 2H), 2.82-2.73 (m, 2H), 2.18-2.11 (m, 2H), 2.09-1.96 (m, 2H), 1.92-1.77 (m, 6H)
LC-MS(ESI): [M+H]+=421.11
Compound 8f (400.0 mg, 0.9 mmol) was dissolved in 50 mL of methanol, 50 mL of tetrahydrofuran, 50 mL of water, and then lithium hydroxide (120.0 mg, 4.5 mmol) was added at room temperature. After the addition was complete, the mixture was stirred at room temperature for 16 h. After the reaction was completed, 1. M dilute hydrochloric acid solution was added to the system to adjust the pH to 6. After extraction, the organic phase was dried, concentrated and purified by column chromatography to obtain compound 8g (350 mg, 94.0%).
1H NMR (600 MHz, DMSO-d6) δ 12.90 (s, 2H), 7.52 (s, 1H), 6.99 (s, 1H), 6.57 (d, J=9.9 Hz, 1H), 5.90 (d, J=9.9 Hz, 1H), 2.92-2.86 (m, 2H), 2.83-2.73 (m, 2H), 2.11 (m, 2H), 2.09-1.97 (m, 2H), 1.94-1.76 (m, 6H)
LC-MS(ESI): [M+H]+=393.08
Compound 8g (300.0 mg, 0.7 mmol) was dissolved in 100 mL of acetic acid, followed by addition of 3-aminopiperidine-2,6-dione hydrochloride (170.0 mg, 1.3 mmol), sodium acetate (360.0 mg, 2.1 mmol). After the addition was completed, the mixture was stirred at 110° C. for 4 h. After the reaction was completed, the solvent was removed from the system under reduced pressure, and the residue was separated and purified by column chromatography to obtain compound 8h (280 mg, 75.0%).
1H NMR (400 MHz, DMSO-d6) δ 11.12 (s, 1H), 7.72 (s, 1H), 7.30 (s, 1H), 6.70 (d, J=8.0 Hz, 1H), 6.03 (d, J=8.8 Hz, 1H), 5.13-5.07 (m, 1H), 2.94-2.84 (m, 3H), 2.84-2.76 (m, 2H), 2.64-2.55 (m, 1H), 2.18-1.98 (m, 6H), 1.95-1.78 (m, 6H)
LC-MS(ESI): [M+H]+=485.11
Compound 8h (280.0 mg, 0.5 mmol) was dissolved in 50 mL of acetonitrile, followed by addition of 50 mL of sodium bicarbonate solution, iodine (1.5 g, 5.0 mmol) at room temperature. After the addition was complete, the mixture was stirred at room temperature for 4 h. After the reaction was completed, the solvent was removed under reduced pressure. After extraction, the organic phase was dried, concentrated and purified by column chromatography to obtain compound 8i (100 mg, 44.0%).
1H NMR (600 MHz, DMSO-d6) δ 11.13 (s, 1H), 7.77 (s, 1H), 7.45 (s, 1H), 6.76 (d, J=10.0 Hz, 1H), 6.09 (d, J=9.9 Hz, 1H), 5.13-5.09 (m, 1H), 2.92-2.84 (m, 1H), 2.78-2.70 (m, 2H), 2.63-2.57 (m, 1H), 2.26-2.15 (m, 4H), 2.12-2.00 (m, 4H)
LC-MS(ESI): [M+H]+=395.12
Compound 8i (280.0 mg, 0.7 mmol) was dissolved in 50 mL of THF, then Pd/C (150.0 mg) was added at room temperature. After the addition was complete, the mixture was stirred at 50° C. for 4 h under hydrogen atmosphere. After the reaction was completed, the reaction solution was filtered through diatomaceous earth, the solvent was removed under reduced pressure, and compound 8 (195 mg, 70.0%) was obtained after purification by preparative liquid chromatography.
1H NMR (600 MHz, DMSO-d6) δ 11.11 (s, 1H), 7.68 (s, 1H), 7.35 (s, 1H), 5.12-5.06 (m, 1H), 2.99-2.93 (m, 2H), 2.92-2.84 (m, 1H), 2.68-2.55 (m, 3H), 2.23-2.15 (m, 2H), 2.13-2.00 (m, 4H), 1.99-1.91 (m, 4H)
LC-MS(ESI): [M+H]+=397.13
Diethylaminosulfur trifluoride (4 mL) was added to compound 1e (2 g, 4.6 mmol), and the reaction solution was stirred at 85° C. for 2 hours. The resulting mixture was poured into ice water, extracted three times with ethyl acetate, the organic phase was concentrated in vacuo, and the mixture was purified by reverse column to obtain compound 9a (0.5 g, 24%) as a yellow oily liquid.
1H NMR (600 MHz, DMSO-d6) δ 8.02 (s, 1H), 7.29 (s, 1H), 3.82 (s, 3H), 3.81 (s, 3H), 3.77-3.68 (m, 2H), 3.22-3.02 (m, 2H), 2.72 (t, J=14.7 Hz, 2H), 1.85 (d, J=13.9 Hz, 2H), 1.70 (td, J=14.0, 11.6, 4.7 Hz, 2H), 1.40 (s, 9H).
LC-MS(ESI): [M-tBu+H]+=400.31.
To a solution of compound 9a (160 mg, 0.4 mmol) in methanol/tetrahydrofuran (2 ml), lithium hydroxide (42 mg, 1.8 mmol) was added slowly. The reaction was stirred overnight at room temperature. The mixture was extracted three times with dichloromethane/methanol (10:1) solvent, and the organic phase was concentrated in vacuo to obtain compound 9b (0.1 g, 67%) as a white solid.
1H NMR (600 MHz, DMSO-d6) δ 13.27 (br s, 2H), 8.12 (s, 1H), 7.36 (s, 1H), 3.21-3.06 (m, 2H), 2.68 (t, J=14.7 Hz, 2H), 2.05-1.93 (m, 2H), 1.84 (d, J=13.8 Hz, 2H), 1.68 (td, J=14.0, 12.9, 4.7 Hz, 2H), 1.40 (s, 9H).
LC-MS(ESI): [M-tBu+H]+=372.27.
To a solution of compound 9b (100 mg, 0.23 mmol) in acetic acid (1 ml) was slowly added 3-aminopiperidine-2,6-dione hydrochloride (77 mg, 0.5 mmol) and sodium acetate (96 mg, 1.2 mmol). The reaction was stirred at 110° C. for 2 hours. After the reaction was completed, the mixture was purified by reverse phase column to obtain a brown solid compound 9 (50 mg, 51%).
1H NMR (600 MHz, DMSO-d6) δ 11.15 (s, 1H), 8.10 (s, 1H), 7.68 (s, 1H), 5.16 (dd, J=13.0, 5.4 Hz, 1H), 3.27-3.15 (m, 4H), 2.92-2.82 (m, 3H), 2.61 (dt, J=17.2, 3.4 Hz, 1H), 2.56-2.51 (m, 1H), 2.11 (d, J=14.4 Hz, 2H), 2.05 (ddt, J=12.9, 5.6, 2.5 Hz, 1H), 1.97-1.90 (m, 2H).
LC-MS(ESI): [M+H]+=420.36.
10b was prepared by referring to the procedure similar to Step 1 in Example 9.
1H NMR (400 MHz, DMSO-d6) δ 8.01 (s, 1H), 7.34 (s, 1H), 4.13 (d, J=9.7 Hz, 2H), 3.91 (d, J=9.7 Hz, 2H), 3.83 (s, 3H), 3.82 (s, 3H), 3.04 (t, J=13.4 Hz, 2H), 1.39 (s, 9H).
LC-MS(ESI): [M-Boc+H]+=328.30.
10c was prepared by referring to the similar procedure of step 2 in Example 9.
LC-MS (ESI): [M+H]+=399.28.
Compound 10 was prepared by referring to the procedure similar to Step 3 in Example 9.
1H NMR (600 MHz, DMSO-d6) δ 11.15 (s, 1H), 8.13 (s, 1H), 7.58 (s, 1H), 5.18 (dd, J=13.0, 5.4 Hz, 1H), 4.33 (d, J=11.7 Hz, 2H), 4.22 (d, J=11.6 Hz, 2H), 3.15 (t, J=13.6 Hz, 2H), 2.94-2.87 (m, 1H), 2.65-2.59 (m, 1H), 2.54-2.51 (m, 1H), 2.10-2.05 (m, 1H).
LC-MS(ESI): [M+H]+=392.30.
Compound 1e (5 g, 11.5 mmol) was dissolved in anhydrous tetrahydrofuran (10 mL). Cool down to −78° C. and add sodium bis(trimethylsilyl)amide (4.23 g, 23.07 mmol) dropwise, keep this temperature for 2 h, then add N-fluorobisbenzenesulfonamide (7.3 g, 23.1 mmol) dropwise at −78° C. The reaction was stirred overnight at −78° C. The resulting mixture was quenched by pouring into saturated ammonium chloride solution, extracted with ethyl acetate, dried over anhydrous sodium sulfate, filtered and concentrated. The crude product was purified by column chromatography (PE:EA=0-40%) to obtain compound 11a (2.5 g, 46%) as a light yellow solid.
1H NMR (400 MHz, DMSO-d6) δ 8.29 (s, 1H), 7.62 (s, 1H), 3.94 (d, J=11.6 Hz, 2H), 3.85 (s, 3H), 3.86 (s, 3H), 3.08 (s, 2H), 2.07 (d, J=13.4 Hz, 2H), 1.74 (td, J=13.5, 4.7 Hz, 2H), 1.41 (s, 9H).
LC-MS(ESI): [M+H]+=470.31.
Compound 11a (2.5 g, 5.3 mmol) was dissolved in ethanol (5 mL), and sodium borohydride (403 mg, 10.7 mmol) was slowly added at 0° C. The reaction was stirred at room temperature for 1 h. The reaction was quenched with acetone. After concentration in vacuo, the crude product was purified by column chromatography (PE:EA=0-40%) to obtain white solid compound 11b (2.4 g, 95%).
1H NMR (400 MHz, DMSO-d6) δ 7.90 (s, 1H), 7.23 (s, 1H), 6.68 (d, J=6.1 Hz, 1H), 5.03 (dt, J=16.1, 7.0 Hz, 1H), 3.97 (d, J=14.1 Hz, 1H), 3.91-3.75 (m, 7H), 3.04 (d, J=55.0 Hz, 2H), 1.99 (d, J=13.3 Hz, 1H), 1.84-1.74 (m, 2H), 1.70-1.56 (m, 1H), 1.41 (s, 9H).
LC-MS(ESI): [M-Boc+H]+=372.34.
Compound 11b (1.5 g, 3.2 mmol) was dissolved in dichloromethane (30 mL), p-toluenesulfonyl chloride (727 mg, 3.8 mmol) and triethylamine (642 mg, 6.4 mmol) were added under ice-cooling conditions. Stir at room temperature for 2 h. After concentration in vacuo, the crude product was purified by column chromatography (PE:EA=0-40%) to obtain a white solid compound 11c (1.0 g, 50%).
1H NMR (600 MHz, DMSO-d6) δ 7.98 (d, J=8.2 Hz, 2H), 7.56 (d, J=8.1 Hz, 2H), 7.41 (s, 1H), 7.35 (s, 1H), 6.25 (dd, J=12.9, 8.1 Hz, 1H), 3.90 (dd, J=21.4, 8.9 Hz, 2H), 3.81 (s, 3H), 3.79 (s, 3H), 3.06 (s, 2H), 2.48 (s, 3H), 1.99-1.91 (m, 2H), 1.67 (dtd, J=39.4, 13.3, 4.7 Hz, 2H), 1.41 (s, 9H).
LC-MS(ESI): [M-Boc+H]+=526.34.
Compound 11c (1.0 g, 1.6 mmol) was dissolved in methanol (20 mL), and palladium on carbon (100 mg) was added. Stir under hydrogen atmosphere at room temperature for 12 h. Suction filtration, wash with methanol, and concentration in vacuo to obtain white solid compound 11d (700 mg, 96%).
1H NMR (600 MHz, DMSO-d6) δ 7.70 (s, 1H), 7.28 (s, 1H), 3.95-3.88 (m, 2H), 3.86-3.74 (m, 8H), 3.51 (t, J=15.8 Hz, 2H), 1.81 (d, J=13.2 Hz, 2H), 1.69 (td, J=13.6, 4.9 Hz, 2H), 1.42 (s, 9H).
LC-MS(ESI): [M-tBu+H]+=400.44.
11e was prepared by referring to the procedure similar to Step 2 in Example 9.
LC-MS (ESI): [M-Boc+H]+=328.32.
Compound 11 was prepared by referring to the procedure similar to Step 3 in Example 9.
1H NMR (400 MHz, CD3OD) δ 7.78 (s, 1H), 7.55 (s, 1H), 5.13 (dd, J=12.6, 5.4 Hz, 1H), 3.60 (t, J=15.5 Hz, 2H), 3.50-3.34 (m, 4H), 2.93-2.82 (m, 1H), 2.80-2.66 (m, 2H), 2.24-2.09 (m, 5H).
LC-MS(ESI): [M+H]+=420.41.
A mixture of 4-bromo-3-methylphenol 12a (5.0 g, 26.7 mmol) and acetyl chloride (10.5 g, 134 mmol) was stirred at 60° C. for 1 hour, then aluminum chloride (5.4 g, 40.1 mmo1) was added at room temperature. After stirring at 160° C. for 2 hours, the mixture was cooled to room temperature, poured into saturated ammonium chloride solution (50 mL), and extracted with EA (50 mL×3). The combined organic layers were dried over anhydrous sodium sulfate, filtered, and concentrated to afford crude compound 12b as an off-white solid (5.5 g, 90%).
1H NMR (600 MHz, DMSO-d6) δ 11.81 (s, 1H), 8.01 (s, 1H), 7.00 (s, 1H), 2.62 (s, 3H), 2.34 (s, 3H).
To a solution of compound 12b (5.5 g, 24.01 mmol) in methanol (50 mL) was added triethylamine (9.72 g, 96.04 mmol) and Pd(dppf)Cl2 (1.76 g, 2.40 mmol). The mixed system was purged with CO three times and then stirred overnight at 60° C. After the reaction, the system was filtered, the filtrate was concentrated under reduced pressure, and the resulting mixture was purified by column chromatography (PE:EA=6:1) to obtain a white solid 12c (3.26 g, 65%).
1H NMR (600 MHz, DMSO-d6) δ 12.14 (s, 1H), 8.34 (s, 1H), 6.90 (s, 1H), 3.82 (s, 3H), 2.64 (s, 3H), 2.53 (s, 3H).
To a solution of 12c (3.1 g, 14.89 mmol) in ethanol (120 mL) was added N-tert-butoxycarbonyl-3-azetidinone (2.8 g, 16.38 mmol) and tetrahydropyrrole (1.59 g, 22.33 mmol). The reaction was stirred overnight at 80° C. After completion of the reaction, the resulting mixture was concentrated in vacuo and purified by column chromatography (PE:EA=0-20%) to give yellow solid 12d (1.9 g, 35%).
1H NMR (600 MHz, DMSO-d6) δ 8.25 (s, 1H), 7.08 (s, 1H), 3.82 (s, 3H), 3.72 (s, 2H), 3.14 (s, 2H), 2.89 (s, 2H), 2.55 (s, 3H), 1.91-1.84 (m, 2H), 1.69-1.61 (m, 2H), 1.40 (s, 9H).
LC-MS(ESI): [M-Boc+H]+=290.27.
To a solution of 12d (1.6 g, 4.43 mmol) in methanol (40 mL) was added sodium borohydride (504 mg, 13.28 mmol). The reaction was stirred at room temperature for 4 hours. After completion of the reaction, the resulting mixture was concentrated in vacuo and purified by column chromatography (PE:EA=0-40%) to give yellow solid 12e (1.5 g, 93.23%).
1H NMR (600 MHz, DMSO-d6) δ 8.01 (s, 1H), 6.73 (s, 1H), 5.52 (d, J=4.7 Hz, 1H), 4.68 (s, 1H), 3.78 (s, 3H), 3.68 (dd, J=22.7, 9.0 Hz, 2H), 3.13 (d, J=77.0 Hz, 2H), 2.46 (s, 3H), 2.13 (dd, J=13.5, 6.0 Hz, 1H), 1.82-1.62 (m, 4H), 1.59-1.50 (m, 1H), 1.41 (s, 9H).
LC-MS(ESI): [M-Boc+H]+=292.30.
To a solution of 12e (1.5 g, 4.13 mmol) in toluene (40 mL) was added p-toluenesulfonic acid (712 mg, 4.13 mmol). The reaction was stirred overnight at 110° C. After the reaction was complete, the system was concentrated under vacuum to obtain a mixture, which was dissolved in tetrahydrofuran (40 mL), and triethylamine (2.23 g, 22.02 mmol) and di-tert-butyl dicarbonate (2.4 g, 11.01 mmol) were added thereto, and the mixture was stirred overnight at room temperature. After the reaction was completed, the system was concentrated under vacuum, and the resulting mixture was purified by column chromatography (PE:EA=0-300%) to give 12f (1.1 g, 77%) as a yellow solid.
1H NMR (600 MHz, DMSO-d) δ 7.64 (s, 1H), 6.80 (s, 1H), 6.55 (d, J=9.9 Hz, 1H), 5.80 (d, J=9.8 Hz, 1H), 3.78 (s, 3H), 3.70 (d, J=13.0 Hz, 2H), 3.30-3.11 (m, 2H), 2.47 (s, 3H), 1.84-1.78 (m, 2H), 1.68-1.60 (m, 2H), 1.41 (s, 9H).
LC-MS(ESI): [M-Boc+H]+=274.26.
Add N-bromosuccinimide (620 mg, 3.47 mmol) and azobisisobutyronitrile (48 mg, 290 mol) to a solution of compound 12f (1 g, 2.90 mmol) in carbon tetrachloride (10 mL). The system was replaced with a nitrogen atmosphere, and stirred overnight at 80° C. After the reaction was completed, the system was concentrated under vacuum, and the obtained crude product was directly used in the next step.
The mixture obtained in the previous step was dissolved in acetonitrile (40 mL), and diisopropylethylamine (1.12 g, 8.70 mmol) and 3-amino-2,6-piperidinedione (446 mg, 3.48 mmol) were added thereto, and the mixture was stirred overnight at 80° C. After concentration, acetic acid (10 mL) was added to continue the reaction for 2 hours. After the reaction was complete, the system was concentrated in vacuo, and the resulting mixture was purified by column chromatography (PE:EA=0-50%) to obtain 12 g (285 mg, 22%) of a black solid.
1H NMR (600 MHz, CD3OD) δ 7.52 (s, 1H), 7.13 (s, 1H), 6.69 (d, J=9.9 Hz, 1H), 5.83 (d, J=9.9 Hz, 1H), 5.13 (dd, J=13.3, 5.1 Hz, 1H), 4.45 (dd, J=35.2, 17.3 Hz, 2H), 3.48-3.40 (m, 2H), 3.38-3.33 (m, 2H), 2.91 (ddd, J=18.2, 11.8, 5.0 Hz, 2H), 2.82-2.77 (m, 1H), 2.49 (ddd, J=17.9, 12.7, 3.8 Hz, 1H), 2.29-2.22 (m, 2H), 2.17 (tdd, J=13.4, 6.7, 4.2 Hz, 1H), 2.03-1.95 (m, 2H).
LC-MS(ESI): [M+H]+=368.38.
Dissolve 12 g of the product from the previous step with methanol (10 mL), add palladium carbon (150 mg) thereto, replace the reaction system with a hydrogen atmosphere, and stir at room temperature for 2 hours. After the reaction is complete, the mixture was filtered, and the filtrate was concentrated under vacuum. Purification by preparative high performance liquid chromatography to obtain compound 12 (120 mg, 35% yield in two steps) as a white solid.
1H NMR (600 MHz, DMSO-d6) δ 10.97 (s, 1H), 7.51 (s, 1H), 7.05 (s, 1H), 5.07 (dd, J=13.3, 5.1 Hz, 1H), 4.34 (d, J=16.9 Hz, 1H), 4.22 (d, J=16.9 Hz, 1H), 3.26-3.20 (m, 2H), 3.16-3.08 (m, 2H), 2.94-2.86 (m, 3H), 2.63-2.57 (m, 1H), 2.41-2.33 (m, 1H), 2.00-1.87 (m, 5H), 1.85-1.77 (m, 2H).
LC-MS(ESI): [M+H]+=370.39.
13a was prepared by referring to the procedure similar to Step 3 in Example 12.
1H NMR (600 MHz, DMSO-d6) δ 8.24 (s, 1H), 7.15 (s, 1H), 4.00 (d, J=8.7 Hz, 2H), 3.89 (d, J=8.7 Hz, 2H), 3.82 (s, 3H), 3.20 (s, 2H), 2.56 (s, 3H), 1.38 (s, 9H).
LC-MS(ESI): [M-Boc+H]+=262.25.
13b was prepared by referring to the procedure similar to Step 4 in Example 12.
1H NMR (600 MHz, CD3OD) δ 8.02 (s, 1H), 6.81 (s, 1H), 4.82 (t, J=5.2 Hz, 1H), 4.28 (d, J=9.6 Hz, 1H), 4.04 (dd, J=19.0, 9.1 Hz, 2H), 3.95 (d, J=9.7 Hz, 1H), 3.87-3.84 (m, 3H), 2.54 (s, 3H), 2.32 (d, J=5.2 Hz, 2H), 1.47 (s, 9H).
LC-MS(ESI): [M-Boc+H]+=264.27.
13c was prepared by referring to the similar procedure of Step 5 in Example 12.
1H NMR (600 MHz, DMSO-d6) δ 7.67 (s, 1H), 6.84 (s, 1H), 6.65 (d, J=9.9 Hz, 1H), 6.16 (d, J=9.9 Hz, 1H), 4.04 (s, 4H), 3.78 (s, 3H), 2.47 (s, 3H), 1.40 (s, 9H).
LC-MS(ESI): [M-Boc+H]+=246.23.
Add N-bromosuccinimide (620 mg, 3.47 mmol) and azobisisobutyronitrile (48 mg, 290 mol) to a solution of compound 13c (1 g, 2.90 mmol) in carbon tetrachloride (10 mL). The reaction system was replaced with a nitrogen atmosphere, and stirred overnight at 80° C. The resulting mixture was concentrated under vacuum, dissolved with acetonitrile (40 mL), to which diisopropylethylamine (1.12 g, 8.70 mmol) and 3-amino-2,6-piperidinedione (446 mg, 3.48 mmol) were added, The mixture was stirred overnight at 80° C. The resulting mixture was concentrated under vacuum and purified by column chromatography (PE:EA=0-50%) to obtain 13d (285 mg, 22%) as a black solid.
1H NMR (600 MHz, CD3OD) δ 7.50 (s, 1H), 7.09 (s, 1H), 6.69 (d, J=9.9 Hz, 1H), 6.17 (d, J=9.8 Hz, 1H), 5.12 (dd, J=13.2, 5.2 Hz, 1H), 4.50-4.40 (m, 2H), 4.24-4.19 (m, 2H), 4.10 (d, J=8.9 Hz, 2H), 2.94-2.86 (m, 1H), 2.82-2.77 (m, 1H), 2.49 (ddd, J=26.5, 13.2, 4.5 Hz, 1H), 2.19-2.15 (m, 1H), 1.48 (s, 9H).
LC-MS(ESI): [M+H]+=440.38.
Compound 13d (285 mg, 648.5 mol) was dissolved in dichloromethane (5 mL) solution, trifluoroacetic acid (636 mg, 6.49 mmol) was added, and the reaction was stirred at room temperature for 2 hours. After the reaction was completed, it was concentrated and used directly in the next step.
LC-MS (ESI): [M+H]+=340.32.
Compound 13 was prepared by referring to the steps similar to Step 8 in Example 12.
1H NMR (400 MHz, CD3OD) δ 7.57 (s, 1H), 7.10 (s, 1H), 5.11 (dd, J=13.3, 5.1 Hz, 1H), 4.50-4.35 (m, 2H), 4.24 (s, 4H), 2.97 (t, J=5.8 Hz, 2H), 2.91-2.84 (m, 1H), 2.77 (ddd, J=17.6, 4.5, 2.3 Hz, 1H), 2.46 (ddd, J=26.4, 13.2, 4.7 Hz, 1H), 2.28 (t, J=6.1 Hz, 2H), 2.14 (dtd, J=12.7, 5.2, 2.3 Hz, 1H).
LC-MS(ESI): [M+H]+=342.32.
14b was prepared by referring to the procedure similar to Step 1 in Example 12.
1H NMR (600 MHz, CDCl3) δ 12.10 (s, 1H), 7.57 (s, 1H), 7.25 (s, 1H), 2.63 (s, 3H), 2.39 (s, 3H).
14c was prepared by referring to the similar procedure of step 2 in Example 12.
1H NMR (600 MHz, CDCl3) δ 11.88 (s, 1H), 7.60 (s, 1H), 7.49 (s, 1H), 3.93 (s, 3H), 2.68 (s, 3H), 2.53 (s, 3H).
14d was prepared by referring to the similar procedure of Step 3 in Example 12.
1H NMR (600 MHz, DMSO-d6) δ 7.64 (s, 1H), 7.45 (s, 1H), 3.85 (s, 3H), 3.71 (s, 2H), 3.12 (d, J=50.1 Hz, 2H), 2.88 (s, 2H), 2.43 (s, 3H), 1.90-1.84 (m, 2H), 1.62 (td, J=12.7, 4.6 Hz, 2H), 1.40 (s, 9H).
LC-MS(ESI): [M-Boc+H]+=290.26.
14e was prepared by referring to the similar procedure of Step 4 in Example 12.
1H NMR (600 MHz, CDCl3) δ 7.40 (s, 1H), 7.32 (s, 1H), 4.83 (t, J=7.0 Hz, 1H), 3.90-3.84 (m, 5H), 3.31-3.05 (m, 2H), 2.93 (s, 1H), 2.49 (s, 3H), 2.11 (dd, J=13.6, 6.1 Hz, 1H), 1.92-1.85 (m, 2H), 1.78-1.72 (m, 1H), 1.63 (td, J=13.3, 4.6 Hz, 1H), 1.52 (ddd, J=24.7, 12.6, 7.9 Hz, 1H), 1.46 (s, 9H).
LC-MS(ESI): [M-Boc+H]+=292.27.
Compound 14e (15.23 g, 38.91 mmol) and p-toluenesulfonic acid (6.7 g, 38.91 mmol) were dissolved in toluene, and the reaction was refluxed overnight at 110° C. The reaction mixture was concentrated in vacuo to obtain the resulting light yellow oily mixture 14f (8.6 g, 80%). The crude product was used directly in the next step.
LC-MS (ESI): [M-Boc+H]+=274.29.
Compound 14f (8.6 g, 31.46 mmol), di-tert-butyl dicarbonate (13.73 g, 62.93 mmol) and triethylamine (9.55 g, 94.39 mmol) were dissolved in dichloromethane (30 mL), and the mixture was stirred at room temperature for 4 hours. The resulting mixture was concentrated under vacuum and purified by column chromatography (PE:EA=0-30%) to obtain 14 g (1.8 g, 83%) of colorless solid.
1H NMR (600 MHz, CDCl3) δ 7.41 (s, 1H), 6.85 (s, 1H), 6.37 (d, J=9.8 Hz, 1H), 5.65 (d, J=9.6 Hz, 1H), 3.98-3.77 (m, 5H), 3.28 (s, 2H), 2.50 (s, 3H), 1.97 (d, J=13.4 Hz, 2H), 1.59 (td, J=13.4, 4.7 Hz, 2H), 1.47 (s, 9H).
LC-MS(ESI): [M-Boc+H]+=274.24.
14h was prepared by referring to the similar procedure of Step 6 in Example 12.
14i was prepared by referring to the similar steps of Step 7 in Example 12.
1H NMR (600 MHz, DMSO-d6) δ 11.00 (s, 1H), 7.36 (s, 1H), 7.25 (s, 1H), 6.68 (d, J=9.8 Hz, 1H), 5.99 (d, J=9.8 Hz, 1H), 5.08 (dd, J=13.3, 5.2 Hz, 1H), 4.37 (d, J=16.9 Hz, 1H), 4.25 (d, J=16.9 Hz, 1H), 3.26-3.19 (m, 4H), 2.90 (ddd, J=17.3, 13.7, 5.5 Hz, 1H), 2.64-2.57 (m, 1H), 2.39 (qd, J=13.2, 4.5 Hz, 1H), 2.10-2.03 (m, 2H), 2.00 (dtd, J=12.8, 5.4, 2.4 Hz, 1H), 1.94-1.86 (m, 2H).
LC-MS(ESI): [M+H]+=368.37.
Compound 14 was prepared by referring to the steps similar to Step 8 in Example 12.
1H NMR (600 MHz, DMSO-d6) δ 10.99 (s, 1H), 7.36 (s, 1H), 7.18 (s, 1H), 5.07 (dd, J=13.3, 5.1 Hz, 1H), 4.34 (d, J=16.6 Hz, 1H), 4.21 (d, J=16.5 Hz, 1H), 3.25-3.11 (m, 4H), 2.94-2.87 (m, 3H), 2.63-2.57 (m, 1H), 2.39 (qd, J=13.2, 4.5 Hz, 1H), 2.01-1.95 (m, 1H), 1.93-1.87 (m, 4H), 1.82-1.74 (m, 2H).
LC-MS(ESI): [M+H]+=370.36.
15a was prepared by referring to the similar steps of Step 3 in Example 12.
1H NMR (600 MHz, CDCl3) δ 7.74 (s, 1H), 7.59 (s, 1H), 4.09 (d, J=9.6 Hz, 2H), 3.97 (d, J=9.5 Hz, 2H), 3.94 (s, 3H), 3.07 (s, 2H), 2.53 (s, 3H), 1.46 (s, 9H).
LC-MS(ESI): [M-Boc+H]+=262.28.
15b was prepared by referring to the similar procedure of Step 4 in Example 12.
1H NMR (600 MHz, CDCl3) δ 7.50 (s, 1H), 7.24 (s, 1H), 4.89 (q, J=5.2 Hz, 1H), 4.36 (q, J=7.2 Hz, 1H), 4.27-4.23 (m, 1H), 4.10 (d, J=9.3 Hz, 1H), 4.00 (dd, J=17.3, 9.6 Hz, 2H), 3.90 (s, 3H), 2.54 (s, 3H), 2.35 (d, J=5.3 Hz, 2H), 1.47 (s, 9H).
LC-MS(ESI): [M-Boc+H]+=264.27.
15c was prepared by referring to the similar step of Step 5 in Example 14.
LC-MS (ESI): [M+H]+=246.22.
15d was prepared by referring to the similar procedure of Step 6 in Example 14.
1H NMR (600 MHz, CDCl3) δ 7.45 (s, 1H), 6.88 (s, 1H), 6.47 (d, J=9.8 Hz, 1H), 6.06 (d, J=9.7 Hz, 1H), 4.24 (d, J=9.5, 2H), 4.01 (d, J=9.5, 2H), 3.89 (s, 3H), 2.51 (s, 3H), 1.48 (s, 9H).
LC-MS(ESI): [M-tBu+H]+=290.20
15e was prepared by referring to the similar procedure of Step 6 in Example 12.
3-amino-2,6-piperidinedione hydrochloride (173 mg, 1.05 mmol) and N,N-diisopropylethylamine (0.4 mL) were added to a solution of compound 15e (2 g, 0.7 mmol) in acetonitrile (15 mL). The reaction solution was stirred at 80° C. for 12 hours. The resulting mixture was concentrated in vacuo, and purified by C18 reverse phase column to obtain compound 15f (120 mg, 38%) as a gray solid.
1H NMR (400 MHz, DMSO-d6) δ 10.97 (s, 1H), 7.35 (s, 1H), 7.13 (s, 1H), 6.71 (d, J=9.9 Hz, 1H), 6.33 (d, J=9.8 Hz, 1H), 5.08 (dd, J=13.3, 5.1 Hz, 1H), 4.41-4.19 (m, 2H), 4.15-3.96 (m, 4H), 2.97-2.85 (m, 1H), 2.66-2.55 (m, 1H), 2.41-2.30 (m, 1H), 2.03-1.95 (m, 1H), 1.40 (s, 9H).
LC-MS(ESI): [M-tBu+H]+=384.35
Compound 15f (120 mg, 0.7 mmol) dichloromethane (3 mL), trifluoroacetic acid (1 mL). The reaction mixture was stirred at room temperature for 2 hours, and the reaction mixture was concentrated in vacuo. The crude product was used directly in the next step without further purification.
LC-MS (ESI): [M+H]+=340.34
Compound 15 was prepared by referring to the step similar to Step 8 in Example 12.
1H NMR (400 MHz, DMSO-d6) δ 10.97 (s, 1H), 7.36 (s, 1H), 7.11 (s, 1H), 5.07 (dd, J=13.3, 5.1 Hz, 1H), 4.34 (d, J=16.8 Hz, 1H), 4.21 (d, J=16.8 Hz, 1H), 4.15-4.04 (m, 4H), 2.91 (q, J=5.6, 4.4 Hz, 2H), 2.67-2.56 (m, 2H), 2.40-2.31 (m, 1H), 2.20 (t, J=6.9 Hz, 2H), 1.98 (m, 1H).
LC-MS(ESI): [M+H]+=342.43
16b was prepared by referring to the procedure similar to Step 1 in Example 1.
1H NMR (600 MHz, DMSO-d6) δ 7.90 (dd, J=8.7, 6.2 Hz, 1H), 7.22 (dd, J=10.1, 2.8 Hz, 1H), 7.15 (td, J=8.5, 2.7 Hz, 1H), 3.82 (s, 3H), 2.53 (s, 3H).
LC-MS(ESI): [M+H]+=169.19.
16c was prepared by referring to the similar procedure of step 2 in Example 5.
1H NMR (600 MHz, DMSO-d6) δ 8.16 (d, J=6.3 Hz, 1H), 7.19 (dd, J=10.4, 3.2 Hz, 1H), 3.83 (s, 3H), 2.55 (s, 3H), 1.30 (s, 12H).
16d was prepared by referring to the similar procedure of Step 3 in Example 5.
1H NMR (600 MHz, DMSO-d6) δ 10.04 (s, 1H), 7.47 (d, J=9.3 Hz, 1H), 7.13 (d, J=12.2 Hz, 1H), 3.80 (s, 3H), 2.41 (s, 3H).
LC-MS(ESI): [M+H]−=183.21.
Compound 16d (1.0 g, 5.4 mmol) was dissolved in 10 ml of ultra-dry N,N-dimethylformamide, followed by the addition of tert-butyl 1-oxa-6-azaspiro[2.5]octane-6-carboxylate ester (1.2 g, 5.3 mmol) and sodium hydride (196 mg, 4.9 mmol), the reaction was heated to 110° C. and stirred overnight. The reaction solution was subjected to silica gel column chromatography to obtain white solid compound 16e (1.4 g, 68%).
1H NMR (600 MHz, DMSO-d6) δ 7.37 (s, 1H), 6.85 (s, 1H), 4.04 (s, 2H), 3.77 (s, 3H), 3.73 (d, J=13.4 Hz, 2H), 3.24-3.04 (m, 2H), 2.42 (s, 3H), 1.66-1.61 (m, 4H), 1.41 (s, 9H).
LC-MS(ESI): [M+H]+=278.29.
To a solution of compound 16e (1.0 g, 2.7 mmol) in carbon tetrachloride (10 mL) was added N-bromosuccinimide (613 mg, 3.4 mmol) and azobisisobutyronitrile (44 mg, 0.3 mmol), After nitrogen replacement three times, the reaction was stirred overnight at 85° C. under nitrogen protection. After the reaction was completed, the reaction solution was filtered, and the filtrate was concentrated to obtain a crude product (1.2 g), which was used in the next reaction without further purification.
To a solution of compound 16f (1.2 g, 2.7 mmol) in acetonitrile (10 mL) was added 3-aminopiperidine-2,6-dione hydrochloride (655 mg, 4.0 mmol) and N,N-diisopropylethylamine (1.0 g, 8.0 mmol), the reaction solution was stirred overnight at 80° C., then the reaction solution was concentrated and dissolved in acetic acid (10 mL), and the reaction solution was stirred at 110° C. for 2 h. The reaction solution was purified by reverse phase to obtain white solid compound 16 (235 mg, 24%).
1H NMR (400 MHz, DMSO-d6) δ 10.96 (s, 1H), 7.22 (s, 1H), 7.17 (s, 1H), 5.06 (dd, J=13.3, 5.1 Hz, 1H), 4.32 (d, J=16.8 Hz, 1H), 4.20 (d, J=16.8 Hz, 1H), 4.19-4.11 (m, 2H), 3.30-3.25 (m, 2H), 3.18-3.08 (m, 2H), 2.90 (ddd, J=17.3, 13.6, 5.4 Hz, 1H), 2.59 (dt, J=16.6, 3.4 Hz, 1H), 2.42-2.30 (m, 1H), 2.00-1.93 (m, 1H), 1.92-1.85 (m, 4H).
LC-MS(ESI): [M+H]+=373.35.
17b was prepared by referring to the same procedure as Step 1 in Example 1.
LC-MS (ESI): [M-OMe+H]+=179.12.
Under nitrogen protection, compound 17b (10 g, 47 mmol) was dissolved in trifluoroacetic acid (100 mL), N-bromosuccinimide (4.24 g, 47 mmol) was added to the reaction system, and the reaction mixture was reacted overnight at room temperature. The resulting reaction system was concentrated under reduced pressure, and after adding water (100 mL), it was extracted with ethyl acetate (100 mL*3 times). Wash with saturated brine (100 ml), combine the organic phases and dry over anhydrous sodium sulfate. The concentrated crude product was purified by reverse column chromatography to obtain light yellow solid compound 17c (4.6 g, 33%).
1H NMR (600 MHz, CD3OD) δ 7.98 (s, 1H), 7.07 (s, 1H), 3.88 (s, 3H), 3.86 (s, 3H).
LC-MS(ESI): [M+H]+=289.04.
Under nitrogen protection conditions, compound 17c (8 g, 27 mmol, 1.0 eq), (1-benzyl-1,2,3,6-tetrahydropyridin-4-yl)methanol (5.64 g, 27 mmol) and triphenyl phosphine (7.2 g, 27 mmol) were dissolved in tetrahydrofuran (80 mL), diethyl azodicarboxylate (4.08 mL, 27 mmol, 1.0 eq) was added, and the reaction mixture was reacted at room temperature for 4 hours. The resulting reaction system was concentrated under reduced pressure, water (50 mL) was added to the system, and the reaction mixture was extracted with ethyl acetate (100 mL*3 times). Wash with saturated brine (100 mL), combine the organic phases and dry over anhydrous sodium sulfate. Compound 17d (6 g, 46%) was obtained as a reddish-brown solid after purification by column chromatography.
1H NMR (600 MHz, CDCl3) δ 8.04 (s, 1H), 7.47 (hd, J=5.4, 1.9 Hz, 5H), 7.09 (s, 1H), 5.90 (tt, J=3.2, 1.5 Hz, 1H), 4.67-4.55 (m, 2H), 4.38 (d, J=12.9 Hz, 1H), 4.28 (d, J=12.8 Hz, 1H), 3.98 (d, J=16.7 Hz, 1H), 3.93 (s, 3H), 3.91 (s, 3H), 3.78 (dd, J=19.0, 12.3 Hz, 1H), 3.48 (d, J=16.5 Hz, 1H), 3.06 (s, 1H), 2.75 (s, 1H), 2.54 (d, J=18.6 Hz, 1H).
LC-MS(ESI): [M+H]+=474.26/476.27.
Under the condition of nitrogen protection, compound 17d (8.4 g, 17 mmol), tri-n-butyltin hydrogen (10.2 g, 34 mmol) and azobisisobutyronitrile (574 mg, 3.4 mmol) were dissolved in toluene (84 mL), heated to 110° C. for reaction 4 hours. The reaction system was cooled to room temperature, concentrated under reduced pressure, added water (100 mL), and extracted with ethyl acetate (100 mL*3 times). Wash with saturated brine (100 mL), combine the organic phases and dry over anhydrous sodium sulfate. After purification by column chromatography, reddish-brown oily compound 17e (5 g, 71%) was obtained.
1H NMR (400 MHz, CDCl3) δ 7.67 (s, 1H), 7.55-7.41 (m, 5H), 6.97 (s, 1H), 4.47 (s, 2H), 4.27 (s, 2H), 3.91 (s, 3H), 3.89 (s, 3H), 3.70 (d, J=12.3 Hz, 2H), 2.78-2.63 (m, 2H), 2.52 (td, J=14.3, 4.0 Hz, 2H), 1.94 (d, J=14.5 Hz, 2H).
LC-MS(ESI): [M+H]+=396.40.
Compound 17e (2.3 g, 5.8 mmol) was dissolved in methanol (20 mL) and water (2 mL), and lithium hydroxide (1.39 g, 58 mmol) was added to the system. React at room temperature for 4 hours. The reaction system was concentrated under reduced pressure, and compound 17f (1.28 g, 60%) was obtained as a light reddish brown solid after reverse purification.
1H NMR (600 MHz, DMSO-d6) δ 7.51 (ddd, J=12.6, 6.6, 3.4 Hz, 6H), 6.98 (s, 1H), 4.64 (s, 2H), 4.36 (s, 2H), 3.39 (s, 2H), 3.13 (t, J=13.4 Hz, 2H), 2.15 (dd, J=20.6, 8.4 Hz, 2H), 1.96 (d, J=14.1 Hz, 2H).
LC-MS(ESI): [M+H]+=368.34.
Under nitrogen protection conditions, compound 17f (3.0 g, 8 mmol), sodium acetate (3.3 g, 24 mmol) and compound 3-amino-2,6-piperidinedione hydrochloride (1.3 g, 10 mmol, 1.25 eq) were dissolved in acetic acid (30 mL), the system was reacted at 110° C. for 4 hours. The reaction system was cooled to room temperature, concentrated under reduced pressure, added water (50 mL), and extracted with ethyl acetate (100 mL*3 times). Wash with saturated brine (80 mL), combine the organic phases and dry over anhydrous sodium sulfate. After purification by column chromatography, 17 g (1.8 g, 48%) of light yellow solid compound was obtained.
1H NMR (600 MHz, CDCl3) δ 7.69 (s, 1H), 7.54-7.43 (m, 5H), 7.25 (s, 1H), 4.96 (dd, J=12.7, 5.4 Hz, 1H), 4.56 (s, 2H), 4.28 (s, 2H), 2.97-2.75 (m, 4H), 2.70 (t, J=13.6 Hz, 2H), 2.63-2.53 (m, 2H), 1.99 (d, J=14.6 Hz, 2H), 1.63-1.57 (m, 2H).
LC-MS(ESI): [M+H]+=460.34.
Under nitrogen protection, compound 17g (3.0 g, 6.5 mmol, 1.0 eq) was dissolved in methanol (30 mL), and 10% wet palladium on carbon (600 mg) was added to the system. After hydrogen replacement for 3 times, the reaction was carried out at room temperature overnight. The reaction system was filtered, the filter cake was washed three times with methanol (15 mL), the obtained filtrate was spin-dried under reduced pressure, and the concentrated crude product was purified by reverse column chromatography to obtain white solid compound 17 (630 mg, 26%).
1H NMR (600 MHz, CD3OD) δ 7.78 (s, 1H), 7.26 (d, J=0.6 Hz, 1H), 5.14-5.10 (m, 1H), 4.74 (s, 2H), 3.38 (dt, J=12.7, 3.2 Hz, 2H), 3.05 (td, J=13.1, 3.0 Hz, 2H), 2.90-2.85 (m, 1H), 2.78 (dd, J=4.4, 2.6 Hz, 1H), 2.77-2.69 (m, 2H), 2.14 (ddt, J=13.2, 11.0, 4.1 Hz, 4H).
LC-MS(ESI): [M+H]+=370.33.
Under nitrogen protection conditions, methyl 5-bromo-4-methoxy-2-methylbenzoate (5 g, 19.30 mmol), NBS (3.61 g, 20.30 mmol) and AIBN (316.9 mg, 1.90 mmol) were dissolved in carbon tetrachloride (50 mL), the temperature was raised to 75° C., then react overnight. After the reaction was completely completed as detected by TLC, the reaction system was cooled to room temperature and then saturated aqueous sodium thiosulfate solution was added. After stirring for 30 minutes, it was extracted with dichloromethane, washed with saturated brine, and the organic phases were combined and dried over anhydrous sodium sulfate. After concentration, the crude product was purified by column chromatography to obtain white solid compound 18a (6.9 g, 96.7%).
LC-MS (ESI): [M+H]+=336.90
Under nitrogen protection conditions, compound 18a (7 g, 20.70 mmol) and 3-amino-2,6-piperidinedione hydrochloride (5.1 g, 31.10 mmol) were dissolved in acetonitrile (100 mL), and N, N-diisopropylethylamine (13.4 g, 103.60 mmol) was added to the reaction system, react overnight at 80° C. The obtained reaction system was concentrated under reduced pressure, and after adding acetic acid (50 mL), the reaction was carried out at 120° C. for 2 hours. After the reaction was completely completed as detected by TLC, the reaction system was concentrated under reduced pressure and purified by reverse column chromatography to obtain solid compound 18b (5.3 g, 72.5%).
LC-MS (ESI): [M+H]+=353.15
Under nitrogen protection, compound 18b (5 g, 14.20 mmol) was dissolved in dichloromethane (10 mL), and a solution of boron tribromide in dichloromethane (1 M, 140 mL) was slowly added at 0° C., and the reaction was carried out overnight at room temperature after the addition was complete. After the reaction was completely completed by TLC detection, the resulting reaction system was concentrated under reduced pressure to remove most of the boron tribromide, then dichloromethane (50 mL) was added to the system, and the reaction system was quenched with a small amount of methanol. The reaction system was concentrated under reduced pressure, and purified by reverse column chromatography to obtain an off-white solid compound 18c (3.6 g, 75.0%).
LC-MS (ESI): [M+H]+=338.99
Compound 18d was prepared by referring to the similar procedure of Step 3 in Example 17.
1H NMR (600 MHz, DMSO-d6) δ 11.00 (s, 1H), 7.90 (s, 1H), 7.55-7.46 (m, 5H), 7.42 (s, 1H), 5.91 (s, 1H), 5.08 (dd, J=13.3, 5.1 Hz, 1H), 4.75 (s, 2H), 4.45-4.34 (m, 3H), 4.27 (d, J=17.5 Hz, 1H), 3.71 (s, 2H), 3.56 (s, 1H), 3.17 (s, 1H), 2.91 (ddd, J=17.3, 13.6, 5.5 Hz, 1H), 2.63-2.57 (m, 1H), 2.41 (ddd, J=21.9, 16.2, 11.7 Hz, 3H), 2.00 (dtd, J=12.7, 5.4, 2.3 Hz, 1H).
LC-MS(ESI): [M+H]+=524.09
Compound 18e was prepared by referring to the similar procedure of Step 4 in Example 17.
1H NMR (600 MHz, DMSO-d6) δ 10.97 (s, 1H), 7.56-7.47 (m, 5H), 7.42 (s, 1H), 7.04 (s, 1H), 5.03 (dd, J=13.2, 4.9 Hz, 1H), 4.64 (q, J=9.3 Hz, 2H), 4.39-4.33 (m, 3H), 4.23 (d, J=17.3 Hz, 1H), 3.33-3.24 (m, 2H), 3.16 (d, J=16.2 Hz, 2H), 2.94-2.84 (m, 1H), 2.59 (d, J=16.7 Hz, 1H), 2.41-2.32 (m, 1H), 2.16 (t, J=12.4 Hz, 2H), 2.00-1.92 (m, 3H).
LC-MS(ESI): [M+H]+=446.35
Under nitrogen protection, compound 18e (25 mg, 0.06 mmol) was dissolved in methanol (2 mL), and 10% wet palladium/carbon (24 mg) was added to the system. After hydrogen replacement for 3 times, the reaction was carried out at room temperature overnight. The reaction system was filtered, the filter cake was washed three times with methanol (5 mL), the obtained filtrate was spin-dried under reduced pressure, and the concentrated crude product was purified by reverse column chromatography to obtain a white solid compound 18 (8.7 mg, 43%).
1H NMR (600 MHz, DMSO-d6) δ 10.97 (s, 1H), 7.47 (s, 1H), 7.04 (s, 1H), 5.06 (dd, J=13.3, 5.1 Hz, 1H), 4.65-4.60 (m, 2H), 4.36 (d, J=17.2 Hz, 1H), 4.24 (d, J=17.2 Hz, 1H), 3.36-3.34 (m, 2H), 3.04 (d, J=8.5 Hz, 2H), 2.95-2.86 (m, 1H), 2.60 (d, J=17.1 Hz, 1H), 2.38 (qd, J=13.2, 4.4 Hz, 1H), 2.10-2.03 (m, 2H), 2.00-1.95 (m, 1H), 1.91-1.85 (m, 2H).
LC-MS(ESI): [M+H]+=356.35
19a was prepared by referring to the similar procedure of Step 1 in Example 1.
1H NMR (600 MHz, CDCl3) δ 11.31 (s, 1H), 7.31-7.28 (m, 1H), 6.87-6.86 (m, 1H), 6.75-6.74 (m, 1H), 3.99 (s, 3H), 2.56 (s, 3H).
LC-MS (ESI): [M+H]+=167.13
Under nitrogen protection, compound 19a (10 g, 60.24 mmol) was dissolved in trifluoroacetic acid (20 mL), NIS (16.2 g, 72.00 mmol) was added, and the mixture was kept at room temperature for 16 h. After the reaction was complete by TLC detection, the reaction was quenched with saturated Na2S2O3 aqueous solution, washed with water, extracted with ethyl acetate, dried over anhydrous sodium sulfate, concentrated to obtain a crude product, which was purified in reverse to obtain compound 19b (3.8 g, 21.6%).
1H NMR (600 MHz, CDCl3) δ 12.19 (s, 1H), 7.77 (d, J=8.0 Hz, 1H), 6.56 (dd, J=8.0, 0.8 Hz, 1H), 4.01 (s, 3H), 2.54 (s, 3H).
Under nitrogen protection condition, compound 19b (3.8 g, 13.02 mmol) and butyl vinyl ether (3.9 g, 39.00 mmol) were dissolved in methanol (20 ml), Pd(dppf)Cl2 (951 mg, 1.30 mmol) and TEA (3.95 g, 39.00 mmol) were added. Nitrogen replacement three times, and the reaction was performed at 60° C. for 16 h. The end of the reaction was detected by TLC, the reaction solution was spin-dried, dissolved by adding dichloromethane, washed with 5M/L HCl, washed with saturated brine, combined the organic phases, dried over anhydrous sodium sulfate, and concentrated through the column to obtain compound 19c (2.0 g, 73.8%).
1H NMR (600 MHz, CDCl3) δ 12.74 (s, 1H), 7.70 (d, J=8.2 Hz, 1H), 6.79 (d, J=8.2 Hz, 1H), 3.97 (s, 3H), 2.64 (s, 3H), 2.38 (s, 3H).
LC-MS (ESI): [M-OCH3+H]+=177.14
19d was prepared by referring to the similar procedure of Step 3 in Example 12.
1H NMR (600 MHz, DMSO-d6) δ 7.72 (d, J=8.0 Hz, 1H), 6.99 (d, J=8.0 Hz, 1H), 3.87 (s, 3H), 3.78 (br, 2H), 2.99 (br, 2H), 2.84 (s, 2H), 2.28 (s, 3H), 1.96-1.78 (m, 2H), 1.59 (td, J=13.3, 4.8 Hz, 2H), 1.40 (s, 9H).
LC-MS (ESI): [M+H]+=390.35
19e was prepared by referring to the similar procedure of Step 4 in Example 12.
1H NMR (600 MHz, DMSO-d6) δ 7.37 (d, J=7.8 Hz, 1H), 6.80 (d, J=7.9 Hz, 1H), 5.44 (d, J=5.5 Hz, 1H), 4.67 (q, J=6.7 Hz, 1H), 3.80 (s, 3H), 3.76 (br, 2H), 2.97 (br, 2H), 2.17 (s, 3H), 2.06 (dd, J=13.5, 6.2 Hz, 1H), 1.85-1.66 (m, 3H), 1.60-1.47 (m, 2H), 1.41 (s, 9H).
LC-MS (ESI): [M+H]+=392.40
Under nitrogen protection, compound 19e (1.5 g, 3.83 mmol) was dissolved in toluene (25 mL), p-toluenesulfonic acid (659.8 mg, 3.83 mmol) was added, and the reaction mixture was carried out at 110° C. for 12 hours. After the reaction was detected by TLC, the obtained reaction system was concentrated under reduced pressure and the crude compound 19f was directly used in the next step (850 mg, 81%).
LC-MS (ESI): [M+H]+=274.25
Under the condition of nitrogen protection, compound 19f (850 mg, 3.11 mmol) was dissolved in dichloromethane (20 mL), Boc2O (1.36 g, 6.22 mmol) and TEA (600 mg, 6.22 mmol) were added to the reaction system, and the reaction mixture was carried out at room temperature to 25° C. for 1 hour. After the reaction was detected by TLC, the reaction system was cooled to room temperature, concentrated under reduced pressure, added water, extracted with ethyl acetate, washed with saturated brine, combined with organic phases and dried over anhydrous sodium sulfate. After purification by column chromatography, 19g (1 g, 86%) of white solid compound was obtained.
1H NMR (600 MHz, DMSO-d6) δ 7.07 (d, J=7.6 Hz, 1H), 6.78 (d, J=7.6 Hz, 1H), 6.48 (d, J=9.8 Hz, 1H), 5.72 (d, J=9.8 Hz, 1H), 3.83 (s, 5H), 3.07 (s, 2H), 2.18 (s, 3H), 1.81 (dq, J=14.3, 2.6 Hz, 2H), 1.57 (td, J=13.1, 4.8 Hz, 2H), 1.41 (s, 9H).
LC-MS(ESI): [M+H]+=374.40
19h was prepared by referring to the similar procedure of Step 6 in Example 12.
LC-MS (ESI): [M+H]+=452.25
Under nitrogen protection, compound 19h (1.2 g, 2.65 mmol), DIPEA (1 g, 7.95 mmol) and 3-amino-2,6-piperidinedione hydrochloride (654.9 mg, 3.98 mmol) were dissolved in acetonitrile (30 mL), the system was reacted at 85° C. for 16 hours. After the reaction was detected by TLC, the reaction system was cooled to room temperature, concentrated under reduced pressure, added water, extracted with ethyl acetate, washed with saturated brine, combined with organic phases and dried over anhydrous sodium sulfate. Compound 19i (400 mg, 32%) was obtained as a gray solid after purification by column chromatography.
1H NMR (600 MHz, DMSO-d6) δ 11.00 (s, 1H), 7.31 (d, J=7.5 Hz, 1H), 7.04 (d, J=7.5 Hz, 1H), 6.57 (d, J=9.9 Hz, 1H), 5.79 (d, J=9.9 Hz, 1H), 4.99 (d, J=5.1 Hz, 1H), 4.44-4.16 (m, 2H), 3.79 (s, 2H), 3.19 (d, J=22.8 Hz, 4H), 2.87 (ddd, J=18.2, 13.6, 5.4 Hz, 1H), 2.73-2.56 (m, 1H), 2.51 (p, J=1.9 Hz, 1H), 2.06-1.56 (m, 3H), 1.41 (s, 9H).
LC-MS(ESI): [M-Boc+H]+=368.19
Under nitrogen protection, compound 19i (400 mg, 0.85 mmol) was dissolved in THF (10 mL), and 10% wet palladium on carbon (250 mg) was added to the system. After hydrogen replacement for 3 times, the reaction was carried out overnight at 50° C. After the reaction was detected by TLC, the reaction system was filtered, the filter cake was washed three times with methanol (15 mL), the obtained filtrate was spin-dried under reduced pressure, and the concentrated crude solid compound 19j was directly used in the next step (380 mg, 94%).
LC-MS (ESI): [M-Boc+H]+=370.29
Under nitrogen protection, compound 19j (380 mg, 0.80 mmol) was dissolved in dichloromethane (5 mL), and TFA (2 mL) was added to the system. React for 1 h at 25° C. After the reaction was detected by TLC, the reaction solution was spin-dried under reduced pressure, and a yellow solid compound 19 (290 mg, 97%) was prepared by pre-HPLC.
1H NMR (600 MHz, DMSO-d6) δ 10.98 (s, 1H), 7.34 (d, J=7.7 Hz, 1H), 7.04 (d, J=7.6 Hz, 1H), 4.97 (dd, J=13.3, 5.2, 1H), 4.43-4.20 (m, 2H), 3.30-3.08 (m, 4H), 2.97-2.78 (m, 3H), 2.61 (dt, J=17.1, 3.7 Hz, 1H), 2.47-2.31 (m, 1H), 2.04-1.88 (m, 5H), 1.80 (tt, J=12.6, 5.0 Hz, 2H).
LC-MS(ESI): [M+H]+=370.39
Compound 20a was prepared by referring to the procedure similar to Step 3 in Example 12.
1H NMR (600 MHz, CDCl3) δ 7.84 (d, J=8.0 Hz, 1H), 6.96 (d, J=8.0 Hz, 1H), 4.09 (d, J=9.5 Hz, 2H), 3.99 (s, 3H), 3.96 (d, J=9.5 Hz, 2H), 3.06 (s, 2H), 2.39 (s, 3H), 1.46 (s, 9H).
LC-MS (ESI): [M-Boc+H]+=262.17
Compound 20b was prepared by referring to the similar steps of Step 4 in Example 12.
LC-MS (ESI): [M-Boc+H]+=264.17
Under nitrogen protection, compound 20b (900.0 mg, 2.48 mmol) was dissolved in toluene (10 mL), TsOH (471 mg, 2.48 mmol) was added, and the reaction mixture was carried out at 110° C. for 2 h. After the reaction was detected by TLC, the reaction solution was spin-dried to obtain the crude compound 20c (1.1 g, 181.1%), which could be directly carried out in the next step without purification.
LC-MS (ESI): [M+H]+=246.11
Under nitrogen protection, compound 20c (1.1 g, 1 eq) was dissolved in dichloromethane (15 mL), (Boc)2O (1.95 g, 2.0 eq) and TEA (1.36 g mg, 3.0 eq) were added, and the reaction mixture was performed at room temperature for 2 h. After the reaction was detected by TLC, the reaction solution was spin-dried and passed through the column to obtain compound 20d (300.0 mg, 19.4%).
1H NMR (600 MHz, CDCl3) δ 6.95 (d, J=7.6 Hz, 1H), 6.76 (d, J=7.6 Hz, 1H), 6.45 (d, J=9.8 Hz, 1H), 5.93 (d, J=9.8 Hz, 1H), 4.24 (d, J=9.5 Hz, 2H), 3.99 (d, J=9.5 Hz, 2H), 3.96 (s, 3H), 2.30 (s, 3H), 1.48 (s, 9H).
LC-MS (ESI): [M-Boc+H]+=256.17
Compound 20e was prepared by referring to the step similar to Step 6 in Example 12.
LC-MS (ESI): [M-Boc+H]+=324.07
Under nitrogen protection, compound 20e (600 mg, 1 eq) was dissolved in CAN (10 mL), 3-aminopiperidine-2,6-dione hydrochloride (287 mg, 1.5 eq) and DIEA (448 mg, 3.0 eq) were added to the reaction solution and reacted at 80° C. for 16 h. After the reaction was detected by TLC, the reaction solution was spin-dried and passed through the column to obtain compound 20f (60 mg, two-step yield 21.4%).
1H NMR (600 MHz, DMSO-d6) δ 11.00 (s, 1H), 7.34 (d, J=7.6 Hz, 1H), 7.08 (d, J=7.6 Hz, 1H), 6.67 (d, J=9.9 Hz, 1H), 6.20 (d, J=9.9 Hz, 1H), 4.99 (dd, J=13.2, 5.2 Hz, 1H), 4.37 (d, J=17.6 Hz, 1H), 4.25 (d, J=17.6 Hz, 1H), 4.04-4.01 (m, 4H), 2.93-2.87 (m, 1H), 2.62-2.57 (m, 1H), 2.41-2.33 (m, 1H), 2.00-1.96 (m, 1H), 1.41 (s, 9H).
LC-MS (ESI): [M-Boc+H]+=340.19
Under nitrogen protection, compound 20f (60.0 mg, 1 eq) was dissolved in THF (10 mL), Pd/C (60 mg, 1 eq) was added, hydrogen was replaced three times, and the reaction was performed at 50° C. for 16 h. After the reaction was detected by TLC, the reaction solution was spin-dried to obtain 20 g (80 mg) of the crude compound, which could be directly carried out to the next reaction without purification.
LC-MS (ESI): [M-Boc+H]+=342.17
Under the condition of nitrogen protection, compound 20g (80 mg, 1 eq) was dissolved in dichloromethane (5 mL), TFA (1 mL) was added, and the reaction mixture was performed at room temperature for 2 h. After the reaction was detected by TLC, the reaction solution was spin-dried to obtain compound 20 (15 mg, two-step yield 24.3%) by separation and lyophilization.
1H NMR (600 MHz, DMSO-d6) δ 10.98 (s, 1H), 7.35 (d, J=7.6 Hz, 1H), 7.08 (d, J=7.6 Hz, 1H), 4.97 (dd, J=13.2, 5.2 Hz, 1H), 4.35 (d, J=17.3 Hz, 1H), 4.23 (d, J=17.3 Hz, 1H), 4.14-4.10 (m, 4H), 2.93-2.85 (m, 3H), 2.62-2.58 (m, 1H), 2.42-2.35 (m, 1H), 2.22-2.19 (m, 2H), 1.99-1.95 (m, 1H).
LC-MS (ESI): [M+H]+=342.29
To a solution of 3-hydroxy-2-methylbenzoic acid (compound 21a) (10 g, 65.73 mmol) in methanol (100 mL) was slowly added thionyl chloride (15.64 g, 131.45 mmol). The reaction solution was stirred at 80° C. for 2 hours. The reaction solution was spin-dried to obtain gray solid 21b (10.0 g, 91.56%).
1H NMR (600 MHz, DMSO-d6) δ 9.71 (s, 1H), 7.19 (dd, J=7.7, 1.3 Hz, 1H), 7.09 (t, J=7.9 Hz, 1H), 7.02 (dd, J=8.0, 1.4 Hz, 1H), 3.80 (s, 3H), 2.29 (s, 3H).
LC-MS(ESI): [M+H]+=167.23.
N-iodosuccinimide (17.6 g, 78.23 mmol) was slowly added to a solution of methyl 3-hydroxy-2-methylbenzoate (compound 21b) (10.0 g, 60.18 mmol) in trifluoroacetic acid (80 mL) and methanol (40 mL). The reaction was stirred at room temperature for 2 hours. The resulting mixture was concentrated in vacuo and purified by C18 reverse phase column to obtain 21c (4 g, 22.76%) as a white solid.
1H NMR (600 MHz, DMSO-d6) δ 9.36 (s, 1H), 7.67 (d, J=8.2 Hz, 1H), 7.03 (d, J=8.2 Hz, 1H), 3.80 (s, 3H), 2.38 (s, 3H).
LC-MS(ESI): [M+H]+=364.22.
Under nitrogen protection, compound 21c (5.4 g, 18.49 mmol), butyl vinyl ether (5.56 g, 55.47 mmol) and Pd(dppf)Cl (1.35 g, 1.85 mmol) were dissolved in methanol (80 mL). Triethylamine (5.61 g, 55.47 mmol) was added to the reaction system. The reaction was raised up to 60° C., and stirred for 4 h. After the reaction, the mixture was filtered and the filtrate was concentrated to obtain a crude product, which was purified by column chromatography (PE:EA=20%) to obtain yellow oil 21d (3 g, 77.93%).
1H NMR (600 MHz, CD3OD) δ 7.28 (d, J=8.2 Hz, 1H), 7.18 (d, J=8.3 Hz, 1H), 3.87 (s, 3H), 2.38 (s, 3H), 1.59 (s, 3H).
LC-MS(ESI): [M+H]+=209.15.
21e was prepared by referring to the procedure similar to Step 3 in Example 12.
1H NMR (600 MHz, DMSO-d6) δ 7.65 (d, J=8.2 Hz, 1H), 7.33 (d, J=8.2 Hz, 1H), 3.86 (s, 3H), 2.89 (s, 2H), 2.38 (s, 3H), 1.92 (d, J=2.5 Hz, 1H), 1.89 (q, J=2.8 Hz, 1H), 1.64 (td, J=13.1, 4.8 Hz, 2H), 1.43 (s, 4H), 1.40 (s, 9H).
LC-MS(ESI): [M-Boc+H]+=290.18.
Compound 21f was prepared by referring to the procedure similar to Step 4 in Example 12.
LC-MS (ESI): [M-Boc+H]+=292.28.
Compound 21g was prepared by referring to the procedure similar to Step 5 in Example 14.
LC-MS (ESI): [M+H]+=274.47.
21h was prepared by referring to the similar procedure of Step 6 in Example 14.
1H NMR (600 MHz, CD3OD) δ 7.37 (d, J=7.9 Hz, 1H), 6.94 (d, J=7.9 Hz, 1H), 6.48 (d, J=9.8 Hz, 1H), 5.77 (d, J=9.7 Hz, 1H), 3.87 (s, 3H), 2.45 (s, 3H), 1.97-1.93 (m, 2H), 1.66 (td, J=13.9, 12.2, 4.8 Hz, 2H), 1.58 (s, 2H), 1.53 (s, 2H), 1.49 (s, 9H).
LC-MS(ESI): [M-Boc+H]+=274.17.
Compound 21i was prepared by referring to the similar procedure of Step 6 in Example 12.
LC-MS (ESI): [M+H]+=316.08.
Compound 21i (4 g, 8.84 mmol) was dissolved in acetonitrile (100 mL) solution, 3-aminopiperidine-2,6-dione hydrochloride (2.18 g, 13.26 mmol) and N,N-diisopropyl Ethylamine (3.43 g, 26.53 mmol) were added, the reaction was stirred overnight at 80° C. After the reaction was completed, the mixture was purified by normal phase column (100% EA) to obtain compound 21j (1 g, 24.19%) as a blue solid.
LC-MS (ESI): [M-tBu+H]+=412.29.
Compound 21j (90 mg, 192.50 mol) was dissolved in tetrahydrofuran (5 mL) solution, palladium on carbon (20 mg) was added, hydrogen was replaced three times, and the reaction was stirred overnight at 50° C. After the reaction was completed, the mixture was filtered to obtain white solid compound 21k (90 mg, 99.57%).
LC-MS (ESI): [M-tBu+H]+=414.39.
Compound 21k (90 mg, 191.68 mol) was dissolved in dichloromethane (2 mL) solution, trifluoroacetic acid (1 mL) was added, and the reaction was stirred at room temperature for 2 hours. After the reaction was completed, it was spin-dried and purified by preparative liquid phase to obtain white solid compound 21 (50 mg, 70.61%).
1H NMR (400 MHz, CD3OD) δ 7.36 (d, J=7.8 Hz, 1H), 7.32 (d, J=7.8 Hz, 1H), 5.18 (dd, J=13.3, 5.2 Hz, 1H), 4.55-4.40 (m, 2H), 3.38 (dd, J=12.5, 3.3 Hz, 4H), 3.03-2.89 (m, 3H), 2.81 (m, J=17.6, 4.6, 2.4 Hz, 1H), 2.51 (m, J=13.3, 4.7 Hz, 1H), 2.24-2.09 (m, 3H), 2.02 (t, J=6.8 Hz, 2H), 1.98-1.87 (m, 2H).
LC-MS(ESI): [M+H]+=370.39
Compound 22a (1 g, 4.69 mmol) was dissolved in triacetic acid (10 mL), and the solution was protected with nitrogen and cooled to 0° C. N-iodosuccinimide (1.16 g, 5.16 mmol) was added to the reaction solution at 0° C. The reaction solution was raised to room temperature overnight. The resulting mixture was poured into ice water and filtered. The filter cake was washed with water and purified by column chromatography (EA:PE=0%-15%) to obtain white solid compound 22b (800 mg, 50.28%).
1H NMR (600 MHz, DMSO-d6) δ 7.94-7.91 (m, 1H), 7.78 (dd, J=8.1, 1.1 Hz, 1H), 5.21 (s, 2H).
LC-MS(ESI): [M−H]−=230.86.
A solution of sodium hydroxide (413.04 mg, 10.33 mmol) in water (7 mL) was added to a solution of compound 22b in N,N-dimethylacetamide (3.5 mL). Cuprous oxide (59.11 mg, 413.07 μmol) was added to the above solution. The reaction was warmed to 80° C. and stirred overnight. The reaction solution was neutralized to neutrality with 1N hydrochloric acid, extracted with ethyl acetate and the organic phase was washed with saturated brine. The organic phase was concentrated under vacuum and the resulting mixture was purified by column chromatography (EA:PE=0%-35%) to give white solid compound 22c (249 mg, 52.64%).
1H NMR (600 MHz, DMSO-d6) δ 10.91 (s, 1H), 7.73 (d, J=8.0 Hz, 1H), 7.24 (d, J=8.0 Hz, 1H), 5.35 (s, 2H).
LC-MS(ESI): [M−H]−=230.86.
Compound 22d was prepared by referring to the procedure similar to Step 3 in Example 17.
1H NMR (600 MHz, DMSO-d6) δ 7.83 (d, J=8.0 Hz, 1H), 7.45 (d, J=8.0 Hz, 1H), 7.37-7.30 (m, 4H), 7.29-7.19 (m, 1H), 5.84 (s, 1H), 5.68 (s, 2H), 4.64 (s, 2H), 3.56 (s, 2H), 2.93 (s, 2H), 2.57 (t, J=5.7 Hz, 2H), 2.25 (s, 2H).
LC-MS(ESI): [M+H]+=414.20.
Compound 22e was prepared by referring to the procedure similar to Step 4 in Example 17.
1H NMR (600 MHz, CDCl3) δ 7.56 (d, J=7.7 Hz, 1H), 7.52-7.46 (m, 6H), 5.26 (s, 2H), 4.54 (s, 2H), 4.29 (s, 2H), 2.73 (t, J=13.1 Hz, 2H), 2.60 (td, J=14.6, 4.0 Hz, 2H), 2.12-1.91 (m, 4H).
LC-MS(ESI): [M−H]−=336.30.
A solution of sodium hydroxide (120.44 mg, 3.01 mmol) in water (2.0 mL) was added to a solution of compound 22e (202 mg, 602.26 μmol) in tetrahydrofuran (2.0 mL). The reaction was stirred overnight at room temperature. The resulting mixture was concentrated under vacuum and purified by reverse column to give compound 22f (98 mg, 46.04%) as a white solid.
LC-MS(ESI): [M−H]−=354.59.
Compound 22f (20 mg, 56.59 mol) and IBX (31.69 mg, 113.18 μmol) were dissolved in tetrahydrofuran and dimethyl sulfoxide=20:1 (0.5 mL), and the mixture was stirred at 80° C. for 2 h. The reaction was filtered and the resulting mixture was concentrated in vacuo, and the crude product was used directly in the next step without purification.
LC-MS (ESI): [M+H]+=352.30.
Compound 22g (48 mg, 136.60 mol) and 3-aminopiperidine-2,6-dione hydrochloride (44.96 mg, 273.19 μmol) were dissolved in tetrahydrofuran and dimethyl sulfoxide=20:1 (1.0 mL), and the reaction mixture was added a drop of N,N-diisopropylethylamine. The reaction was stirred at 50° C. for 1 h. The mixed solution was cooled to room temperature, and sodium triacetoxyborohydride (86.85 mg, 409.79 mol) and a drop of acetic acid were added, and the temperature was raised to 50° C. for 1 h. The mixture was concentrated under vacuum and purified by reverse-phase HPLC to give compound 22h (16 mg, 26.29%) as a white solid.
1H NMR (600 MHz, DMSO-d6) δ 11.00 (s, 1H), 7.46-7.56 (m, 5H), 7.34 (d, J=7.6 Hz, 1H), 7.26 (d, J=7.7 Hz, 1H), 5.09 (dd, J=13.3, 5.1 Hz, 1H), 4.67 (t, J=6.3 Hz, 2H), 4.43-4.34 (m, 3H), 4.24 (d, J=17.1 Hz, 1H), 3.16 (d, J=14.4 Hz, 2H), 2.91 (ddd, J=18.0, 13.7, 5.4 Hz, 1H), 2.64-2.56 (m, 1H), 2.44 (td, J=13.2, 4.6 Hz, 2H), 2.12 (d, J=15.4 Hz, 2H), 2.03-1.89 (m, 4H).
LC-MS(ESI): [M+H]+=446.35.
Compound 22h (150 mg, 336.69 mol) was dissolved in hexafluoroisopropanol (4.5 mL) solution, 20% palladium hydroxide/carbon (75 mg) was added, hydrogen was replaced three times, and the reaction mixture was stirred at room temperature under hydrogen for 5 hours. After the reaction was completed, the reaction solution was filtered, concentrated under reduced pressure and purified by reverse-phase HPLC to obtain compound 22 (35 mg, 29.25%) as a white solid.
1H NMR (600 MHz, CD3OD) δ7.46-7.40 (m, 2H), 5.16 (dd, J=13.4, 5.2 Hz, 1H), 4.70 (s, 2H), 4.52-4.40 (m, 2H), 3.54-3.47 (m, 2H), 3.24-3.14 (m, 2H), 2.96-2.87 (m, 2H), 2.80 (m, J=17.7, 4.6, 2.2 Hz, 1H), 2.52 (m, J=13.3, 4.6 Hz, 1H), 2.25-2.16 (m, 3H), 2.05-2.01 (m, 1H).
LC-MS(ESI): [M+H]+=356.29.
Under nitrogen protection conditions, dimethyl 4-bromo-3-hydroxyphthalate (10 g, 34.59 mmol) was dissolved in MeOH (100 mL), and 1-(vinyloxy)butane (13.86 g, 138.87 mmol), Pd(dppf)Cl2 (2.53 g, 3.46 mmol), and triethylamine (10.50 g, 103.78 mmol) were added. The reaction was stirred at 70° C. for 12 h. After the reaction was completed, HCl-1,4-dioxane (100 mL) was added and stirred at room temperature for 30 minutes. Then it was extracted three times with EA, the organic phases were combined, washed with saturated sodium chloride solution, dried over anhydrous sodium sulfate and concentrated to give the crude compound 23b (8 g, 91.6%) as a yellow oil, which was directly used in the next step.
LC-MS (ESI): [M−32+H]+=211.06
Compound 23c was prepared by referring to the procedure similar to Step 3 in Example 12.
1H NMR (600 MHz, CDCl3) δ 7.93 (d, J=8.2 Hz, 1H), 7.68 (d, J=8.2 Hz, 1H), 4.00 (s, 3H), 3.95 (s, 3H), 3.30 (t, J=9.7 Hz, 2H), 3.17 (t, J=9.7 Hz, 2H), 3.14 (s, 2H), 2.47 (t, J=9.7 Hz, 2H), 2.06 (t, J=9.7 Hz, 2H).
LC-MS(ESI): [M+H]+=334.19
Compound 23d was prepared by referring to the similar procedure of Step 4 in Example 12.
LC-MS (ESI): [M+H]+=336.29
Compound 23d (2.80 g, 6.43 mmol) was dissolved in 3 mL of toluene, p-toluenesulfonic acid (2.21 g, 12.86 mmol) was added, and the mixture was refluxed at 110° C. overnight. After the reaction was completed, the reaction mixture was concentrated, added water, extracted with ethyl ester and spin-dried, separated and purified by column chromatography to obtain yellow oily compound 23e (1.4 g, 68.6%).
LC-MS (ESI): [M+H]+=318.38
Compound 23e (1.40 g, 4.41 mmol) was dissolved in MeOH, Pd/C (700 mg) was added and the reaction was replaced by hydrogen stirred under hydrogen for 4 hours. After the reaction was completed, compound 23f (1.3 g, 93.8%) was obtained as a yellow oily compound after filtration and concentration, and the obtained crude product was directly sent to the next step.
LC-MS (ESI): [M+H]+=320.58
Compound 23f (1.4 g, 4.38 mmol) was dissolved in a mixed solution of THF:H2O (4:1), LiOH (629.87 mg, 26.30 mmol) was added and stirred overnight at room temperature. The crude product (1.35 g, 96.84%) was obtained after the reaction was completed and concentrated under reduced pressure.
LC-MS (ESI): [M+H]+=306.58
Compound 23g (1.50 g, 4.91 mmol), 3-aminopiperidine-2,6-dione hydrochloride (970.30 mg, 5.90 mmol) and sodium acetate (806.02 mg, 9.83 mmol) were dissolved in acetic acid (5 mL). The mixture was stirred at 110° C. for 5 h. After the reaction, suction filtration, the filtrate was concentrated under reduced pressure and purified by prep-HPLC (phase A=water, phase B=ACN, 0.1% TFA, B %=00% 50% in 30 min) to obtain a white solid compound 23 (0.55 g, 31.3%).
1H NMR (600 MHz, DMSO-d6) δ 11.11 (s, 1H), 7.62 (d, J=7.5 Hz, 1H), 7.39 (d, J=7.4 Hz, 1H), 5.08 (dd, J=12.9, 5.5 Hz, 1H), 3.28 (t, J=11.8 Hz, 2H), 3.09 (t, J=11.8 Hz, 2H), 2.96 (t, J=12.4 Hz, 2H), 2.91-2.78 (m, 3H), 2.63-2.52 (m, 2H), 2.05-1.94 (m, 3H), 1.91-1.81 (m, 2H).
LC-MS(ESI): [M+H]+=384.19
Compound 24b was prepared by referring to the same procedure as Step 1 in Example 23.
1H NMR (600 MHz, DMSO-d6) δ 10.70 (s, 1H), 7.48 (d, J=7.5 Hz, 1H), 7.12 (d, J=7.4 Hz, 1H), 3.88 (s, 3H), 3.87 (s, 3H), 2.42 (s, 3H).
LC-MS(ESI): [M−32+H]+=221.16
Compound 24c was prepared by referring to the similar step of Step 3 in Example 12.
1H NMR (600 MHz, CDCl3) δ 7.99 (dd, J=8.2, 0.9 Hz, 1H), 7.71 (dd, J=8.2, 1.0 Hz, 1H), 7.42-7.32 (m, 5H), 5.13 (s, 2H), 4.20 (d, J=9.7 Hz, 2H), 4.06 (d, J=9.7 Hz, 2H), 4.00 (s, 3H), 3.95 (s, 3H), 3.14 (s, 2H).
LC-MS(ESI): [M+H]+=438.20
Compound 24d was prepared by referring to the similar step of Step 4 in Example 12.
1H NMR (600 MHz, DMSO-d6) δ 7.63 (d, J=7.5 Hz, 1H), 7.44 (d, J=7.0 Hz, 1H), 7.42-7.30 (m, 5H), 5.11 (s, 2H), 4.50 (s, 1H), 4.25-4.15 (m, 1H), 4.10 (d, J=9.7 Hz, 4H), 3.88 (s, 3H), 3.86 (s, 3H), 2.60 (d, J=17.6, 2H).
LC-MS(ESI): [M+H]+=442.29
Compound 24d (180 mg, 407.76 μmol) was dissolved in trifluoroacetic acid, then triethylsilane (330 mg, 2.84 mmol) was added. The reaction mixture was stirred in an oil bath at 100° C. for 6 hours and then cooled to room temperature. Then another batch of triethylsilane (330 mg, 2.84 mmol) was added, the reaction was completed and spin-dried, the crude product was dissolved in methanol and stirred for 1 h and then spin-dried, the supernatant was poured off, the lower layer was the product, and no purification was required to obtain yellow oily compound 24e (160 mg, 96.6%), directly used in the next step.
LC-MS (ESI): [M+H]+=289.98
Under nitrogen protection, compound 24e (160 mg, 409.28 mol) was dissolved in methanol. Pd/C (50 mg, 10%) was added. The reaction was replaced by hydrogen, and stirred overnight at room temperature under hydrogen. After the reaction was completed and filtered, the filtrate was spin-dried to obtain compound 24f (130 mg, 81.3%) as a yellow oil, which was directly used in the next step without purification.
LC-MS (ESI): [M+H]+=292.18
Compound 24f (130 mg, 410.57 μmol) was dissolved in a mixed solvent of tetrahydrofuran/methanol/water=1:1:5, and lithium hydroxide (150 mg, 6.26 mmol) was added in an ice-water bath. The reaction mixture was stirred overnight at room temperature. After the reaction was completed, the pH was adjusted to 7 with aqueous hydrochloric acid, and the filtrate was spin-dried to obtain 24 g (140 mg, 105%) of the yellow oil product, which was directly used in the next step without purification.
LC-MS (ESI): [M+H]+=264.17
Compound 24g (200 mg, 1.22 mmol) and sodium acetate (200 mg, 2.44 mmol) were dissolved in acetic acid. The reaction mixture was stirred overnight in an oil bath at 120° C., after spin-drying, it was dissolved with methanol, purified in reverse phase, ACN/H2O (0-20% 40 min), to obtain a white solid compound 24 (80 mg, 18.4%).
1H NMR (600 MHz, DMSO-d6) δ 11.13 (s, 1H), 7.63 (d, J=7.5 Hz, 1H), 7.44 (d, J=7.0 Hz, 1H), 5.09 (dd, J=12.9, 5.5 Hz, 1H), 4.15 (s, 4H), 2.96 (t, J=6.5 Hz, 2H), 2.94-2.87 (m, 1H), 2.60-2.54 (m, 2H), 2.31-2.20 (m, 2H), 2.03-1.94 (m, 1H).
LC-MS(ESI): [M+H]+=356.30
Compound 26b was prepared by referring to the similar procedure of step 3 in Example 17.
1H NMR (600 MHz, DMSO-d6) δ 7.95 (d, J=8.4 Hz, 1H), 7.70 (d, J=8.4 Hz, 1H), 7.58-7.45 (m, 5H), 5.87-5.83 (m, 1H), 4.45 (s, 2H), 3.84 (s, 3H), 3.83 (s, 3H), 3.81 (s, 2H), 3.47-3.30 (m, 2H), 3.25-3.10 (m, 2H), 2.47-2.34 (m, 2H).
LC-MS(ESI): [M+H]+=474.09
Compound 25c was prepared by referring to the similar procedure of step 4 in Example 17.
1H NMR (600 MHz, DMSO-d6) δ 7.55-7.47 (m, 5H), 7.46 (d, J=8.4 Hz, 1H), 7.31 (d, J=8.4 Hz, 1H), 4.65 (s, 2H), 4.35 (s, 2H), 3.80 (s, 3H), 3.78 (s, 3H), 3.43-3.30 (m, 2H), 3.15-3.06 (m, 2H), 2.17-2.07 (m, 2H), 2.01-1.94 (m, 2H).
LC-MS(ESI): [M+H]+=360.29
Compound 25d was prepared by referring to the procedure similar to Step 5 in Example 17.
LC-MS (ESI): [M+H]+=367.99
Compound 25e was prepared by referring to the procedure similar to Step 6 in Example 17.
1H NMR (600 MHz, DMSO-d6) δ 11.10 (s, 1H), 7.62 (d, J=7.4 Hz, 1H), 7.55-7.47 (m, 5H), 7.43 (d, J=7.3 Hz, 1H), 5.11 (dd, J=13.0, 5.4 Hz, 1H), 4.81 (s, 2H), 3.96 (s, 2H), 3.38 (d, J=13.0 Hz, 2H), 3.09-3.01 (m, 2H), 2.91-2.84 (m, 1H), 2.63-2.58 (m, 2H), 2.08-2.00 (m, 3H), 1.98-1.90 (m, 2H).
LC-MS(ESI): [M+H]+=460.49
Compound 25 was prepared by referring to the similar procedure of Step 7 in Example 17.
1H NMR (600 MHz, DMSO-d6) δ 11.12 (s, 1H), 7.61 (d, J=7.4 Hz, 1H), 7.49 (d, J=7.3 Hz, 1H), 5.11 (dd, J=13.0, 5.4 Hz, 1H), 4.81 (s, 2H), 3.38 (d, J=13.0 Hz, 2H), 3.09-3.00 (m, 2H), 2.92-2.85 (m, 1H), 2.62-2.59 (m, 2H), 2.09-2.01 (m, 3H), 1.98-1.91 (m, 2H).
LC-MS(ESI): [M+H]+=370.09
Compound 26a (2.0 g, 10.6 mmol) was subsequently added to a solution of lithium diisopropylamide (1.3M in THF/hexane, 6.9 mL, 13 mmol) in tetrahydrofuran (10.6 mL) at −78° C., then react at this temperature for 2 hours, and quickly pour into dry ice. After it was warmed to room temperature, the solvent was spin-dried to obtain a white solid compound 26b (2.84 g, 115%), which was directly used in the next step.
LC-MS (ESI): [M−H]−=230.95
Iodomethane (2.5 g, 17.8 mmol), potassium carbonate (3.7 g, 26.7 mmol) were added to a solution of compound 26b (2.1 g, 8.9 mmol) in N,N-dimethylformamide (12 mL). The reaction was carried out at 80° C. for 1 hour. After the reaction was completed, water (50 mL) was added, the reaction mixture was extracted with ethyl acetate (100 mL*3 times), washed with saturated brine (100 mL), and the combined organic phases were dried over anhydrous sodium sulfate. After spin-drying, the obtained solution was purified by column chromatography (PE:EA=0-20%) to obtain compound 26c (1.9 g, 86%) as a yellow oil.
1H NMR (600 MHz, CDCl3) δ 7.27 (d, J=9.2 Hz, 1H), 7.14-7.09 (m, 1H), 3.97 (s, 3H), 2.25 (d, J=1.9 Hz, 3H).
LC-MS(ESI): [M+H]+=247.66.
Under nitrogen atmosphere, N-bromosuccinimide (1.6 g, 9.2 mmol) and azobisisobutyronitrile (62.8 mg, 0.38 mmol) were added to a solution of compound 26c (1.9 g, 7.6 mmol) in carbon tetrachloride (98 mL), and the reaction was performed overnight at 85° C. The solvent was spin-dried and purified by column chromatography (PE:EA=0-30%) to obtain compound 26d (219.9 mg, 8%) as a yellow oil.
1H NMR (600 MHz, CDCl3) δ 7.39 (d, J=8.4 Hz, 1H), 7.35-7.31 (m, 1H), 4.45 (s, 2H), 3.98 (s, 3H).
Under a nitrogen atmosphere, at −30° C., potassium tert-butoxide (75.7 mg, 0.67 mmol) was added to a solution of 1-tert-butoxycarbonylpiperidine-4-carbaldehyde (158.4 mg, 0.74 mmol) in tetrahydrofuran (4 mL). The reaction was performed at this temperature for 1 hour, then slowly added a solution of compound 26d (219.9 mg, 0.67 mmol) in tetrahydrofuran (3 mL) to it, and reacted at this temperature for 2 hours, then the reaction mixture was moved to room temperature for overnight reaction. The solvent was spin-dried and purified by column chromatography (PE:EA=0-40%) to obtain compound 26e (147.6 mg, 48%) as a colorless oil.
1H NMR (600 MHz, CDCl3) δ 9.55 (s, 1H), 7.30 (d, J=8.3 Hz, 1H), 7.00-6.95 (m, 1H), 3.96 (s, 3H), 4.00-3.75 (m, 2H), 2.95-2.70 (m, 2H), 2.77 (s, 2H), 1.94 (d, J=13.0 Hz, 2H), 1.58-1.46 (m, 2H), 1.43 (s, 9H).
LC-MS(ESI): [M-Boc+H]+=358.19.
At 0° C., sodium borohydride (1.9 g, 51.4 mmol) was slowly added in portions to a solution of compound 26e (4.7 g, 10.3 mmol) in methanol (50 mL), and then moved to room temperature for 2 hours. Water was added slowly to quench the reaction. The reaction mixture was spin-dried and purified by column chromatography (PE:EA=0-50%) to obtain colorless oily liquid compound 26f (4.14 g, 88%).
1H NMR (600 MHz, DMSO-d6) δ 7.51 (d, J=8.3 Hz, 1H), 7.41-7.36 (m, 1H), 4.78 (t, J=5.0 Hz, 1H), 3.90 (s, 3H), 3.46-3.39 (m, 2H), 3.20 (d, J=4.5 Hz, 4H), 2.66 (s, 2H), 1.99 (s, 2H), 1.42-1.32 (m, 11H).
LC-MS(ESI): [M-Boc+H]+=360.29.
Under nitrogen atmosphere, compound 26f (125 mg, 0.27 mmol) was slowly added to a solution of sodium hydride (11.9 mg, 0.27 mmol, 60% dispersed in liquid paraffin) in N,N-dimethylformamide (1 mL), and the reaction was moved to 110° C. for 1 hour. After cooling to room temperature, water (2 mL) was added slowly to quench the reaction. After adding water (10 mL), the reaction solution was extracted with ethyl acetate (20 ml*3 times), washed with saturated brine (40 mL), combined the organic phases, and purified by column chromatography (PE:EA=0-40%) to obtain a white solid compound 26g (73.0 mg, 61%).
1H NMR (600 MHz, CDCl3) δ 7.03 (d, J=8.2 Hz, 1H), 6.92 (d, J=8.2 Hz, 1H), 3.92 (s, 3H), 3.90 (s, 2H), 3.59-3.47 (m, 2H), 3.37-3.28 (m, 2H), 2.61 (s, 2H), 1.50-1.40 (m, 13H).
LC-MS(ESI): [M-Boc+H]+=339.99
Under a nitrogen atmosphere, compound 26g (73.0 mg, 0.17 mmol) was dissolved in a mixed solvent of tetrahydrofuran/water (10:1, 1.1 mL), and then potassium vinyl trifluoroborate (33.5 mg, 0.25 mmol), palladium acetate (7.5 mg, 0.033 mmol), triphenylphosphine (6.1 mmol, 0.023 mmol) and cesium carbonate (108.2 mg, 0.33 mmol) were added and reacted overnight in an oil bath at 85° C. The solvent was spin-dried and purified by column chromatography (PE:EA=0-40%) to obtain white solid compound 26h (38.9 mg, 45%).
1H NMR (600 MHz, CDCl3) δ 7.08 (d, J=7.9 Hz, 1H), 7.04 (d, J=7.8 Hz, 1H), 6.63 (dd, J=17.4, 11.0 Hz, 1H), 5.69 (d, J=17.4 Hz, 1H), 5.28 (d, J=11.0 Hz, 1H), 3.96-3.88 (m, 5H), 3.60-3.50 (m, 2H), 3.38-3.29 (m, 2H), 2.67 (s, 2H), 1.52-1.42 (m, 13H).
LC-MS(ESI): [M-Boc+H]+=288.18
Sodium periodate (64.9 mg, 0.3 mmol) and potassium osmate dihydrate (1.4 mg, 0.003 mmol) were added to compound 26h (29.4 mg, 0.076 mmol) in acetone/water mixed solution (5:1,2.7 mL), react overnight at room temperature. The reaction mixture was spin-dried of the solvent and purified by column chromatography (PE:EA=0-40%) to obtain compound 26i (8.9 mg, 30%) as a white solid.
1H NMR (600 MHz, CDCl3) δ 9.89 (s, 1H), 7.37 (d, J=7.7 Hz, 1H), 7.25 (d, J=6.4 Hz, 1H), 4.00-3.94 (m, 5H), 3.59-3.48 (m, 2H), 3.42-3.33 (m, 2H), 2.75 (s, 2H), 1.54-1.48 (m, 2H), 1.48-1.41 (m, 11H).
LC-MS(ESI): [M-Boc+H]+=290.18
Compound 26i (500 mg, 1.3 mmol) and 3-amino-2,6-piperidinedione hydrochloride (316 mg, 1.9 mmol) were mixed in a mixed solvent of dichloromethane/methanol (1/2, 39 mL), After reacting at room temperature for 1 hour, sodium cyanoborohydride (241 mg, 3.8 mmol) was added, and the reaction was carried out overnight at room temperature. The solvent was spin-dried, and the mixture was purified by a reverse-phase column to obtain compound 26j (233 mg, 39%) as a white solid.
1H NMR (600 MHz, DMSO-d6) δ 10.95 (s, 1H), 7.28 (d, J=7.6 Hz, 1H), 7.00 (d, J=7.6 Hz, 1H), 5.02 (dd, J=13.3, 5.1 Hz, 1H), 4.30 (d, J=17.2 Hz, 1H), 4.17 (d, J=17.1 Hz, 1H), 4.01 (s, 2H), 3.50-3.43 (m, 2H), 3.30-3.25 (m, 2H), 2.94-2.87 (m, 1H), 2.73 (s, 2H), 2.60-2.55 (m, 1H), 2.38-2.30 (m, 1H), 1.99-1.91 (m, 1H), 1.45-1.35 (m, 11H), 1.35-1.29 (m, 2H).
LC-MS(ESI): [M+H]+=470.40
Trifluoroacetic acid (2 mL) was added to a solution of compound 26j (230 mg, 0.49 mmol) in dichloromethane (5 mL), and the reaction was carried out at room temperature for 1.5 hours. The solvent was spin-dried, and the mixture was purified by a preparative column to obtain compound 26 (151.2 mg, 84%) as a white solid.
1H NMR (600 MHz, DMSO-d6) δ 10.96 (s, 1H), 7.30 (d, J=7.6 Hz, 1H), 7.04 (d, J=7.4 Hz, 1H), 5.01 (dd, J=13.2, 4.9 Hz, 1H), 4.31 (d, J=17.1 Hz, 1H), 4.18 (d, J=17.2 Hz, 1H), 4.11-4.04 (m, 2H), 3.20-3.06 (m, 4H), 2.94-2.86 (m, 1H), 2.77 (s, 2H), 2.60-2.53 m, 1H), 2.39-2.30 (m, 1H), 1.98-1.91 (m, 1H), 1.69-1.53 (m, 4H).
LC-MS(ESI): [M+H]+=370.29.
Compound 1b (33.6 g, 160 mmol, 1.0 eq) was dissolved in trifluoroacetic acid (100 mL), N-bromosuccinimide (34.2 g, 192 mmol, 1.2 eq) was added to the reaction system, and reacted overnight at room temperature. The resulting reaction system was concentrated under reduced pressure, water (100 mL) was added and extracted with ethyl acetate (100 ml*3 times). The mixture was washed with saturated brine (100 mL). The organic phases was combined and dried over anhydrous sodium sulfate. The concentrated crude product was purified by reverse column chromatography to obtain light yellow solid compound 27a (9.8 g, 21.3%).
1H NMR (600 MHz, DMSO-d6) δ11.68 (s, 1H), 7.87 (d, J=12.0 Hz, 1H), 7.10 (d, J=12.0 Hz, 1H), 3.84 (s, 3H), 3.79 (s, 3H).
LCMS(ESI): [M−H]−=287.09.
Compound 27b was prepared by referring to the procedure similar to step 3 in Example 17.
1H NMR (600 MHz, DMSO-d6) δ8.00 (d, J=8.8 Hz, 1H), 7.40-7.29 (m, 5H), 7.27-7.24 (m, 1H), 5.84 (tt, J=3.4, 1.6 Hz, 1H), 4.68 (s, 2H), 3.86 (s, 3H), 3.81 (s, 3H), 3.54 (s, 2H), 2.93-2.92 (m, 2H), 2.54 (t, J=5.7 Hz, 2H), 2.16 (s, 2H).
LCMS (ESI): [M+H]+=474.09.
27c was prepared by referring to the similar procedure of step 4 in Example 17.
1H NMR (600 MHz, DMSO-d6) δ7.84 (d, J=8.5 Hz, 1H), 7.38-7.29 (m, 4H), 7.29-7.23 (m, 1H), 7.00 (d, J=8.5 Hz, 1H), 4.52 (s, 2H), 3.86 (s, 3H), 3.78 (s, 3H), 3.48 (s, 2H), 2.75 (dd, J=11.6, 3.7 Hz, 2H), 2.08-1.93 (m, 4H), 1.58 (d, J=12.2 Hz, 2H).
LCMS (ESI): [M+H]+=395.71
27d was prepared by referring to the similar procedure of step 5 in Example 17.
1H NMR (400 MHz, DMSO-d6) δ7.81 (d, J=8.5 Hz, 1H), 7.60-7.65 (m, 2H), 7.44-7.39 (m, 3H), 6.94 (d, J=8.5 Hz, 1H), 4.61 (s, 2H), 4.25 (s, 2H), 3.16 (s, 4H), 2.39 (s, 2H), 1.78 (d, J=12.2 Hz, 2H).
LCMS (ESI) [M+H]+=368.30
Compound 27e was prepared by referring to the procedure similar to step 6 in Example 17.
1H NMR (400 MHz, CD3OD) δ7.79 (d, J=8.2 Hz, 1H), 7.69-7.44 (m, 5H), 7.19 (d, J=8.1 Hz, 1H), 5.13 (dd, J=12.6, 5.4 Hz, 1H), 4.80 (s, 2H), 4.38 (s, 2H), 3.55 (d, J=13.0 Hz, 2H), 3.35-3.36 (m, 1H), 3.17 (t, J=13.4 Hz, 2H), 2.97-2.85 (m, 2H), 2.81-2.65 (m, 2H), 2.17-1.98 (m, 3H).
LCMS (ESI) [M+H]+=460.29
Compound 27 was prepared by referring to the steps similar to step 7 in Example 17.
1H NMR (600 MHz, CD3OD) δ7.79 (dd, J=8.2, 1.2 Hz, 1H), 7.19 (dd, J=8.1, 1.3 Hz, 1H), 5.15 (dd, J=12.9, 5.4 Hz, 1H), 4.78 (s, 2H), 3.49 (dt, J=13.3, 2.7 Hz, 2H), 3.14 (td, J=13.7, 2.8 Hz, 2H), 2.94-2.87 (m, 3H), 2.80-2.70 (m, 2H), 2.18-2.14 (m, 1H), 2.04 (d, J=14.3 Hz, 2H).
LCMS (ESI) [M+H]+=370.39
Compound 28b was prepared by referring to the procedure similar to step 3 in Example 26.
1H NMR (600 MHz, CDCl3) δ7.39 (d, J=8.1 Hz, 1H), 7.21-7.15 (m, 1H), 7.08-7.03 (m, 1H), 4.65 (d, J=1.7 Hz, 2H).
Compound 28c was prepared by referring to the similar procedure of step 4 in Example 26.
1H NMR (600 MHz, CDCl3) δ9.65 (d, J=2.9 Hz, 1H), 7.37 (d, J=8.0 Hz, 1H), 7.14-7.08 (m, 1H), 7.04-6.98 (m, 1H), 4.15-3.70 (m, 2H), 3.02 (d, J=2.4 Hz, 2H), 2.90-2.60 (m, 2H), 2.18-1.94 (m, 2H), 1.76-1.62 (m, 2H), 1.44 (s, 9H).
LC-MS (ESI): [M-Boc+H]+=300.06.
Compound 28d was prepared by referring to the procedure similar to step 5 in Example 26.
1H NMR (600 MHz, CDCl3) δ7.39 (d, J=8.0 Hz, 1H), 7.12-7.07 (m, 1H), 7.05-6.99 (m, 1H), 3.80-3.65 (m, 2H), 3.62 (d, J=6.2 Hz, 2H), 3.11 (t, J=11.0 Hz, 2H), 2.94 (d, J=2.6 Hz, 2H), 1.67-1.62 (m, 2H), 1.62-1.59 (m, 1H), 1.55-1.47 (m, 2H), 1.44 (s, 9H).
LC-MS (ESI): [M-Boc+H]+=301.98.
Compound 28e was prepared by referring to the procedure similar to step 6 in Example 26.
1H NMR (400 MHz, CDCl3) δ7.15 (dd, J=7.9, 1.1 Hz, 1H), 7.02-6.95 (m, 1H), 6.78 (dd, J=8.2, 1.1 Hz, 1H), 3.85 (s, 2H), 3.64-3.51 (m, 2H), 3.45-3.32 (m, 2H), 2.64 (s, 2H), 1.52-1.44 (m, 13H).
LC-MS (ESI): [M-tBu+H]+=325.98.
Triethylamine (2.5 g, 24.6 mmol) and Pd(dppf)2Cl2·CH2Cl2 (670.7 mg, 0.8 mmol) were added to a methanol solution (60 mL) of compound 28e (3.1 g, 8.2 mmol) under the carbon monoxide atmosphere, and subsequently shifted to 65° C. to react overnight. After the reaction was completed, it was spin-dried and purified by column chromatography (PE:EA=0-20%) to obtain colorless oily liquid compound 28f (2.7 g, 90%).
1H NMR (400 MHz, CDCl3) δ7.52 (d, J=7.5 Hz, 1H), 7.20-7.11 (m, 1H), 6.99 (d, J=8.0 Hz, 1H), 3.90 (s, 2H), 3.88 (s, 3H), 3.60-3.46 (m, 2H), 3.46-3.34 (m, 2H), 3.00 (s, 2H), 1.55-1.40 (m, 13H).
LC-MS (ESI): [M-Boc+H]+=262.17.
An aqueous solution (20 mL) of potassium hydroxide (1.5 g, 37 mmol) was added to compound 28f in methanol/tetrahydrofuran (1/1, 40 mL) at 0° C., and shifted to a 60° C. oil bath to react overnight. The solution was spin-dried, added 1M hydrochloric acid solution (50 mL), extracted with ethyl acetate (40 mL*3 times), washed with saturated brine (80 mL), combined the organic phases, and spin-dried to obtain the white solid compound 28g (2.5 g, 97%), which was directly used for the next step.
LC-MS (ESI): [M−H]−=346.20.
To a solution of compound 28g (84.9 mg, 0.244 mmol) in N, N-dimethylformamide (1 mL) was added iodine simple substance (62.0 mg, 0.244 mmol), iodobenzenediethyl ester (78.7 mg, 0.244 mmol) and palladium acetate (2.7 mg, 0.012 mmol) under air atmosphere. The mixture was reacted in an oil bath at 80° C. for half an hour. The resulting mixture was extracted with ethyl acetate (10 mL*3 times), washed with saturated brine (20 mL), combined the organic phases, spin-dried to undergo the preparative liquid phase, and freeze-dried to obtain white solid compound 28h (72.3 mg, 65%).
1H NMR (400 MHz, CDCl3) δ7.54 (d, J=8.6 Hz, 1H), 6.63 (d, J=8.5 Hz, 1H), 3.89 (s, 2H), 3.64-3.45 (m, 2H), 3.45-3.26 (m, 2H), 2.71 (s, 2H), 1.52-1.36 (m, 13H).
LC-MS (ESI): [M−H]−=472.10.
Compound 28i was prepared by referring to the procedure similar to step 2 in Example 26.
1H NMR (600 MHz, CDCl3) δ7.51 (d, J=8.7 Hz, 1H), 6.62 (d, J=8.7 Hz, 1H), 3.95 (s, 3H), 3.88 (s, 2H), 3.57-3.47 (m, 2H), 3.40-3.27 (m, 2H), 2.59 (s, 2H), 1.49-1.41 (m, 13H).
LC-MS (ESI): [M-Boc+H]+=388.19.
Compound 28j was prepared by referring to the procedure similar to step 7 in Example 26.
LC-MS (ESI): [M+H]+=388.09.
Compound 28k was prepared by referring to the procedure similar to step 8 in Example 26.
1H NMR (600 MHz, CDCl3) δ9.83 (s, 1H), 7.65 (d, J=8.5 Hz, 1H), 6.99 (d, J=8.5 Hz, 1H), 4.05-3.95 (m, 5H), 3.58-3.47 (m, 2H), 3.40-3.31 (m, 2H), 2.64 (s, 2H), 1.55-1.29 (m, 13H).
LC-MS (ESI): [M-tBu+H]+=334.19.
Compound 281 was prepared by referring to the similar procedure of step 9 in Example 26.
1H NMR (600 MHz, DMSO-d6) δ10.99 (s, 1H), 7.29 (d, J=8.2 Hz, 1H), 7.02 (d, J=8.2 Hz, 1H), 5.01 (dd, J=13.2, 5.1 Hz, 1H), 4.30 (d, J=16.9 Hz, 1H), 4.19 (d, J=16.9 Hz, 1H), 3.98 (s, 2H), 3.47-3.38 (m, 2H), 3.31-3.24 (m, 2H), 3.12-3.02 (m, 2H), 2.92-2.85 (m, 1H), 2.63-2.56 (m, 1H), 2.42-2.32 (m, 1H), 2.00-1.94 (m, 1H), 1.51-1.30 (m, 11H), 1.38-1.30 (m, 2H).
LC-MS (ESI): [M-Boc+H]+=370.29.
Compound 28 was prepared by referring to the procedure similar to step 10 in Example 26.
1H NMR (600 MHz, DMSO-d6) δ11.00 (s, 1H), 7.31 (d, J=8.2 Hz, 1H), 7.04 (d, J=8.2 Hz, 1H), 5.01 (dd, J=13.3, 5.1 Hz, 1H), 4.31 (d, J=17.0 Hz, 1H), 4.20 (d, J=17.0 Hz, 1H), 4.05 (s, 2H), 3.21-3.03 (m, 6H), 2.94-2.84 (m, 1H), 2.64-2.56 (m, 1H), 2.38 (qd, J=13.0, 4.2 Hz, 1H), 2.02-1.91 (m, 1H), 1.70-1.52 (m, 4H).
LC-MS (ESI): [M+H]+=370.29.
To a solution of compound 12 (15 mg, 31.03 mol) and diisopropylethylamine (13 mg, 100.58 μmol) in N,N-dimethylformamide (1 mL) at room temperature was added the compound P-1 (17 mg, 32.76 mol) and HATU (15 mg, 39.45 mol). The reaction was stirred at room temperature for 1.5 hours, and then purified by preparative high performance liquid chromatography to obtain the yellow solid compound PROTAC 1 (8.7 mg, 32.22%).
1H NMR (600 MHz, CD3OD) δ8.66 (s, 1H), 8.49 (d, J=8.5 Hz, 1H), 7.93 (d, J=8.4 Hz, 1H), 7.87 (s, 1H), 7.63 (dd, J=15.4, 8.1 Hz, 1H), 7.54 (d, J=9.5 Hz, 1H), 7.49 (s, 1H), 7.41 (d, J=6.6 Hz, 1H), 7.23 (dd, J=37.6, 8.1 Hz, 1H), 7.08-6.88 (m, 3H), 5.17-5.07 (m, 1H), 4.52-4.32 (m, 3H), 4.29 (s, 1H), 3.93 (d, J=5.4 Hz, 3H), 3.58-3.44 (m, 3H), 3.41-3.35 (m, 2H), 3.02-2.86 (m, 3H), 2.80 (t, J=15.9 Hz, 1H), 2.53-2.43 (m, 1H), 2.40-2.28 (m, 1H), 2.24-2.12 (m, 1H), 2.02-1.89 (m, 3H), 1.79 (s, 3H), 1.69-1.56 (m, 1H).
LC-MS(ESI): [M+H]+=870.45/872.45.
To a solution of compound 16 (10 mg, 20.60 mol) and diisopropylethylamine (8 mg, 61.90 mol) in N, N-dimethylformamide (1 mL) at room temperature, compound P-1 (11 mg, 21.20 μmol) and HATU (10 mg, 26.30 μmol) were added. The reaction was stirred at room temperature for 1.5 hours, and then purified by preparative high performance liquid chromatography to obtain the yellow solid compound PROTAC 2 (4.2 mg, 23.37%).
1H NMR (600 MHz, CD3OD) δ8.64 (s, 1H), 8.45 (d, J=8.5 Hz, 1H), 7.92-7.84 (m, 2H), 7.57 (dd, J=15.4, 8.1 Hz, 1H), 7.43 (s, 1H), 7.40 (d, J=7.8 Hz, 1H), 7.29 (s, 1H), 7.28-7.19 (m, 1H), 7.15 (d, J=15.4 Hz, 1H), 6.92 (s, 2H), 5.16-5.07 (m, 1H), 4.55-4.46 (m, 1H), 4.46-4.31 (m, 2H), 4.25-4.14 (m, 1H), 4.13-4.05 (m, 2H), 3.92 (d, J=8.4 Hz, 3H), 3.54 (s, 2H), 3.50-3.35 (m, 2H), 2.97-2.85 (m, 1H), 2.85-2.74 (m, 1H), 2.53-2.42 (m, 1H), 2.20-2.12 (m, 1H), 1.96 (s, 1H), 1.91-1.77 (m, 2H), 1.74-1.58 (m, 1H).
LC-MS(ESI): [M+H]+=872.09/873.97.
To a solution of compound 13 (10 mg, 21.96 μmol) and diisopropylethylamine (9 mg, 69.63 μmol) in N, N-dimethylformamide (1 mL) at room temperature, compound P-1 (12 mg, 23.12 μmol) and HATU (10 mg, 26.30 μmol) were added. The reaction was stirred at room temperature for 1.5 hours, and then purified by preparative high performance liquid chromatography to obtain the yellow solid compound PROTAC 3 (6.2 mg, 33.52%).
1H NMR (600 MHz, CD3OD) δ8.67 (s, 1H), 8.49 (d, J=8.5 Hz, 1H), 7.94 (d, J=8.4 Hz, 1H), 7.91 (s, 1H), 7.63 (dd, J=15.3, 8.2 Hz, 1H), 7.55 (s, 1H), 7.50 (s, 1H), 7.40 (s, 2H), 7.07 (s, 1H), 7.05-6.88 (m, 2H), 5.12 (td, J=12.9, 5.1 Hz, 1H), 4.47-4.34 (m, 2H), 4.27-4.17 (m, 4H), 4.14 (d, J=10.1 Hz, 1H), 3.96 (s, 3H), 3.78-3.53 (m, 3H), 3.04-2.86 (m, 4H), 2.82-2.75 (m, 1H), 2.53-2.43 (m, 1H), 2.28-2.21 (m, 2H), 2.19-2.12 (m, 1H).
LC-MS(ESI): [M+H]+=841.98/843.98.
To a solution of compound 15 (10 mg, 21.96 μmol) and diisopropylethylamine (9 mg, 69.63 μmol) in N, N-dimethylformamide (1 mL), compound P-1 (12 mg, 23.12 mol) and HATU (10 mg, 26.30 μmol) were added at room temperature. The reaction was stirred at room temperature for 1.5 hours, and then purified by preparative high performance liquid chromatography to obtain the yellow solid compound PROTAC 4 (7.8 mg, 42.17%).
1H NMR (600 MHz, CD3OD) δ8.65 (s, 1H), 8.45 (d, J=8.5 Hz, 1H), 7.92 (s, 1H), 7.88 (d, J=8.5 Hz, 1H), 7.60-7.55 (m, 1H), 7.44 (s, 1H), 7.39 (s, 2H), 7.31 (s, 1H), 7.28 (s, 1H), 6.86 (s, 2H), 5.14 (td, J=13.0, 5.1 Hz, 1H), 4.45-4.33 (m, 2H), 4.26-4.16 (m, 4H), 4.12 (d, J=10.0 Hz, 1H), 3.96 (s, 3H), 3.06-2.86 (m, 4H), 2.83-2.75 (m, 1H), 2.53-2.43 (m, 1H), 2.28-2.14 (m, 3H).
LC-MS(ESI): [M+H]+=841.98/843.98.
At room temperature, to a solution of tert-butyl 3-(2-aminoethoxy)propionate (20 mg, 105.68 μmol) and diisopropylethylamine (38 mg, 294.01 μmol) in N,N-dimethylformamide (1 mL), compound P-1 (50 mg, 96.35 μmol) and HATU (45 mg, 118.35 mol) were added. The reaction was stirred at room temperature for 1.5 hours. Compound P-2 (54 mg, 81.20%) was obtained as a yellow solid by reverse phase purification.
1H NMR (600 MHz, CD3OD) δ8.67 (s, 1H), 8.45 (d, J=8.5 Hz, 1H), 8.04 (s, 1H), 7.97 (d, J=8.7 Hz, 1H), 7.86 (d, J=8.5 Hz, 1H), 7.56 (dd, J=15.2, 8.4 Hz, 1H), 7.43 (s, 1H), 7.38 (dd, J=8.7, 2.0 Hz, 1H), 6.92 (s, 2H), 4.05 (s, 3H), 3.77 (t, J=6.0 Hz, 2H), 3.65 (t, J=5.5 Hz, 2H), 3.61 (t, J=5.5 Hz, 2H), 2.54 (t, J=6.1 Hz, 2H), 1.44 (s, 9H).
LC-MS(ESI): [M+H]+=690.11/692.04.
To a solution of compound P-2 (54 mg, 78.24 mol) in dichloromethane (3 mL) was added trifluoroacetic acid (1 mL) under stirring at room temperature. After the reaction was stirred at room temperature for 3 hours, the resulting mixture was concentrated in vacuo, dissolved by adding N, N-dimethylformamide (1 mL), to which was added diisopropylethylamine (52 mg, 402.33 mol), then added compound 1 (15 mg, 30.16 μmol) and HATU (15 mg, 39.45 μmol), and then purified by preparative high performance liquid chromatography to obtain the white solid compound PROTAC 5 (8.4 mg, 27.87%).
1H NMR (600 MHz, CD3OD) δ8.64 (s, 1H), 8.37 (d, J=8.4 Hz, 1H), 8.07-7.95 (m, 2H), 7.78 (d, J=8.4 Hz, 1H), 7.52-7.43 (m, 2H), 7.36 (s, 2H), 7.20 (s, 1H), 6.87 (s, 2H), 5.08-5.01 (m, 1H), 4.31 (d, J=11.9 Hz, 1H), 4.13-4.07 (m, 1H), 4.02 (d, J=3.2 Hz, 3H), 3.89-3.79 (m, 3H), 3.72-3.66 (m, 2H), 3.64-3.59 (m, 2H), 3.52-3.44 (m, 2H), 3.13 (t, J=12.6 Hz, 2H), 2.86-2.62 (m, 5H), 2.11-2.04 (m, 1H), 1.76 (d, J=11.4 Hz, 3H), 1.66-1.58 (m, 1H), 1.52 (d, J=13.0 Hz, 1H).
LC-MS(ESI): [M+H]+=999.13/1001.01.
To a solution of compound P-2 (54 mg, 78.24 mol) in dichloromethane (3 mL) was added trifluoroacetic acid (1 mL) with stirring at room temperature. After the reaction was stirred at room temperature for 3 hours, the resulting mixture was concentrated in vacuo, dissolved by adding N,N-dimethylformamide (1 mL), to which was added diisopropylethylamine (52 mg, 402.33 mol), and then added compound 4 (15 mg, 30.16 mol) and HATU (15 mg, 39.45 μmol). The resulting mixture was quenched with water and purified by preparative high performance liquid chromatography to obtain the white solid compound PROTAC 6 (5.4 mg, 17.92%).
1H NMR (600 MHz, CD3OD) δ8.65 (s, 1H), 8.40 (d, J=8.3 Hz, 1H), 8.07 (s, 1H), 8.00 (dd, J=8.7, 2.9 Hz, 1H), 7.81 (d, J=8.3 Hz, 1H), 7.52 (dd, J=15.5, 7.6 Hz, 1H), 7.48 (d, J=8.3 Hz, 1H), 7.44-7.31 (m, 2H), 7.06 (d, J=6.5 Hz, 1H), 6.92 (s, 2H), 5.07-5.02 (m, 1H), 4.13 (s, 2H), 4.03 (d, J=2.5 Hz, 2H), 4.01-3.94 (m, 2H), 3.89-3.80 (m, 2H), 3.72-3.54 (m, 6H), 2.88-2.63 (m, 6H), 2.10-2.04 (m, 1H), 1.63 (s, 1H), 1.55-1.33 (m, 5H).
LC-MS(ESI): [M+H]+=999.17/1001.17.
To a solution of compound P-2 (54 mg, 78.24 mol) in dichloromethane (3 mL) was added trifluoroacetic acid (1 mL) with stirring at room temperature. After the reaction was stirred at room temperature for 3 hours, the resulting mixture was concentrated in vacuo, dissolved by adding N, N-dimethylformamide (1 mL), to which was added diisopropylethylamine (52 mg, 402.33 mol), and then added compound 17 (15 mg, 31.03 mol) and HATU (15 mg, 39.45 μmol). The resulting mixture was quenched with water, and purified by preparative high performance liquid chromatography to obtain compound PROTAC 7 (8.2 mg, 26.82%) as a light yellow solid.
1H NMR (600 MHz, CD3OD) δ8.64 (d, J=2.9 Hz, 1H), 8.39 (s, 1H), 8.03-7.94 (m, 2H), 7.82 (d, J=7.6 Hz, 1H), 7.60 (d, J=10.9 Hz, 1H), 7.53 (dd, J=15.3, 8.3 Hz, 1H), 7.43-7.33 (m, 2H), 7.05 (s, 1H), 6.91 (s, 2H), 5.09 (dd, J=12.7, 5.4 Hz, 1H), 4.61 (s, 2H), 4.53 (d, J=13.6 Hz, 1H), 4.23-4.05 (m, 3H), 4.04 (s, 3H), 3.90-3.81 (m, 2H), 3.74-3.59 (m, 4H), 3.22-3.14 (m, 2H), 2.95-2.81 (m, 2H), 2.81-2.65 (m, 5H), 2.16-2.07 (m, 1H), 1.97-1.71 (m, 5H).
LC-MS(ESI): [M+H]+=984.98/986.98.
To a solution of compound P-2 (54 mg, 78.24 μmol) in dichloromethane (3 mL) was added trifluoroacetic acid (1 mL) with stirring at room temperature. After the reaction was stirred at room temperature for 3 hours, the resulting mixture was concentrated in vacuo, dissolved by adding N,N-dimethylformamide (1 mL), to which was added diisopropylethylamine (52 mg, 402.33 mol), and then added compound 3 (15 mg, 31.96 mol) and HATU (15 mg, 39.45 μmol). The resulting mixture was quenched with water and purified by preparative HPLC to obtain the yellow solid compound PROTAC 8 (7.7 mg, 24.80%).
1H NMR (600 MHz, CD3OD) δ8.64 (s, 1H), 8.39 (d, J=8.3 Hz, 1H), 8.06-7.99 (m, 1H), 7.97 (dd, J=8.6, 2.4 Hz, 1H), 7.80 (d, J=8.4 Hz, 1H), 7.50 (dd, J=15.2, 8.2 Hz, 1H), 7.46 (s, 1H), 7.40-7.28 (m, 2H), 7.16 (s, 1H), 6.89 (s, 2H), 5.09-5.01 (m, 1H), 4.39-4.28 (m, 2H), 4.06-4.00 (m, 4H), 3.83 (t, J=5.8 Hz, 2H), 3.71-3.66 (m, 2H), 3.66-3.59 (m, 2H), 2.89-2.79 (m, 3H), 2.76-2.60 (m, 3H), 2.56-2.44 (m, 2H), 2.19-2.04 (m, 3H).
LC-MS(ESI): [M+H]+=970.97/972.97.
To a dichloromethane (40 mL) solution of compound P-3 (1.5 g, 7.88 mmol) and pyridine (1.37 g, 17.35 mmol) was added Dess-Martin oxidizing agent (4.35 g, 10.25 mmol) at 0° C. under stirring. The reaction was stirred at room temperature for 14 hours. The resulting mixture was quenched with saturated aqueous sodium thiosulfate (50 mL) and saturated aqueous sodium bicarbonate (100 mL), and extracted three times by adding dichloromethane (150 mL). The organic phase was concentrated under vacuum, and the resulting mixture was purified by normal phase to obtain the colorless oily compound P-4 (1.2 g, 80.86%).
1H NMR (600 MHz, DMSO-d6) δ9.56 (s, 1H), 4.17 (s, 2H), 3.67 (t, J=6.2 Hz, 2H), 2.48 (t, J=6.2 Hz, 2H), 1.41 (s, 9H).
To a solution of N, N-dimethylformamide (15 mL) of N-tert-butoxycarbonylpiperazine (230 mg, 1.23 mmol) and diisopropylethylamine (380 mg, 2.94 mmol) was added compound P-5 (400 mg, 1.01 mmol).
The reaction was stirred at 110° C. for 14 hours. The resulting mixture was quenched with water (150 mL), and extracted three times with the addition of ethyl acetate (150 mL), The organic phase was concentrated under vacuum and the resulting mixture was purified by normal phase to obtain the yellow solid compound P-6 (379 mg, 68.66%).
1H NMR (600 MHz, DMSO-d6) δ11.57 (s, 1H), 9.90 (s, 1H), 8.24 (d, J=7.7 Hz, 1H), 7.41 (d, J=7.8 Hz, 1H), 7.30 (d, J=6.9 Hz, 1H), 7.26 (t, J=7.7 Hz, 1H), 6.11 (s, 1H), 3.55 (s, 4H), 3.43 (s, 4H), 2.43 (s, 3H), 2.24 (s, 3H), 1.43 (s, 9H).
LC-MS (ESI): [M+H]+=544.12.
To a solution of compound P-6 (380 mg, 78.24 μmol) in dichloromethane (20 mL) was added trifluoroacetic acid (4 mL) under stirring at room temperature. The reaction was stirred at room temperature for 3 hours, and the resulting mixture was concentrated in vacuo, dissolved by adding tetrahydrofuran (20 mL), to which was added diisopropylethylamine (420 mg, 3.25 mmol), and then added acetic acid (100 mg, 1.67 mmol) and titanic acid tetraisopropyl ester (0.5 mL). Finally compound P-4 (264 mg, 1.40 mmol) and sodium cyanoborohydride (176 mg, 2.80 mmol) were added. The reaction was stirred at room temperature for 3 hours, the resulting mixture was concentrated in vacuo, and light yellow oily compound P-7 (287 mg, 86.16%) was obtained by normal phase purification.
1H NMR (600 MHz, DMSO-d6) δ11.65 (s, 1H), 9.92 (s, 1H), 8.25 (s, 1H), 7.41 (d, J=7.8 Hz, 1H), 7.30 (d, J=7.0 Hz, 1H), 7.27 (t, J=7.7 Hz, 1H), 6.15 (s, 1H), 4.34 (s, 2H), 3.77-3.74 (m, 2H), 3.68-3.54 (m, 4H), 3.36 (s, 2H), 3.30-3.23 (m, 2H), 3.12 (s, 2H), 2.53 (d, J=6.1 Hz, 2H), 2.45 (s, 3H), 2.24 (s, 3H), 1.42 (s, 9H).
To a solution of compound P-7 (35 mg, 56.80 mol) in dichloromethane (2 mL) was added trifluoroacetic acid (0.5 mL) under stirring at room temperature. The reaction was stirred at room temperature for 3 hours. The resulting mixture was concentrated in vacuo, added N, N-dimethylformamide (1 mL) to dissolve, then added diisopropylethylamine (50 mg, 286.85 μmol) thereto, and then added compound 1 (30 mg, 60.31 μmol) and HATU (28 mg, 73.64 μmol). The reaction was stirred at room temperature for 3 hours, and then purified by preparative high performance liquid chromatography to obtain the white solid compound PROTAC 9 (16 mg, 28.67%).
1H NMR (600 MHz, CD3OD) δ8.18 (s, 1H), 7.64 (s, 1H), 7.38 (d, J=7.4 Hz, 1H), 7.29 (d, J=1.3 Hz, 1H), 7.27 (d, J=5.3 Hz, 1H), 7.25 (d, J=7.6 Hz, 1H), 6.18 (s, 1H), 5.10 (dd, J=12.8, 5.5 Hz, 1H), 4.62 (s, 2H), 4.38 (d, J=13.3 Hz, 1H), 3.92 (t, J=5.2 Hz, 2H), 3.89-3.80 (m, 3H), 3.74 (s, 2H), 3.59 (t, J=11.9 Hz, 1H), 3.53-3.48 (m, 2H), 3.44 (s, 1H), 3.27-3.16 (m, 3H), 2.99 (t, J=6.7 Hz, 2H), 2.92-2.67 (m, 6H), 2.53 (s, 3H), 2.34 (s, 3H), 2.16-2.09 (m, 1H), 1.98-1.87 (m, 4H), 1.81-1.72 (m, 1H), 1.69-1.58 (m, 1H).
LC-MS (ESI): [M+H]+=925.45.
To a solution of compound P-7 (35 mg, 56.80 mol) in dichloromethane (2 mL) was added trifluoroacetic acid (0.5 mL) under stirring at room temperature. The reaction was stirred at room temperature for 3 hours. The resulting mixture was concentrated in vacuo, dissolved by adding N, N-dimethylformamide (1 mL), added diisopropylethylamine (50 mg, 286.85 μmol) thereto, and then added compound 4 (30 mg, 60.31 mol) and HATU (28 mg, 73.64 mol). The reaction was stirred at room temperature for 3 hours, and then purified by preparative high performance liquid chromatography to obtain the white solid compound PROTAC 10 (9.7 mg, 17.38%).
1H NMR (600 MHz, CD3OD) δ8.18 (s, 1H), 7.62 (s, 1H), 7.38 (d, J=7.7 Hz, 1H), 7.30-7.23 (m, 2H), 7.21 (d, J=2.9 Hz, 1H), 6.18 (s, 1H), 5.10 (dd, J=12.8, 5.4 Hz, 1H), 4.62 (s, 3H), 4.12 (s, 2H), 3.91 (t, J=5.2 Hz, 2H), 3.87-3.80 (m, 3H), 3.78-3.64 (m, 3H), 3.63-3.55 (m, 2H), 3.54-3.47 (m, 2H), 3.46-3.41 (m, 1H), 3.26-3.16 (m, 1H), 2.94-2.84 (m, 3H), 2.79-2.68 (m, 4H), 2.53 (s, 3H), 2.34 (s, 3H), 2.15-2.09 (m, 1H), 1.67-1.46 (m, 5H).
LC-MS(ESI): [M+H]+=925.35/927.35.
To a solution of compound P-7 (35 mg, 56.80 mol) in dichloromethane (2 mL) at room temperature was added trifluoroacetic acid (0.5 mL) under stirring. The reaction was stirred at room temperature for 3 h. The resulting mixture was concentrated under vacuum, dissolved by adding N, N-dimethylformamide (1 mL), to which diisopropylethylamine (50 mg, 286.85 μmol) was added, followed by adding the compound 3 (30 mg, 63.92 μmol) and HATU (28 mg, 73.64 mol). The reaction was stirred at room temperature for 3 hours. The white solid compound PROTAC 11 (16 mg, 27.90%) was obtained by preparative HPLC purification.
1H NMR (600 MHz, CD3OD) δ8.18 (s, 1H), 7.64 (d, J=5.8 Hz, 1H), 7.38 (d, J=7.6 Hz, 1H), 7.30-7.22 (m, 3H), 6.15 (d, J=5.9 Hz, 1H), 5.08 (dd, J=12.8, 5.4 Hz, 1H), 4.61 (s, 2H), 4.38 (s, 2H), 4.13 (q, J=10.9 Hz, 2H), 3.94-3.88 (m, 2H), 3.88-3.78 (m, 3H), 3.71 (s, 2H), 3.36-3.34 (m, 1H), 3.50 (t, J=4.4 Hz, 2H), 3.26-3.20 (m, 1H), 3.04 (t, J=12.8 Hz, 2H), 2.90-2.82 (m, 1H), 2.78-2.59 (m, 4H), 2.49 (s, 4H), 2.35 (s, 3H), 2.31-2.22 (m, 2H), 2.17-2.09 (m, 1H).
LC-MS(ESI): [M+H]+=897.35/899.35.
To a solution of compound P-7 (35 mg, 56.80 mol) in dichloromethane (2 mL) was added trifluoroacetic acid (0.5 mL) under stirring at room temperature. The reaction was stirred at room temperature for 3 hours, the resulting mixture was concentrated in vacuo, N, N-dimethylformamide (1 mL) was added to dissolve, diisopropylethylamine (50 mg, 286.85 mol) was added thereto, and then compound 17 (30 mg, 62.06 μmol) and HATU (28 mg, 73.64 μmol) were added. The reaction was stirred at room temperature for 3 hours, and then purified by preparative high performance liquid chromatography to obtain the white solid compound PROTAC 12 (20 mg, 35.36%).
1H NMR (600 MHz, CD3OD) δ8.18 (s, 1H), 7.69 (s, 1H), 7.38 (d, J=7.4 Hz, 1H), 7.30-7.21 (m, 3H), 6.18 (s, 1H), 5.10 (dd, J=12.9, 5.4 Hz, 1H), 4.75 (s, 2H), 4.69-4.56 (m, 3H), 4.05 (d, J=14.2 Hz, 1H), 3.96-3.91 (m, 2H), 3.86 (t, J=5.1 Hz, 2H), 3.75 (s, 2H), 3.55-3.41 (m, 4H), 3.31-3.20 (m, 3H), 2.95-2.82 (m, 3H), 2.82-2.68 (m, 3H), 2.52 (s, 3H), 2.34 (s, 3H), 2.17-2.10 (m, 1H), 2.07-1.99 (m, 1H), 1.94-1.86 (m, 3H).
LC-MS(ESI): [M+H]+=911.35/913.25.
To a solution of compound P-7 (35 mg, 56.80 mol) in dichloromethane (2 mL) was added trifluoroacetic acid (0.5 mL) under stirring at room temperature. The reaction was stirred at room temperature for 3 hours, the resulting mixture was concentrated under vacuum, N, N-dimethylformamide (1 mL) was added to dissolve, diisopropylethylamine (50 mg, 286.85 mol) was added thereto, and then compound 14 (30 mg, 62.05 μmol) and HATU (28 mg, 73.64 μmol) were added. The reaction was stirred at room temperature for 3 hours, and then purified by preparative high performance liquid chromatography to obtain the white solid compound PROTAC 13 (14 mg, 24.75%).
1H NMR (600 MHz, CD3OD) δ8.18 (s, 1H), 7.38 (d, J=7.4 Hz, 1H), 7.31-7.22 (m, 4H), 6.18 (s, 1H), 5.14 (dd, J=13.3, 5.1 Hz, 1H), 4.61 (s, 2H), 4.45-4.32 (m, 3H), 3.92 (t, J=5.0 Hz, 2H), 3.88-3.80 (m, 3H), 3.79-3.67 (m, 2H), 3.63-3.55 (m, 1H), 3.52-3.41 (m, 3H), 3.28-3.17 (m, 3H), 2.98-2.92 (m, 2H), 2.92-2.87 (m, 1H), 2.87-2.71 (m, 3H), 2.55-2.42 (m, 4H), 2.36 (s, 3H), 2.20-2.14 (m, 1H), 1.96-1.84 (m, 4H), 1.77-1.68 (m, 1H), 1.65-1.55 (m, 1H), 1.41-1.37 (m, 1H).
LC-MS(ESI): [M+H]+=911.45/913.25.
To a solution of compound P-7 (35 mg, 56.80 mol) in dichloromethane (2 mL) was added trifluoroacetic acid (0.5 mL) under stirring at room temperature. The reaction was stirred at room temperature for 3 hours, the resulting mixture was concentrated in vacuo, N,N-dimethylformamide (1 mL) was added to dissolve, and diisopropylethylamine (50 mg, 286.85 mol) was added thereto, followed by adding the compound 15 (20 mg, 43.92 μmol) and HATU (20 mg, 52.60 μmol). The reaction was stirred at room temperature for 3 hours, and the purified by preparative high performance liquid chromatography to obtain the white solid compound PROTAC 14 (8.7 mg, 22.42%).
1H NMR (600 MHz, CD3OD) δ8.18 (d, J=2.8 Hz, 1H), 7.38 (d, J=7.6 Hz, 1H), 7.32-7.18 (m, 4H), 6.15 (d, J=27.6 Hz, 1H), 5.12 (dd, J=13.3, 5.1 Hz, 1H), 4.61 (s, 2H), 4.44-4.28 (m, 4H), 4.16-4.01 (m, 2H), 3.95-3.78 (m, 4H), 3.73 (s, 2H), 3.50 (dd, J=10.0, 5.0 Hz, 2H), 3.46-3.36 (m, 2H), 3.30-3.18 (m, 2H), 3.06-2.94 (m, 2H), 2.94-2.83 (m, 2H), 2.83-2.73 (m, 1H), 2.71-2.60 (m, 1H), 2.52-2.40 (m, 4H), 2.35 (s, 3H), 2.31-2.11 (m, 3H).
LC-MS(ESI): [M+H]+=883.36/885.26.
To a solution of compound P-7 (35 mg, 56.80 mol) in dichloromethane (2 mL) at room temperature was added trifluoroacetic acid (0.5 mL) under stirring. The reaction was stirred at room temperature for 3 h. The resulting mixture was concentrated under vacuum, dissolved by adding N, N-dimethylformamide (1 mL) to which diisopropylethylamine (50 mg, 286.85 μmol) was added, followed by adding the compound 16 (30 mg, 61.80 μmol) and HATU (28 mg, 73.64 mol). The reaction was stirred at room temperature for 3 hours. The white solid compound PROTAC 15 (24 mg, 42.51%) was obtained by preparative HPLC purification.
1H NMR (600 MHz, CD3OD) δ8.18 (s, 1H), 7.38 (d, J=7.4 Hz, 1H), 7.29 (s, 1H), 7.27 (d, J=5.2 Hz, 1H), 7.25 (d, J=7.7 Hz, 1H), 7.10 (s, 1H), 6.19 (s, 1H), 5.11 (dt, J=13.2, 5.2 Hz, 1H), 4.63 (s, 2H), 4.43-4.31 (m, 3H), 4.11-4.02 (m, 2H), 3.95-3.80 (m, 5H), 3.74 (s, 2H), 3.61-3.53 (m, 2H), 3.53-3.47 (m, 2H), 3.44 (s, 1H), 3.29-3.17 (m, 3H), 2.95-2.85 (m, 1H), 2.85-2.74 (m, 3H), 2.53 (s, 3H), 2.50-2.41 (m, 1H), 2.34 (s, 3H), 2.19-2.13 (m, 1H), 1.97-1.83 (m, 2H), 1.83-1.75 (m, 1H), 1.74-1.64 (m, 1H).
LC-MS(ESI): [M+H]+=913.45/915.35.
To a solution of compound P-7 (35 mg, 56.80 mol) in dichloromethane (2 mL) was added trifluoroacetic acid (0.5 mL) under stirring at room temperature. The reaction was stirred at room temperature for 3 hours, the resulting mixture was concentrated in vacuo, N, N-dimethylformamide (1 mL) was added to dissolve, diisopropylethylamine (50 mg, 286.85 mol) was added thereto, and then compound 13 (20 mg, 43.92 μmol) and HATU (20 mg, 52.60 μmol) were added. The reaction was stirred at room temperature for 3 hours, and the purified by preparative high performance liquid chromatography to obtain the white solid compound PROTAC 16 (9.0 mg, 23.20%).
1H NMR (600 MHz, CD3OD) δ8.18 (s, 1H), 7.55 (s, 1H), 7.38 (d, J=7.4 Hz, 1H), 7.30-7.23 (m, 2H), 6.99 (d, J=19.3 Hz, 1H), 6.16 (d, J=9.1 Hz, 1H), 5.14-5.05 (m, 1H), 4.61 (s, 2H), 4.43-4.29 (m, 4H), 4.15-4.04 (m, 2H), 3.93-3.86 (m, 2H), 3.86-3.77 (m, 2H), 3.72 (s, 2H), 3.53-3.46 (m, 2H), 3.46-3.39 (m, 2H), 3.27-3.17 (m, 2H), 3.03-2.95 (m, 2H), 2.95-2.85 (m, 2H), 2.81-2.74 (m, 1H), 2.66-2.57 (m, 1H), 2.56-2.46 (m, 3H), 2.46-2.37 (m, 1H), 2.35 (s, 3H), 2.24 (t, J=6.4 Hz, 2H), 2.19-2.11 (m, 1H).
LC-MS(ESI): [M+H]+=883.36/885.36.
To a solution of compound P-8 (100 mg, 224.25 mol) and compound P-4 (85 mg, 451.59 mol) in methanol (4 mL) was added acetic acid (1 mL) and tetraisopropyl titanate (1 mL), and finally sodium cyanoborohydride (46 mg, 732.02 mol) was added with stirring at room temperature. The reaction was stirred at room temperature for 14 hours. The resulting mixture was concentrated under vacuum and purified by preparative high performance liquid chromatography to obtain the light yellow oily compound P-9 (82 mg, 59.15%).
1H NMR (600 MHz, CD3OD) δ8.76 (s, 1H), 8.00 (s, 1H), 7.96 (dd, J=6.6, 2.6 Hz, 1H), 7.68 (ddd, J=8.9, 4.1, 2.7 Hz, 1H), 7.39 (t, J=8.9 Hz, 1H), 7.29 (s, 1H), 4.38 (t, J=5.6 Hz, 2H), 4.11 (s, 3H), 3.85-3.80 (m, 2H), 3.75 (t, J=5.9 Hz, 2H), 3.72 (t, J=6.2 Hz, 1H), 3.68-3.64 (m, 1H), 3.56-3.48 (m, 4H), 3.43-3.36 (m, 2H), 3.35 (d, J=4.9 Hz, 1H), 3.25 (t, J=7.1 Hz, 2H), 2.56 (t, J=5.9 Hz, 2H), 2.54-2.49 (m, 1H), 2.38-2.31 (m, 2H), 1.48 (s, 9H).
LC-MS (ESI): [M+H]+=618.02.
To a solution of compound P-9 (25 mg, 40.44 mol) in dichloromethane (2 mL) was added trifluoroacetic acid (0.5 mL) under stirring at room temperature. The reaction was stirred at room temperature for 3 h. The resulting mixture was concentrated under vacuum, dissolved by adding N, N-dimethylformamide (1 mL) to which diisopropylethylamine (60 mg, 464.22 μmol) was added, followed by adding the compound 14 (20 mg, 41.37 μmol) and HATU (20 mg, 52.60 mol). The reaction was stirred at room temperature for 1.5 hours. The white solid compound PROTAC 17 (6.0 mg, 19.06%) was obtained by preparative HPLC purification.
1H NMR (600 MHz, CD3OD) δ8.75 (d, J=0.9 Hz, 1H), 7.99 (d, J=4.0 Hz, 1H), 7.95 (dd, J=6.6, 2.6 Hz, 1H), 7.70-7.65 (m, 1H), 7.39 (t, J=8.9 Hz, 1H), 7.32 (s, 1H), 7.28-7.24 (m, 1H), 7.22 (s, 1H), 5.13 (ddd, J=21.6, 13.3, 5.1 Hz, 1H), 4.45-4.33 (m, 5H), 4.07 (d, J=20.7 Hz, 2H), 3.90-3.77 (m, 5H), 3.71-3.64 (m, 1H), 3.62-3.53 (m, 2H), 3.48-3.36 (m, 5H), 3.27-3.14 (m, 4H), 3.05 (s, 2H), 2.97 (t, J=6.6 Hz, 2H), 2.93-2.87 (m, 2H), 2.86-2.76 (m, 2H), 2.76-2.66 (m, 1H), 2.56-2.44 (m, 1H), 2.31-2.24 (m, 2H), 2.20-2.14 (m, 1H), 1.98-1.84 (m, 4H), 1.79-1.68 (m, 1H), 1.68-1.57 (m, 1H).
LC-MS(ESI): [M+H]+=913.35/915.35.
To a solution of compound P-9 (25 mg, 40.44 μmol) in dichloromethane (2 mL) was added trifluoroacetic acid (0.5 mL) under stirring at room temperature. The reaction was stirred at room temperature for 3 h. The resulting mixture was concentrated under vacuum, dissolved by adding N,N-dimethylformamide (1 mL), to which diisopropylethylamine (60 mg, 464.22 μmol) was added, followed by adding the compound 16 (20 mg, 41.20 mol) and HATU (20 mg, 52.60 μmol). The reaction solution was stirred at room temperature for 1.5 hours. The white solid compound PROTAC 18 (5.8 mg, 18.48%) was obtained by preparative HPLC purification.
1H NMR (600 MHz, CD3OD) δ8.75 (d, J=2.7 Hz, 1H), 7.98 (d, J=6.0 Hz, 1H), 7.95 (dd, J=6.6, 2.6 Hz, 1H), 7.70-7.65 (m, 1H), 7.39 (t, J=8.9 Hz, 1H), 7.30-7.23 (m, 2H), 7.13 (s, 1H), 5.12 (ddd, J=23.3, 13.4, 5.1 Hz, 1H), 4.44-4.31 (m, 5H), 4.12-4.00 (m, 5H), 3.92-3.78 (m, 5H), 3.64-3.54 (m, 2H), 3.53-3.36 (m, 5H), 3.30-3.22 (m, 3H), 3.20-3.13 (m, 1H), 3.06 (s, 2H), 2.96-2.86 (m, 2H), 2.83-2.75 (m, 3H), 2.53-2.42 (m, 1H), 2.28 (s, 2H), 2.21-2.12 (m, 1H), 1.96-1.77 (m, 3H), 1.75-1.66 (m, 1H).
LC-MS(ESI): [M+H]+=915.45/917.35.
To a solution of compound P-9 (25 mg, 40.44 μmol) in dichloromethane (2 mL) was added trifluoroacetic acid (0.5 mL) under stirring at room temperature. The reaction was stirred at room temperature for 3 h. The resulting mixture was concentrated under vacuum, dissolved by adding N, N-dimethylformamide (1 mL) to which diisopropylethylamine (60 mg, 464.22 μmol) was added, followed by adding the compound 15 (20 mg, 43.92 mol) and HATU (20 mg, 52.60 mol). The reaction was stirred at room temperature for 1.5 hours. The white solid compound PROTAC 19 (7.3 mg, 22.53%) was obtained by preparative HPLC purification.
1H NMR (600 MHz, CD3OD) δ8.75 (d, J=2.2 Hz, 1H), 7.98 (d, J=7.6 Hz, 1H), 7.96 (dd, J=6.6, 2.6 Hz, 1H), 7.70-7.66 (m, 1H), 7.54 (d, J=11.0 Hz, 1H), 7.39 (t, J=8.9 Hz, 1H), 7.26 (d, J=5.4 Hz, 1H), 7.06 (d, J=2.2 Hz, 1H), 5.12 (ddd, J=13.3, 10.7, 5.1 Hz, 1H), 4.46-4.38 (m, 2H), 4.37-4.31 (m, 4H), 4.13-4.06 (m, 5H), 3.85-3.76 (m, 4H), 3.47-3.36 (m, 4H), 3.32-3.29 (m, 1H), 3.25-3.10 (m, 4H), 3.05 (s, 2H), 3.00-2.95 (m, 2H), 2.94-2.86 (m, 1H), 2.81-2.75 (m, 1H), 2.60-2.54 (m, 1H), 2.54-2.42 (m, 3H), 2.30-2.19 (m, 4H), 2.19-2.12 (m, 1H).
LC-MS(ESI): [M+H]+=885.46/887.36.
To a solution of compound P-9 (25 mg, 40.44 μmol) in dichloromethane (2 mL) was added trifluoroacetic acid (0.5 mL) under stirring at room temperature. The reaction was stirred at room temperature for 3 h. The resulting mixture was concentrated under vacuum, dissolved by adding N,N-dimethylformamide (1 mL) to which diisopropylethylamine (60 mg, 464.22 μmol) was added, followed by adding the compound 13 (20 mg, 43.92 μmol) and HATU (20 mg, 52.60 mol). The reaction was stirred at room temperature for 1.5 hours, and purified by preparative HPLC to obtain the white solid compound PROTAC 20 (8.0 mg, 24.69%)
1H NMR (600 MHz, CD3OD) δ8.75 (d, J=1.9 Hz, 1H), 7.98 (d, J=4.5 Hz, 1H), 7.95 (dd, J=6.6, 2.6 Hz, 1H), 7.70-7.65 (m, 1H), 7.39 (t, J=8.9 Hz, 1H), 7.33 (s, 1H), 7.25 (s, 1H), 7.23 (s, 1H), 5.13-5.05 (m, 1H), 4.45-4.28 (m, 6H), 4.13-4.03 (m, 5H), 3.87-3.72 (m, 4H), 3.47-3.35 (m, 3H), 3.31-3.26 (m, 2H), 3.22-3.12 (m, 3H), 3.06 (s, 2H), 3.03-2.98 (m, 2H), 2.90-2.82 (m, 1H), 2.81-2.74 (m, 1H), 2.65-2.58 (m, 1H), 2.51-2.40 (m, 3H), 2.30-2.18 (m, 5H), 2.17-2.11 (m, 1H).
LC-MS(ESI): [M+H]+=885.36/887.36.
To a solution of compound P-9 (40 mg, 64.71 μmol) in dichloromethane (2 mL) was added trifluoroacetic acid (0.5 mL) under stirring at room temperature. The reaction was stirred at room temperature for 3 h. The resulting mixture was concentrated under vacuum, dissolved by adding N,N-dimethylformamide (1 mL) to which diisopropylethylamine (60 mg, 464.22 μmol) was added, followed by adding the compound 1 (40 mg, 80.41 mol) and HATU (40 mg, 105.20 mol). The reaction was stirred at room temperature for 1.5 hours. The white solid compound PROTAC 21 (14 mg, 18.77%) was obtained by preparative HPLC purification.
1H NMR (600 MHz, DMSO-d6) δ8.73 (s, 1H), 7.96 (s, 1H), 7.93 (d, J=6.5 Hz, 1H), 7.70-7.61 (m, 2H), 7.37 (t, J=8.9 Hz, 1H), 7.28 (s, 1H), 7.24 (s, 1H), 5.08 (dd, J=12.8, 5.3 Hz, 1H), 4.39 (d, J=12.7 Hz, 1H), 4.33 (t, J=5.2 Hz, 2H), 4.06 (s, 3H), 3.89-3.75 (m, 5H), 3.59-3.53 (m, 1H), 3.46-3.34 (m, 6H), 3.26-3.18 (m, 2H), 3.17-3.06 (m, 2H), 3.04-2.95 (m, 4H), 2.89-2.65 (m, 6H), 2.30-2.20 (m, 2H), 2.14-2.05 (m, 1H), 2.00-1.86 (m, 4H), 1.79-1.71 (m, 1H), 1.69-1.61 (m, 1H).
LC-MS (ESI): [M+H]+=927.60.
To a solution of compound P-9 (40 mg, 64.71 μmol) in dichloromethane (2 mL) was added trifluoroacetic acid (0.5 mL) under stirring at room temperature. The reaction was stirred at room temperature for 3 h. The resulting mixture was concentrated under vacuum, dissolved by adding N, N-dimethylformamide (1 mL) to which diisopropylethylamine (60 mg, 464.22 μmol) was added, followed by adding the compound 4 (40 mg, 80.41 mol) and HATU (40 mg, 105.20 μmol). The reaction was stirred at room temperature for 1.5 hours. The white solid compound PROTAC 22 (16 mg, 21.45%) was obtained by preparative HPLC purification.
1H NMR (600 MHz, DMSO-d6) δ8.74 (s, 1H), 7.98-7.92 (m, 2H), 7.69-7.61 (m, 2H), 7.38 (t, J=8.9 Hz, 1H), 7.24 (s, 1H), 7.20 (d, J=6.2 Hz, 1H), 5.11-5.04 (m, 1H), 4.35-4.29 (m, 2H), 4.16-4.09 (m, 2H), 4.05 (d, J=4.3 Hz, 3H), 3.89-3.81 (m, 3H), 3.78 (t, J=5.2 Hz, 2H), 3.70-3.55 (m, 4H), 3.47-3.35 (m, 4H), 3.18-3.07 (m, 2H), 3.03-2.97 (m, 2H), 2.93 (s, 2H), 2.90-2.80 (m, 2H), 2.77-2.65 (m, 5H), 2.28-2.20 (m, 2H), 2.13-2.06 (m, 1H), 1.66-1.46 (m, 5H).
LC-MS (ESI): [M+H]+=927.55.
To a solution of compound P-9 (40 mg, 64.71 μmol) in dichloromethane (2 mL) was added trifluoroacetic acid (0.5 mL) under stirring at room temperature. The reaction was stirred at room temperature for 3 h. The resulting mixture was concentrated under vacuum, dissolved by adding N,N-dimethylformamide (1 mL) to which diisopropylethylamine (60 mg, 464.22 μmol) was added, followed by adding the compound 3 (40 mg, 85.22 mol) and HATU (40 mg, 105.20 mol). The reaction was stirred at room temperature for 1.5 hours. The white solid compound PROTAC 23 (5.4 mg, 7.05%) was obtained by preparative HPLC purification.
1H NMR (600 MHz, CD3OD) δ8.75 (s, 1H), 7.97 (s, 1H), 7.96 (dd, J=6.6, 2.5 Hz, 1H), 7.70-7.65 (m, 2H), 7.39 (t, J=8.9 Hz, 1H), 7.32 (d, J=1.9 Hz, 1H), 7.26 (d, J=8.9 Hz, 1H), 5.09 (dd, J=12.8, 5.4 Hz, 1H), 4.38-4.33 (m, 3H), 4.16-4.10 (m, 2H), 4.09 (d, J=2.0 Hz, 3H), 3.86-3.74 (m, 4H), 3.42-3.35 (m, 2H), 3.30 (s, 3H), 3.14 (s, 4H), 3.07-2.98 (m, 4H), 2.89-2.81 (m, 2H), 2.77-2.65 (m, 3H), 2.62-2.56 (m, 1H), 2.51-2.44 (m, 1H), 2.32-2.22 (m, 4H), 2.13-2.07 (m, 1H).
LC-MS (ESI): [M+H]+=899.35.
To a solution of compound P-9 (40 mg, 64.71 μmol) in dichloromethane (2 mL) was added trifluoroacetic acid (0.5 mL) under stirring at room temperature. The reaction was stirred at room temperature for 3 h. The resulting mixture was concentrated under vacuum, dissolved by adding N,N-dimethylformamide (1 mL) to which diisopropylethylamine (60 mg, 464.22 μmol) was added, followed by adding the compound 17 (40 mg, 82.75 μmol) and HATU (40 mg, 105.20 mol). The reaction was stirred at room temperature for 1.5 hours. The white solid compound PROTAC 24 (20 mg, 26.46%) was obtained by preparative HPLC purification.
1H NMR (600 MHz, DMSO-d6) δ8.73 (s, 1H), 7.95 (s, 1H), 7.93 (dd, J=6.6, 2.5 Hz, 1H), 7.69 (s, 1H), 7.68-7.63 (m, 1H), 7.37 (t, J=8.9 Hz, 1H), 7.24 (s, 1H), 7.21 (d, J=2.5 Hz, 1H), 5.11-5.04 (m, 1H), 4.74 (s, 2H), 4.61 (d, J=13.3 Hz, 1H), 4.32 (t, J=5.5 Hz, 2H), 4.07 (s, 3H), 4.04 (d, J=14.0 Hz, 1H), 3.89-3.79 (m, 4H), 3.52-3.35 (m, 6H), 3.28 (d, J=13.8 Hz, 1H), 3.19 (s, 2H), 3.03 (s, 2H), 2.94-2.80 (m, 4H), 2.78-2.65 (m, 4H), 2.29-2.22 (m, 2H), 2.14-2.07 (m, 1H), 2.07-1.98 (m, 1H), 1.94-1.84 (m, 3H).
LC-MS (ESI): [M+H]+=913.55.
To a solution of compound P-9 (40 mg, 64.71 μmol) in dichloromethane (2 mL) was added trifluoroacetic acid (0.5 mL) with stirring at room temperature. The reaction was stirred at room temperature for 3 hours. The resulting mixture was concentrated in vacuo, and dissolved by adding N,N-dimethylformamide (1 mL). Diisopropylethylamine (60 mg, 464.22 mol) was added thereto, and then compound 17 and HATU (40 mg, 105.20 μmol) were added (40 mg, 82.75 μmol). The reaction was stirred at room temperature for 1.5 hours. Purified by preparative high performance liquid chromatography to obtain white solid compound PROTAC 24 (20 mg, 26.46%).
1H NMR (600 MHz, DMSO-d6) δ8.73 (s, 1H), 7.95 (s, 1H), 7.93 (dd, J=6.6, 2.5 Hz, 1H), 7.69 (s, 1H), 7.68-7.63 (m, 1H), 7.37 (t, J=8.9 Hz, 1H), 7.24 (s, 1H), 7.21 (d, J=2.5 Hz, 1H), 5.11-5.04 (m, 1H), 4.74 (s, 2H), 4.61 (d, J=13.3 Hz, 1H), 4.32 (t, J=5.5 Hz, 2H), 4.07 (s, 3H), 4.04 (d, J=14.0 Hz, 1H), 3.89-3.79 (m, 4H), 3.52-3.35 (m, 6H), 3.28 (d, J=13.8 Hz, 1H), 3.19 (s, 2H), 3.03 (s, 2H), 2.94-2.80 (m, 4H), 2.78-2.65 (m, 4H), 2.29-2.22 (m, 2H), 2.14-2.07 (m, 1H), 2.07-1.98 (m, 1H), 1.94-1.84 (m, 3H).
LC-MS (ESI): [M+H]+=913.55.
At room temperature, to a solution of compound P-10 (100 mg, 179.17 mol) and compound P-4 (68 mg, 361.27 μmol) in methanol (5 mL) was added acetic acid (1 mL) and tetraisopropyl titanate (1 mL), and finally sodium cyanoborohydride (34 mg, 541.04 mol) was added with further stirring. The reaction was stirred at room temperature for 14 hours. The resulting mixture was concentrated in vacuo and purified by high performance liquid chromatography to obtain light yellow oily compound P-11 (84 mg, 64.19%).
1H NMR (600 MHz, CD3OD) δ8.41 (d, J=8.2 Hz, 1H), 8.22 (s, 1H), 7.98 (dd, J=8.0, 1.4 Hz, 1H), 7.76-7.68 (m, 2H), 7.46 (t, J=7.7 Hz, 1H), 6.84 (s, 1H), 4.60 (dt, J=12.1, 6.0 Hz, 1H), 3.90-3.83 (m, 2H), 3.81-3.74 (m, 3H), 3.44-3.40 (m, 1H), 3.40-3.34 (m, 1H), 3.27-3.20 (m, 2H), 3.17-3.10 (m, 1H), 2.60 (t, J=5.8 Hz, 2H), 2.20 (s, 3H), 2.09-1.95 (m, 5H), 1.50 (s, 9H), 1.49-1.47 (m, 1H), 1.35 (d, J=6.1 Hz, 6H), 1.28 (d, J=6.8 Hz, 6H).
LC-MS (ESI): [M+H]+=730.15.
At room temperature, to a solution of compound P-11 (25 mg, 34.23 μmol) in dichloromethane (2 mL) was added trifluoroacetic acid (0.5 mL) with stirring. The reaction was stirred at room temperature for 3 hours. The resulting mixture was concentrated in vacuo, and dissolved by adding N,N-dimethylformamide (1 mL). Diisopropylethylamine (60 mg, 464.22 mol) was added thereto, and then compound 12 (20 mg, 41.37 μmol) and HATU (20 mg, 52.60 μmol) were added. The reaction was stirred at room temperature for 1.5 hours. Purified by preparative high performance liquid chromatography to obtain white solid compound PROTAC 25 (5.5 mg, 15.55%).
1H NMR (600 MHz, CD3OD) δ8.37 (d, J=8.3 Hz, 1H), 8.21 (d, J=2.8 Hz, 1H), 8.00-7.94 (m, 1H), 7.72-7.63 (m, 2H), 7.56 (d, J=8.7 Hz, 1H), 7.44 (td, J=7.6, 2.9 Hz, 1H), 7.06-6.97 (m, 1H), 6.84 (d, J=9.4 Hz, 1H), 5.13 (td, J=13.4, 5.1 Hz, 1H), 4.52-4.31 (m, 4H), 3.98-3.74 (m, 6H), 3.62 (dd, J=22.7, 10.1 Hz, 1H), 3.56-3.47 (m, 1H), 3.47-3.40 (m, 1H), 3.37 (dd, J=13.8, 7.0 Hz, 1H), 3.28-3.12 (m, 4H), 2.98-2.87 (m, 3H), 2.87-2.74 (m, 3H), 2.52-2.42 (m, 1H), 2.21 (s, 3H), 2.19-2.03 (m, 4H), 1.99-1.85 (m, 4H), 1.83-1.74 (m, 1H), 1.63 (ddd, J=26.4, 13.4, 4.7 Hz, 1H), 1.41-1.37 (m, 1H), 1.37-1.30 (m, 1H), 1.25 (t, J=8.8 Hz, 6H), 1.18-1.10 (m, 6H).
LC-MS(ESI): [M+H]+=1025.45/1027.45.
At room temperature, to a solution of compound P-11 (25 mg, 34.23 μmol) in dichloromethane (2 mL) was added trifluoroacetic acid (0.5 mL) with stirring. The reaction was stirred at room temperature for 3 hours. The resulting mixture was concentrated in vacuo, and dissolved by adding N,N-dimethylformamide (1 mL). Diisopropylethylamine (60 mg, 464.22 mol) was added thereto, and then compound 14 (20 mg, 41.37 μmol) and HATU (20 mg, 52.60 μmol) were added. The reaction was stirred at room temperature for 1.5 hours. Purified by preparative high performance liquid chromatography to obtain white solid compound PROTAC 26 (6.4 mg, 18.10%).
1H NMR (600 MHz, CD3OD) δ8.38 (d, J=8.2 Hz, 1H), 8.22 (s, 1H), 7.95 (td, J=8.1, 1.3 Hz, 1H), 7.70-7.62 (m, 2H), 7.43 (dd, J=14.6, 7.2 Hz, 1H), 7.32 (d, J=8.0 Hz, 1H), 7.25 (d, J=8.3 Hz, 1H), 6.83 (d, J=8.3 Hz, 1H), 5.19-5.08 (m, 1H), 4.54-4.32 (m, 4H), 3.99-3.73 (m, 6H), 3.67-3.47 (m, 3H), 3.47-3.39 (m, 1H), 3.39-3.34 (m, 1H), 3.28-3.11 (m, 4H), 2.99-2.88 (m, 3H), 2.87-2.74 (m, 3H), 2.56-2.44 (m, 1H), 2.23 (s, 3H), 2.16-2.02 (m, 4H), 1.98-1.83 (m, 4H), 1.81-1.72 (m, 1H), 1.60 (ddd, J=26.2, 12.9, 4.7 Hz, 1H), 1.36-1.32 (m, 1H), 1.26 (d, J=6.8 Hz, 6H), 1.20-1.12 (m, 6H).
LC-MS(ESI): [M+H]+=1025.45/1027.35.
To a solution of compound P-11 (25 mg, 34.23 μmol) in dichloromethane (2 mL) was added trifluoroacetic acid (0.5 mL) with stirring at room temperature. The reaction was stirred at room temperature for 3 hours. The resulting mixture was concentrated in vacuo, and dissolved by adding N,N-dimethylformamide (1 mL), diisopropylethylamine (60 mg, 464.22 μmol) was added thereto, and then compound 13 (20 mg, 43.92 μmol) and HATU (20 mg, 52.60 mol) were added. The reaction was stirred at room temperature for 1.5 hours. Purified by preparative high performance liquid chromatography to obtain white solid compound PROTAC 27 (7.5 mg, 20.54%).
1H NMR (600 MHz, CD3OD) δ8.37 (d, J=8.3 Hz, 1H), 8.21 (d, J=4.7 Hz, 1H), 7.98-7.92 (m, 1H), 7.68 (t, J=6.1 Hz, 2H), 7.55 (s, 1H), 7.44 (dt, J=10.6, 4.6 Hz, 1H), 6.88 (d, J=5.0 Hz, 1H), 6.87 (d, J=5.0 Hz, 1H), 5.10 (ddd, J=13.4, 5.1, 2.3 Hz, 1H), 4.58-4.51 (m, 1H), 4.41-4.24 (m, 4H), 4.14 (d, J=10.5 Hz, 1H), 4.09-4.03 (m, 1H), 3.95-3.84 (m, 4H), 3.83-3.73 (m, 3H), 3.49-3.40 (m, 2H), 3.40-3.34 (m, 1H), 3.28-3.20 (m, 1H), 3.19-3.12 (m, 1H), 2.98 (t, J=6.3 Hz, 2H), 2.95-2.85 (m, 1H), 2.80-2.73 (m, 1H), 2.67-2.61 (m, 1H), 2.56-2.50 (m, 1H), 2.48-2.38 (m, 1H), 2.27-2.20 (m, 4H), 2.16-2.05 (m, 5H), 1.37-1.30 (m, 1H), 1.27-1.24 (m, 6H), 1.24-1.21 (m, 6H).
LC-MS(ESI): [M+H]+=997.35/999.25.
To a solution of compound P-11 (25 mg, 34.23 μmol) in dichloromethane (2 mL) was added trifluoroacetic acid (0.5 mL) with stirring at room temperature. The reaction was stirred at room temperature for 3 hours. The resulting mixture was concentrated under vacuum, and dissolved by adding N,N-dimethylformamide (1 mL). Diisopropylethylamine (60 mg, 464.22 mol) was added thereto, and then compound 15 (20 mg, 43.92 μmol) and HATU (20 mg, 52.60 μmol) were added. The reaction was stirred at room temperature for 1.5 hours. Purified by preparative high performance liquid chromatography to obtain white solid compound PROTAC 28 (7.2 mg, 19.72%).
1H NMR (600 MHz, CD3OD) δ8.38 (d, J=8.2 Hz, 1H), 8.21 (s, 1H), 7.98-7.93 (m, 1H), 7.71-7.64 (m, 1H), 7.63 (d, J=4.3 Hz, 1H), 7.47-7.39 (m, 1H), 7.34 (s, 1H), 7.25 (s, 1H), 6.87 (d, J=4.0 Hz, 1H), 5.17-5.08 (m, 1H), 4.57-4.49 (m, 1H), 4.48-4.38 (m, 2H), 4.38-4.28 (m, 2H), 4.16-4.02 (m, 2H), 3.96-3.83 (m, 3H), 3.83-3.73 (m, 3H), 3.51-3.39 (m, 2H), 3.39-3.35 (m, 1H), 3.28-3.18 (m, 2H), 3.17-3.10 (m, 1H), 2.99 (dd, J=15.2, 8.5 Hz, 2H), 2.95-2.86 (m, 1H), 2.79 (d, J=17.4 Hz, 1H), 2.65 (ddd, J=16.4, 8.1, 3.7 Hz, 1H), 2.57-2.43 (m, 2H), 2.29-2.22 (m, 2H), 2.22-2.17 (m, 3H), 2.14-1.99 (m, 4H), 1.33 (dt, J=12.6, 6.7 Hz, 1H), 1.26 (d, J=6.7 Hz, 6H), 1.28-1.19 (m, 6H).
LC-MS(ESI): [M+H]+=997.45/999.35.
To a solution of compound P-11 (40 mg, 54.77 μmol) in dichloromethane (2 mL) was added trifluoroacetic acid (0.5 mL) with stirring at room temperature. The reaction was stirred at room temperature for 3 hours. The resulting mixture was concentrated under vacuum, and dissolved by adding N,N-dimethylformamide (1 mL). Diisopropylethylamine (60 mg, 464.22 mol) was added thereto, and then compound 1 (35 mg, 70.36 μmol) and HATU (32 mg, 84.16 mol) were added. The reaction was stirred at room temperature for 1.5 hours. Purified by preparative high performance liquid chromatography to obtain white solid compound PROTAC 29 (7.2 mg, 9.84%).
1H NMR (600 MHz, CD3OD) δ8.37 (d, J=8.2 Hz, 1H), 8.22 (d, J=10.1 Hz, 1H), 7.98 (d, J=9.9 Hz, 1H), 7.72-7.63 (m, 3H), 7.45 (t, J=7.7 Hz, 1H), 7.31 (d, J=10.8 Hz, 1H), 6.85 (d, J=1.7 Hz, 1H), 5.11 (dd, J=12.7, 5.5 Hz, 1H), 4.52-4.45 (m, 2H), 3.98-3.84 (m, 4H), 3.84-3.73 (m, 3H), 3.61 (t, J=12.4 Hz, 1H), 3.54-3.48 (m, 1H), 3.48-3.40 (m, 1H), 3.40-3.34 (m, 1H), 3.27-3.18 (m, 3H), 3.18-3.11 (m, 1H), 2.99 (t, J=6.3 Hz, 2H), 2.91-2.68 (m, 5H), 2.21 (s, 3H), 2.17-2.03 (m, 5H), 2.00-1.91 (m, 3H), 1.88 (d, J=14.0 Hz, 1H), 1.85-1.76 (m, 1H), 1.69-1.60 (m, 1H), 1.26 (d, J=6.8 Hz, 6H), 1.22-1.16 (m, 6H).
LC-MS (ESI): [M+H]+=1039.44.
To a solution of compound P-11 (40 mg, 54.77 mol) in dichloromethane (2 mL) was added trifluoroacetic acid (0.5 mL) with stirring at room temperature. The reaction was stirred at room temperature for 3 hours. The resulting mixture was concentrated under vacuum, and dissolved by adding N,N-dimethylformamide (1 mL). Diisopropylethylamine (60 mg, 464.22 mol) was added thereto, and then compound 4 (35 mg, 70.36 μmol) and HATU (32 mg, 84.16 μmol) were added. The reaction was stirred at room temperature for 1.5 hours. Purified by preparative high performance liquid chromatography to obtain white solid compound PROTAC 30 (7.6 mg, 10.39%).
1H NMR (600 MHz, CD3OD) δ8.37 (d, J=7.5 Hz, 1H), 8.21 (d, J=9.7 Hz, 1H), 7.98 (d, J=10.1 Hz, 1H), 7.75-7.61 (m, 3H), 7.45 (t, J=7.6 Hz, 1H), 7.24 (d, J=1.7 Hz, 1H), 6.83 (d, J=1.5 Hz, 1H), 5.11 (dd, J=12.8, 5.4 Hz, 1H), 4.46 (dt, J=12.1, 5.9 Hz, 1H), 4.17 (d, J=11.1 Hz, 1H), 4.10 (d, J=11.0 Hz, 1H), 3.99-3.88 (m, 3H), 3.83 (t, J=5.1 Hz, 2H), 3.77 (d, J=12.0 Hz, 2H), 3.75-3.70 (m, 1H), 3.66-3.58 (m, 2H), 3.46 (dd, J=13.2, 3.1 Hz, 2H), 3.40-3.35 (m, 1H), 3.22 (dd, J=15.4, 6.8 Hz, 2H), 3.14 (dd, J=13.6, 10.0 Hz, 1H), 3.01-2.93 (m, 2H), 2.93-2.84 (m, 1H), 2.83-2.68 (m, 4H), 2.21 (s, 3H), 2.16-2.03 (m, 5H), 1.68-1.53 (m, 3H), 1.53-1.46 (m, 1H), 1.26 (d, J=6.8 Hz, 6H), 1.22-1.14 (m, 6H).
LC-MS (ESI): [M+H]+=1039.54.
To a solution of compound P-11 (40 mg, 54.77 μmol) in dichloromethane (2 mL) was added trifluoroacetic acid (0.5 mL) with stirring at room temperature. The reaction was stirred at room temperature for 3 hours. The resulting mixture was concentrated in vacuo, and dissolved by adding N,N-dimethylformamide (1 mL). Diisopropylethylamine (60 mg, 464.22 mol) was added thereto, and then compound 17 (35 mg, 74.57 mol) and HATU (32 mg, 84.16 mol) were added. The reaction was stirred at room temperature for 1.5 hours. Purified by preparative high performance liquid chromatography to obtain white solid compound PROTAC 31 (10.5 mg, 13.92%).
1H NMR (600 MHz, CD3OD) δ8.39 (d, J=8.2 Hz, 1H), 8.19 (s, 1H), 7.95 (d, J=7.9 Hz, 1H), 7.73 (s, 1H), 7.71-7.66 (m, 2H), 7.43 (t, J=7.6 Hz, 1H), 7.25 (s, 1H), 6.85 (s, 1H), 5.09 (dd, J=12.9, 5.4 Hz, 1H), 4.75 (s, 2H), 4.65 (d, J=13.8 Hz, 1H), 4.56-4.49 (m, 1H), 4.07 (d, J=14.3 Hz, 1H), 3.96-3.90 (m, 2H), 3.89-3.84 (m, 2H), 3.84-3.75 (m, 2H), 3.52-3.43 (m, 2H), 3.38-3.34 (m, 1H), 3.29-3.20 (m, 2H), 3.17-3.11 (m, 1H), 3.00-2.93 (m, 1H), 2.90-2.69 (m, 6H), 2.21 (s, 3H), 2.15-2.03 (m, 6H), 1.95-1.90 (m, 2H), 1.87-1.82 (m, 1H), 1.29-1.21 (m, 12H).
LC-MS (ESI): [M+H]+=1025.35.
To a solution of compound P-12 (100 mg, 258.76 μmol) and compound P-4 (97.41 mg, 517.52 mol) in methanol (5 mL) were added acetic acid (1 mL) at room temperature, finally sodium cyanoborohydride (49 mg, 776.28 mol) was added with stirring. The reaction was stirred at room temperature for 14 hours. The resulting mixture was concentrated in vacuo and purified by high performance liquid chromatography to obtain compound P-13 (112 mg, 77.47%) as a colorless oil.
1H NMR (600 MHz, CD3OD) δ8.35 (s, 1H), 7.78-7.68 (m, 2H), 7.44 (t, J=7.9 Hz, 2H), 7.26-7.17 (m, 3H), 7.16-7.10 (m, 2H), 5.38-5.22 (m, 1H), 3.98-3.82 (m, 3H), 3.79-3.68 (m, 3H), 3.57-3.47 (m, 2H), 3.28-3.16 (m, 1H), 2.59-2.52 (m, 1H), 2.44-2.37 (m, 1H), 2.36-2.15 (m, 3H), 2.13-2.03 (m, 1H), 2.01-1.94 (m, 1H), 1.36 (d, J=38.3 Hz, 9H).
LC-MS (ESI): [M+H]+=559.38.
To a solution of compound P-13 (40 mg, 71.60 μmol) in dichloromethane (2 mL) was added trifluoroacetic acid (0.5 mL) with stirring at room temperature. The reaction was stirred at room temperature for 3 hours. The resulting mixture was concentrated in vacuo, and dissolved by adding N,N-dimethylformamide (1 mL). Diisopropylethylamine (50 mg, 386.86 mol) was added thereto, and then compound 1 (35 mg, 74.57 μmol) and HATU (20 mg, 52.60 μmol) were added. The reaction was stirred at room temperature for 1.5 hours. Purified by preparative high performance liquid chromatography to obtain white solid compound PROTAC 32 (10.5 mg, 13.25%).
1H NMR (600 MHz, CD3OD) δ8.41-8.35 (m, 1H), 7.79-7.63 (m, 3H), 7.46-7.34 (m, 2H), 7.30 (d, J=13.4 Hz, 1H), 7.26-7.14 (m, 3H), 7.14-7.06 (m, 2H), 5.46-5.35 (m, 1H), 5.11 (dd, J=10.4, 5.1 Hz, 1H), 4.55-4.36 (m, 1H), 4.14-3.97 (m, 1H), 3.95-3.70 (m, 7H), 3.65-3.49 (m, 3H), 3.28-3.17 (m, 2H), 3.04-2.93 (m, 2H), 2.92-2.69 (m, 5H), 2.40-2.09 (m, 5H), 2.03-1.65 (m, 6H).
LC-MS (ESI): [M+H]+=868.46.
To a solution of compound P-13 (40 mg, 71.60 mol) in dichloromethane (2 mL) was added trifluoroacetic acid (0.5 mL) with stirring at room temperature. The reaction was stirred at room temperature for 3 hours. The resulting mixture was concentrated under vacuum, and dissolved by adding N,N-dimethylformamide (1 mL). Diisopropylethylamine (50 mg, 386.86 mol) was added thereto, and then compound 3 (35 mg, 74.57 μmol) and HATU (20 mg, 52.60 mol) were added thereto. The reaction was stirred at room temperature for 1.5 hours. Purified by preparative high performance liquid chromatography to obtain white solid compound PROTAC 33 (9.4 mg, 15.01%).
1H NMR (600 MHz, CD3OD) δ8.43-8.32 (m, 1H), 7.80-7.62 (m, 3H), 7.46-7.38 (m, 2H), 7.30-7.04 (m, 6H), 5.46-5.34 (m, 1H), 5.11 (dd, J=12.7, 5.2 Hz, 1H), 4.37-4.22 (m, 2H), 4.11 (dd, J=27.1, 13.2 Hz, 2H), 3.98-3.68 (m, 7H), 3.61-3.46 (m, 2H), 3.29-3.17 (m, 1H), 3.05-2.93 (m, 2H), 2.92-2.84 (m, 1H), 2.80-2.68 (m, 2H), 2.63-2.46 (m, 1H), 2.40-1.97 (m, 8H).
LC-MS (ESI): [M+H]+=840.56.
To a solution of compound P-13 (40 mg, 71.60 μmol) in dichloromethane (2 mL) was added trifluoroacetic acid (0.5 mL) with stirring at room temperature. The reaction was stirred at room temperature for 3 hours. The resulting mixture was concentrated under vacuum, and dissolved by adding N,N-dimethylformamide (1 mL). Diisopropylethylamine (50 mg, 386.86 mol) was added thereto, and then compound 4 (35 mg, 70.36 μmol) and HATU (20 mg, 52.60 mol) were added (35 mg, 70.36 μmol). The reaction was stirred at room temperature for 1.5 hours. Purified by preparative high performance liquid chromatography to obtain white solid compound PROTAC 34 (10.6 mg, 17.36%).
1H NMR (600 MHz, CD3OD) δ8.37 (d, J=8.8 Hz, 1H), 7.80-7.60 (m, 3H), 7.43 (t, J=7.9 Hz, 2H), 7.29-7.07 (m, 6H), 5.47-5.36 (m, 1H), 5.11 (dd, J=12.8, 5.4 Hz, 1H), 4.30-3.46 (m, 15H), 3.28-3.13 (m, 1H), 3.00-2.67 (m, 6H), 2.41-1.96 (m, 6H), 1.67-1.29 (m, 4H).
LC-MS (ESI): [M+H]+=868.46.
To a solution of compound P-13 (40 mg, 71.60 mol) in dichloromethane (2 mL) was added trifluoroacetic acid (0.5 mL) with stirring at room temperature. The reaction was stirred at room temperature for 3 hours. The resulting mixture was concentrated in vacuo, and dissolved by adding N,N-dimethylformamide (1 mL). Diisopropylethylamine (50 mg, 386.86 mol) was added thereto, and then compound 12 (30 mg, 60.05 μmol) and HATU (20 mg, 52.60 μmol) were added. The reaction was stirred at room temperature for 1.5 hours. Purified by preparative high performance liquid chromatography to obtain white solid compound PROTAC 35 (7.0 mg, 13.21%).
1H NMR (600 MHz, CD3OD) δ8.42-8.35 (m, 1H), 7.81-7.66 (m, 2H), 7.56 (d, J=13.4 Hz, 1H), 7.48-7.35 (m, 2H), 7.25-6.93 (m, 6H), 5.47-5.34 (m, 1H), 5.17-5.07 (m, 1H), 4.53-4.28 (m, 3H), 4.12-3.48 (m, 11H), 3.29-3.15 (m, 2H), 2.98-2.74 (m, 5H), 2.58-2.12 (m, 6H), 2.07-1.46 (m, 7H).
LC-MS (ESI): [M+H]+=854.56.
To a solution of compound P-13 (40 mg, 71.60 mol) in dichloromethane (2 mL) was added trifluoroacetic acid (0.5 mL) with stirring at room temperature. The reaction was stirred at room temperature for 3 hours. The resulting mixture was concentrated in vacuo, and dissolved by adding N,N-dimethylformamide (1 mL). Diisopropylethylamine (50 mg, 386.86 mol) was added thereto, and then compound 14 (30 mg, 62.05 μmol) and HATU (20 mg, 52.60 mol) were added. The reaction was stirred at room temperature for 1.5 hours. Purified by preparative high performance liquid chromatography to obtain white solid compound PROTAC 36 (10.0 mg, 18.87%).
1H NMR (600 MHz, CD3OD) δ8.40-8.34 (m, 1H), 7.78-7.66 (m, 2H), 7.46-7.27 (m, 3H), 7.26-7.06 (m, 6H), 5.46-5.33 (m, 1H), 5.14 (dd, J=13.0, 4.8 Hz, 1H), 4.51-4.26 (m, 3H), 3.96-3.48 (m, 10H), 3.29-3.16 (m, 2H), 3.00-2.72 (m, 5H), 2.56-2.13 (m, 7H), 2.07-1.59 (m, 7H).
LC-MS (ESI): [M+H]+=854.46.
To a solution of compound P-13 (40 mg, 71.60 mol) in dichloromethane (2 mL) was added trifluoroacetic acid (0.5 mL) with stirring at room temperature. The reaction was stirred at room temperature for 3 hours. The resulting mixture was concentrated in vacuo, and dissolved by adding N,N-dimethylformamide (1 mL). Diisopropylethylamine (50 mg, 386.86 mol) was added thereto, and then compound 16 (30 mg, 61.80 μmol) and HATU (20 mg, 52.60 μmol) were added. The reaction was stirred at room temperature for 1.5 hours. Purified by preparative high performance liquid chromatography to obtain white solid compound PROTAC 37 (6.4 mg, 12.10%).
1H NMR (600 MHz, CD3OD) δ8.39-8.34 (m, 1H), 7.80-7.67 (m, 2H), 7.48-7.35 (m, 2H), 7.34-7.27 (m, 1H), 7.25-7.01 (m, 6H), 5.46-5.34 (m, 1H), 5.18-5.09 (m, 1H), 4.54-4.27 (m, 3H), 4.12-3.48 (m, 12H), 3.28-3.14 (m, 2H), 2.96-2.75 (m, 4H), 2.61-2.41 (m, 2H), 2.40-2.13 (m, 4H), 2.06-1.63 (m, 5H).
LC-MS (ESI): [M+H]+=856.36.
To a solution of compound P-14 (110 mg, 245.79 μmol) and compound P-4 (92.53 mg, 491.57 mol) in methanol (3 mL) were added acetic acid (1 mL) and tetraisopropyl titanate (1 mL) at room temperature, sodium triacetoxyborohydride (156.28 mg, 737.36 μmol) was added with stirring. The reaction was stirred at 40° C. for 2 hours. The resulting mixture was concentrated under vacuum and purified by preparative high performance liquid chromatography to obtain colorless oily compound P-15 (131 mg, 86.00%).
1H NMR (600 MHz, CD3OD) δ8.96 (s, 1H), 8.12-8.05 (m, 2H), 7.59 (dd, J=9.1, 2.7 Hz, 1H), 6.03-5.95 (m, 1H), 3.91-3.86 (m, 2H), 3.79 (t, J=5.8 Hz, 2H), 3.71-3.22 (m, 10H), 2.60 (t, J=5.8 Hz, 2H), 2.51 (s, 3H), 2.40 (s, 3H), 2.37-2.30 (m, 2H), 2.08-2.00 (m, 2H), 1.94-1.87 (m, 2H), 1.74-1.66 (m, 2H), 1.54-1.45 (m, 9H).
LC-MS (ESI): [M+H]+=620.48.
To a solution of compound P-15 (45 mg, 72.61 μmol) in dichloromethane (2 mL) was added trifluoroacetic acid (0.5 mL) with stirring at room temperature. The reaction was stirred at room temperature for 3 hours, The resulting mixture was concentrated in vacuo, and dissolved by adding N,N-dimethylformamide (1 mL). Diisopropylethylamine (50 mg, 386.86 mol) was added thereto, and then compound 1 (35 mg, 70.36 μmol) and HATU (20 mg, 52.60 μmol) were added. The reaction was stirred at room temperature for 1.5 hours. Purified by preparative high performance liquid chromatography to obtain yellow solid compound PROTAC 38 (10.0 mg, 15.30%).
1H NMR (600 MHz, CD3OD) δ9.06 (s, 1H), 8.08-8.02 (m, 2H), 7.75 (d, J=9.5 Hz, 1H), 7.64 (s, 1H), 7.29 (s, 1H), 6.04-5.96 (m, 1H), 5.11-5.06 (m, 1H), 4.38 (d, J=13.2 Hz, 1H), 3.99-3.80 (m, 7H), 3.64-3.50 (m, 4H), 3.48-3.37 (m, 3H), 3.26-3.18 (m, 1H), 2.99 (t, J=6.7 Hz, 2H), 2.91-2.66 (m, 6H), 2.52 (s, 3H), 2.43 (s, 3H), 2.37-2.28 (m, 2H), 2.14-2.03 (m, 3H), 1.99-1.85 (m, 6H), 1.82-1.61 (m, 5H).
LC-MS (ESI): [M+H]+=929.55.
To a solution of compound P-15 (45 mg, 72.61 μmol) in dichloromethane (2 mL) was added trifluoroacetic acid (0.5 mL) with stirring at room temperature. The reaction was stirred at room temperature for 3 hours. The resulting mixture was concentrated in vacuo, and dissolved by adding N,N-dimethylformamide (1 mL). Diisopropylethylamine (50 mg, 386.86 mol) was added thereto, and then compound 3 (35 mg, 74.57 mol) and HATU (20 mg, 52.60 mol) were added. The reaction was stirred at room temperature for 1.5 hours. Purified by preparative high performance liquid chromatography to obtain yellow solid compound PROTAC 39 (10.0 mg, 14.88%).
1H NMR (600 MHz, CD3OD) δ9.07 (s, 1H), 8.11-7.99 (m, 2H), 7.74-7.64 (m, 2H), 7.28 (d, J=10.2 Hz, 1H), 6.01 (dt, J=17.8, 8.8 Hz, 1H), 5.10-5.03 (m, 1H), 4.39 (d, J=1.6 Hz, 2H), 4.12 (q, J=10.9 Hz, 2H), 3.97-3.88 (m, 3H), 3.87-3.79 (m, 3H), 3.53 (t, J=4.8 Hz, 3H), 3.06-3.00 (m, 2H), 2.90-2.81 (m, 2H), 2.76-2.61 (m, 3H), 2.54-2.48 (m, 4H), 2.44 (s, 3H), 2.37-2.30 (m, 2H), 2.27 (t, J=6.4 Hz, 2H), 2.09 (d, J=5.3 Hz, 3H), 1.94-1.87 (m, 2H), 1.75-1.66 (m, 2H).
LC-MS (ESI): [M+H]+=901.45.
To a solution of compound P-15 (45 mg, 72.61 μmol) in dichloromethane (2 mL) was added trifluoroacetic acid (0.5 mL) with stirring at room temperature. The reaction was stirred at room temperature for 3 hours. The resulting mixture was concentrated under vacuum, and dissolved by adding N,N-dimethylformamide (1 mL). Diisopropylethylamine (50 mg, 386.86 mol) was added thereto, and then compound 4 (35 mg, 70.36 μmol) and HATU (20 mg, 52.60 mol) were added. The reaction was stirred at room temperature for 1.5 hours. Purified by preparative high performance liquid chromatography to obtain yellow solid compound PROTAC 40 (10.0 mg, 15.30%).
1H NMR (400 MHz, CD3OD) δ9.07 (s, 1H), 7.98 (s, 2H), 7.62 (d, J=12.9 Hz, 2H), 7.17 (s, 1H), 5.99 (dt, J=17.5, 8.7 Hz, 1H), 5.06 (dd, J=12.7, 5.3 Hz, 1H), 4.09 (s, 2H), 3.96-3.89 (m, 2H), 3.86-3.79 (m, 3H), 3.71-3.51 (m, 7H), 2.99 (s, 3H), 2.91 (s, 2H), 2.86 (s, 3H), 2.80-2.69 (m, 4H), 2.50 (s, 3H), 2.45-2.38 (m, 3H), 2.36-2.28 (m, 2H), 2.13-2.02 (m, 3H), 1.95-1.84 (m, 2H), 1.74-1.40 (m, 7H).
LC-MS (ESI): [M+H]+=929.12.
To a solution of compound P-15 (45 mg, 72.61 μmol) in dichloromethane (2 mL) was added trifluoroacetic acid (0.5 mL) with stirring at room temperature. The reaction was stirred at room temperature for 3 hours. The resulting mixture was concentrated in vacuo, and dissolved by adding N,N-dimethylformamide (1 mL). Diisopropylethylamine (50 mg, 386.86 mol) was added thereto, and then compound 17 (35 mg, 70.36 mol) and HATU (20 mg, 52.60 μmol) were added. The reaction was stirred at room temperature for 1.5 hours. Purified by preparative high performance liquid chromatography to obtain yellow solid compound PROTAC 41 (10.0 mg, 15.30%).
1H NMR (600 MHz, CD3OD) δ9.05 (s, 1H), 8.08-8.01 (m, 2H), 7.75-7.69 (m, 2H), 7.22 (s, 1H), 6.00 (dt, J=17.8, 8.9 Hz, 1H), 5.13-5.07 (m, 1H), 4.75 (s, 2H), 4.60 (d, J=13.4 Hz, 1H), 4.06 (d, J=14.1 Hz, 1H), 4.01-3.73 (m, 8H), 3.57 (t, J=5.0 Hz, 2H), 3.53-3.36 (m, 3H), 2.95-2.83 (m, 3H), 2.82-2.67 (m, 4H), 2.52 (d, J=4.8 Hz, 3H), 2.43 (s, 3H), 2.37-2.28 (m, 2H), 2.16-2.01 (m, 5H), 1.95-1.83 (m, 5H), 1.74-1.66 (m, 2H).
LC-MS (ESI): [M+H]+=916.45.
To a solution of compound P-15 (45 mg, 72.61 mol) in dichloromethane (2 mL) was added trifluoroacetic acid (0.5 mL) with stirring at room temperature. The reaction was stirred at room temperature for 3 hours. The resulting mixture was concentrated under vacuum, and dissolved by adding N,N-dimethylformamide (1 mL). Diisopropylethylamine (50 mg, 386.86 mol) was added thereto, and then compound 14 (30 mg, 62.05 μmol) and HATU (20 mg, 52.60 μmol) were added. The reaction was stirred at room temperature for 1.5 hours. Purified by preparative high performance liquid chromatography to obtain yellow solid compound PROTAC 42 (10.0 mg, 15.30%).
1H NMR (600 MHz, CD3OD) δ9.06 (s, 1H), 8.06-8.00 (m, 2H), 7.77-7.72 (m, 1H), 7.31 (s, 1H), 7.23 (s, 1H), 5.97 (td, J=17.1, 8.7 Hz, 1H), 5.12 (dt, J=13.3, 4.7 Hz, 1H), 4.44-4.33 (m, 3H), 3.97-3.80 (m, 8H), 3.61-3.50 (m, 4H), 3.48-3.37 (m, 2H), 3.24-3.18 (m, 1H), 2.98-2.72 (m, 7H), 2.53-2.41 (m, 7H), 2.36-2.27 (m, 2H), 2.23-2.12 (m, 1H), 2.07-2.01 (m, 2H), 1.96-1.84 (m, 6H), 1.78-1.57 (m, 5H).
LC-MS (ESI): [M+H]+=916.45.
To a solution of compound P-15 (45 mg, 72.61 mol) in dichloromethane (2 mL) was added trifluoroacetic acid (0.5 mL) with stirring at room temperature. The reaction was stirred at room temperature for 3 hours. The resulting mixture was concentrated in vacuo, and was dissolved by adding N,N-dimethylformamide (1 mL). Diisopropylethylamine (50 mg, 386.86 mol) was added thereto, and then compound 12 (30 mg, 62.05 μmol) and HATU (20 mg, 52.60 μmol) were added. The reaction was stirred at room temperature for 1.5 hours. Purified by preparative high performance liquid chromatography to obtain yellow solid compound PROTAC 43 (10.0 mg, 15.30%).
1H NMR (600 MHz, CD3OD) δ9.05 (s, 1H), 8.03 (t, J=6.9 Hz, 2H), 7.78-7.73 (m, 1H), 7.53 (d, J=5.7 Hz, 1H), 7.01 (s, 1H), 6.03-5.96 (m, 1H), 5.11 (dd, J=13.3, 5.2 Hz, 1H), 4.44-4.33 (m, 3H), 3.98-3.81 (m, 7H), 3.62-3.53 (m, 3H), 3.46-3.35 (m, 3H), 3.24-3.17 (m, 1H), 2.96-2.88 (m, 3H), 2.84-2.75 (m, 3H), 2.54-2.41 (m, 7H), 2.36-2.28 (m, 2H), 2.23-2.12 (m, 2H), 2.11-2.02 (m, 3H), 1.97-1.84 (m, 6H), 1.79-1.58 (m, 5H).
LC-MS (ESI): [M+H]+=916.45.
To a solution of compound P-15 (45 mg, 72.61 μmol) in dichloromethane (2 mL) was added trifluoroacetic acid (0.5 mL) with stirring at room temperature. The reaction was stirred at room temperature for 3 hours. The resulting mixture was concentrated in vacuo, and dissolved by adding N,N-dimethylformamide (1 mL). Diisopropylethylamine (50 mg, 386.86 μmol) was added thereto, and then compound 15 (35 mg, 65.88 μmol) and HATU (20 mg, 52.60 μmol) were added. The reaction was stirred at room temperature for 1.5 hours. Purified by preparative high performance liquid chromatography to obtain yellow solid compound PROTAC 44 (10.0 mg, 15.30%).
1H NMR (600 MHz, CD3OD) δ9.06 (s, 1H), 8.06-8.01 (m, 2H), 7.71 (t, J=9.5 Hz, 1H), 7.33 (d, J=2.8 Hz, 1H), 7.23 (d, J=6.1 Hz, 1H), 6.03-5.95 (m, 1H), 5.09 (dt, J=13.2, 5.4 Hz, 1H), 4.45-4.32 (m, 4H), 4.13-4.03 (m, 2H), 3.96-3.78 (m, 6H), 3.57-3.50 (m, 3H), 3.48-3.37 (m, 2H), 3.01 (t, J=5.6 Hz, 2H), 2.91-2.83 (m, 1H), 2.81-2.74 (m, 1H), 2.69-2.62 (m, 1H), 2.54-2.42 (m, 8H), 2.36-2.20 (m, 5H), 2.18-2.02 (m, 4H), 1.94-1.85 (m, 2H), 1.73-1.60 (m, 3H).
LC-MS (ESI): [M+H]+=888.46.
To a solution of compound P-15 (45 mg, 72.61 mol) in dichloromethane (2 mL) was added trifluoroacetic acid (0.5 mL) with stirring at room temperature. The reaction was stirred at room temperature for 3 hours. The resulting mixture was concentrated in vacuo, and dissolved by adding N,N-dimethylformamide (1 mL). Diisopropylethylamine (50 mg, 386.86 mol) was added thereto, and then compound 13 (30 mg, 65.88 μmol) and HATU (20 mg, 52.60 μmol) were added. The reaction was stirred at room temperature for 1.5 hours. Purified by preparative high performance liquid chromatography to obtain yellow solid compound PROTAC 45 (10.0 mg, 15.30%).
1H NMR (600 MHz, CD3OD) δ9.05 (s, 1H), 8.05-7.97 (m, 2H), 7.75 (t, J=10.1 Hz, 1H), 7.55 (s, 1H), 7.01 (s, 1H), 6.04-5.96 (m, 1H), 5.13-5.07 (m, 1H), 4.43-4.31 (m, 4H), 4.11 (d, J=10.7 Hz, 1H), 4.07 (d, J=10.7 Hz, 1H), 3.96-3.80 (m, 6H), 3.57-3.48 (m, 3H), 3.26-3.20 (m, 2H), 2.99 (t, J=6.4 Hz, 2H), 2.94-2.84 (m, 1H), 2.80-2.73 (m, 1H), 2.66-2.59 (m, 1H), 2.56-2.49 (m, 4H), 2.49-2.39 (m, 4H), 2.37-2.19 (m, 5H), 2.17-2.03 (m, 4H), 1.96-1.85 (m, 2H), 1.74-1.59 (m, 3H).
LC-MS (ESI): [M+H]+=888.46.
To a solution of compound 13 (15 mg, 32.94 μmol) and diisopropylethylamine (13 mg, 100.58 μmol) in N,N-dimethylformamide (1 mL) at room temperature, was added compound P-16 and HATU (15 mg, 39.45 mol). The reaction was stirred at room temperature for 1.5 hours. Purified by preparative high performance liquid chromatography to obtain white solid compound PROTAC 46 (8.4 mg, 35.21%).
1H NMR (600 MHz, CD3OD) δ7.60 (d, J=3.0 Hz, 1H), 7.52-7.44 (m, 4H), 7.10 (d, J=7.4 Hz, 1H), 5.13 (dd, J=13.0, 4.6 Hz, 1H), 4.71-4.64 (m, 2H), 4.58-4.54 (m, 1H), 4.51-4.39 (m, 2H), 4.21-4.11 (m, 2H), 3.48-3.35 (m, 2H), 3.08-3.02 (m, 2H), 2.96-2.87 (m, 1H), 2.83-2.77 (m, 1H), 2.73 (s, 3H), 2.54-2.48 (m, 1H), 2.47 (s, 3H), 2.40-2.27 (m, 2H), 2.21-2.14 (m, 1H), 1.72 (d, J=5.0 Hz, 3H).
LC-MS(ESI): [M+H]+=724.10/726.05.
To a solution of compound 12 (15 mg, 31.03 μmol) and diisopropylethylamine (13 mg, 100.58 μmol) in N,N-dimethylformamide (1 mL) at room temperature, was added compound P-16 (13 mg, 32.43 μmol) and HATU (15 mg, 39.45 mol). The reaction was stirred at room temperature for 1.5 hours. Purified by preparative high-performance liquid chromatography to obtain PROTAC 47 (10.20 mg, 43.70%) as a light yellow solid.
1H NMR (600 MHz, CD3OD) δ7.58 (d, J=11.5 Hz, 1H), 7.52-7.46 (m, 4H), 7.08 (d, J=3.9 Hz, 1H), 5.13 (dd, J=13.3, 5.2 Hz, 1H), 4.80 (t, J=6.9 Hz, 1H), 4.46 (dd, J=17.0, 8.7 Hz, 1H), 4.40 (dd, J=20.5, 6.0 Hz, 2H), 4.11 (t, J=14.8 Hz, 1H), 3.76-3.63 (m, 3H), 3.26 (dd, J=17.6, 8.4 Hz, 1H), 2.99 (dd, J=11.9, 5.9 Hz, 2H), 2.96-2.88 (m, 1H), 2.83-2.75 (m, 4H), 2.54-2.45 (m, 4H), 2.20-2.15 (m, 1H), 2.06-1.83 (m, 5H), 1.76-1.65 (m, 4H).
LC-MS (ESI): [M+H]+=752.12/754.01.
To a solution of compound 14 (30 mg, 62.05 μmol) and diisopropylethylamine (25 mg, 193.43 μmol) in N,N-dimethylformamide (1 mL) at room temperature, was added compound P-16 (25 mg, 62.36 μmol) and HATU (29 mg, 76.26 mol). The reaction was stirred at room temperature for 1.5 hours. Purified by preparative high performance liquid chromatography to obtain light yellow solid compound PROTAC 48 (13.2 mg, 28.28%).
1H NMR (600 MHz, CD3OD) δ7.52-7.45 (m, 4H), 7.34 (s, 1H), 7.29 (d, J=6.0 Hz, 1H), 5.15 (dd, J=13.3, 4.8 Hz, 1H), 4.79 (t, J=6.9 Hz, 1H), 4.44 (d, J=16.4 Hz, 1H), 4.41-4.35 (m, 2H), 4.14-4.05 (m, 1H), 3.72 (dd, J=16.4, 7.8 Hz, 2H), 3.69-3.61 (m, 1H), 3.27 (t, J=11.5 Hz, 1H), 3.00 (dd, J=11.6, 6.2 Hz, 2H), 2.97-2.88 (m, 1H), 2.80 (d, J=18.0 Hz, 1H), 2.77 (d, J=3.2 Hz, 3H), 2.54-2.46 (m, 4H), 2.22-2.15 (m, 1H), 2.04-1.83 (m, 5H), 1.73 (s, 3H), 1.72-1.62 (m, 1H).
LC-MS(ESI): [M+H]+=752.17/754.27.
To a solution (1 mL) of compound 16 (15 mg, 30.90 μmol) and diisopropylethylamine (12 mg, 92.85 mol) in N,N-dimethylformamide at room temperature, was added compound P-16 (13 mg, 32.43 μmol) and HATU (15 mg, 39.45 mol). The reaction was stirred at room temperature for 1.5 hours. Purified by preparative high performance liquid chromatography to obtain white solid compound PROTAC 49 (3.5 mg, 15.02%).
1H NMR (600 MHz, CD3OD) δ7.50-7.44 (m, 4H), 7.33 (s, 1H), 7.17 (d, J=3.7 Hz, 1H), 5.13 (dd, J=13.3, 5.1 Hz, 1H), 4.76 (t, J=6.9 Hz, 1H), 4.46-4.34 (m, 2H), 4.19-4.08 (m, 2H), 3.76-3.64 (m, 3H), 3.31-3.25 (m, 1H), 2.96-2.88 (m, 1H), 2.84-2.76 (m, 1H), 2.74 (s, 3H), 2.54-2.49 (m, 1H), 2.47 (d, J=9.6 Hz, 3H), 2.21-2.15 (m, 1H), 2.05 (d, J=5.2 Hz, 3H), 2.04-1.86 (m, 2H), 1.81-1.75 (m, 1H), 1.73 (s, 3H).
LC-MS(ESI): [M+H]+=754.11/755.98.
To a solution of compound 1 (15 mg, 30.90 mol) and diisopropylethylamine (12 mg, 92.8 mol) in N,N-dimethylformamide (1 mL) at room temperature, was added compound P-16 (13 mg, 32.43 mol) and HATU (15 mg, 39.45 μmol). The reaction was stirred at room temperature for 1.5 hours. Purified by preparative high performance liquid chromatography to obtain white solid compound PROTAC 50 (3.5 mg, 15.02%).
1H NMR (400 MHz, CD3OD) δ7.69 (s, 1H), 7.54-7.46 (m, 4H), 7.36 (d, J=2.5 Hz, 1H), 5.11 (dd, J=12.6, 5.4 Hz, 1H), 4.84 (t, J=7.0 Hz, 1H), 4.39 (d, J=9.1 Hz, 1H), 4.11 (t, J=12.8 Hz, 1H), 3.79-3.67 (m, 4H), 3.04 (d, J=2.7 Hz, 2H), 2.91-2.83 (m, 1H), 2.80 (t, J=3.0 Hz, 4H), 2.74 (dd, J=13.8, 4.0 Hz, 1H), 2.50 (s, 3H), 2.18-2.08 (m, 1H), 2.04-1.89 (m, 5H), 1.77-1.70 (m, 4H).
LC-MS(ESI): [M+H]+=766.26/768.26.
To a solution of compound 1 (15 mg, 30.90 μmol) and diisopropylethylamine (12 mg, 92.8 s mol) in N,N-dimethylformamide (1 mL) at room temperature, was added compound P-16 (13 mg, 32.43 mol) and HATU (15 mg, 39.45 mol). The reaction was stirred at room temperature for 1.5 hours. Purified by preparative high performance liquid chromatography to obtain white solid compound PROTAC 51 (3.5 mg, 15.02%).
1H NMR (400 MHz, CD3OD) δ7.71 (d, J=1.7 Hz, 1H), 7.54-7.45 (m, 4H), 7.37 (d, J=4.7 Hz, 1H), 5.12 (dd, J=12.4, 5.4 Hz, 1H), 4.76-4.70 (m, 1H), 4.70-4.54 (m, 2H), 4.23-4.14 (m, 2H), 3.47-3.37 (m, 2H), 3.10 (t, J=6.5 Hz, 2H), 2.94-2.83 (m, 1H), 2.81-2.70 (m, 5H), 2.48 (s, 3H), 2.40-2.29 (m, 2H), 2.18-2.10 (m, 1H), 1.73 (d J=2.2 Hz, 3H).
LC-MS(ESI): [M+H]+=738.27/740.27.
Under the condition of nitrogen protection, 3-(4-((1R,2S)-6-(benzyloxy)-2-phenyl-1,2,3,4-tetrahydronaphthalene-1-yl) phenyl)-3-azaspiro[5.5]undecane-9-one (120 mg, 0.21 mmol), 3 mL DCM, 3 mL DMF and TEA (0.5 ml) were added to the reaction flask in order, after stirring at room temperature for half an hour. NaBH3CN (40 mg, 0.63 mmol) and compound 13 (88 mg, 0.21 mmol) were added, heated to 50° C. for 12 hours. The reaction solution was prepared by reverse preparation to obtain white solid P-18 (100 mg, 540).
1H NMR (400 MHz, DMSO-d6) δ10.96 (s, 1H), 7.55-7.30 (m, 6H), 7.20-7.02 (m, 4H), 6.93-6.66 (m, 7H), 6.32 (s, 2H), 5.08 (s, 2H), 4.50-4.31 (m, 4H), 4.25 (d, J=4.0 Hz, 3H), 3.39-3.30 (m, 2H), 3.09 (d, J=23.4 Hz, 5H), 2.95-2.83 (m, 3H), 2.65-2.55 (m, 1H), 2.42-2.27 (m, 2H), 2.25-2.05 (m, 3H), 2.02-1.94 (m, 1H), 1.78 (d, J=19.7 Hz, 5H), 1.53 (d, J=44.4 Hz, 4H), 1.39-1.21 (m, 3H), 1.11 (t, J=7.3 Hz, 2H).
LC-MS (ESI): [M+H]+=881.46
The second step: 3-(1-(3-(4-((1R,2S)-6-hydroxy-2-phenyl-1,2,3,4-tetrahydronaphthalen-1-yl)phenyl)-3-azaspiro[5.5]undec-9-yl)-6′-oxo-3′,4′,6′,8′-tetrahydro-7′H-spiro[azetidine-3,2′-pyrano[2,3]isoindol]-7′-yl)piperidine-2,6-dione (Compound PROTAC 52)
Under the condition of nitrogen protection, compound P-18 (100 mg, 0.11 mmol) was dissolved in 5 mL methanol solution, 10% Pd/C (20 mg) was added, after being replaced with hydrogen three times, and heated to 50° C. under hydrogen atmosphere for 16 hours. After the reaction was completed, the reaction solution was prepared by reverse preparation to obtain PROTAC 52 (70 mg, 80%) as a white solid.
1H NMR (400 MHz, DMSO-d) δ10.96 (s, 1H), 9.15 (s, 1H), 7.53 (s, 1H), 7.22-7.00 (m, 4H), 6.94-6.70 (m, 4H), 6.67-6.59 (m, 2H), 6.48 (dd, J=8.3, 2.6 Hz, 1H), 6.31 (d, J=8.1 Hz, 2H), 5.06 (dd, J=13.2, 5.1 Hz, 1H), 4.44-4.16 (m, 7H), 3.38-3.30 (m, 1H), 3.17 (d, J=41.0 Hz, 4H), 3.01-2.82 (m, 5H), 2.66-2.55 (m, 1H), 2.42-2.30 (m, 1H), 2.25-1.95 (m, 4H), 1.89-1.66 (m, 5H), 1.54 (d, J=45.2 Hz, 4H), 1.39-1.25 (m, 2H), 1.11 (dd, J=11.3, 9.6 Hz, 3H).
LC-MS (ESI): [M+H]+=791.41
Under the condition of nitrogen protection, 2-(4-(6-(benzyloxy)-2-phenyl-3,4-dihydronaphthalen-1-yl)phenyl)-2-azaspiro[3.5]non-7-one (60 mg, 0.11 mmol) and compound 15 (38.8 mg, 0.11 mmol) were dissolved in DCE:DMF=2:1 (6 mL). 3 drops of acetic acid (catalytic amount) were added, stirred at room temperature for 1 h. And sodium cyanoborohydride (21.5 mg, 0.33 mmol) was added to the above reaction solution, and the reaction was carried out at room temperature for 2 h. Purified by preparative HPLC to obtain compound P-19 (20 mg, 20.6%).
1H NMR (600 MHz, DMSO-d6) δ10.98 (d, J=2.3 Hz, 1H), 7.46 (d, J=7.0 Hz, 2H), 7.43-7.37 (m, 3H), 7.35 (d, J=7.3 Hz, 1H), 7.18-7.08 (m, 4H), 6.89-6.82 (m, 3H), 6.77-6.70 (m, 2H), 6.18 (d, J=8.2 Hz, 2H), 6.06 (d, J=8.1 Hz, 2H), 5.08 (s, 3H), 4.44-4.39 (m, 1H), 4.34 (td, J=11.5, 10.7, 5.3 Hz, 2H), 4.25-4.16 (m, 3H), 3.42-3.37 (m, 4H), 3.33-3.27 (m, 2H), 3.04 (d, J=5.7 Hz, 1H), 3.01-2.83 (m, 5H), 2.63-2.56 (m, 1H), 2.40-2.35 (m, 1H), 2.15 (ddt, J=31.6, 12.8, 6.9 Hz, 3H), 1.99-1.92 (m, 3H), 1.87-1.82 (m, 1H), 1.74 (ddt, J=12.3, 6.1 Hz, 1H), 1.46 (dt, J=24.2, 13.1 Hz, 3H), 1.18 (t, J=11.8 Hz, 2H).
LC-MS(ESI): [M+H]+=853.43.
Under the condition of nitrogen protection, compound P-19 (25 mg, 1 eq) was dissolved in methanol (5 mL), 10% Pd/C (5 mg, 20% wt) was added. After three replacements with hydrogen, heated to 50° C. under hydrogen atmosphere for 16 hours. After the reaction was completed, the reaction solution was prepared by reverse preparation to obtain white solid PROTAC 53 (3.5 mg, 19.6%).
1H NMR (400 MHz, DMSO-d6) δ10.97 (s, 1H), 9.10 (s, 1H), 7.54 (s, 1H), 7.19-7.03 (m, 4H), 6.89-6.80 (m, 2H), 6.65-6.57 (m, 2H), 6.48 (dd, J=8.3, 2.5 Hz, 1H), 6.18 (d, J=8.2 Hz, 2H), 6.05 (d, J=8.3 Hz, 2H), 5.09-5.04 (m, 1H), 4.47-4.32 (m, 4H), 4.27-4.19 (m, 2H), 4.12 (d, J=4.9 Hz, 1H), 3.43-3.37 (m, 3H), 3.28 (d, J=12.5 Hz, 2H), 3.01-2.83 (m, 5H), 2.69-2.57 (m, 2H), 2.42-2.31 (m, 2H), 2.18 (t, J=6.2 Hz, 2H), 2.09 (dd, J=12.4, 6.4 Hz, 1H), 1.96 (d, J=14.0 Hz, 3H), 1.85 (d, J=11.4 Hz, 1H), 1.70 (d, J=11.9 Hz, 1H), 1.51-1.41 (m, 2H), 1.17 (d, J=12.2 Hz, 2H).
LC-MS (ESI): [M+H]+=763.38.
Under the condition of nitrogen protection, 3-(4-((1R,2S)-6-(benzyloxy)-2-phenyl-1,2,3,4-tetrahydronaphthalene-1-yl) phenyl)-3-azaspiro[5.5]undecane-9-one (90 mg, 0.16 mmol), 3 mL DCM, 3 mL DMF and TEA (0.5 ml) were added to the reaction flask in order. After stirring at room temperature for half an hour, NaBH3CN (30 mg, 0.49 mmol) and compound 17 (88 mg, 0.21 mmol) were added, heated to 50° C. 12 hours. White solid compound P-20 (100 mg, 52%) was obtained by reverse preparation of the reaction solution.
1H NMR (400 MHz, DMSO-d6) δ11.11 (s, 1H), 7.66 (s, 1H), 7.48-7.29 (m, 6H), 7.11 (t, J=7.4 Hz, 2H), 7.07-6.98 (m, 3H), 6.95-6.82 (m, 5H), 6.72 (dd, J=8.6, 2.7 Hz, 1H), 6.56 (dd, J=8.6, 2.8 Hz, 1H), 5.09 (s, 2H), 4.73 (d, J=10.6 Hz, 2H), 3.46 (d, J=11.8 Hz, 2H), 3.14 (d, J=36.2 Hz, 5H), 2.99-2.79 (m, 8H), 2.72-2.57 (m, 6H), 2.22 (dd, J=19.5, 11.3 Hz, 4H), 2.11-1.93 (m, 5H), 1.80 (s, 1H), 1.75-1.50 (m, 4H), 0.87 (t, J=7.3 Hz, 1H).
LC-MS (ESI): [M+H]+=909.45.
Under the condition of nitrogen protection, compound P-20 (30 mg, 0.03 mmol) was dissolved in methanol (5 mL), and 10% Pd/C (6 mg, 20% wt) was added. After being replaced with hydrogen three times, the reaction was heated to 50° C. under hydrogen atmosphere for 16 hours. After the reaction was completed, the reaction solution was prepared in reverse to obtain PROTAC 54 (6.5 mg, 26.6%) as a white solid.
1H NMR (600 MHz, DMSO-d6) δ11.12 (s, 1H), 9.80 (s, 1H), 8.24 (s, 1H), 7.65 (s, 1H), 7.36 (d, J=13.7 Hz, 1H), 7.16 (ddt, J=19.8, 12.5, 6.7 Hz, 3H), 6.84 (d, J=7.2 Hz, 2H), 6.73-6.56 (m, 3H), 6.49 (dd, J=8.3, 2.5 Hz, 1H), 6.26 (d, J=8.1 Hz, 2H), 5.12 (ddd, J=12.9, 7.7, 5.3 Hz, 1H), 4.72 (d, J=13.8 Hz, 2H), 4.17 (d, J=5.0 Hz, 1H), 3.46-3.19 (m, 8H), 3.13-2.75 (m, 9H), 2.60 (dt, J=17.0, 3.4 Hz, 1H), 2.30-1.92 (m, 10H), 1.78-1.50 (m, 5H), 1.34-1.19 (m, 1H).
LC-MS (ESI): [M+H]+=819.40.
Under the condition of nitrogen protection, 2-(4-(6-(benzyloxy)-2-phenyl-3,4-dihydronaphthalen-1-yl)phenyl)-2-azaspiro[3.5]non-7-ketone (90 mg, 0.17 mmol) and compound 13 (38.8 mg, 0.17 mmol) were dissolved in DCE:DMF=2:1 (6 mL). 3 drops of acetic acid (catalytic amount) was added, stirred at room temperature for 1 h. Sodium cyanoborohydride (21.5 mg, 0.51 mmol) was added to the above reaction solution, and the reaction was carried out at room temperature for 2 h. Purified by preparative HPLC to obtain the product P-21 (30 mg, 20.6%).
1H NMR (600 MHz, DMSO-d6) δ10.97 (d, J=2.5 Hz, 1H), 7.54 (d, J=7.7 Hz, 1H), 7.46 (d, J=7.0 Hz, 2H), 7.41 (t, J=7.5 Hz, 2H), 7.36-7.32 (m, 1H), 7.16-7.04 (m, 4H), 6.89-6.83 (m, 3H), 6.77-6.72 (m, 2H), 6.18 (d, J=8.1 Hz, 2H), 6.06 (d, J=8.1 Hz, 2H), 5.10-5.04 (m, 3H), 4.42 (q, J=5.6 Hz, 1H), 4.36 (dd, J=17.5, 7.5 Hz, 3H), 4.23 (dt, J=13.6, 7.0 Hz, 2H), 4.17 (d, J=5.0 Hz, 1H), 3.40-3.37 (m, 4H), 3.31-3.29 (m, 1H), 3.07-3.02 (m, 1H), 3.01-2.85 (m, 5H), 2.62-2.57 (m, 1H), 2.39-2.35 (m, 1H), 2.15 (ddt, J=34.0, 12.7, 6.5 Hz, 4H), 2.00-1.90 (m, 4H), 1.87-1.82 (m, 1H), 1.75-1.72 (m, 1H), 1.48-1.42 (m, 2H), 1.18 (d, J=12.1 Hz, 2H).
LC-MS(ESI): [M+H]+=853.43.
Under the condition of nitrogen protection, compound P-21 (25 mg, 0.03 mmol) was dissolved in methanol (5 mL). 10% Pd/C (5 mg, 20% wt) was added. After three times of hydrogen replacements, and heated to 50° C. under hydrogen atmosphere for 16 hours. After the reaction was completed, the reaction solution was prepared in reverse to obtain PROTAC 55 (7.5 mg, 32.7%) as a white solid.
1H NMR (600 MHz, DMSO-d6) δ10.98 (d, J=2.1 Hz, 1H), 9.11 (s, 1H), 7.38 (d, J=3.1 Hz, 1H), 7.23-7.02 (m, 4H), 6.88-6.79 (m, 2H), 6.66-6.57 (m, 2H), 6.48 (dd, J=8.3, 2.5 Hz, 1H), 6.18 (d, J=8.2 Hz, 2H), 6.05 (d, J=8.1 Hz, 2H), 5.08 (dd, J=13.3, 5.2 Hz, 1H), 4.44-4.32 (m, 4H), 4.25-4.20 (m, 2H), 4.12 (d, J=5.0 Hz, 1H), 3.48-3.32 (m, 4H), 3.29-3.18 (m, 2H), 3.02-2.83 (m, 5H), 2.63-2.56 (m, 1H), 2.38 (dq, J=13.3, 9.2, 7.0 Hz, 1H), 2.19 (dt, J=20.4, 7.1 Hz, 2H), 2.09 (dt, J=12.5, 6.2 Hz, 1H), 2.03-1.88 (m, 4H), 1.84 (d, J=11.7 Hz, 1H), 1.70 (dd, J=12.9, 6.1 Hz, 1H), 1.46 (dt, J=24.0, 13.2 Hz, 2H), 1.19 (d, J=13.9 Hz, 2H).
LC-MS (ESI): [M+H]+=763.38.
Under the condition of nitrogen protection, 7-(4-((1R, 2S)-6-(benzyloxy)-2-phenyl-1,2,3,4-tetrahydronaphthalen-1-yl)phenyl)-7-azaspiro[3.5]non-2-one (100 mg, 0.19 mmol) and compound 13 (64.8 mg, 0.19 mmol) were dissolved in DCE:DMF=2:1 (6 mL), and 3 drops of acetic acid (catalytic amount) were added, stirred at room temperature for 1 h. Sodium cyanoborohydride (35.9 mg, 0.57 mmol) was added into the above reaction solution, and the reaction was performed at room temperature for 2 h. Purified by preparative HPLC to obtain the product compound P-22 (32 mg, 19.7%).
1H NMR (600 MHz, DMSO-d6) δ10.97 (s, 1H), 7.53 (s, 1H), 7.46 (d, J=7.3 Hz, 2H), 7.41 (t, J=7.5 Hz, 2H), 7.34 (t, J=7.3 Hz, 1H), 7.14 (dq, J=14.2, 7.0 Hz, 3H), 7.08 (d, J=8.8 Hz, 1H), 6.90 (d, J=2.0 Hz, 1H), 6.84 (d, J=7.0 Hz, 2H), 6.78-6.71 (m, 2H), 6.64 (s, 2H), 6.26 (d, J=7.0 Hz, 2H), 5.11-5.04 (m, 3H), 4.42-4.33 (m, 2H), 4.30-4.19 (m, 4H), 4.10 (dd, J=30.4, 9.5 Hz, 2H), 3.09-3.01 (m, 3H), 2.99-2.91 (m, 4H), 2.87 (d, J=6.0 Hz, 2H), 2.60 (d, J=18.5 Hz, 1H), 2.40-2.33 (m, 1H), 2.21-2.09 (m, 5H), 2.01-1.95 (m, 1H), 1.88-1.81 (m, 2H), 1.78-1.72 (m, 1H), 1.64 (s, 4H).
LC-MS(ESI): [M+H]+=853.43.
Under the condition of nitrogen protection, compound P-22 (30 mg, 0.03 mmol) was dissolved in methanol (5 mL). 10% Pd/C (6 mg, 20% wt) was added, and after being replaced with hydrogen three times, heated to 50° C. under hydrogen atmosphere for 16 hours. After the reaction was completed, white solid PROTAC 56 (9.5 mg, 41.4%) was obtained by revers preparation of reaction solution.
1H NMR (600 MHz, DMSO-d6) δ10.97 (s, 1H), 9.16 (s, 1H), 7.53 (s, 1H), 7.19-7.10 (m, 3H), 7.08 (d, J=7.8 Hz, 1H), 6.83 (d, J=7.1 Hz, 2H), 6.69-6.60 (m, 4H), 6.48 (dd, J=8.3, 2.3 Hz, 1H), 6.26 (d, J=7.5 Hz, 2H), 5.06 (dd, J=13.2, 5.0 Hz, 1H), 4.37 (dd, J=23.1, 6.9 Hz, 2H), 4.25 (dd, J=18.6, 12.8 Hz, 3H), 4.18-4.07 (m, 5H), 3.31 (d, J=14.1 Hz, 1H), 3.04 (s, 2H), 2.99-2.85 (m, 7H), 2.60 (d, J=17.2 Hz, 1H), 2.41-2.32 (m, 1H), 2.19 (s, 2H), 2.16-2.06 (m, 2H), 1.99 (d, J=7.0 Hz, 1H), 1.84 (d, J=8.4 Hz, 2H), 1.72 (d, J=5.9 Hz, 1H), 1.65 (s, 3H).
LC-MS (ESI): [M+H]+=763.38.
Under the condition of nitrogen protection, 7-(4-((1R, 2S)-6-(benzyloxy)-2-phenyl-1,2,3,4-tetrahydronaphthalen-1-yl)phenyl)-7-azaspiro[3.5]nonan-2-one (90 mg, 0.17 mmol) and compound 12 (62.8 mg, 0.17 mmol) were dissolved in DCE:DMF=2:1 (6 mL), and 3 drops of acetic acid (catalytic amount) were added, stirred at room temperature for 1 h. Sodium cyanoborohydride (32.1 mg, 0.51 mmol) was added into the above reaction solution, and the reaction was performed at room temperature for 2 h.
Purified by preparative HPLC to obtain the product compound P-23 (24 mg, 16.60%).
1H NMR (600 MHz, DMSO-d6) δ10.97 (s, 1H), 7.51 (d, J=7.3 Hz, 1H), 7.46 (d, J=7.3 Hz, 2H), 7.41 (t, J=7.5 Hz, 2H), 7.34 (t, J=7.3 Hz, 1H), 7.14 (dq, J=14.5, 7.6, 7.1 Hz, 3H), 7.02 (s, 1H), 6.90 (d, J=1.9 Hz, 1H), 6.84 (d, J=7.2 Hz, 2H), 6.79-6.72 (m, 2H), 6.66 (s, 2H), 6.26 (d, J=7.3 Hz, 2H), 5.10-5.03 (m, 3H), 4.35 (d, J=16.9 Hz, 1H), 4.22 (dd, J=10.7, 6.0 Hz, 2H), 3.32 (d, J=11.2 Hz, 3H), 3.09-2.90 (m, 11H), 2.60 (d, J=17.0 Hz, 1H), 2.36 (s, 1H), 2.19 (s, 2H), 2.12 (dd, J=12.6, 5.9 Hz, 1H), 2.00 (dt, J=20.6, 11.4 Hz, 5H), 1.90 (t, J=6.8 Hz, 2H), 1.86-1.80 (m, 3H), 1.75 (d, J=5.6 Hz, 1H), 1.64 (d, J=20.5 Hz, 4H).
LC-MS (ESI): [M+H]+=881.46.
Under the condition of nitrogen protection, compound P-23 (30 mg, 0.03 mmol) was dissolved in methanol (5 mL). 10% Pd/C (6 mg, 20% wt) was added, and after three replacements with hydrogen, and heated to 50° C. under hydrogen atmosphere for 16 hours. After the reaction was completed, the reaction solution was prepared by reverse preparation to obtain PROTAC 57 (10.5 mg, 50.6%) as a white solid.
1H NMR (600 MHz, DMSO-d6) δ10.97 (s, 1H), 9.84 (s, 1H), 7.51 (d, J=6.7 Hz, 1H), 7.14 (dt, J 11.5, 6.7 Hz, 3H), 7.02 (s, 1H), 6.86-6.74 (m, 4H), 6.66-6.60 (m, 2H), 6.49 (dd, J=8.3, 2.3 Hz, 1H), 6.31 (d, J=8.1 Hz, 2H), 5.07 (dd, J=13.2, 5.1 Hz, 1H), 4.35 (d, J=16.9 Hz, 1H), 4.25-4.18 (m, 2H), 3.32 (d, J=11.4 Hz, 3H), 3.13 (s, 2H), 3.08-2.97 (m, 6H), 2.95-2.85 (m, 5H), 2.60 (d, J=16.7 Hz, 1H), 2.36 (dd, J=13.2, 4.6 Hz, 1H), 2.22 (s, 2H), 2.11-1.95 (m, 6H), 1.92-1.84 (m, 3H), 1.70 (d, J=23.7 Hz, 5H).
LC-MS (ESI): [M+H]+=791.41.
Under the condition of nitrogen protection, 7-(4-((1R, 2S)-6-(benzyloxy)-2-phenyl-1,2,3,4-tetrahydronaphthalene-1-yl)phenyl)-7-azaspiro[3.5]nonan-2-one (60 mg, 0.11 mmol) and compound 16 (40.8 mg, 0.11 mmol) were dissolved in DCE:DMF=2:1 (6 mL). 3 drops of acetic acid (catalytic amount) were added, stirred at room temperature for 1 h. Sodium cyanoborohydride (21.5 mg, 0.33 mmol) was added into the above reaction solution, and the reaction was performed at room temperature. Purified by preparative HPLC to obtain the product P-24 (22 mg, 21.60%).
1H NMR (600 MHz, DMSO-d6) δ10.98 (s, 1H), 7.46 (d, J=7.3 Hz, 2H), 7.41 (t, J=7.5 Hz, 2H), 7.34 (t, J=7.3 Hz, 1H), 7.24 (s, 1H), 7.14 (dt, J=12.4, 5.4 Hz, 4H), 6.91-6.89 (m, 1H), 6.84 (d, J=7.2 Hz, 2H), 6.78-6.72 (m, 2H), 6.65 (s, 2H), 6.26 (d, J 7.3 Hz, 2H), 5.07 (d, J=15.1 Hz, 3H), 4.33 (d, J=17.0 Hz, 2H), 4.24-4.13 (m, 4H), 3.41-3.31 (m, 4H), 2.99 (ddt, J=50.0, 25.9, 11.9 Hz, 10H), 2.60 (d, J=16.8 Hz, 1H), 2.40-2.34 (m, 1H), 2.22-2.12 (m, 3H), 1.99 (t, J=14.4 Hz, 4H), 1.92-1.85 (m, 2H), 1.75 (d, J=7.7 Hz, 1H), 1.64 (d, J=18.9 Hz, 4H).
LC-MS (ESI): [M+H]+=883.44.
Under the condition of nitrogen protection, compound P-24 (30 mg, 0.03 mmol) was dissolved in methanol (5 mL). 10% Pd/C (6 mg, 20% wt) was added, and after three replacement with hydrogen, then heated to 50° C. in hydrogen atmosphere for 16 hours. After the reaction was completed, white solid PROTAC 58 (8.5 mg, 35.7%) was obtained by reverse preparation of the reaction solution.
1H NMR (600 MHz, DMSO-d6) δ10.97 (s, 1H), 9.54 (s, 1H), 7.24 (s, 1H), 7.14 (dd, J=12.2, 4.7 Hz, 3H), 6.83 (d, J=7.2 Hz, 2H), 6.69-6.57 (m, 4H), 6.48 (dd, J=8.3, 2.2 Hz, 1H), 6.24 (d, J=8.0 Hz, 2H), 7.24 (s, 1H), 5.07 (dd, J=13.3, 4.9 Hz, 1H), 4.36-4.30 (m, 1H), 4.30-4.19 (m, 2H), 4.18-4.13 (m, 2H), 3.84 (dd, J=16.4, 8.1 Hz, 1H), 2.96 (dtd, J=38.2, 20.2, 13.1 Hz, 8H), 2.60 (d, J=16.8 Hz, 1H), 2.40-2.33 (m, 5H), 2.19 (s, 3H), 2.09 (dd, J=11.7, 5.2 Hz, 2H), 1.98 (d, J=11.5 Hz, 4H), 1.87 (t, J=13.7 Hz, 2H), 1.71 (d, J=5.7 Hz, 1H), 1.63 (d, J=18.2 Hz, 3H).
LC-MS (ESI): [M+H]+=793.39.
Under the condition of nitrogen protection, 3-(4-((1R,2S)-6-hydroxyl-2-phenyl-1,2,3,4-tetrahydronaphthalene-1-yl)phenyl)-3-azaspiro[5.5]undecane-9-one (30 mg, 0.06 mmol) and compound 26 (36 mg, 0.09 mmol) were dissolved in DCE:DMSO=1:1 (4 mL), triethylamine (15 mg, 0.18 mmol) and acetic acid (15 mg, 0.18 mmol) were added, stirred at room temperature for 1 h. Sodium triacetoxyborohydride (42 mg, 0.18 mmol) was added to the above reaction solution, and the reaction was performed at room temperature for 24 h. Purified by preparative HPLC to obtain the product PROTAC 59 (13 mg, 23.6%).
1H NMR (600 MHz, DMSO-d6) δ10.98 (s, 1H), 9.07 (s, 1H), 7.30 (dd, J=12.9, 7.7 Hz, 1H), 7.14 (dd, J=11.7, 7.0 Hz, 3H), 7.05 (t, J=8.2 Hz, 1H), 6.84 (d, J=7.3 Hz, 4H), 6.66-6.61 (m, 2H), 6.51-6.47 (m, 1H), 6.35 (s, 2H), 5.02 (dt, J=11.8, 5.4 Hz, 1H), 4.32 (d, J=17.3 Hz, 1H), 4.23 (dd, J=19.4, 8.0 Hz, 2H), 3.95 (s, 1H), 63.37-3.30 (m, 3H), 3.26-3.07 (m, 7H), 2.94 (ddq, J=30.1, 12.9, 8.7, 6.2 Hz, 5H), 2.68-2.55 (m, 2H), 2.34 (td, J=13.3, 4.7 Hz, 1H), 2.08 (q, J=7.2, 5.9 Hz, 1H), 1.94 (ddt, J=34.1, 13.4, 7.0 Hz, 4H), 1.69 (dddd, J=63.4, 41.3, 26.0, 13.2 Hz, 9H), 1.48 (d, J=22.9 Hz, 2H), 1.25-1.12 (m, 3H).
LC-MS (ESI): [M+H]+=819.44.
Intermediate P-25 (20 g, 92.9 mmol) was dissolved in a one-necked flask containing tetrahydrofuran (100 mL), cooled to 0° C., sodium hydrogen (8.36 g, 209.02 mmol) was slowly added, and stirred at 0° C. for 1 h. A solution of 2-chloro-4-fluorobenzonitrile (20 g, 92.9 mmol) in tetrahydrofuran (100 mL) was added into the one-necked flask, and stirred at 0° C. for 0.5 h. Transfer to room temperature and continue stirring for 3 h. After the reaction was completed, the reaction mixture was cooled to 0° C., quenched with ice water, and extracted three times with ethyl acetate (100 mL). And the organic phase was washed with saturated brine, dried over anhydrous sodium sulfate, filtered and concentrated to obtain a crude product. The crude product was purified by column (0-25% ethyl acetate) to obtain white solid intermediate P-26 (21.92 g).
1H NMR (600 MHz, CD3OD) δ7.69 (d, J=8.8 Hz, 1H), 7.18 (d, J=2.4 Hz, 1H), 7.03 (dd, J=8.8, 2.4 Hz, 1H), 4.43 (tt, J=10.2, 4.1 Hz, 1H), 3.42 (dq, J=10.9, 5.8 Hz, 1H), 2.18-2.10 (m, 2H), 2.07-1.97 (m, 2H), 1.61-1.51 (m, 2H), 1.46 (s, 9H), 1.43-1.40 (m, 2H).
LC-MS (ESI): [M+H]+=351.11
Intermediate P-26 (21.92 g, 62.48 mmol) was added to a one-necked flask containing dioxane (25 mL). A solution (of hydrochloric acid in dioxane (25 mL) was added to the reaction flask. Stir at room temperature for 2 h after addition. After the reaction was completed, it was directly concentrated to obtain the crude white solid intermediate P-27 (20.05 g).
LC-MS (ESI): [M+H]+=251.13
The intermediate P-27 (20.05 g, 79.97 mmol), triethylamine (32.37 g, 319.87 mmol), 1-propyl phosphoric anhydride (12.2 g, 96 mmol) and dichloromethane (200 mL) were added to a single-necked flask in order, stirred at room temperature for 5 min after addition. 6-Chloropyridazine-3-carboxylic acid (16.48 g, 103.96 mmol) was slowly added into the one-necked flask, and the temperature was raised to 30° C. and stirring was continued for 2.5 h. After the reaction was completed, the reaction mixture was concentrated and purified by pulping (petroleum ether: ethyl acetate=1:1) to obtain intermediate 18 (13.45 g) as a brown solid.
1H NMR (600 MHz, DMSO-d6) δ9.17 (d, J=8.2 Hz, 1H), 8.23 (d, J=8.9 Hz, 1H), 8.11 (d, J=8.9 Hz, 1H), 7.87 (d, J=8.7 Hz, 1H), 7.41 (d, J=2.4 Hz, 1H), 7.15 (dd, J=8.8, 2.4 Hz, 1H), 4.54 (td, J=10.5, 5.1 Hz, 1H), 3.91 (dtd, J=11.5, 7.7, 4.1 Hz, 1H), 2.13 (d, J=12.3 Hz, 2H), 1.90 (s, 2H), 1.71 (q, J=13.4 Hz, 2H), 1.57-1.48 (m, 2H).
LC-MS (ESI): [M+H]+=391.33
Compound 3 (100 mg, 0.281 mmol) and tert-butyl 4-formylpiperidine-1-carboxylate (90 mg, 0.422 mmol) were added the solvent DCE (2 mL), AcOH (4 drop), reacted at room temperature for 1 h. Then NaBH(OAc)3 (179 mg, 0.844 mmol) was added, and the reaction was continued for 1 h at room temperature. After the reaction solution was concentrated, it was washed with 5 mL of saturated aqueous ammonium chloride solution, extracted with ethyl acetate, concentrated and purified by preparative thin-layer plate chromatography (DCM:MeOH=10:1) to obtain white solid compound P-28 (120 mg).
LC-MS (ESI): [M+H]+=497.40
Compound P28 (100 mg, 0.181 mmol), dioxane (1 mL) and 4M dioxane hydrochloride solution (1 mL) were reacted at room temperature for 0.5 h, and concentrated to obtain white solid compound P-29 (40 mg).
Compound P-29 (40 mg, 0.088 mmol), intermediate 18 (69.17 mg, 0.176 mmol), DIEA (22.85 mg, 0.177 mmol), and DMSO (1 mL) were sequentially added into a 8 mL sealed tube. Under the protection of N2, react at 80° C. for 2 h. After the reaction was completed, it was cooled to room temperature, concentrated, and purified by Prep-HPL=C (15-50% acetonitrile) to obtain white solid compound PROTAC 60 (1.23 mg).
1H NMR (400 MHz, DMSO-d) δ11.11 (s, 1H), 8.57 (d, J=8.2 Hz, 1H), 7.84 (dd, J=11.0, 9.1 Hz, 2H), 7.78 (s, 1H), 7.41-7.34 (m, 2H), 7.29 (s, 1H), 7.13 (dd, J=8.8, 2.5 Hz, 1H), 5.32 (s, 1H), 5.10 (dd, J=12.8, 5.5 Hz, 2H), 4.43 (m, J=65.0 Hz, 8H), 3.85 (s, 3H), 2.60 (d, J=17.5 Hz, 3H), 2.25 (s, 3H), 2.15-1.86 (m, 8H), 1.78 (d, J=12.5 Hz, 3H), 1.63 (q, J=12.3 Hz, 3H).
LC-MS (ESI): [M+H]+=807.65
Compound 1 (200 mg, 0.521 mmol) and tert-butyl 2-oxo-7-azaspiro[3,5]nonane-7-tert-butyl carboxylate (249.67 mg, 1.04 mmol) were added in a glass vial in order, solvent THF (2 mL) and Ti(O-iPr)4 (296.5 mg, 1.04 mmol) was added, stirred and reacted at 60° C. for 1 h, then NaBH3CN (98.3 mg, 1.56 mmol) was added, the reaction was continued to react at 60° C. for 1 h, after the reaction was completed, it was concentrated, and then purified by preparative thin-layer plate chromatography (DCM:MeOH=10:1) to obtain white solid compound P-30 (200 mg).
LC-MS: [M+H]+=607.53
Compound P-30 (200 mg, 0.346 mmol), DCM (1 mL) solution and TFA (1 mL) were sequentially added into a one-necked bottle in order. Stir at room temperature for 1 h. After the reaction was completed, it was directly concentrated to obtain white solid compound P-31 (150 mg).
LC-MS: [M+H]+=507.50
The preparation refers to the third step of Example 89.
1H NMR (600 MHz, DMSO-d6) δ11.11 (s, 1H), 9.84 (q, J=9.5 Hz, 1H), 8.57 (d, J=8.2 Hz, 1H), 7.84 (dd, J=21.0, 9.1 Hz, 2H), 7.75 (d, J=4.2 Hz, 1H), 7.42-7.33 (m, 3H), 7.14 (dd, J=8.8, 2.4 Hz, 1H), 5.10 (dd, J=12.9, 5.5 Hz, 1H), 4.54 (tt, J=9.9, 4.4 Hz, 1H), 3.92-3.81 (m, 2H), 3.74 (t, J=5.5 Hz, 2H), 3.66 (t, J=5.4 Hz, 2H), 3.33 (d, J=11.4 Hz, 2H), 3.08 (q, J=11.8 Hz, 2H), 2.97 (t, J=7.0 Hz, 2H), 2.89 (ddd, J=17.7, 13.9, 5.5 Hz, 1H), 2.64-2.52 (m, 2H), 2.27 (dd, J=12.0, 7.9 Hz, 2H), 2.14-2.01 (m, 7H), 1.92 (dddd, J=27.9, 23.3, 16.9, 5.0 Hz, 6H), 1.72-1.57 (m, 6H), 1.57-1.47 (m, 2H).
LC-MS: [M+H]+=861.66
Compound 12 (100 mg, 0.271 mmol) and tert-butyl 3-oxoazetidine-1-carboxylate (139.03 mg, 0.812 mmol) added the solvent DCE (1 mL), AcOH (4 drops), reacted 1 h at room temperature. NaBH(OAc)3 (172.11 mg, 0.812 mmol) was added, and the reaction was continued for 1 h at room temperature. After the reaction solution was concentrated, it was washed with 5 mL of saturated aqueous ammonium chloride solution, extracted with ethyl acetate. After concentration, the white solid compound P-32 (109 mg) was purified by thin-layer plate chromatography (DCM:MeOH=10:1).
Refer to the second step of Example 89 for preparation.
Refer to the third step of Example 89 for preparation.
1H NMR (400 MHz, DMSO-d6) δ10.95 (s, 1H), 8.60 (d, J=8.1 Hz, 1H), 7.93 (d, J=9.2 Hz, 1H), 7.85 (d, J=8.8 Hz, 1H), 7.51 (s, 1H), 7.37 (d, J=2.4 Hz, 1H), 7.12 (dd, J=8.8, 2.4 Hz, 1H), 7.06-6.96 (m, 2H), 5.05 (dd, J=13.3, 5.1 Hz, 1H), 4.57-4.29 (m, 6H), 4.21 (d, J=17.0 Hz, 1H), 3.32-3.12 (s, 5H), 2.89 (m, 2H), 2.58 (d, J=16.7 Hz, 2H), 2.36 (d, J=13.2 Hz, 2H), 2.17-1.77 (m, 11H), 1.64 (q, J=11.5 Hz, 2H), 1.57-1.43 (m, 2H).
LC-MS (ESI): [M+H]+=779.63
Compound 17 (20 mg, 0.054 mmol), tert-butyl 2-oxo-7-azaspiro[3,5]nonane-7-carboxylate (25.9 mg, 0.108 mmol) and Ti(O-iPr)4 (30 mg, 0.10 mmol) were added in a glass vial in order. THF (1 mL) was added dropwise, and the reaction was stirred at 60° C. with for 2 h. And then NaBH3CN (11 mg, 0.162 mmol) was added. The reaction was stirred at 60° C. for 1 h. After the reaction was completed, it was concentrated and purified by TLC (DCM:MeOH=10:1) to obtain white solid compound P-34 (20 mg).
Refer to the second step of PROTAC 60 for preparation.
Refer to the third step of Example 89 for preparation.
1H NMR (600 MHz, DMSO-d) δ11.13 (s, 1H), 8.59 (t, J=6.9 Hz, 1H), 7.90-7.80 (m, 2H), 7.66 (s, 1H), 7.44-7.34 (m, 3H), 7.14 (dd, J=8.8, 2.4 Hz, 1H), 5.16-5.08 (m, 1H), 4.74 (d, J=12.9 Hz, 3H), 4.55 (dd, J=9.3, 5.2 Hz, 2H), 4.02-3.99 (m, 1H), 3.93-3.61 (m, 6H), 3.47 (d, J=10.8 Hz, 1H), 3.28 (dd, J=29.5, 23.9 Hz, 2H), 2.98-2.85 (m, 2H), 2.61 (dd, J=9.5, 7.6 Hz, 1H), 2.32-2.22 (m, 2H), 2.09 (ddd, J=27.3, 21.5, 20.0 Hz, 7H), 1.90 (d, J=12.2 Hz, 3H), 1.79-1.46 (m, 7H).
LC-MS (ESI): [M+H]+=847.65
Compound 4 (200 mg, 0.522 mmol) and tert-butyl 3-oxoazetidine-1-carboxylate (178.61 mg, 1.04 mmol) were successively added into a glass vial. Solvent THF (2 mL) and Ti(O-iPr)4 (296.52 mg, 1.04 mmol) were added and reacted 1 h at 60° C. with stirring. And then NaBH3CN (98.34 mg, 1.56 mmol) was added, continued to react 1 h at 60° C. with stirring. After the reaction was completed, it was concentrated, and then purified by preparing thin layer chromatography (DCM:MeOH=10:1) to obtain compound P-36 (170 mg) as a white solid.
1H NMR (600 MHz, CD3OD) δ7.68 (s, 1H), 7.30 (s, 1H), 5.14-5.10 (m, 1H), 4.36-4.24 (m, 4H), 4.14 (t, J=11.1 Hz, 4H), 3.52 (s, 2H), 3.14 (s, 4H), 2.96-2.82 (m, 4H), 2.79-2.69 (m, 3H), 1.48 (s, 9H).
LC-MS: [M+H]+=539.43
Refer to the second step of Example 89 for preparation.
LC-MS: [M+H]+=439.36
Refer to the third step of Example 89 for preparation.
1H NMR (400 MHz, DMSO-d6) δ11.09 (s, 1H), 8.55 (d, J=8.2 Hz, 1H), 7.84 (dd, J=9.0, 7.2 Hz, 2H), 7.69 (s, 1H), 7.38 (d, J=2.4 Hz, 1H), 7.25 (s, 1H), 7.13 (dd, J=8.8, 2.4 Hz, 1H), 6.85 (d, J=9.3 Hz, 1H), 5.08 (dd, J=12.8, 5.4 Hz, 1H), 4.53 (dt, J=10.3, 5.6 Hz, 1H), 4.26-4.12 (m, 2H), 4.07 (s, 2H), 3.95 (dd, J=9.0, 5.0 Hz, 2H), 3.86 (d, J=11.6 Hz, 1H), 2.86 (s, 3H), 2.69-2.51 (m, 3H), 2.46 (d, J=6.1 Hz, 2H), 2.32 (d, J=11.0 Hz, 2H), 2.16-2.06 (m, 2H), 2.06-1.93 (m, 2H), 1.90 (dt, J=8.1, 4.1 Hz, 2H), 1.71-1.58 (m, 2H), 1.52 (td, J=9.7, 4.6 Hz, 3H), 1.42 (d, J=14.3 Hz, 2H).
LC-MS: [M+H]+=793.8
Compound 16 (150 mg, 0.404 mmol) and tert-butyl 4-formylpiperidine-1-carboxylate (172.1 mg, 0.808 mmol) were successively added into a reaction flask, added solvent DCE (2 mL), AcOH (4 drop) in drops, and reacted 1 h at room temperature with stirring. Then NaBH(OAc)3 (257 mg, 1.21 mmol) was added, and continued to react 1 h at room temperature with stirring. After reaction was completed, it was concentrated, and then purified by preparing thin layer chromatography to obtain white solid compound P-38 (130 mg).
1H NMR (600 MHz, DMSO-d6) δ10.95 (s, 1H), 7.16 (s, 1H), 7.09 (s, 1H), 5.05 (dd, J=13.4, 5.1 Hz, 1H), 4.46 (t, J=5.3 Hz, 1H), 4.30 (d, J=16.6 Hz, 1H), 4.17 (d, J=16.7 Hz, 1H), 4.09-4.00 (m, 2H), 3.91 (s, 3H), 3.27-3.18 (m, 1H), 2.94-2.83 (m, 2H), 2.63-2.53 (m, 3H), 2.40-2.24 (m, 3H), 2.17-2.12 (m, 2H), 2.04-1.91 (m, 2H), 1.74-1.57 (m, 6H), 1.38 (s, 9H).
LC-MS: [M+H]+=469.50
Refer to the second step of Example 89 for preparation.
LC-MS: [M+H]+=469.50
Refer to the third step of Example 89 for preparation.
1H NMR (400 MHz, DMSO-d6) δ10.96 (s, 1H), 8.57 (d, J=8.2 Hz, 1H), 7.90-7.80 (m, 2H), 7.44-7.35 (m, 2H), 7.24 (s, 1H), 7.18-7.10 (m, 2H), 5.07 (dd, J=13.3, 5.0 Hz, 1H), 4.60-4.46 (m, 2H), 4.39-4.28 (m, 1H), 4.26-4.10 (m, 2H), 3.92-3.80 (m, 1H), 3.63-3.45 (m, 10H), 3.23-3.00 (m, 5H), 2.98-2.83 (m, 1H), 2.68-2.56 (m, 1H), 2.43-2.28 (m, 1H), 2.26-2.07 (m, 3H), 2.04-1.81 (m, 6H), 1.71-1.44 (m, 4H).
LC-MS: [M+H]+=823.77
Compound 6 (80 mg, 0.216 mmol) and tert-butyl 4-formylpiperidine-1-carboxylate (92.4 mg, 433 mmol) were added successively into a glass vial, added solvent DCM (2 mL) and AcOH (2 drops) in drops, and reacted 1 h at room temperature with stirring. Then NaBH(OAc)3 (138 mg, 648 mmol) was added, and continued to react 1 h at room temperature with stirring. After the reaction was completed, the reaction mixture was concentrated, and then purified by TLC (DCM:MeOH=10:1) to obtain white solid compound P-40 (67.3 mg).
Refer to the second step of Example 89 for preparation.
Refer to the third step of Example 89 for preparation.
1H NMR (600 MHz, CD3OD) δ7.99 (d, J=9.7 Hz, 1H), 7.71 (d, J=8.8 Hz, 1H), 7.63 (s, 1H), 7.47 (d, J=9.7 Hz, 1H), 7.28 (s, 1H), 7.21 (d, J=2.3 Hz, 1H), 7.06 (dd, J=8.8, 2.3 Hz, 1H), 5.11 (dd, J=12.8, 5.4 Hz, 1H), 4.62-4.49 (m, 3H), 4.05-3.96 (m, 1H), 3.75 (dd, J=59.4, 9.9 Hz, 2H), 3.25-3.03 (m, 6H), 2.94-2.60 (m, 8H), 2.39-2.28 (m, 1H), 2.27-1.97 (m, 8H), 1.66 (dd, J=13.0, 7.1 Hz, 4H), 1.42 (ddd, J=77.8, 40.4, 18.7 Hz, 3H).
LC-MS (ESI): [M+H]+=821.68
Compound 6 (60 mg, 0.162 mmol) and tert-butyl 3-oxoazetidine-1-carboxylate (41.71 mg, 0.243 mmol) were added successively in a glass vial. Solvent THF (2 mL) and Ti(O-iPr)4 (92.33 mg, 0.324 mmol) were added, stirred and reacted at 60° C. for 1 h. Then NaBH3CN (30.62 mg, 0.487 mmol) was added and stirring was continued to react at 60° C. for 1 h. After the reaction was completed, the reaction mixture was subjected to concentrated treatment, and then purified by prepared thin layer chromatography (DCM:MeOH=10:1) to obtain the white solid compound P-42 (30 mg).
LC-MS: [M+H]+=525.43
Refer to the second step of PROTAC 60 for preparation.
LC-MS: [M+H]+=425.43
Refer to the third step of Example 89 for preparation.
1H NMR (400 MHz, CD3OD) δ 7.97 (d, J=8.4 Hz, 1H), 7.69 (d, J=8.7 Hz, 1H), 7.60 (s, 1H), 7.25 (s, 1H), 7.20 (d, J=2.4 Hz, 1H), 7.05 (dd, J=8.8, 2.4 Hz, 1H), 6.95 (d, J=8.2 Hz, 1H), 5.34 (s, 1H), 4.42 (m, 4H), 4.24 (s, 2H), 3.98 (s, 1H), 3.89-3.58 (m, 3H), 3.04-2.51 (m, 9H), 2.27-1.96 (m, 6H), 1.63 (q, J=13.6, 11.5 Hz, 4H).
LC-MS: [M+H]+=779.59
4-bromo-2-fluorobenzoic acid (5 g, 22.83 mmol), 2-methylalanine (7.06 g, 68.49 mmol), copper powder (2.90 g, 45.66 mmol), copper iodide (8.70 g, 45.66 mmol), potassium carbonate (15.78 g, 114.15 mmol) and N,N-dimethylglycine (1.17 g, 11.41 mmol) were added. Solvent DMF (100 mL) and water (10 mL) were added, and the reaction was stirred at 110° C. for 16 h. After cooling the reaction solution to room temperature, ice water (200 mL) was added, and ice hydrochloric acid was added dropwise to adjust the pH to 3-4. The mixture was extracted with ethyl acetate 3 times (500 mL*3). The organic phase was concentrated and was then purified by positive phase column (Ethyl acetate: petroleum ether=35%) to obtain a yellow solid (P-45) (4 g).
LC-MS: [M+H]+=242.2
P-45 (3 g, 12.44 mmol) and solvent methanol (50 mL) were sequentially added into a one-necked flask. Thionyl chloride (3.70 g, 31.09 mmol) was added dropwise under ice-bath conditions, and the reaction was stirred at 80° C. for 16 h. After the reaction was completed, it was directly concentrated to obtain a yellow solid (P-46) (3 g).
LC-MS: [M+H]+=270.07
P-46 (1 g, 3.71 mmol), 4-isothiocyanate-2-(trifluoromethyl)benzonitrile (1.02 g, 4.46 mmol), isopropyl acetate (10 mL) and DMSO (5 mL) were sequentially added to a single-necked flask. The reaction was carried out at 95° C. for 60 h under the protection of nitrogen. After the reaction was completed, it was concentrated until no solvent was evaporated, methanol was added dropwise, stirred and crystallized at 0-5° C., and intermediate P-47 (500 mg) was obtained by filtration.
LC-MS (ESI): [M+H]+=466.08
Intermediate P-47 (0.5 g, 1.07 mmol), sodium hydroxide (214.84 mg, 5.37 mmol), solvent methanol (5 mL), tetrahydrofuran (5 mL) and water (2.5 mL) were sequentially added into a one-necked flask. The reaction mixture was reacted at 40° C. for 2 h. After the reaction is complete, water (10 mL) was added and glacial hydrochloric acid was added dropwise to adjust the pH to 3-4. The reaction mixture was extracted 3 times with ethyl acetate (10 mL*3). The organic phase was concentrated, and then purified by reverse phase column (acetonitrile: water=60%) to obtain a white solid (Intermediate 30) (300 mg).
1H NMR (600 MHz, DMSO-d6) δ13.55 (s, 1H), 8.41 (d, J=8.2 Hz, 1H), 8.30 (d, J=1.9 Hz, 1H), 8.13-8.01 (m, 2H), 7.47 (dd, J=11.0, 1.9 Hz, 1H), 7.38 (dd, J=8.3, 1.9 Hz, 1H), 1.56 (s, 6H).
LC-MS (ESI): [M+H]+=450.22
Compound P-48 (40 mg, 0.08 mmol), intermediate 30 (52.2 mg, 0.12 mmol), DIEA (3 eq), BOP (3 eq) and DMF (2 mL) were sequentially added into an 8 mL sealed tube. The reaction mixture was reacted at room temperature for 3 h under N2 conditions. After the reaction was completed, it was cooled to room temperature, concentrated, and then purified by preparation to obtain white solid compound PROTAC 68 (5 mg).
1H NMR (600 MHz, MeOD) δ 8.21 (t, J=20.7 Hz, 2H), 8.02 (dd, J=8.2, 1.8 Hz, 1H), 7.63 (s, 1H), 7.45-7.37 (m, 1H), 7.29 (s, 1H), 7.23 (s, 1H), 5.13 (dd, J=13.3, 5.2 Hz, 1H), 4.57 (d, J=13.2 Hz, 1H), 4.39 (q, J=16.5 Hz, 2H), 3.64 (d, J=8.6 Hz, 2H), 3.56 (d, J=12.9 Hz, 1H), 3.47 (dd, J=26.8, 4.7 Hz, 3H), 3.24 (td, J=12.9, 2.4 Hz, 1H), 3.00-2.87 (m, 5H), 2.82-2.75 (m, 1H), 2.49 (ddd, J=26.5, 13.3, 4.6 Hz, 1H), 2.22 (t, J=6.5 Hz, 2H), 2.18-2.13 (m, 1H), 2.12-2.05 (m, 1H), 1.96 (d, J=4.7 Hz, 1H), 1.78 (ddd, J=19.9, 19.0, 5.4 Hz, 2H), 1.63 (s, 6H).
LC-MS (ESI): [M+H]+=890.27
Refer to the first step of Example 98 for preparation.
1H NMR (600 MHz, DMSO-d6) δ11.11 (s, 1H), 8.42 (d, J=8.2 Hz, 1H), 8.30 (d, J=1.3 Hz, 1H), 8.09 (dd, J=8.2, 1.5 Hz, 1H), 7.73 (s, 1H), 7.61 (t, J=7.7 Hz, 1H), 7.46 (dd, J=9.9, 1.4 Hz, 1H), 7.37-7.29 (m, 2H), 5.10 (dd, J=12.9, 5.4 Hz, 1H), 4.54 (d, J=11.9 Hz, 1H), 4.32 (s, 1H), 4.04 (s, 1H), 3.45 (d, J=13.0 Hz, 4H), 3.21-3.02 (m, 7H), 2.88 (dd, J=20.1, 9.0 Hz, 2H), 2.80 (s, 1H), 2.57 (dd, J=38.4, 10.8 Hz, 2H), 2.16 (s, 1H), 2.06-2.00 (m, 1H), 1.91 (t, J=5.7 Hz, 1H), 1.81-1.68 (m, 4H), 1.62 (d, J=13.6 Hz, 1H), 1.57 (s, 6H).
LC-MS (ESI): [M+H]+=914.29
Refer to the first step of Example 98 for preparation.
1H NMR (600 MHz, DMSO-d6) δ11.11 (s, 1H), 8.42 (dd, J=8.2, 2.6 Hz, 1H), 8.30 (s, 1H), 8.09 (d, J=8.2 Hz, 1H), 7.65 (ddd, J=28.3, 20.7, 13.8 Hz, 2H), 7.52-7.44 (m, 1H), 7.39-7.34 (m, 1H), 7.30 (dd, J=13.4, 5.0 Hz, 1H), 5.10 (dt, J=12.3, 6.0 Hz, 1H), 4.31 (d, J=32.9 Hz, 1H), 4.04 (d, J=16.9 Hz, 1H), 3.89 (dd, J=12.0, 7.7 Hz, 1H), 3.67 (d, J=8.8 Hz, 1H), 3.40 (s, 3H), 3.30 (s, 2H), 3.24-2.98 (m, 5H), 2.92-2.83 (m, 1H), 2.77 (dd, J=32.9, 11.9 Hz, 2H), 2.63-2.56 (m, 1H), 2.21-1.98 (m, 3H), 1.81-1.60 (m, 5H), 1.56 (s, 5H).
LC-MS (ESI): [M+H]+=890.27
Refer to the first step of Example 98 for preparation.
1H NMR (600 MHz, DMSO-d6) δ11.10 (s, 1H), 8.42 (d, J=8.2 Hz, 1H), 8.29 (d, J=1.2 Hz, 1H), 8.10-8.07 (m, 1H), 7.77-7.71 (m, 2H), 7.47 (dd, J=10.4, 1.4 Hz, 1H), 7.42-7.34 (m, 2H), 5.10 (dd, J=12.9, 5.4 Hz, 1H), 4.26 (t, J=9.1 Hz, 2H), 3.96 (m, 3H), 3.52 (s, 2H), 3.37 (dd, J=24.5, 10.7 Hz, 2H), 3.21 (dd, J=13.3, 7.9 Hz, 3H), 2.99-2.84 (m, 3H), 2.60 (d, J=17.4 Hz, 2H), 2.09-1.95 (m, 3H), 1.90 (dd, J=18.2, 11.2 Hz, 3H), 1.55 (s, 6H).
LC-MS (ESI): [M+H]+=886.26
Refer to the first step of Example 98 for preparation.
1H NMR (600 MHz, DMSO-d6) δ10.97 (s, 1H), 8.42 (d, J=8.2 Hz, 1H), 8.30 (d, J=1.4 Hz, 1H), 8.09 (dd, J=8.3, 1.5 Hz, 1-), 7.61 (t, J=7.8 Hz, 1H), 7.52 (d, J=5.6 Hz, 1H), 7.46 (dd, J=10.0, 1.3 Hz, 1H), 7.37-7.32 (m, 1H), 7.03 (d, J=17.1 Hz, 1H), 5.07 (dd, J=13.3, 5.2 Hz, 1H), 4.55 (d, J=12.8 Hz, 1H), 4.34 (t, J=13.7 Hz, 2H), 4.26-4.19 (m, 2H), 3.46 (d, J=10.5 Hz, 3H), 3.16 (d, J=29.2 Hz, 5H), 2.94-2.82 (m, 4H), 2.60 (d, J=17.0 Hz, 1H), 2.37 (dd, J=13.2, 4.6 Hz, 1H), 2.19 (s, 1H), 2.09-2.03 (m, 1H), 1.96 (m, 7H), 1.76 (s, 1H), 1.57 (s, 6H).
LC-MS (ESI): [M+H]+=890.31
Refer to the first step of Example 98 for preparation.
1H NMR (600 MHz, DMSO-d6) δ11.11 (d, J=4.8 Hz, 1H), 8.42 (d, J=8.3 Hz, 1H), 8.30 (s, 1H), 8.09 (d, J=8.3 Hz, 1H), 7.71 (ddd, J=62.7, 35.5, 12.2 Hz, 2H), 7.46 (ddd, J=10.4, 4.8, 1.6 Hz, 1H), 7.39-7.33 (m, 1H), 7.29 (dd, J=11.7, 3.9 Hz, 1H), 5.12-5.07 (m, 1H), 4.31 (d, J=19.1 Hz, 1H), 4.02 (d, J=17.4 Hz, 1H), 3.78 (dd, J=39.2, 11.7 Hz, 4H), 3.40-3.29 (m, 2H), 3.14 (ddd, J=50.3, 31.4, 16.0 Hz, 4H), 2.93-2.83 (m, 1H), 2.77 (d, J=17.9 Hz, 1H), 2.63-2.55 (m, 3H), 2.12-1.86 (m, 5H), 1.78-1.40 (m, 13H).
LC-MS (ESI): [M+H]+=940.30
Refer to the first step of Example 98 for preparation 1H NMR (600 MHz, DMSO) δ11.11 (s, 1H), 8.42 (d, J=8.2 Hz, 1H), 8.33-8.26 (m, 1H), 8.08 (t, J=10.5 Hz, 1H), 7.78-7.58 (m, 2H), 7.45 (d, J=10.2 Hz, 1H), 7.35-7.27 (m, 2H), 5.15-5.03 (m, 1H), 4.33 (d, J=8.5 Hz, 1H), 4.03 (t, J=12.5 Hz, 1H), 3.49-3.30 (m, 6H), 3.27-3.03 (m, 6H), 2.93-2.83 (m, 1H), 2.77 (dd, J=28.0, 9.5 Hz, 1H), 2.64-2.55 (m, 1H), 2.09-1.94 (m, 3H), 1.91-1.66 (m, 7H), 1.63-1.53 (m, 7H), 1.50 (d, J=2.8 Hz, 1H), 1.45-1.34 (m, 2H).
LC-MS (ESI): [M+H]+=954.35
Refer to the first step of Example 98 for preparation.
1H NMR (600 MHz, DMSO-d6) δ11.12 (s, 1H), 8.41 (t, J=13.1 Hz, 1H), 8.30-8.24 (m, 1H), 8.10 (t, J=11.0 Hz, 1H), 7.79 (d, J=13.4 Hz, 1H), 7.64-7.57 (m, 1H), 7.46 (d, J=9.8 Hz, 1H), 7.36 (d, J=7.8 Hz, 1H), 7.29 (s, 1H), 5.11 (dd, J=12.9, 5.4 Hz, 1H), 4.61-4.21 (m, 6H), 3.66 (s, 1H), 3.56 (s, 1H), 3.26 (d, J=21.9 Hz, 2H) 3.23 (s, 3H), 3.11 (d, J=9.6 Hz, 1H), 3.00-2.83 (m, 4H), 2.63-2.54 (m, 2H), 2.29 (s, 2H), 2.08-1.99 (m, 1H), 1.87 (d, J=9.9 Hz, 1H), 1.76 (s, 1H), 1.59 (s, 6H).
LC-MS (ESI): [M+H]+=916.92
Refer to the first step of Example 98 for the preparation.
1H NMR (600 MHz, DMSO-d6) δ 10.97 (s, 1H), 8.42 (d, J=8.3 Hz, 1H), 8.29 (d, J=1.4 Hz, 1H), 8.09 (dd, J=8.2, 1.6 Hz, 1H), 7.61 (t, J=7.8 Hz, 1H), 7.52 (s, 1H), 7.46 (d, J=9.9 Hz, 1H), 7.36 (d, J=8.1 Hz, 1H), 7.05 (t, J=22.5 Hz, 1H), 5.07 (dd, J=13.3, 5.0 Hz, 1H), 4.55 (s, 1H), 4.44 (s, 1H), 4.41-4.20 (m, 4H), 3.68-3.51 (m, 3H), 3.28 (s, 2H), 3.22 (s, 3H), 3.11 (d, J=9.5 Hz, 1H), 2.95-2.85 (m, 3H), 2.63-2.56 (m, 1H), 2.42-2.32 (m, 1H), 2.25 (s, 2H), 2.03-1.94 (m, 1H), 1.87 (d, J=10.6 Hz, 1H), 1.76 (s, 1H), 1.68-1.58 (m, 1H), 1.56 (s, 6H).
LC-MS (ESI): [M+H]+=902.93
Refer to the first step of Example 98 for preparation.
1H NMR (600 MHz, DMSO-d6) δ 11.12 (s, 1H), 8.42 (d, J=8.3 Hz, 1H), 8.28 (t, J=19.0 Hz, 1H), 8.16-7.98 (m, 1H), 7.81-7.67 (m, 2H), 7.48 (dt, J=26.3, 13.1 Hz, 1H), 7.41-7.29 (m, 1H), 7.30 (s, 1H), 5.11 (dd, J=12.9, 5.4 Hz, 1H), 4.45-3.99 (m, 8H), 2.95 (t, J=6.0 Hz, 2H), 2.89 (ddd, J=17.1, 14.1, 5.4 Hz, 1H), 2.61 (m, 2H), 2.22 (t, J=6.2 Hz, 2H), 2.07-1.98 (m, 2H), 1.55 (s, 6H).
LC-MS(ESI): [M+H]+=902.93
Refer to the first step of Example 98 for preparation.
1H NMR (600 MHz, DMSO-d6) δ 11.11 (s, 1H), 8.42 (d, J=8.3 Hz, 1H), 8.30 (s, 1H), 8.09 (dd, J=8.2, 1.3 Hz, 1H), 7.75-7.60 (m, 2H), 7.48 (dd, J=10.0, 1.4 Hz, 1H), 7.36 (dt, J=12.7, 6.3 Hz, 1H), 7.30 (d, J=3.9 Hz, 1H), 5.10 (dd, J=12.9, 5.4 Hz, 1H), 4.70 (d, J=12.3 Hz, 1H), 4.33 (s, 1H), 4.04 (s, 1H), 3.63-3.51 (m, 3H), 3.18 (d, J=11.1 Hz, 3H), 3.08 (t, J=21.2 Hz, 1H), 2.95-2.83 (m, 2H), 2.80 (s, 1H), 2.66-2.56 (m, 2H), 2.22 (s, 1H), 2.09 (d, J=12.4 Hz, 1H), 2.01 (ddd, J=19.3, 11.2, 6.0 Hz, 2H), 1.69 (dd, J=33.8, 13.8 Hz, 6H), 1.57 (s, 6H).
LC-MS(ESI): [M+H]+=900.92
Refer to the first step of Example 98 for preparation.
1H NMR (600 MHz, DMSO-d6) δ 11.14-11.08 (m, 1H), 8.42 (d, J=8.2 Hz, 1H), 8.29 (t, J=9.1 Hz, 1H), 8.13-8.03 (m, 1H), 7.74 (dt, J=7.8, 4.7 Hz, 1H), 7.66 (d, J=13.0 Hz, 1H), 7.47 (t, J=10.4 Hz, 1H), 7.35 (dd, J=22.0, 13.7 Hz, 2H), 5.12 (ddd, J=12.6, 11.9, 5.6 Hz, 1H), 4.72 (dd, J=18.7, 3.4 Hz, 2H), 4.25-4.16 (m, 2H), 4.10 (s, 2H), 3.33-3.17 (m, 2H), 2.97-2.82 (m, 3H), 2.68-2.56 (m, 3H), 2.55-2.45 (m, 1H), 2.44-2.36 (m, 2H), 2.28-2.08 (m, 2H), 2.02 (dd, J=23.3, 18.5 Hz, 3H), 1.85 (s, 1H), 1.55 (s, 6H).
LC-MS(ESI): [M+H]+=898.90
Refer to the first step of Example 98 for preparation.
1H NMR (600 MHz, DMSO-d6) δ 11.12 (s, 1H), 8.42 (d, J=8.3 Hz, 1H), 8.28 (t, J=14.1 Hz, 1H), 8.09 (dd, J=8.2, 1.6 Hz, 1H), 7.65 (d, J=12.6 Hz, 2H), 7.52-7.45 (m, 1H), 7.43-7.34 (m, 2H), 5.16-5.07 (m, 1H), 4.79-4.68 (m, 3H), 3.59 (d, J=10.8 Hz, 4H), 3.18 (dd, J=57.5, 11.3 Hz, 3H), 2.94-2.83 (m, 2H), 2.69-2.56 (m, 2H), 2.23 (s, 3H), 2.06 (dd, J=20.0, 6.9 Hz, 3H), 1.97-1.85 (m, 1H), 1.72 (d, J=47.8 Hz, 2H), 1.57 (s, 6H).
LC-MS(ESI): [M+H]+=886.89
Refer to the first step of Example 98 for preparation.
1H NMR (600 MHz, DMSO-d6) δ 11.12 (s, 1H), 8.41 (d, J=8.3 Hz, 1H), 8.30 (d, J=1.6 Hz, 1H), 8.19-8.04 (m, 1H), 7.76 (t, J=7.7 Hz, 2H), 7.48 (d, J=10.0 Hz, 1H), 7.41-7.25 (m, 2H), 5.10 (dd, J=13.0, 5.3 Hz, 1H), 4.66 (s, 2H), 4.07 (m, 7H), 2.93-2.83 (m, 2H), 2.64-2.55 (m, 2H), 2.02 (ddd, J=20.5, 19.7, 14.6 Hz, 4H), 1.76 (s, 2H), 1.55 (s, 6H).
LC-MS(ESI): [M+H]+=858.84
The first step 4-(3-(4-(4-(7′-(2,6-dioxopiperidin-3-yl)-6′,8′dioxo-4′,6′,7′,8′-tetrahydro-3′H-spiro[piperidine-4,2′-pyrano[2,3-f]isoindol]-1-yl)methyl)-4-fluoropiperidine-1-carbonyl)-3-fluorophenyl)-4,4-dimethyl-5-oxo-2-thioimidazolidin-1-yl)-2-(trifluoromethyl)benzonitrile (Compound PROTAC 82)
Refer to the first step of Example 98 for preparation
1H NMR (600 MHz, DMSO-d6) δ 11.10 (s, 1H), 8.42 (d, J=8.2 Hz, 1H), 8.30 (s, 1H), 8.09 (d, J=8.2 Hz, 1H), 7.70 (s, 1H), 7.64 (t, J=7.8 Hz, 1H), 7.45 (dd, J=10.0, 1.4 Hz, 1H), 7.35 (d, J=8.1 Hz, 1H), 7.22 (s, 1H), 5.09 (dd, J=12.9, 5.5 Hz, 1H), 4.31 (d, J=13.0 Hz, 2H), 3.20-3.12 (m, 2H), 2.95-2.87 (m, 2H), 2.71-2.54 (m, 6H), 2.06-1.94 (m, 3H), 1.92-1.81 (m, 3H), 1.75-1.62 (m, 5H), 1.54 (d, J=36.8 Hz, 5H).
LC-MS(ESI): [M+H]+=932.01
Refer to the first step of Example 98 for preparation.
1H NMR (600 MHz, DMSO-d6) δ 11.10 (s, 1H), 8.41 (d, J=8.3 Hz, 1H), 8.30 (d, J=1.5 Hz, 1H), 8.09 (dd, J=8.2, 1.7 Hz, 1H), 7.69 (d, J=8.4 Hz, 1H), 7.60 (td, J=7.8, 4.9 Hz, 1H), 7.43 (ddd, J=9.9, 4.4, 1.7 Hz, 1H), 7.38-7.29 (m, 1H), 7.20 (d, J=14.6 Hz, 1H), 5.12-5.04 (m, 1H), 3.62 (d, J=26.6 Hz, 2H), 3.23 (d, J=7.3 Hz, 5H), 2.88 (m, 3H), 2.66-2.56 (m, 4H), 2.32 (d, J=10.9 Hz, 1H), 2.02 (dd, J=11.3, 5.2 Hz, 1H), 1.79 (m, 5H), 1.64 (d, J=27.9 Hz, 5H), 1.56 (s, 5H), 1.47-1.21 (m, 6H), 1.17-1.06 (m, 2H).
LC-MS(ESI): [M+H]+=968.34
Refer to the first step of Example 98 for preparation.
1H NMR (600 MHz, DMSO-d6) δ 11.10 (s, 1H), 8.41 (d, J=8.2 Hz, 1H), 8.30 (d, J=1.6 Hz, 1H), 8.09 (dd, J=8.2, 1.6 Hz, 1H), 7.67 (d, J=30.4 Hz, 1H), 7.61 (d, J=6.9 Hz, 1H), 7.44 (dd, J=9.9, 1.7 Hz, 1H), 7.37-7.30 (m, 1H), 7.20 (s, 1H), 5.09 (dd, J=12.9, 5.5 Hz, 1H), 4.52 (d, J=12.9 Hz, 1H), 3.42 (d, J=21.6 Hz, 3H), 3.08 (d, J=35.2 Hz, 2H), 2.93-2.78 (m, 4H), 2.66-2.53 (m, 3H), 2.40-2.29 (m, 2H), 2.26-2.15 (m, 2H), 2.02 (dd, J=14.2, 8.9 Hz, 1H), 1.83 (dd, J=23.5, 17.1 Hz, 3H), 1.76-1.61 (m, 5H), 1.58-1.38 (m, 7H).
LC-MS(ESI): [M+H]+=914.29
Refer to the first step of Example 98 for preparation.
1H NMR (600 MHz, DMSO-d6) δ 11.08 (t, J=22.1 Hz, 1H), 8.40 (dt, J=49.2, 24.6 Hz, 1H), 8.27 (d, J=36.8 Hz, 1H), 8.09 (d, J=8.2 Hz, 1H), 7.77-7.62 (m, 2H), 7.47-7.39 (m, 1H), 7.37-7.28 (m, 1H), 7.28 (d, J=9.9 Hz, 1H), 5.07 (ddd, J=64.2, 34.7, 29.5 Hz, 1H), 4.15 (s, 2H), 4.04 (s, 2H), 3.59 (dd, J=13.2, 7.4 Hz, 2H), 2.89 (m, 4H), 2.64-2.55 (m, 2H), 2.48-2.41 (m, 2H), 2.00 (m, 1H), 1.93-1.76 (m, 4H), 1.59-1.39 (m, 11H).
LC-MS(ESI): [M+H]+=912.27
Refer to the first step of Example 98 for preparation.
1H NMR (600 MHz, MeOD) δ8.19 (m, 2H), 8.02 (d, J=8.2 Hz, 1H), 7.71 (s, 1H), 7.65 (s, 1H), 7.44 (m, 3H), 5.12 (dd, J=12.8, 5.5 Hz, 1H), 4.95 (d, J=12.6 Hz, 1H), 3.88 (d, J=12.8 Hz, 1H), 3.63 (d, J=35.9 Hz, 3H), 3.52-3.42 (m, 2H), 3.02 (dt, J=23.9, 8.8 Hz, 3H), 2.88 (m, 2H), 2.80-2.69 (m, 3H), 2.39 (s, 1H), 2.24 (d, J=13.4 Hz, 2H), 2.13 (dd, J=8.8, 3.7 Hz, 2H), 2.01 (t, J=29.6 Hz, 3H), 1.84 (s, 2H), 1.63 (s, 6H).
LC-MS(ESI): [M+H]+=900.27
Refer to the first step of Example 98 for preparation.
1H NMR (600 MHz, MeOD) δ 8.20-8.18 (m, 2H), 8.01 (d, J=8.3 Hz, 1H), 7.79 (t, J=7.7 Hz, 1H), 7.70 (s, 1H), 7.45 (dd, J=16.7, 9.3 Hz, 1H), 7.38 (s, 1H), 5.11 (dd, J=12.9, 5.4 Hz, 1H), 4.58-4.52 (m, 2H), 4.45 (d, J=36.2 Hz, 2H), 4.25 (s, 1H), 3.52-3.37 (m, 2H), 3.29-3.16 (m, 2H), 3.04 (t, J=6.4 Hz, 2H), 2.94-2.67 (m, 4H), 2.24-2.09 (m, 3H), 2.03 (d, J=6.3 Hz, 3H), 1.62 (s, 6H).
LC-MS(ESI): [M+H]+=872.24
Refer to the first step of Example 98 for preparation.
1H NMR (600 MHz, MeOD) δ 8.19 (d, J=5.7 Hz, 2H), 8.02 (d, J=8.3 Hz, 1H), 7.70 (s, 1H), 7.60 (dd, J=14.3, 7.2 Hz, 1H), 7.45-7.35 (m, 3H), 5.16-5.07 (m, 1H), 3.83 (t, J=42.4 Hz, 3H), 3.56-3.36 (m, 4H), 3.21 (dd, J=6.4, 4.8 Hz, 2H), 3.04 (s, 2H), 2.93-2.67 (m, 4H), 2.47 (m, 2H), 2.19-2.13 (m, 4H), 2.01 (t, J=29.6 Hz, 4H), 1.77 (m, 4H), 1.63 (s, 6H).
LC-MS(ESI): [M+H]+=940.30
Refer to the first step of Example 98 for preparation.
1H NMR (600 MHz, MeOD) δ 8.19 (dd, J=4.9, 3.1 Hz, 2H), 8.01 (dd, J=8.3, 1.7 Hz, 1H), 7.62 (dd, J=14.7, 6.7 Hz, 2H), 7.45-7.39 (m, 2H), 7.10 (d, J=35.3 Hz, 1H), 5.13 (dd, J=13.3, 5.1 Hz, 1H), 4.41 (dt, J=33.6, 16.8 Hz, 3H), 3.71-3.35 (m, 10H), 3.04-2.97 (m, 2H), 2.92 (m, 2H), 2.85-2.72 (m, 1H), 2.49 (ddd, J=26.4, 13.3, 4.3 Hz, 2H), 2.14 (m, 6H), 1.99 (t, J=6.7 Hz, 2H), 1.79-1.58 (m, 9H).
LC-MS(ESI): [M+H]+=914.32
Refer to the first step of Example 98 for preparation.
1H NMR (600 MHz, MeOD) δ 8.22-8.13 (m, 2H), 8.01 (d, J=8.2 Hz, 1H), 7.80 (t, J=7.8 Hz, 1H), 7.52-7.41 (m, 2H), 7.36 (s, 1H), 7.32 (s, 1H), 5.14 (dd, J=13.2, 4.8 Hz, 1H), 4.63-4.55 (m, 2H), 4.52 (m, 1H), 4.47-4.33 (m, 2H), 4.48-4.33 (m, 4H), 3.63-3.35 (m, 4H), 3.01 (t, J=6.5 Hz, 2H), 2.95-2.87 (m, 1H), 2.84-2.75 (m, 1H), 2.52-2.43 (m, 1H), 2.25-2.11 (m, 3H), 2.06-1.91 (m, 4H), 1.62 (s, 6H).
LC-MS(ESI): [M+H]+=858.26
Refer to the first step of Example 98 for preparation.
1H NMR (600 MHz, DMSO-d6) δ 11.11 (s, 1H), 8.42 (d, J=8.2 Hz, 1H), 8.30 (s, 1H), 8.09 (d, J=8.2 Hz, 1H), 7.75 (s, 1H), 7.61 (t, J=7.8 Hz, 1H), 7.48-7.32 (m, 3H), 5.10 (dd, J=12.9, 5.4 Hz, 1H), 4.08 (s, 2H), 3.22 (m, 10H), 2.96 (s, 2H), 2.88 (m, 1H), 2.66-2.52 (m, 5H), 2.17-1.83 (m, 7H), 1.67-1.46 (m, 8H).
LC-MS(ESI): [M+H]+=928.03
Refer to the first step of Example 98 for preparation.
1H NMR (600 MHz, DMSO-d6) δ 11.12 (s, 1H), 8.42 (d, J=8.3 Hz, 1H), 8.30 (t, J=4.3 Hz, 1H), 8.09 (dd, J=8.2, 1.6 Hz, 1H), 7.63 (dd, J=19.8, 11.9 Hz, 2H), 7.46 (dd, J=14.3, 12.8 Hz, 1H), 7.39-7.30 (m, 2H), 5.15-5.08 (m, 1H), 4.74 (d, J=19.1 Hz, 2H), 4.56 (d, J=12.3 Hz, 1H), 3.59 (d, J=13.2 Hz, 2H), 3.37-3.30 (m, 2H), 3.22-3.01 (m, 5H), 2.94-2.82 (m, 2H), 2.65-2.56 (m, 2H), 2.23 (m, 3H), 2.10-1.88-1.79 (m, 6H), 1.57 (s, 6H).
LC-MS(ESI): [M+H]+=857.69
Refer to the first step of Example 98 for preparation.
1H NMR (600 MHz, DMSO-d6) δ 11.12 (s, 1H), 8.42 (d, J=8.3 Hz, 1H), 8.29 (t, J=7.9 Hz, 1H), 8.09 (dd, J=8.3, 1.4 Hz, 1H), 7.77-7.66 (m, 1H), 7.64 (s, 1H), 7.49 (dd, J=10.5, 1.6 Hz, 1H), 7.37 (dd, J=13.9, 5.7 Hz, 2H), 5.15-5.07 (m, 1H), 4.78-4.65 (m, 2H), 4.29 (dd, J=16.0, 8.6 Hz, 2H), 4.01-3.90 (m, 2H), 3.34 (s, 2H), 3.25-3.03 (m, 3H), 2.94-2.84 (m, 1H), 2.64-2.52 (m, 2H), 2.24 (s, 1H), 2.14 (t, J=13.2 Hz, 2H), 2.03 (dd, J=17.3, 10.0 Hz, 3H), 1.93-1.84 (m, 1H), 1.55 (s, 6H).
LC-MS(ESI): [M+H]+=872.09
Refer to the first step of Example 98 for preparation.
1H NMR (600 MHz, DMSO-d6) δ 10.98 (s, 1H), 8.42 (d, J=8.3 Hz, 1H), 8.30 (d, J=1.7 Hz, 1H), 8.09 (dd, J=8.2, 1.7 Hz, 1H), 7.61 (t, J=7.8 Hz, 1H), 7.46 (dd, J=10.0, 1.5 Hz, 1H), 7.36 (dd, J=8.1, 1.5 Hz, 1H), 7.23 (d, J=7.4 Hz, 1H), 7.12 (d, J=25.8 Hz, 1H), 5.11-5.04 (m, 1H), 4.55 (d, J=12.3 Hz, 1H), 4.38-4.26 (m, 2H), 4.18 (m, 4H), 3.60-3.40 (m, 4H), 3.20 (d, J=60.7 Hz, 5H), 2.91 (dt, J=24.4, 8.1 Hz, 2H), 2.60 (d, J=16.8 Hz, 1H), 2.42-2.30 (m, 1H), 2.16 (s, 2H), 2.02-1.89 (m, 5H), 1.76 (s, 1H), 1.57 (s, 5H).
LC-MS(ESI): [M+H]+=902.17
Refer to the first step of Example 98 for preparation.
1H NMR (600 MHz, MeOD) δ 8.19 (d, J=7.6 Hz, 2H), 8.02-7.98 (m, 1H), 7.79 (t, J=7.8 Hz, 1H), 7.67 (s, 1H), 7.45 (ddd, J=9.8, 9.4, 1.5 Hz, 2H), 7.29 (s, 1H), 5.11 (dd, J=12.8, 5.4 Hz, 1H), 4.61-4.54 (m, 2H), 4.51 (dd, J=10.7, 4.9 Hz, 1H), 4.44 (dd, J=11.8, 4.8 Hz, 1H), 4.33-4.11 (m, 4H), 3.42 (m, 1H), 3.01 (s, 3H), 2.92-2.66 (m, 4H), 2.14-2.07 (m, 1H), 1.82 (t, J=48.7 Hz, 4H), 1.62 (s, 6H).
LC-MS(ESI): [M+H]+=872.36
Refer to the first step of Example 98 for preparation.
1H NMR (600 MHz, DMSO-d6) δ 11.11 (d, J=4.4 Hz, 1H), 8.42 (d, J=8.3 Hz, 1H), 8.30 (s, 1H), 8.09 (d, J=8.4 Hz, 1H), 7.64 (ddd, J=31.3, 21.8, 7.8 Hz, 2H), 7.49-7.40 (m, 1H), 7.38-7.25 (m, 2H), 5.14-5.06 (m, 1H), 4.32 (d, J=11.2 Hz, 1H), 4.03 (d, J=14.1 Hz, 1H), 3.43-3.32 (m, 7H), 3.28-3.10 (m, 5H), 3.07 (d, J=12.8 Hz, 1H), 2.89 (dd, J=20.1, 10.2 Hz, 1H), 2.79 (d, J=13.4 Hz, 1H), 2.58 (m, 2H), 2.08-1.98 (m, 1H), 1.89 (s, 3H), 1.79-1.61 (m, 4H), 1.53 (m, 7H), 1.47 (d, J=27.9 Hz, 1H), 1.40 (s, 1H), 1.32 (s, 1H), 1.26-1.12 (m, 2H).
LC-MS(ESI): [M+H]+=968.45
Refer to the first step of Example 98 for preparation.
1H NMR (600 MHz, DMSO-d6) δ 11.11 (s, 1H), 8.42 (d, J=8.2 Hz, 1H), 8.29 (s, 1H), 8.09 (d, J=8.2 Hz, 1H), 7.75-7.61 (m, 2H), 7.46 (d, J=9.9 Hz, 1H), 7.36 (d, J=8.0 Hz, 1H), 7.30 (s, 1H), 5.10 (dd, J=12.9, 5.4 Hz, 1H), 4.30 (s, 1H), 4.10 (s, 1H), 4.05 (s, 1H), 3.32 (s, 11H), 3.17 (s, 2H), 3.07 (s, 1H), 2.95-2.84 (m, 1H), 2.81 (s, 1H), 2.58 (m, 1H), 2.08-1.99 (m, 1H), 1.85 (dd, J=34.1, 28.7 Hz, 3H), 1.71 (s, 1H), 1.64-1.42 (m, 10H).
LC-MS(ESI): [M+H]+=928.45
Refer to the first step of Example 98 for preparation.
1H NMR (600 MHz, DMSO-d6) δ 11.10 (s, 1H), 8.42 (d, J=8.2 Hz, 1H), 8.29 (s, 1H), 8.09 (d, J=8.2 Hz, 1H), 7.72 (dd, J=15.9, 5.9 Hz, 1H), 7j61 (d, J=7.4 Hz, 1H), 7.46 (dd, J=10.4, 3.4 Hz, 1H), 7.35 (d, J=6.8 Hz, 1H), 7.30 (d, J=7.2 Hz, 1H), 5.10 (dd, J=12.8, 5.2 Hz, 1H), 4.30 (d, J=5.9 Hz, 1H), 4.18 (s, 1H), 4.09 (s, 1H), 4.05 (d, J=5.0 Hz, 1H), 3.68 (m, 2H), 3.32-3.22 (m, 3H), 3.05 (d, J=6.5 Hz, 1H), 3.02-2.83 (m, 3H), 2.80 (d, J=5.8 Hz, 1H), 2.60 (d, J=16.0 Hz, 2H), 2.48-2.35 (m, 2H), 2.05-2.00 (m, 1H), 1.78-1.72 (m, 1H), 1.62 (m, 8H).
LC-MS(ESI): [M+H]+=912.45
Refer to the first step of Example 98 for preparation.
1H NMR (600 MHz, DMSO-d6) δ 10.98 (s, 1H), 8.42 (d, J=8.2 Hz, 1H), 8.30 (s, 1H), 8.09 (d, J=8.3 Hz, 1H), 7.61 (d, J=7.7 Hz, 1H), 7.46 (d, J=9.9 Hz, 1H), 7.35 (d, J=8.3 Hz, 2H), 7.24 (s, 1H), 5.07 (dd, J=13.2, 5.2 Hz, 1H), 5.07 (dd, J=13.2, 5.2 Hz, 1H), 4.55 (d, J=12.6 Hz, 1H), 4.26 m, 2H), 3.46 (m, 3H), 3.30-3.07 (m, 6H), 2.89 (dd, J=19.3, 6.0 Hz, 4H), 2.64-2.55 (m, 1H), 2.42-2.34 (m, 1H), 2.19 (s, 1H), 2.02-1.81 (m, 8H), 1.76 (s, 1H), 1.57 (s, 6H).
LC-MS(ESI): [M+H]+=900.45
Refer to the first step of Example 98 for preparation.
1H NMR (600 MHz, DMSO-d6) δ 11.11 (d, J=6.2 Hz, 1H), 8.42 (d, J=8.2 Hz, 1H), 8.29 (s, 1H), 8.09 (d, J=8.2 Hz, 1H), 7.80-7.67 (m, 2H), 7.49-7.42 (m, 1H), 7.40 (s, 1H), 7.38-7.32 (m, 1H), 5.14-5.02 (m, 1H), 3.85-3.72 (m, 5H), 3.44-3.15 (m, 7H), 3.00-2.79 (m, 4H), 2.60 (d, J=18.0 Hz, 1H), 2.48-2.42 (m, 1H), 2.12-1.80 (m, 10H), 1.63-1.40 (m, 9H).
LC-MS(ESI): [M+H]+=940.45
The first step: 4-(3-(4-(4-(7′-(2,6-dioxopiperidin-3-yl)-6′,8′-dioxo-4′,6′, 7′,8′-tetrahydro-3′H-spiro [azetidine-3,2′-pyrano[2,3-f]isoindol]-1-yl)methyl) piperidin-1-carbonyl)-3-fluorophenyl)-4,4-dimethyl-5-oxo-2-thioimidazolidin-1-yl)-2-(trifluoromethyl)benzonitrile (Compound PROTAC 101)
Refer to the first step of Example 98 for preparation.
1H NMR (600 MHz, DMSO-d6) δ 11.12 (s, 1H), 8.42 (d, J=8.2 Hz, 1H), 8.29 (d, J=1.3 Hz, 1H), 8.09 (dd, J=8.2, 1.4 Hz, 1H), 7.79 (s, 1H), 7.60 (t, J=7.7 Hz, 1H), 7.46 (dd, J=9.9, 1.3 Hz, 1H), 7.35 (dd, J=8.1, 1.3 Hz, 1H), 7.30 (d, J=12.7 Hz, 1H), 5.11 (dd, J=12.9, 5.4 Hz, 1H), 4.56-4.29 (m, 5H), 3.33 (s, 2H), 3.25 (s, 2H), 3.12 (s, 1H), 3.00 (d, J=5.9 Hz, 1H), 2.96-2.79 (m, 3H), 2.64-2.54 (m, 2H), 2.25 (d, J=5.8 Hz, 2H), 2.09-1.88 (m, 3H), 1.84 (d, J=11.4 Hz, 1H), 1.69 (s, 1H), 1.56 (s, 6H).
LC-MS (ESI): [M+H]+=886.11
Refer to the first step of Example 98 for preparation.
1H NMR (600 MHz, DMSO-d6) δ 11.12 (d, J=4.6 Hz, 1H), 8.42 (d, J=8.2 Hz, 1H), 8.29 (d, J=7.1 Hz, 1H), 8.09 (d, J=8.3 Hz, 1H), 7.68-7.52 (m, 2H), 7.48-7.42 (m, 1H), 7.40-7.30 (m, 2H), 5.16-5.06 (m, 1H), 4.81-4.68 (m, 2H), 3.68 (s, 2H), 3.62-3.53 (m, 1H), 3.25 (dd, J=12.5, 5.2 Hz, 4H), 3.17-3.02 (m, 2H), 2.96-2.83 (m, 1H), 2.64-2.56 (m, 2H), 2.26-2.14 (m, 2H), 2.09-1.98 (m, 3H), 1.90 (d, J=9.5 Hz, 4H), 1.69 (d, J=11.3 Hz, 1H), 1.63-1.48 (m, 8H), 1.42-1.27 (m, 5H).
LC-MS(ESI): [M+H]+=954.11
Refer to the first step of Example 98 for preparation.
1H NMR (600 MHz, DMSO-d6) δ11.12 (d, J=5.1 Hz, 1H), 8.42 (d, J=8.3 Hz, 1H), 8.30 (s, 1H), 8.09 (d, J=8.3 Hz, 1H), 7.74 (dt, J=19.1, 7.9 Hz, 1H), 7.62 (d, J=11.4 Hz, 1H), 7.50-7.42 (m, 1H), 7.36 (dd, J=12.1, 7.9 Hz, 2H), 5.20-5.02 (m, 1H), 4.83-4.67 (m, 2H), 3.81 (t, J=16.5 Hz, 2H), 3.79-3.70 (m, 2H), 3.54 (d, J=11.1 Hz, 2H), 3.26 (s, 2H), 3.10 (m, 2H), 2.97-2.82 (m, 1H), 2.60 (dd, J=24.1, 8.5 Hz, 1H), 2.26-1.81 (m, 10H), 1.66-1.41 (m, 9H).
LC-MS (ESI): [M+H]+=926.15
Refer to the first step of Example 116 for preparation.
1H NMR (600 MHz, DMSO-d6) δ 11.15-11.09 (m, 1H), 8.42 (d, J=8.2 Hz, 1H), 8.30 (s, 1H), 8.09 (d, J=8.4 Hz, 1H), 7.66 (t, J=11.8 Hz, 1H), 7.62 (td, J=8.0, 3.4 Hz, 1H), 7.46 (t, J=9.9 Hz, 1H), 7.36 (dd, J=12.8, 10.0 Hz, 2H), 5.16-5.07 (m, 1H), 4.73 (dd, J=15.6, 9.0 Hz, 2H), 3.79-3.56 (m, 3H), 3.32-3.14 (m, 4H), 2.90 (dd, J=16.5, 14.0 Hz, 3H), 2.63 (t, J=24.7 Hz, 2H), 2.29 (m, 3H), 2.17-1.98 (m, 6H), 1.66 (s, 2H), 1.57 (s, 8H).
LC-MS(ESI): [M+H]+=926.11
Refer to the first step of Example 98 for preparation.
1H NMR (400 MHz, DMSO-d6) δ 11.11 (s, 1H), 8.42 (d, J=8.3 Hz, 1H), 8.30 (s, 1H), 8.09 (d, J=8.3 Hz, 1H), 7.71 (t, J=5.5 Hz, 1H), 7.64-7.55 (m, 1H), 7.45 (dd, J=9.9, 2.5 Hz, 1H), 7.37-7.22 (m, 2H), 5.10 (dd, J=12.7, 5.3 Hz, 1H), 4.31 (s, 1H), 4.06 (s, 1H), 3.79 (m, 2H), 3.37-3.13 (m, 5H), 3.09-2.77 (m, 6H), 2.68-2.55 (m, 2H), 2.25 (d, J=9.4 Hz, 2H), 2.13-1.95 (m, 3H), 1.77 (d, J=11.7 Hz, 1H), 1.66-1.56 (s, 12H).
LC-MS(ESI): [M+H]+=940.35
Refer to the first step of Example 98 for preparation.
1H NMR (600 MHz, DMSO-d6) δ 10.97 (s, 1H), 8.42 (d, J=8.2 Hz, 1H), 8.29 (s, 1H), 8.09 (d, J=8.1 Hz, 1H), 7.60 (t, J=7.5 Hz, 1H), 7.53 (s, 1H), 7.45 (d, J=10.0 Hz, 1H), 7.35 (dd, J=8.0, 1.2 Hz, 1H), 7.07 (d, J=19.3 Hz, 1H), 5.07 (dd, J=13.2, 4.8 Hz, 1H), 4.53-4.42 (m, 2H), 4.37-4.19 (m, 6H), 3.11 (s, 2H), 2.95-2.77 (m, 4H), 2.60 (d, J=17.8 Hz, 1H), 2.40-2.28 (m, 1H), 2.27-2.13 (m, 2H), 2.03-1.88 (m, 2H), 1.83 (s, 1H), 1.69 (s, 1H), 1.56 (s, 6H), 1.32-1.11 (m, 3H).
LC-MS(ESI): [M+H]+=872.36
Refer to the first step of Example 98 for preparation.
1H NMR (600 MHz, DMSO-d6) δ 11.12 (d, J=5.7 Hz, 1H), 8.42 (d, J=8.2 Hz, 1H), 8.30 (s, 1H), 8.09 (d, J=8.2 Hz, 1H), 7.74-7.59 (m, 2H), 7.48 (dd, J=12.6, 7.5 Hz, 1H), 7.36 (dd, J=16.4, 9.9 Hz, 2H), 5.17-5.03 (m, 1H), 4.80-4.67 (m, 2H), 3.91 (m, 1H), 3.39-3.31 (m, 4H), 3.23-3.04 (m, 3H), 2.96-2.83 (m, 2H), 2.83-2.65 (m, 1H), 2.61 (d, J=16.4 Hz, 1H), 2.57-2.52 (m, 1H), 2.35-2.11 (m, 3H), 2.10-1.97 (m, 3H), 1.97-1.68 (m, 2H), 1.56 (s, 6H).
LC-MS(ESI): [M+H]+=886.36
The binding activity of compounds disclosed in the invention and positive control samples lenalidomide and pomalidomide with E3 ubiquitin ligase CRBN was determined by the way of compounds disclosed by the present invention, lenalidomide and pomalidomide with thalidomide marked with XL665 competitively binding to CRBN protein, which prevented the occurrence of fluorescence resonance energy transfer (FRET). From this, the binding force of the compounds to the CRBN protein was determined.
CRBN binding activity test kit (CEREBLON BINDING KITS (PE, 64BDCRBNPEG).
The detection is based on homogeneous time-resolved fluorescence (HTRF) technology. When the donor and the acceptor are close, the donor can transfer energy to the acceptor, exciting it to emit light at 665 nm.
The donor used in this kit was Europium-labeled GST antibody (GST Eu Cryptate Antibody), and the acceptor was thalidomide labeled with XL665 (Thalidomide-Red reagent). The donor binded to the CRBN protein containing the GST tag. The disclosed compounds in the present, along with lenalidomide and pomalidomide, competitively binded to the CRBN protein with thalidomide. The binding activity of the compounds was determined based on the fluorescence value emitted at 665 nm. The stronger the binding affinity of the compounds, the weaker the signal.
The donor and acceptor were diluted 50 times using the binding buffer from the kit (PROTAC binding buffer 1). The CRBN protein was diluted 45 times. The disclosed compound solution at 8 mM was diluted 10 times with the diluent from the kit (1× diluent), followed by a five-fold gradient dilution, performed 7 times sequentially.
5 μl of compound, 5 μl of diluted CRBN protein and 10 μl of donor-receptor mixture in equal volume were sequentially added to wells of a white 384-well plate (PE, Part number: 6008280). Incubate at room temperature for 3 hours.
The ratio of the emission signals from the acceptor and donor in each individual well is calculated.
Ratio=665 nm signal value/620 nm signal value*104
The coefficient of variation (CV)=standard deviation (SD)/mean Ratio*100
Using GraphPad Prism, IC50 values were calculated based on compound concentration, Ratio and coefficient of variation.
Table 1 shows the IC50 values of the disclosed compounds, lenalidomide, and pomalidomide.
Conclusion: The compounds disclosed in the present invention can well bind to CRBN protein, and the binding ability of some compounds to CRBN is equivalent to or even better than that of positive control samples lenalidomide and pomalidomide.
Inhibitory Effect of the Compounds Disclosed in the Present Invention on Cell Proliferation in HeLa Cells (Sourced from ATCC)
The complete medium required for HeLa cells is DMEM high glucose (Gibco, Product number: 11995-065) basal medium containing 10% fetal bovine serum. HeLa cells were seeded in a 96-well plate at a density of 4,000 cells/well, and the next day, different concentrations of the compound of the present invention and the positive control sample Ceritinib (source: Bide Pharmaceuticals) were prepared to treat the cells. The cells were incubated at 37° C., 5% CO2 cell culture incubator for 3 days. Cell viability was detected using the CCK-8 kit (Beyotime, Product number: C0040) according to the manufacturer's instructions. According to the inhibition rate, GraphPad Prism software was used to calculate the IC50 value of each compound for 3 days, and the results are shown in Table 2.
Conclusion: the compounds of the present invention can effectively inhibit the proliferation of HeLa cells, and the inhibitory effect is better than that of the positive control.
Inhibitory Effect of the Compounds Disclosed in the Present Invention on Cell Proliferation in A549 Cells (Sourced from ATCC)
The complete medium required for A549 cells is F-12K (ATCC, Product number: 30-2004) basal medium containing 10% fetal bovine serum. A549 cells were seeded in a 96-well plate at a density of 2,800 cells/well, and the next day, different concentrations of the compound of the present invention and a positive control sample Ceritinib (source: Bide Pharmaceuticals) were prepared to treat the cells. The cells were incubated at 37° C., 5% CO2 cell incubator for 4 days. Cell viability was detected using the CCK-8 kit (Beyotime, Product number: C0040) according to the manufacturer's instructions. According to the inhibition rate, the GraphPad Prism software was used to calculate the IC50 value of each compound for 4 days, and the results are shown in Table 3.
Conclusion: The compounds of the present invention can effectively inhibit the proliferation of A549 cells, and the inhibitory effect of some compounds is equivalent to that of the positive control.
Inhibition of Cell Proliferation by Compounds Disclosed in the Present Invention in 22RV1 Cells (Sourced from ATCC)
The complete medium required for 22RV1 cells is RPM11640 (ATCC, Product number: 30-2001) basal medium containing 10% fetal bovine serum. 22RV1 cells were seeded in a 96-well plate at a density of 4,000 cells/well, and the next day, different concentrations of the compound of the present invention and the positive control sample JQ1 (source: Bide Pharmaceuticals) were prepared to treat the cells, and the cells were cultured at 37° C. and 5% CO2 for 7 days in the incubator. Cell viability was detected using the CCK-8 kit (Beyotime, Product number: C0040) according to the manufacturer's instructions. According to the inhibition rate, the GraphPad Prism software was used to calculate the IC50 value of each compound for 7 days, and the results are shown in Table 4.
Conclusion: The compounds in the present invention can effectively inhibit the proliferation of 22RV1 cells, and the inhibitory effect of some compounds is better than that of the positive control.
Inhibitory Effect of the Compounds Disclosed in the Present Invention on Cell Proliferation in NCI-H358 Cells (Sourced from ATCC)
The complete medium required for NCI-H358 cells is RPMI1640 (ATCC, Product number: 30-2001) basal medium containing 10% fetal bovine serum. NCI-H358 cells were seeded in a 96-well plate at a density of 2,800 cells/well, and the next day, different concentrations of the compounds of the present invention were prepared to treat the cells, and the cells were treated for 4 days in a 37° C., 5% CO2 cell culture incubator. Cell viability was detected using a CCK-8 kit (Beyotime, Product number: C0040) according to the manufacturer's instructions. According to the inhibition rate, the GraphPad Prism software was used to calculate the IC50 value of each compound for 4 days, and the results are shown in Table 5.
Conclusion: the compounds of the present invention can effectively inhibit the proliferation of NCI-H358 cells.
Inhibition of Cell Proliferation by the Compounds Disclosed in the Present Invention in MV-4-11 Cells (Sourced from ATCC)
The complete medium required for MV-4-11 cells is IMDM (ATCC, Product number: 30-2005) basal medium containing 10% fetal bovine serum. MV-4-11 cells were seeded in a 96-well plate at a density of 7,500 cells/well, and different concentrations of the compound of the present invention and positive control samples Alisertib (source: Bide Pharmaceuticals), Palbociclib (source: Bide Pharmaceuticals), Ibrutinib (source Bide Pharmaceuticals) treated cells. The cells were incubated in a 37° C., 5% CO2 cell incubator for 4 days. Cell viability was detected using the CCK-8 kit (Beyotime, Product number: C0040) according to the manufacturer's instructions. According to the inhibition rate, GraphPad Prism software was used to calculate the IC50 value of each compound for 4 days, and the results are shown in Table 6-8.
Conclusion: The compounds of the present invention can effectively inhibit the proliferation of MV-4-11 cells, wherein the inhibitory effect of some compounds is equivalent to or even better than that of the positive control.
The complete medium required for MOLM-13 cells is 1640 (ATCC, Product number: 30-2001) basal medium containing 20% fetal bovine serum. MOLM-13 cells were seeded in a 96-well plate at a density of 7,500 cells/well, and different concentrations of the compounds of the present invention were prepared to treat the cells at the same time. The cells were incubated in a 37° C., 5% CO2 cell culture incubator for 4 days. Cell viability was detected using the CCK-8 kit (Beyotime, Product number: C0040) according to the manufacturer's instructions. According to the inhibition rate, GraphPad Prism software was used to calculate the IC50 value of each compound for 4 days, and the results are shown in Table 9-10.
Conclusion: the compounds of the present invention can effectively inhibit the proliferation of MOLM-13 cells.
Inhibitory Effect of Compounds Disclosed by the Invention on Cell Proliferation in K562 Cells (Sourced from ATCC)
The complete medium required for K562 cells is IMDM (ATCC, Product number: 30-2005) basal medium containing 10% fetal bovine serum. K562 cells were seeded in a 96-well plate at a density of 7,500 cells/well, and different concentrations of the compound of the present invention and positive control samples Alisertib (source: Bide Pharmaceuticals) and dBET6 (source: Bide Pharmaceuticals) were prepared to treat the cells. The cells were incubated at a 37° C., 5% CO2 cell culture incubator for 4 days.
Cell viability was detected using the CCK-8 kit (Beyotime, Product number: C0040) according to the manufacturer's instructions. According to the inhibition rate, GraphPad Prism software was used to calculate the IC50 value of each compound for 4 days, and the results are shown in Table 11-12.
Conclusion: The compounds of the present invention can effectively inhibit the proliferation of K562 cells, wherein the inhibitory effect of some compounds is equivalent to or even better than that of the positive control.
Inhibitory Effect of Compound Disclosed by the Invention on Cell Proliferation in JeKo-1 Cells (Sourced from ATCC)
The complete medium required for JeKo-1 cells is RPMI1640 (ATCC, Product number: 30-2001) basal medium containing 20% fetal bovine serum. The JeKo-1 cells were seeded in a 96-well plate at a density of 10,000 cells/well, and different concentrations of the compounds of the present invention were prepared to treat the cells at the same time. The cells were incubated at a 37° C., 5% CO2 cell culture incubator for 4 days. Cell viability was detected using the CCK-8 kit (Beyotime, Product number: C0040) according to the manufacturer's instructions. According to the inhibition rate, the GraphPad Prism software was used to calculate the IC50 value of each compound for 4 days, and the results are shown in Table 13.
Conclusion: the compounds of the present invention can effectively inhibit the proliferation of Jeko-1 cells.
Inhibitory Effect of Compounds Disclosed in the Present Invention on Cell Proliferation in Z138 Cells (Sourced from ATCC)
The complete medium required for Z138 cells is IMDM (ATCC, Product number: 30-2005) basal medium containing 10% horse serum. Z138 cells were seeded in a 96-well plate at a density of 10,000 cells/well, and different concentrations of the compound of the present invention and a positive control sample Ibrutinib (source: Bide Pharmaceuticals) were prepared to treat the cells. The cells were incubated in a 37° C., 5% CO2 cell incubator for 4 days. Cell viability was detected using the CCK-8 kit (Beyotime, Product number: C0040) according to the manufacturer's instructions. According to the inhibition rate, the GraphPad Prism software was used to calculate the IC50 value of each compound for 4 days, and the results are shown in Table 14.
Conclusion: The compounds of the present invention can effectively inhibit the proliferation of Z138 cells, wherein the inhibitory effect of some compounds is equivalent to or even better than that of the positive control.
Inhibitory Effect of Compounds Disclosed by the Invention on Cell Proliferation in VCaP Cells (Sourced from ATCC)
The complete medium required for VCaP cells is DMEM high glucose (Gibco, Product number: 11995-065) basal medium containing 10% fetal bovine serum. The VCaP cells were seeded in a 96-well plate at a density of 15,000 cells/well, and the next day, different concentrations of the compound of the present invention and the positive control sample ARV-110 (source: Anaiji) were prepared to treat the cells, and the cells were incubated at 37° C. and 5% CO2 for 7 days in the cell culture incubator. Cell viability was detected using the CCK-8 kit (Beyotime, Product number: C0040) according to the manufacturer's instructions. According to the inhibition rate, the GraphPad Prism software was used to calculate the IC50 value of each compound for 7 days, and the results are shown in Table 15.
Conclusion: The compounds of the present invention can effectively inhibit the proliferation of VCaP cells, and the inhibitory effect of some compounds is equivalent to or even better than that of the positive control.
Inhibitory Effect of Compounds Disclosed in the Present Invention on Cell Proliferation in LNCaP FGC Cells (Sourced from ATCC)
The complete medium required for LNCaP FGC cells is RPMI 1640 (ATCC, Product number: 30-2001) basal medium containing 10% fetal calf serum. Seed LNCAP FGC cells in a 96-well plate at a density of 5,000 cells/well, prepare different concentrations of the compound of the present invention and the positive control sample ARV-110 (source: Energy chemical) to treat the cells on the next day, and store them at 37° C., 5% CO2 for 4 days in a cell incubator. Cell viability was detected using the CCK-8 kit (Beyotime, Product number: C0040) according to the manufacturer's instructions. According to the inhibition rate, the GraphPad Prism software was used to calculate the IC50 value of each compound for 4 days, and the results are shown in Table 16.
Conclusion: The compounds of the present invention can effectively inhibit the proliferation of LNCaP cells, wherein the inhibitory effect of some compounds is equivalent to or even better than that of the positive control.
Inhibition of Cell Proliferation by Compounds Disclosed in the Present Invention in T-47D Cells (Sourced from ATCC)
The complete medium required for T-47D cells is 1640 (ATCC, Product number: A10491-01) basal medium containing 10% fetal bovine serum. T-47D cells were seeded in a 96-well plate at a density of 3,000/well, and different concentrations of the compound of the present invention and the positive control sample ARV-471 (source: Innochem) were prepared to treat the cells. The cells were incubated at 37° C., 5% for 6 days in a CO2 cell incubator. Cell viability was detected using the CCK-8 kit (Beyotime, Product number: C0040) according to the manufacturer's instructions. According to the inhibition rate, the GraphPadPrism software was used to calculate the IC50 value of each compound for 6 days, and the results are shown in Table 17.
Conclusion: The compounds of the present invention can effectively inhibit the proliferation of T-47D cells, wherein the inhibitory effect of some compounds is equivalent to or even better than that of the positive control.
Inhibitory Effect of Compounds Disclosed by the Invention on Cell Proliferation in MCF-7 Cells (Sourced from ATCC)
The complete medium required for MCF-7 cells is DMEM (gibco, Product number: 11995-065) basal medium containing 10% fetal bovine serum. MCF-7 cells were seeded in a 96-well plate at a density of 800/well, and different concentrations of the compound of the present invention and the positive control sample ARV-471 (source: Enochem) were prepared to treat the cells. The cells were incubated at 37° C., 5% 9 days in a CO2 cell incubator. Cell viability was detected using the CCK-8 kit (Beyotime, Product number: C0040) according to the manufacturer's instructions. According to the inhibition rate, GraphPadPrism software was used to calculate the IC50 value of each compound for 9 days, and the results are shown in Table 18.
Conclusion: The compounds of the present invention can effectively inhibit the proliferation of MCF-7 cells, wherein the inhibitory effect of some compounds is equivalent to or even better than that of the positive control.
| Number | Date | Country | Kind |
|---|---|---|---|
| 202210109488.6 | Jan 2022 | CN | national |
| 202310005966.3 | Jan 2023 | CN | national |
| Filing Document | Filing Date | Country | Kind |
|---|---|---|---|
| PCT/CN2023/073745 | 1/29/2023 | WO |
