Spirocyclic compounds and uses thereof

Abstract

The present application provides a spiro compound having a binding activity with E3 ligase protein CRBN shown in formula I, and a use thereof in preparing a medicine for treating a cell abnormal proliferation disease.

Description

Technical Field

[0001] This invention belongs to the field of medicine, specifically relating to a novel ligand compound that binds to the cereblon E3 ubiquitin ligase protein. Background Technology

[0002] Protein degradation is a highly regulated process essential for maintaining cellular homeostasis. The ubiquitin-proteasome pathway (UPP) enables the selective identification and removal of damaged, misfolded, or excess proteins. UPP removes defective proteins and is characterized by ATP dependence, high efficiency, and high selectivity. Its catalytic component is the ubiquitin-derived E3 ligase, but it requires the prior recruitment of the protein to be degraded. PROTACs technology is designed based on the UPP principle, linking the target protein ligand and the E3 ligase ligand with appropriate chemical bonds. This allows for the recognition of the target protein and enhances the binding affinity of the E3 ligase to the target protein, thereby targeting ubiquitination and forcing the degradation of the target protein. It also features high catalytic activity, high efficiency, and high selectivity.

[0003] Multiple ubiquitin molecules are covalently linked to terminal lysine residues via E3 ubiquitin ligase to label proteins for proteasomal degradation. The protein is then digested into small peptides and ultimately into its constituent amino acids, which serve as building blocks for new proteins. Defective proteasomal degradation is associated with a variety of clinical conditions, including Alzheimer's disease, Parkinson's disease, Huntington's disease, muscular dystrophy, cardiovascular disease, and cancer.

[0004] Cereblon is a thalidomide-binding protein and part of the E3 ubiquitin ligase protein complex. It acts as a substrate receptor, selectively targeting ubiquitinated proteins. Encoded by the human CRBN gene, cereblon, along with DNA damage binding protein 1 (DDBl), Cullin-4A (CUL4A), and the Cullin-1 regulator (ROCI), forms the E3 ubiquitin ligase complex. This complex can ubiquitinate a range of proteins, but the specific mechanism is not yet fully understood. Cereblon is currently a commonly used E3 ligase in PROTACs technology.

[0005] This invention discloses a novel class of spirocyclic compounds that can serve as effective CRBN ligands. Furthermore, they can be used to synthesize corresponding bifunctional PROTACs that target protein degradation chimeras, which can be used to treat various medical conditions, especially abnormal cell proliferation. Summary of the Invention

[0006] This invention provides a compound of Formula I, or a stereoisomer thereof, or a deuterated compound thereof, or a pharmaceutically acceptable salt thereof:

[0007]

[0008] in,

[0009] Ring A is selected from 3-12 membered cycloalkyl, 4-12 membered heterocycloalkyl, 6-10 membered aromatic ring, and 5-10 membered heteroaromatic ring; wherein, the cycloalkyl, heterocycloalkyl, aromatic ring, and heteroaromatic ring may be further optionally surrounded by one, two, three, or four independent R. A1 replace;

[0010] Each R A1 Each group is independently selected from hydrogen, halogen, cyano, nitro, =O, =S, =CR A2 R A3 -C 1～6 Alkyl, -C 2～6 alkenyl, -C 2～6 Alkyne- or halogen-substituted -C 1～6 Alkyl, halogen-substituted -C 2～6 Alkenyl, halogen-substituted -C 2～6 alkynyl group, -C 0～4 Alkylene-OR A2 -C 0～4 Alkylene-OC(O)R A2 -C 0～4 Alkylene-SR A2 -C 0～4 Alkylene-S(O)2R A2 -C 0～4 Alkylene-S(O)R A2 -C 0～4 Alkylene-S(O)2NR A2 R A3 -C 0～4 Alkylene-S(O)NR A2 R A3 -C 0～4 Alkylene-C(O)R A2 -C 0～4 Alkylene-C(O)OR A2 -C 0～4 Alkylene-C(O)NR A2 R A3 -C 0～4 Alkylene-NR A2 R A3 -C 0～4 Alkylene-NR A2 C(O)R A3 -C 0～4 Alkylene-NRA2 S(O)2R A3 -C 0～4 Alkylene-NR A2 S(O)R A3 -C 0～4 Alkylene (3- to 10-membered cycloalkyl), -C 0～4 Alkylene (4- to 10-membered heterocyclic alkyl), -C 0～4 alkylene-(6- to 10-membered aromatic ring), -C 0～4 Alkylene (5- to 10-membered heteroaryl ring); wherein the alkylene, cycloalkyl, heterocycloalkyl, aromatic ring, and heteroaryl ring may be optionally surrounded by one, two, three, or four independent R... A4 replace;

[0011] Each R A4 Each group is independently selected from hydrogen, halogen, cyano, nitro, =O, =S, =CR A2 R A3 -C 1～6 Alkyl, -C 2～6 alkenyl, -C 2～6 Alkyne- or halogen-substituted -C 1～6 Alkyl, halogen-substituted -C 2～6 Alkenyl, halogen-substituted -C 2～6 alkynyl group, -C 0～4 Alkylene-OR A2 -C 0～4 Alkylene-OC(O)R A2 -C 0～4 Alkylene-SR A2 -C 0～4 Alkylene-S(O)2R A2 -C 0～4 Alkylene-S(O)R A2 -C 0～4 Alkylene-S(O)2NR A2 R A3 -C 0～4 Alkylene-S(O)NR A2 R A3 -C 0～4 Alkylene-C(O)R A2 -C 0～4 Alkylene-C(O)OR A2 -C 0～4 Alkylene-C(O)NR A2 R A3 -C 0～4 Alkylene-NR A2 R A3 -C 0～4 Alkylene-NR A2 C(O)RA3 -C 0～4 Alkylene-NR A2 S(O)2R A3 -C 0～4 Alkylene-NR A2 S(O)R A3 ;

[0012] R A2 R A3 Selected independently from hydrogen and -C 1-6 alkyl, -C 2～6 alkenyl, -C 2～6 Alkyne- or halogen-substituted -C 1～6 Alkyl, halogen-substituted -C 2～6 Alkenyl, halogen-substituted -C 2～6 alkynyl group;

[0013] R 2 Independently selected from hydrogen, halogen, cyano, nitro, =O, =S, =CR 21 R 22 -C 1～6 Alkyl, -C 2～6 alkenyl, -C 2～6 Alkyne- or halogen-substituted -C 1～6 Alkyl, halogen-substituted -C 2～6 Alkenyl, halogen-substituted -C 2～6 alkynyl group, -C 0～4 Alkylene-OR 21 -C 0～4 Alkylene-OC(O)R 21 -C 0～4 Alkylene-SR 21 -C 0～4 Alkylene-S(O)2R 21 -C 0～4 Alkylene-S(O)R 21 -C 0～4 Alkylene-S(O)2NR 21 R 22 -C 0～4 Alkylene-S(O)NR 21 R 22 -C 0～4 Alkylene-C(O)R 21 -C 0～4 Alkylene-C(O)OR 21 -C 0～4 Alkylene-C(O)NR 21 R 22 -C 0～4 Alkylene-NR 21 R 22 -C0～4 Alkylene-NR 21 C(O)R 22 -C 0～4 Alkylene-NR 21 S(O)2R 22 -C 0～4 Alkylene-NR 21 S(O)R 22 -C 0～4 Alkylene (3- to 10-membered cycloalkyl), -C 0～4 Alkylene (4- to 10-membered heterocyclic alkyl), -C 0～4 alkylene-(6- to 10-membered aromatic ring), -C 0～4 Alkylene (5- to 10-membered heteroaryl ring); wherein the alkylene, cycloalkyl, heterocycloalkyl, aromatic ring, and heteroaryl ring may be optionally surrounded by one, two, three, or four independent R... 23 replace;

[0014] Each R 23 Each group is independently selected from hydrogen, halogen, cyano, nitro, =O, =S, =CR 21 R 22 -C 1～6 Alkyl, -C 2～6 alkenyl, -C 2～6 Alkyne- or halogen-substituted -C 1～6 Alkyl, halogen-substituted -C 2～6 Alkenyl, halogen-substituted -C 2～6 alkynyl group, -C 0～4 Alkylene-OR 21 -C 0～4 Alkylene-OC(O)R 21 -C 0～4 Alkylene-SR 21 -C 0～4 Alkylene-S(O)2R 21 -C 0～4 Alkylene-S(O)R 21 -C 0～4 Alkylene-S(O)2NR 21 R 22 -C 0～4 Alkylene-S(O)NR 21 R 22 -C 0～4 Alkylene-C(O)R 21 -C 0～4 Alkylene-C(O)OR 21 -C 0～4 Alkylene-C(O)NR 21 R 22 -C 0～4 Alkylene-NR21 R 22 -C 0～4 Alkylene-NR 21 C(O)R 22 -C 0～4 Alkylene-NR 21 S(O)2R 22 -C 0～4 Alkylene-NR 21 S(O)R 22 -C 0～4 Alkylene (3- to 10-membered cycloalkyl), -C 0～4 Alkylene (4- to 10-membered heterocyclic alkyl), -C 0～4 alkylene-(6- to 10-membered aromatic ring), -C 0～4 Alkylene (5- to 10-membered heteroaryl ring); wherein the alkylene, cycloalkyl, heterocycloalkyl, aromatic ring, and heteroaryl ring may be optionally surrounded by one, two, three, or four independent R... 26 replace;

[0015] R 21 R 22 Selected independently from hydrogen and -C 1～6 Alkyl, -C 2～6 alkenyl, -C 2～6 Alkyne- or halogen-substituted -C 1～6 Alkyl, halogen-substituted -C 2～6 Alkenyl, halogen-substituted -C 2～6 alkynyl group, -C 1～4 Alkylene-OR 24 -C 1～4 Alkylene-OC(O)R 24 -C 1～4 Alkylene-SR 24 -C 1～4 Alkylene-S(O)2R 24 -C 1～4 Alkylene-S(O)R 24 -C 1～4 Alkylene-S(O)2NR 24 R 25 -C 1～4 Alkylene-S(O)NR 24 R 25 -C 1～4 Alkylene-C(O)R 24 -C 1～4 Alkylene-C(O)OR 24 -C 1～4 Alkylene-C(O)NR 24 R 25 -C 1～4 Alkylene-NR24 R 25 -C 1～4 Alkylene-NR 24 C(O)R 25 -C 1～4 Alkylene-NR 24 S(O)2R 25 -C 1～4 Alkylene-NR 24 S(O)R 25 -C 0～4 Alkylene (3- to 10-membered cycloalkyl), -C 0～4 Alkylene (4- to 10-membered heterocyclic alkyl), -C 0～4 alkylene-(6- to 10-membered aromatic ring), -C 0～4 Alkylene (5- to 10-membered heteroaryl ring); wherein the alkylene, cycloalkyl, heterocycloalkyl, aromatic ring, and heteroaryl ring may be optionally surrounded by one, two, three, or four independent R... 26 replace;

[0016] Each R 26 Each group is independently selected from hydrogen, halogen, cyano, nitro, =O, =S, =CR 24 R 25 -C 1～6 Alkyl, -C 2～6 alkenyl, -C 2～6 Alkyne- or halogen-substituted -C 1～6 Alkyl, halogen-substituted -C 2～6 Alkenyl, halogen-substituted -C 2～6 alkynyl group, -C 0～4 Alkylene-OR 24 -C 0～4 Alkylene-OC(O)R 24 -C 0～4 Alkylene-SR 24 -C 0～4 Alkylene-S(O)2R 24 -C 0～4 Alkylene-S(O)R 24 -C 0～4 Alkylene-S(O)2NR 24 R 25 -C 0～4 Alkylene-S(O)NR 24 R 25 -C 0～4 Alkylene-C(O)R 24 -C 0～4 Alkylene-C(O)OR 24 -C 0～4 Alkylene-C(O)NR 24 R25 -C 0～4 Alkylene-NR 24 R 25 -C 0～4 Alkylene-NR 24 C(O)R 25 -C 0～4 Alkylene-NR 24 S(O)2R 25 -C 0～4 Alkylene-NR 24 S(O)R 25 -C 0～4 Alkylene (3- to 10-membered cycloalkyl), -C 0～4 Alkylene (4- to 10-membered heterocyclic alkyl), -C 0～4 alkylene-(6- to 10-membered aromatic ring), -C 0～4 Alkylene (5- to 10-membered heteroaryl ring); wherein the alkylene, cycloalkyl, heterocycloalkyl, aromatic ring, and heteroaryl ring may be optionally surrounded by one, two, three, or four independent R... 27 replace;

[0017] R 24 R 25 Selected independently from hydrogen and -C 1-6 alkyl, -C 2～6 alkenyl, -C 2～6 Alkyne- or halogen-substituted -C 1～6 Alkyl, halogen-substituted -C 2～6 Alkenyl, halogen-substituted -C 2～6 alkynyl group;

[0018] Each R 27 Each group is independently selected from hydrogen, halogen, cyano, nitro, =O, =S, and -C. 1～6 Alkyl, -C 2～6 alkenyl, -C 2～6 Alkyne- or halogen-substituted -C 1～6 Alkyl, halogen-substituted -C 2～6 Alkenyl, halogen-substituted -C 2～6 Alkyne group.

[0019] Preferably, the A ring is selected from a 10-membered nitrogen-containing aromatic heterocycle.

[0020] Furthermore,

[0021] The A ring is selected from The B ring is a 6-membered nitrogen-containing aromatic heterocycle. In Formula I, R is connected to the A ring. 2 The N atom can be attached to any substituted position of the A ring described above; preferably, the R atom attached to the A ring in Formula I... 2The N atom can be attached to any substituted position in the B ring described above.

[0022] Preferably,

[0023] The A ring is selected from Among them, the ring A selected from the rings can be further arbitrarily selected by one, two, three or four independent R. A1 Replace. R, which is connected to ring A in equation I. 2 The N atom can be attached to any substituted position in the A ring described above;

[0024] As a preferred option

[0025] Each R A1 Selected independently from hydrogen, halogen, and -NR A2 R A3 -OR A2 -C 1～3 Alkyl, halogen-substituted -C 1～3 alkyl;

[0026] R A2 R A3 Each is independently selected from hydrogen, methyl, ethyl, and propyl.

[0027] As a preferred option

[0028] The A ring is selected from

[0029] As a preferred option

[0030] R 2 Selected from hydrogen, halogen, cyano, nitro, =O, =S, -C 1～6 Alkyl, -C 2～6 alkenyl, -C 2～6 Alkyne- or halogen-substituted -C 1～6 Alkyl, halogen-substituted -C 2～6 Alkenyl, halogen-substituted -C 2～6 alkynyl group, -C 0～4 Alkylene-OR 21 -C 0～4 Alkylene-OC(O)R 21 -C 0～4 Alkylene-C(O)R 21 -C 0～4 Alkylene-C(O)OR 21 -C 0～4 Alkylene-C(O)NR 21 R 22 -C 0～4 Alkylene-NR 21 R 22;-C 0～4 Alkylene (3- to 10-membered cycloalkyl), -C 0～4 Alkylene (4- to 10-membered heterocyclic alkyl), -C 0～4 alkylene-(6- to 10-membered aromatic ring), -C 0～4 Alkylene (5- to 10-membered heteroaryl ring); wherein the alkylene, cycloalkyl, heterocycloalkyl, aromatic ring, and heteroaryl ring may be optionally surrounded by one, two, three, or four independent R... 23 replace;

[0031] Each R 23 Each group is independently selected from hydrogen, halogen, cyano, nitro, =O, =S, and -C. 1～6 Alkyl, -C 2～6 alkenyl, -C 2～6 Alkyne- or halogen-substituted -C 1～6 Alkyl, halogen-substituted -C 2～6 Alkenyl, halogen-substituted -C 2～6 alkynyl group, -C 0～4 Alkylene-OR 21 -C 0～4 Alkylene-OC(O)R 21 -C 0～4 Alkylene-C(O)R 21 ;-C 0～4 Alkylene (3- to 10-membered cycloalkyl), -C 0～4 Alkylene (4- to 10-membered heterocyclic alkyl), -C 0～4 alkylene-(6- to 10-membered aromatic ring), -C 0～4 Alkylene (5- to 10-membered heteroaryl ring); wherein the alkylene, cycloalkyl, heterocycloalkyl, aromatic ring, and heteroaryl ring may be optionally surrounded by one, two, three, or four independent R... 26 replace;

[0032] R 21 R 22 Selected independently from hydrogen and -C 1～6 Alkyl, -C 2～6 alkenyl, -C 2～6 Alkyne- or halogen-substituted -C 1～6 Alkyl, halogen-substituted -C 2～6 Alkenyl, halogen-substituted -C 2～6 alkynyl group, -C 1～4 Alkylene-OR 24 -C 1～4 Alkylene-OC(O)R 24 -C 1～4 Alkylene-C(O)R 24 -C 1～4 Alkylene-C(O)OR 24 -C1～4 Alkylene-C(O)NR 24 R 25 -C 1～4 Alkylene-NR 24 R 25 -C 1～4 Alkylene-NR 24 C(O)R 25 -C 0～4 Alkylene (3- to 10-membered cycloalkyl), -C 0～4 Alkylene (4- to 10-membered heterocyclic alkyl), -C 0～4 alkylene-(6- to 10-membered aromatic ring), -C 0～4 Alkylene (5- to 10-membered heteroaryl ring); wherein the alkylene, cycloalkyl, heterocycloalkyl, aromatic ring, and heteroaryl ring may be optionally surrounded by one, two, three, or four independent R... 26 replace;

[0033] Each R 26 Each group is independently selected from hydrogen, halogen, cyano, nitro, =O, =S, =CR 24 R 25 -C 1～6 Alkyl, -C 2～6 alkenyl, -C 2～6 Alkyne- or halogen-substituted -C 1～6 Alkyl, halogen-substituted -C 2～6 Alkenyl, halogen-substituted -C 2～6 alkynyl group, -C 0～4 Alkylene-OR 24 -C 0～4 Alkylene-OC(O)R 24 -C 0～4 Alkylene-C(O)R 24 -C 0～4 Alkylene-C(O)OR 24 -C 0～4 Alkylene-C(O)NR 24 R 25 -C 0～4 Alkylene-NR 24 R 25 -C 0～4 Alkylene-NR 24 C(O)R 25 -C 0～4 Alkylene (3- to 10-membered cycloalkyl), -C 0～4 Alkylene (4- to 10-membered heterocyclic alkyl), -C 0～4 alkylene-(6- to 10-membered aromatic ring), -C 0～4 Alkylene rings (5- to 10-membered heteroaryl rings);

[0034] R 24 R 25 Selected independently from hydrogen and -C 1-3 Alkyl, halogen-substituted -C 1～3 alkyl.

[0035] As a preferred option

[0036] The R 2 Selected from hydrogen, -C(O)NR 21 R 22 -C(O)R 21 -C 0～2 Alkylene-NR 21 R 22 -C(O)OR 21 -OR 21 ;

[0037] R 21 R 22 Selected independently from hydrogen and -C 1～3 Alkyl, -C 0～1 alkylene-(6-membered aromatic ring), -C 0～1 Alkylene rings are defined as -(10-membered heteroaryl rings), -(4-6-membered heterocyclic alkyl rings), or -(3-6-membered cycloalkyl rings), wherein the aromatic ring, heteroaryl ring, heterocyclic alkyl ring, or cycloalkyl ring may be optionally surrounded by one, two, three, or four independent R groups. 26 replace;

[0038] Each R 26 Selected independently from hydrogen and -C 1～3 Alkyl, -(4- to 6-membered heterocyclic alkyl), -C(O)R 24 -C(O)OR 24 -OC(O)R 24 ;

[0039] R 24 Selected from hydrogen, methyl, ethyl, and propyl.

[0040] Furthermore,

[0041] The R 2 Selected from hydrogen, hydroxyl,

[0042] In some specific embodiments of the invention, the compound represented by Formula I of the present invention is specifically:

[0043]

[0044]

[0045] The present invention also provides the use of the above-mentioned compounds, or stereoisomers thereof, or deuterated compounds thereof, or pharmaceutically acceptable salts thereof, in the treatment of diseases related to abnormal cell proliferation.

[0046] Furthermore, the disease in question is cancer.

[0047] The present invention also provides the use of the above-mentioned compounds, or their stereoisomers, or their deuterated compounds, or their pharmaceutically acceptable salts, in the preparation of targeted protein degradation drugs.

[0048] The present invention also provides the use of the above-mentioned compound, or its stereoisomer, or its deuterated compound, or its pharmaceutically acceptable salt as an intermediate in the preparation of a targeted protein degradation drug.

[0049] The compounds and derivatives provided in this invention can be named according to the IUPAC (International Union of Pure and Applied Chemistry) or CAS (Chemical Abstracts Service, Columbus, OH) nomenclature system.

[0050] Regarding the definition of terms used in this invention: Unless otherwise stated, the initial definitions provided for groups or terms herein apply to the groups or terms used throughout this specification; for terms not specifically defined herein, the meanings that a person skilled in the art would give them should be given based on the disclosure and context.

[0051] "Substitution" refers to the replacement of hydrogen atoms in a molecule by other different atoms or molecules. "Substitution" can also refer to the replacement of lone pairs of electrons in atoms in a molecule by "=O", "=S", etc.

[0052] "Optionally replaceable" means that "replacement" may but does not have to occur, and this statement includes situations in which it may or may not occur.

[0053] The minimum and maximum carbon atom content in hydrocarbon groups are indicated by a prefix, for example, the prefix C. a～b Alkyl indicates any alkyl group containing "a" to "b" carbon atoms. Therefore, for example, "C 1～4 "Alkyl" refers to an alkyl group containing 1 to 4 carbon atoms.

[0054] "Alkyl" refers to a saturated hydrocarbon chain having a specified number of member atoms. For example, C1-6 alkyl refers to an alkyl group having 1 to 6 member atoms, such as 1 to 4 member atoms. Alkyl groups can be straight-chain or branched. Representative branched alkyl groups have one, two, or three branches. Alkyl groups may optionally be substituted by one or more substituents as defined herein. Alkyl groups include methyl, ethyl, propyl (n-propyl and isopropyl), butyl (n-butyl, isobutyl, and tert-butyl), pentyl (n-pentyl, isopentyl, and neopentyl), and hexyl. Alkyl groups may also be part of other groups, such as C1-6 alkoxy groups.

[0055] In this invention, "alkylene" refers to a divalent saturated aliphatic hydrocarbon group having a specified number of carbon atoms. "C" a ~ b "Alkylene" refers to an alkylene group having a to b carbon atoms. Alkylene groups include branched and straight-chain hydrocarbon groups. For example, "C 1～6 The term "alkylene" is intended to include methylene, ethylene, propylene, 2-methylpropylene, dimethylethylene, pentylene, etc. Therefore, the term "propylene" can be exemplified by the following structures: Similarly, the term "dimethylbutylene" can be exemplified, for example, by any of the following structures: Furthermore, the term "(C1-6)alkylene" is intended to include such branched hydrocarbon groups, such as cyclopropylmethylene, which can be exemplified by the following structures: For example, -C0 to -4 alkylene groups can be C0 alkylene, C1 alkylene (e.g., -CH2-), C2 alkylene (e.g., -CH2CH2-), C3 alkylene, or C4 alkylene; C0 alkylene refers to the absence of a group here, which is connected by a chemical bond. For example, A-C0 alkylene-B refers to AB, that is, the A group and the B group are directly connected by a chemical bond.

[0056] "Alkenyl" refers to a straight-chain or branched hydrocarbon group having a specified number of carbon atoms and, in some embodiments, 2 to 6 carbon atoms or 2 to 4 carbon atoms and having at least one vinyl unsaturated site (>C=C<). For example, C a-b Alkenyl refers to an alkenyl group having a to b carbon atoms and is intended to include, for example, vinyl, propenyl, isopropenyl, 1,3-butadienyl, etc.

[0057] In this invention, "alkenyl" refers to a hydrocarbon chain having 2 to 10 carbon atoms, at least one double bond, and two unsaturated valences. For example, (C3-C6)alkenyl groups include >C=CH-CH2-, -CH-CH=CH-CH2-, etc.

[0058] "Alynyl" refers to a straight-chain monovalent hydrocarbon group or a branched monovalent hydrocarbon group containing at least one triple bond. The term "alkynyl" is also intended to include those hydrocarbon groups having one triple bond and one double bond. For example, (C2-C6) alkynyl is intended to include ethynyl, propynyl, etc.

[0059] "Halogen" refers to fluorine, chlorine, bromine, or iodine.

[0060] "Halogenated alkyl" or "halogen-substituted alkyl" refers to an alkyl group in which one or more hydrogen atoms can be replaced by one or more halogen atoms. For example, C 1～4 Halogenated alkyl refers to an alkyl group containing 1 to 4 carbon atoms in which one or more hydrogen atoms are replaced by one or more halogen atoms.

[0061] In this invention, "-OR", "-NRR", etc., refer to the R group being connected to an oxygen atom or a nitrogen atom by a single bond.

[0062] In this invention, the oxygen atom in “-C(O)R”, “-S(O)2R”, etc., is connected to the carbon atom or sulfur atom by a double bond, and the R group is connected to the oxygen atom or sulfur atom by a single bond.

[0063] In this invention, "cycloalkyl" and "cycloalkane" refer to saturated or partially saturated cyclic groups having multiple carbon atoms and no heterocyclic atoms, and having a single ring or multiple rings (including fused, bridged, spirocyclic, and adamantane systems). For polycyclic systems having aromatic and non-aromatic rings without heteroatoms, the term "cycloalkyl" (e.g., 5,6,7,8-tetrahydronaphthalene-5-yl) is used when the connecting point is located on a non-aromatic carbon atom. The term "cycloalkyl" includes cycloalkenyl groups, such as cyclohexenyl. Examples of cycloalkyl groups include, for example, adamantyl, cyclopropyl, cyclobutyl, cyclohexyl, cyclopentyl, cyclooctyl, cyclopentenyl, and cyclohexenyl. Examples of cycloalkyl groups including polycycloalkyl ring systems are dicyclohexyl, dicyclopentyl, dicyclooctyl, etc. adamantyl groups include, but are not limited to, the following structures:

[0064] In this invention, "heterocyclic," "heterocyclic alkyl," and "heterocyclic alkane" refer to a saturated ring or a non-aromatic unsaturated ring containing at least one heteroatom; where heteroatoms refer to nitrogen, oxygen, sulfur, etc. Generally, it represents a monovalent saturated or partially unsaturated monocyclic or bicyclic ring system with multiple ring atoms, preferably a monovalent saturated or partially unsaturated monocyclic or bicyclic ring system with 3 to 9 ring atoms, containing 1, 2, or 3 cyclic heteroatoms selected from N, O, and S, with the remaining ring atoms being carbon. A bicyclic ring represents two rings consisting of two ring atoms, i.e., the bridge separating the two rings is a single bond or a chain of one or two ring atoms. Examples of monocyclic saturated heterocyclic alkyl groups are oxobutyl, aziridine, pyrrolidinyl, 2-oxo-pyrrolidin-3-yl, tetrahydrofuranyl, tetrahydro-thiophenyl, pyrazolyl, imidazoalkyl, thiazoalkyl, piperidinyl, tetrahydropyranyl, tetrahydrothiaranyl, piperazine, morpholinyl, etc. Thiomorpholinyl, 1,1-dioxo-thiomorpholin-4-yl, azacycloheptyl, diazacycloheptyl, periperazinyl, or oxazacycloheptyl. Examples of bicyclic saturated heterocyclic alkyl groups are 8-aza-bicyclo[3.2.1]octyl, quininecycloyl, 8-oxa-3-aza-bicyclo[3.2.1]octyl, 9-aza-bicyclo[3.3.1]nonyl, Examples of partially unsaturated heterocyclic alkyl groups are dihydrofuranyl, imidazolinyl, tetrahydropyridyl, or dihydropyranyl.

[0065] "Spirocycloiden" and "spirocycloheterocycle" are used interchangeably. They refer to non-aromatic saturated rings or non-aromatic unsaturated ring systems with two monocyclic rings sharing a single carbon atom, consisting of a carbon atom and heteroatoms selected from nitrogen, oxygen, sulfur, and phosphorus. For example, "5- to 12-membered spirocycloheterocycles" refer to spirocycloheterocycles with 5 to 12 ring atoms, of which 1, 2, or 3 ring atoms are heteroatoms.

[0066] "Bridged ring or bridged ring group" refers to a saturated or unsaturated cyclic group formed by two or more cyclic structures sharing two non-adjacent atoms. Specific examples include, but are not limited to:

[0067] "Bridged heterocyclic group" and "bridged heterocycle" are used interchangeably, referring to a saturated or unsaturated cyclic group formed by two or more cyclic structures sharing two non-adjacent atoms, composed of carbon atoms and heteroatoms selected from nitrogen, oxygen, sulfur, and phosphorus. Specific embodiments include, but are not limited to:

[0068] In this invention, "aromatic ring" and "aryl" refer to aromatic hydrocarbon groups having multiple carbon atoms. Aryl groups are typically monocyclic, bicyclic, or tricyclic aryl groups having 5-20 carbon atoms. Furthermore, the term "aryl" as used herein refers to an aromatic substituent that can be a single aromatic ring or multiple aromatic rings fused together. Non-limiting examples include phenyl, naphthyl, or tetrahydronaphthyl.

[0069] In this invention, "heteroaromatic ring" and "heteroaromatic cyclic group" refer to an aromatic unsaturated ring containing at least one heteroatom; wherein the heteroatom refers to a nitrogen atom, oxygen atom, sulfur atom, etc. Typically, it refers to an aromatic monocyclic or bicyclic hydrocarbon containing multiple ring atoms, one or more of which are selected from O, N, and S heteroatoms. Preferably, it has one to three heteroatoms. Examples of heterocyclic aryl groups include: pyridyl, indolyl, quinoxalinyl, quinolinyl, isoquinolinyl, benzothiopheneyl, benzofuranyl, benzothiopheneyl, benzopyranyl, benzothiapyranyl, furanyl, pyrroleyl, thiazolyl, oxazolyl, isoxazolyl, triazolyl, tetrazolyl, pyrazolyl, imidazoleyl, thiopheneyl, oxadiazolyl, benzimidazoleyl, benzothiazolyl, and benzoxazolyl.

[0070] "Stereoisomers" include enantiomers and diastereomers;

[0071] The "deuterated compound" of this invention refers to a molecule or group in which one or more hydrogen atoms are replaced by deuterium atoms, wherein the proportion of deuterium atoms is greater than the abundance of deuterium in nature.

[0072] The term "pharmaceutically acceptable" means that a carrier, delivery substance, diluent, excipient, and / or the salt formed therefrom is generally chemically or physically compatible with other components constituting a drug dosage form and physiologically compatible with receptors.

[0073] The terms "salt" and "pharmaceutically acceptable salt" refer to acidic and / or basic salts formed by the above-described compounds or their stereoisomers with inorganic and / or organic acids and bases, including zwitterionic salts (internal salts) and quaternary ammonium salts, such as alkylammonium salts. These salts can be obtained directly during the final separation and purification of the compounds. Alternatively, they can be obtained by mixing the above-described compounds, or their stereoisomers, with an appropriate amount (e.g., equimolar amounts) of an acid or base. These salts may be obtained by precipitating in solution and collecting by filtration, or by recovery after solvent evaporation, or by freeze-drying after reaction in an aqueous medium. The salts described in this invention can be hydrochlorides, sulfates, citrates, benzenesulfonates, hydrobromides, hydrofluoric acids, phosphates, acetates, propionates, succinates, oxalates, malates, succinates, fumarates, maleates, tartrates, or trifluoroacetates of the compounds.

[0074] Obviously, based on the above description of the present invention, and according to common technical knowledge and conventional methods in the field, various other modifications, substitutions, or alterations can be made without departing from the basic technical concept of the present invention. Detailed Implementation

[0075] The following detailed embodiments further illustrate the above-described content of the present invention. However, this should not be construed as limiting the scope of the present invention to the following examples. All technologies implemented based on the above-described content of the present invention fall within the scope of the present invention.

[0076] The known starting materials of this invention can be synthesized using or according to methods known in the art, or can be purchased from companies such as Anaiji Chemical, Chengdu Kelong Chemical, Shaoyuan Chemical Technology, and Bailingwei Technology.

[0077] Unless otherwise specified in the examples, the reaction is carried out under a nitrogen atmosphere. Unless otherwise specified in the examples, the solution refers to an aqueous solution. Unless otherwise specified in the examples, the reaction temperature is room temperature. Room temperature is the optimal reaction temperature, which is 20°C. □ ~30℃ □ Unless otherwise specified in the examples, M represents moles per liter.

[0078] The structure of the compound was determined by nuclear magnetic resonance (NMR) and mass spectrometry (MS). NMR shifts (δ) are given in units of 10⁻⁶ (ppm). NMR measurements were performed using a Bruker Avance III 400 and Bruker Avance 600 NMR spectrometer, with deuterated dimethyl sulfoxide (DMSO-d6), deuterated chloroform (CDCl₃), and deuterated methanol (Methol-d4) as solvents, and tetramethylsilane (TMS) as the internal standard. LC-MS measurements were performed using a Shimadzu liquid chromatography-mass spectrometry (LC-MS) system.

[0079] (Shimadzu LC-MS 2020(ESI)). HPLC determination was performed using a Shimadzu LC-20A high-performance liquid chromatograph. MPLC (medium-pressure preparative chromatography) was performed using a Gilson GX-281 reversed-phase preparative chromatograph. Thin-layer chromatography used Yantai Huanghai HSGF254 or Qingdao GF254 silica gel plates, with a thickness of 0.4mm to 0.5mm for product separation and purification. Column chromatography generally used Yantai Huanghai 200-300 mesh silica gel as the carrier.

[0080] The reagents described in the examples are abbreviated as follows: Pd2(dba)3: tridibenzylacetone dipalladium; DIPEA: N,N-diisopropylethylamine; DMSO: dimethyl sulfoxide; AcOH: acetic acid; Sphos: 2-bicyclohexylphosphine-2',6'-dimethoxy-1,1'-diphenyl; LC-MS: liquid chromatography-mass spectrometry; NaCl: sodium chloride; MPLC: medium-pressure liquid chromatography-preparative; TMEDA: tetramethylethylenediamine.

[0081] Example 1: Preparation of compound A1

[0082]

[0083] Step 1: Preparation of compound A-3

[0084]

[0085] Compounds A-1 (100.00 mg, 502.42 μmol), A-2 (22.65 mg, 502.42 μmol), and DIPEA (194.80 mg, 1.51 mmol, 262.53 μL) were added sequentially to a 50 mL reaction flask in 5 mL of tetrahydrofuran. The reaction mixture was reacted at room temperature for 15 hours (monitored by LC-MS). The reaction solution was then extracted with 10 mL of saturated NaCl solution and 3 × 20 mL of ethyl acetate. The organic phases were combined and dried over anhydrous sodium sulfate. The solvent was evaporated and concentrated under reduced pressure to remove the solvent, yielding compound A-3 (crude product). The crude product was then purified by MPLC to obtain compound A-3 (86.00 mg, 414.14 μmol, 82.43% yield).

[0086] Step 2: Synthesis of Compound A1

[0087]

[0088] Compounds A-3 (28.00 mg, 134.84 μmol), A-4 (18.90 mg, 134.84 μmol), AcOH (2 mL), and DMSO (2 mL) were added sequentially to a 50 mL reaction flask. The reaction was stirred at 100 °C for 2 hours to quench the reaction (monitored by LC-MS). Extraction was completed using saturated NaCl solution (10 mL) and ethyl acetate (3 × 20 mL). The organic phases were combined, dried over anhydrous sodium sulfate, and the solvent was evaporated. After concentration under reduced pressure, compound A1 (4.00 mg, 12.85 μmol, 9.53% yield, 99.9% purity) was analyzed by MPLC. LC-MS: C 16 H 18 N6O2,[M+H] + :312.1,found 312.2.1 H NMR (600MHz, DMSO-d6) δ7.95 (dd, J=8.5, 1.4Hz, 1H), 7.53 (ddd, J=8.5, 6.8, 1.4Hz, 1H), 7.37 (dd, J=8.4, 1.2Hz, 1H), 7.09 (ddd, J=8.4, 6.8, 1.2Hz, 1H), 4.23 (d, J=8.6Hz, 2H), 4.02 (d, J=8.6Hz, 2H), 3.24 (s, 6H), 3.04 (s, 2H).

[0089] Following the synthetic method of compound A1, replace compound A-2 with raw material 1 from the table below, while keeping other raw materials and procedures unchanged, to obtain compound A2.

[0090]

[0091] Example 2, Preparation of compound A3

[0092]

[0093] Step 1: Synthesis of Compound A3

[0094]

[0095] Compounds A-6 (20.00 mg, 110.75 μmol), A-4 (15.52 mg, 110.75 μmol), DIPEA (42.94 mg, 332.24 μmol, 57.87 μL), and DMSO (2 mL) were added sequentially to a 50 mL reaction flask. The reaction was stirred at 150 °C for 15 hours, and the reaction was quenched (monitored by LC-MS). Extraction was completed using saturated NaCl solution (10 mL) and ethyl acetate (3 × 20 mL). The organic phases were combined, dried over anhydrous sodium sulfate, and the solvent was evaporated. After concentration under reduced pressure, compound A3 (8.00 mg, 28.14 μmol, 25.14% yield, 99.1% purity) was extracted by MPLC. LC-MS: C 14 H 13 N4O3,[M+H] + :285.1,found 285.2. 1 HNMR (400MHz, Methanol-d4) δ8.02 (d, J = 8.0 Hz, 1H), 7.69 (t, J = 8.0 Hz, 1H), 7.49-7.23 (m, 2H), 4.54 (d, J = 9.5 Hz, 2H), 4.33 (d, J = 9.5 Hz, 2H), 3.02 (s, 2H).

[0096] Following the synthetic method of compound A3, by replacing compound A-6 with raw material 1 from the table below, while keeping other raw materials and procedures unchanged, compound A4 can be obtained.

[0097]

[0098] Example 3: Preparation of compound A5

[0099]

[0100] Step 1: Preparation of compound A-9

[0101]

[0102] Compounds A-8 (150.00 mg, 753.63 μmol), A-4 (34.95 mg, 753.63 μmol), and DIPEA (292.20 mg, 1.51 mmol, 262.53 μL) were added sequentially to a 50 mL reaction flask in 8 mL of tetrahydrofuran. The reaction system was kept at room temperature for 12 hours (monitored by LC-MS). The reaction solution was then extracted with 10 mL of saturated NaCl solution and 3 × 20 mL of ethyl acetate. The organic phases were combined and dried over anhydrous sodium sulfate. The solvent was evaporated and concentrated under reduced pressure to remove the solvent, yielding compound A-9 (crude product). The crude product was then purified by MPLC to obtain compound A-9 (100.00 mg, 481.40 μmol, 63.90% yield).

[0103] Step 2: Synthesis of Compound A5

[0104]

[0105] Compounds A-9 (20.00 mg, 66.07 μmol), A-2 (2.98 mg, 66.07 μmol), AcOH (2 mL), and DMSO (2 mL) were added sequentially to a 50 mL reaction flask. The reaction was stirred at 100 °C for 15 hours, and then quenched (monitored by LC-MS). Extraction was completed using saturated NaCl solution (10 mL) and ethyl acetate (3 × 20 mL). The organic phases were combined, dried over anhydrous sodium sulfate, and the solvent was evaporated. After concentration under reduced pressure, compound A5 (2.20 mg, 7.07 μmol, 10.70% yield, 99.9% purity) was determined by MPLC. LC-MS: C 16 H 18 N5O2,[M+H] + :312.1,found 312.2. 1H NMR(400MHz,DMSO-d6)δ11.65and 11.37(s,2H),8.16-7.68(m,3H),7.45-7.40(m,1H),5.08-4.88(m,2H),4.61-4.26(m,2H),3.25(s,6H),3.08(s,2H).

[0106] Following the synthetic method of compound A5, replacing compound A-2 with raw material 1 from the table below, while keeping other raw materials and procedures unchanged, yields compounds A6 and A7.

[0107]

[0108] Example 4: Preparation of compound A8

[0109]

[0110] Step 1: Preparation of compound A-13

[0111]

[0112] Compounds A-12 (300.00 mg, 1.51 mmol), A-2 (271.91 mg, 6.03 mmol), and potassium carbonate (1.25 g, 9.04 mmol) were added sequentially to a 50 mL reaction flask in 10 mL of N-methylpyrrolidone. The reaction system was heated at 180 °C for 4 hours (monitored by LC-MS). The reaction solution was then extracted with 10 mL of saturated NaCl solution and 3 × 20 mL of ethyl acetate. The organic phases were combined and dried over anhydrous sodium sulfate. The solvent was evaporated and concentrated under reduced pressure to remove the solvent, yielding compound A-13 (crude product). The crude product was then purified by MPLC to obtain compound A-13 (170.00 mg, 818.65 μmol, 54.31% yield).

[0113] Step 2: Synthesis of Compound A8

[0114]

[0115] Compounds A-13 (100.00 mg, 481.56 μmol), A-4 (80.98 mg, 577.87 μmol), cesium carbonate (313.98 mg, 963.12 μmol), Sphos (19.74 mg, 48.16 μmol), and Pd2(dba)3 (22.03 mg, 24.08 μmol) were added sequentially to a 50 mL reaction flask and dissolved in 1,4-dioxane (5 mL). The reaction was stirred at 120 °C in the absence of air for 2 hours, after which the reaction was quenched (monitored by LC-MS). Extraction was completed using saturated NaCl solution (10 mL) and ethyl acetate (3 × 20 mL). The organic phases were combined, dried over anhydrous sodium sulfate, and the solvent was evaporated. The solution was concentrated under reduced pressure and used to prepare MPLC compound A8 (1.50 mg, 4.82 μmol, 1.00% yield, 92.0% purity). LC-MS: C 16 H 18 N5O2,[M+H] + :312.1,found312.2. 1 H NMR (400MHz, DMSO-d6) δ 8.17-8.03 (m, 1H), 7.97-7.79 (m, 3H), 4.51 (d, J = 8.4Hz, 2H), 4.33 (d, J = 8.4Hz, 2H), 3.11 (s, 2H), 2.92 (s, 6H).

[0116] Example 5: Preparation of compound A9

[0117]

[0118] Step 1: Preparation of compound A-15

[0119]

[0120] Compounds A-14 (398.00 mg, 2.00 mmol), A-2 (90.15 mg, 2.00 mmol), and DIPEA (516.88 mg, 4.00 mmol, 696.60 μL) were added sequentially to a 50 mL reaction flask in 8 mL of DMSO. The reaction system was heated at 80 °C for 10 hours (monitored by LC-MS). The reaction solution was then extracted with 10 mL of saturated NaCl solution and 3 × 20 mL of ethyl acetate. The organic phases were combined and dried over anhydrous sodium sulfate. The solvent was evaporated and concentrated under reduced pressure to remove the solvent, yielding compound A-15 (crude product). The crude product was then purified by MPLC to obtain compound A-15 (300.00 mg, 1.44 mmol, 72.25% yield).

[0121] Step 2: Synthesis of Compound A9

[0122]

[0123] Compounds A-15 (120.00 mg, 577.87 μmol), A-4 (121.47 mg, 866.81 μmol), cesium carbonate (379.08 mg, 1.16 mmol), Sphos (23.69 mg, 57.79 μmol), and Pd2(dba)3 (26.44 mg, 28.89 μmol) were added sequentially to a 50 mL reaction flask and dissolved in 1,4-dioxane (3 mL). The reaction was stirred at 140 °C in the absence of air for 4 hours, after which the reaction was quenched (monitored by LC-MS). Extraction was completed using saturated NaCl solution (10 mL) and ethyl acetate (3 × 20 mL). The organic phases were combined, dried over anhydrous sodium sulfate, and the solvent was evaporated. The solution was concentrated under reduced pressure and used to prepare compound A9 (19.50 mg, 59.00 μmol, 10.21% yield, 94.2% purity) by MPLC. LC-MS: C 16 H 18 N5O2,[M+H] + :312.1,found312.1. 1 H NMR (400MHz, DMSO-d6) δ11.22(s,1H),7.86-7.50(m,2H),7.45-7.11(m,2H),4.30(d,J=9.0Hz,2H),4.14(d,J=9.0Hz,2H),3.02(s,2H),2.50(s,6H).

[0124] Following the synthetic method of compound A9, by replacing compound A-2 with raw material 1 from the table below, while keeping other raw materials and procedures unchanged, compound A10 can be obtained.

[0125]

[0126] Example 6: Preparation of compounds A11 and A12

[0127]

[0128] Step 1: Preparation of compound A-16

[0129]

[0130] Compounds A-14 (100.00 mg, 2.00 mmol), A-4 (70.41 mg, 502.42 μmol), and DIPEA (194.80 mg, 1.51 mmol, 262.54 μL) were added sequentially to a 50 mL reaction flask and dissolved in 5 mL of acetonitrile. The reaction system was heated at 80 °C for 15 hours (monitored by LC-MS). The reaction solution was then extracted with 10 mL of saturated NaCl solution and 3 × 20 mL of ethyl acetate. The organic phases were combined, dried over anhydrous sodium sulfate, and the solvent was evaporated. The solvent was removed by concentration under reduced pressure to obtain compound A-16 (crude product). The crude product was then purified by MPLC to obtain compound A-16 (120.00 mg, 396.41 μmol, 78.90% yield).

[0131] Step 2: Synthesis of Compound A11

[0132]

[0133] Compound A-16 (20.00 mg, 66.07 μmol), 6M HCl (66.07 μmol), and 2 mL of water were added sequentially to a 50 mL reaction flask. The reaction was heated at 100 °C and stirred in the absence of air for 2 hours, after which the reaction was quenched (monitored by LC-MS). Extraction was completed with 10 mL of saturated NaCl solution and 3 × 20 mL of ethyl acetate. The organic phases were combined, dried over anhydrous sodium sulfate, and the solvent was evaporated. The solution was concentrated under reduced pressure and used to prepare compound A11 (8.00 mg, 28.14 μmol, 42.60% yield, 90.0% purity) by MPLC. LC-MS: C 14 H 13 N4O3,[M+H] + :285.1,found 285.2. 1 H NMR (400MHz, Methanol-d4) δ7.59-7.38(m,1H),7.35-7.06(m,3H),4.60-4.40(m,4H),3.12(s,2H).

[0134] Step 3: Synthesis of Compound A12

[0135]

[0136] Compound A-16 (20.00 mg, 66.07 μmol), AcOH (660.69 μmol), Zn (660.69 μmol), and TMEDA (76.77 mg, 660.69 μmol) were added sequentially to a 50 mL reaction flask. The mixture was dissolved in 3 mL of ethanol. The reaction was carried out at 45 °C under air-free conditions with stirring for 15 hours, after which the reaction was quenched (monitored by LC-MS). Extraction was performed using 10 mL of saturated NaCl solution and 3 × 20 mL of ethyl acetate. The organic phases were combined, dried over anhydrous sodium sulfate, and the solvent was evaporated. The solution was concentrated under reduced pressure and used to prepare MPLC compound A12 (2.00 mg, 7.46 μmol, 11.28% yield, 91.2% purity). LC-MS: C 14 H 13 N4O2,[M+H] + :269.1,found 269.1. 1 H NMR (400MHz, DMSO-d6) δ 11.27 (s, 1H), 8.37 (s, 1H), 8.00-7.30 (m, 4H), 4.40 (d, J = 8.8Hz, 2H), 4.24 (d, J = 8.8Hz, 2H), 3.07 (s, 2H).

[0137] The technical effects of this invention are illustrated by the following experimental examples:

[0138] Example 1: Detection of the inhibitory effect of compounds on CRBN / DDB1 activity (FRET)

[0139] 1. Experimental materials and reagents

[0140] Microplate reader (BMG PHERAstar FSX), ECHO (LABCYTE Echo 665), microplate constant temperature shaker (Hangzhou Ruicheng Instrument Co., Ltd.), disodium hydrogen phosphate (Sigma), sodium dihydrogen phosphate (Sigma), bovine serum albumin (Sigma), Anti-6His-Tb crypate Gold (CISBIO), CRBN / DDB1 protein (HitGen), 384-well plate (Grenier Bio-one).

[0141] 2. Experimental Methods

[0142] The compound powder was dissolved in DMSO, and the compound was serially diluted with ECHO and added to a 384-well reaction plate to make the final concentration of DMSO in the entire reaction system (10.0 μL) 1.0%. An equal amount of DMSO was added as a control.

[0143] The CRBN / DDB1 protein was diluted twice to the desired final concentration (5.0 nM) using 20 mM disodium hydrogen phosphate, 20 mM sodium dihydrogen phosphate, 0.08% bovine serum albumin, and pH 7.0 buffer. 5.0 μL of the diluted CRBN / DDB1 protein was added to a 384-well plate containing the added compounds. The plate was centrifuged at 1000 rpm for 1 minute and then placed on a microplate shaker at 25°C and 250 rpm for 15 minutes. Anti-6His-Tb crypate Gold and FITC-labeled thalidomide analogue were diluted twice to the desired final concentration using 20mM disodium hydrogen phosphate, 20mM sodium dihydrogen phosphate, 0.08% bovine serum albumin, and pH 7.0 buffer. The final concentration of Anti-6His-Tb crypate Gold was 0.2 nM, and the final concentration of FITC-labeled thalidomide analogue was 50.0 nM, resulting in a mixture of Anti-6His-Tb crypate Gold and FITC-labeled thalidomide analogue. 5.0 μL of the Anti-6His-Tb crypate Gold / FITC-labeled thalidomide analogue mixture was added to a 384-well plate, centrifuged at 1000 rpm for 1 minute, and then incubated on a microplate shaker at 25°C and 250 rpm for 30 minutes. After the reaction was completed, the microplate reader read the fluorescence signal values ​​in the 384-well plate (Ex = 337nm Em = 520 / 490nm).

[0144] 3. Data Analysis

[0145] The solvent group (containing 5.0 nM CRBN / DDB1, 0.2 nM Anti-6His-Tb crypate Gold, 50.0 nM FITC-labeled thalidomide analogue and 1.0% DMSO) served as the negative control, and the reaction buffer group (containing 0.2 nM Anti-6His-Tb crypate Gold, 50.0 nM FITC-labeled thalidomide analogue and 1.0% DMSO) served as the blank control.

[0146] The formula for calculating the percentage of remaining activity at each concentration is as follows:

[0147] Remaining vitality (%) = 100% × (Flu) 化合物组 -Flu 空白对照 ) / (Flu 阴性对照 -Flu 空白对照 )

[0148] Then, the IC was calculated by fitting the dose-effect curve using GraphPad 6.0. 50 value.

[0149] Table 1: Relationship between compounds and CRBN / DDB1 protein inhibition

[0150]

[0151] Where + represents 200μM > IC 50 >100μM, ++ means 100μM>IC 50 >10μM, +++ indicates 10μM>IC 50 >1μM, ++++ indicates IC 50 <1μM.

Claims

1. The compound represented by Formula I, or its stereoisomer, or its pharmaceutically acceptable salt: Formula I in, Ring A is selected from , , , ; R 2 Selected from hydroxyl, , , .

2. The compound according to claim 1, characterized in that: The R 2 Selected from hydroxyl, , .

3. The compound according to claim 1 or claim 2, characterized in that: The compound is specifically: 、 、 、 、 、 。 4. Use of the compound according to any one of claims 1 to 3, or its stereoisomer, or its pharmaceutically acceptable salt, in the preparation of a medicament for treating diseases related to abnormal cell proliferation.

5. The use according to claim 4, characterized in that: The disease in question is cancer.

6. Use of the compound of any one of claims 1 to 3, or its stereoisomer, or its pharmaceutically acceptable salt, in the preparation of a drug for targeting protein degradation.

7. Use of the compound of any one of claims 1 to 3, or its stereoisomer, or its pharmaceutically acceptable salt as an intermediate in the preparation of a targeted protein degradation drug.

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