Spirocyclic compounds and uses

Abstract

The present application provides a new compound having a binding activity with E3 ligase protein CRBN, and a use thereof in the preparation of a medicament 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] Cerebellum protein (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. Cereblon is 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] Formula I

[0009] in,

[0010] Indicates whether oxygen substitution is present or absent;

[0011] 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;

[0012] 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-NR A2 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-(6-10 membered aromatic ring), -C 0~4 Alkylene (5-10 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;

[0013] 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 0~4Alkylene-NR A2 R A3 -C 0~4 Alkylene-NR A2 C(O)R A3 -C 0~4 Alkylene-NR A2 S(O)2R A3 -C 0~4 Alkylene-NR A2 S(O)R A3 ;

[0014] 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;

[0015] 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)NR21 R 22 -C 0~4 Alkylene-NR 21 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-(6-10 membered aromatic ring), -C 0~4 Alkylene (5-10 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;

[0016] 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)OR21 -C 0~4 Alkylene-C(O)NR 21 R 22 -C 0~4 Alkylene-NR 21 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-(6-10 membered aromatic ring), -C 0~4 Alkylene (5-10 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;

[0017] 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)OR24 -C 1~4 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 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-(6-10 membered aromatic ring), -C 0~4 Alkylene (5-10 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;

[0018] 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)R24 -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-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-(6-10 membered aromatic ring), -C 0~4 Alkylene (5-10 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;

[0019] 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;

[0020] 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.

[0021] Furthermore,

[0022] Ring A is selected from , , , , , , , , , , , , , , , , , , , , , , , , , , , , , , , , , , , , , , , , , , , ; wherein, the ring A selected from the rings can be further arbitrarily selected by one, two, three or four independent Rs. A1 Substitution. In Formula I, the R2 group and the carbonyl group attached to ring A can be attached to any substituted position on ring A.

[0023] As a preferred option, each R A1 Each group is independently selected from hydrogen, halogen, cyano, nitro, =O, =S, and -C. 1~3 Alkyl, halogen-substituted -C 1~3 alkyl.

[0024] Furthermore,

[0025] Ring A is selected from , , , , , , , , .

[0026] As a preferred option

[0027] The R 2Selected 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-(6-10 membered aromatic ring), -C 0~4 Alkylene (5-10 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;

[0028] 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-(6-10 membered aromatic ring), -C 0~4Alkylene (5-10 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;

[0029] 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 -C 1~4 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-(6-10 membered aromatic ring), -C 0~4 Alkylene (5-10 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;

[0030] 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-(6-10 membered aromatic ring), -C 0~4 alkylene rings (5-10 membered heteroaryl rings);

[0031] R 24 R 25 Selected independently from hydrogen and -C 1-3 Alkyl, halogen-substituted -C 1~3 alkyl.

[0032] Furthermore,

[0033] R 2 Selected from -C(O)NR 21 R 22 -C(O)R 21 -C 0~2 Alkylene-NR 21 R 22 -C(O)OR 21 ;

[0034] 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;

[0035] Each R 26 Selected independently from hydrogen and -C 1~3 Alkyl, -(4-6 membered heterocyclic alkyl), -C(O)R24 -C(O)OR 24 -OC(O)R 24 ;

[0036] R 24 Selected from hydrogen, methyl, and ethyl.

[0037] More specifically,

[0038] R 2 Selected from hydrogen, , , , , , , , , , , , , .

[0039] In some specific embodiments of the present invention, the compound is specifically:

[0040] , , , , , , , , , , , , , , , , , , , , , , ,

[0041] , , , .

[0042] The present invention also provides the use of any of the compounds described above, or their stereoisomers, or their deuterated compounds, or their pharmaceutically acceptable salts, in the treatment of diseases related to abnormal cell proliferation.

[0043] Furthermore, the disease in question is cancer.

[0044] The present invention also provides the use of any of the compounds described above, or their stereoisomers, or their deuterated compounds, or their pharmaceutically acceptable salts, in the preparation of targeted protein degradation drugs.

[0045] Furthermore, the use of the said 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 is provided.

[0046] Furthermore, the targeted protein degradation drug is a drug that relies on the E3 ligase CRBN for protein degradation.

[0047] 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 systems.

[0048] 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.

[0049] "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.

[0050] "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.

[0051] 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.

[0052] "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.

[0053] 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: or 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, -C0~4 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 means AB, that is, group A and group B are directly connected by a chemical bond.

[0054] "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.

[0055] 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.

[0056] "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.

[0057] "Halogen" refers to fluorine, chlorine, bromine, or iodine.

[0058] "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.

[0059] 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.

[0060] 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.

[0061] 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: .

[0062] 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, aziridine heptyl, diazacyclic heptyl, periperazinyl, or oxazacyclic heptyl. 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.

[0063] "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.

[0064] "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: , , , .

[0065] "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: , , , , .

[0066] 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.

[0067] 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.

[0068] "Stereoisomers" include enantiomers and diastereomers;

[0069] 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.

[0070] 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.

[0071] 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 (e.g., equimolar) amount of 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.

[0072] 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

[0073] 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.

[0074] 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.

[0075] The reagents described in the examples are abbreviated as follows: DIPEA: N,N-diisopropylethylamine; HATU: 2-(7-azobenzotriazole)-N,N,N',N'-tetramethylurea hexafluorophosphate; DIPEA: N,N-diisopropylethylamine; HATU: 2-(7-azobenzotriazole)-N,N,N',N'-tetramethylurea hexafluorophosphate; HOBT: 1-hydroxybenzotriazole; DMSO: dimethyl sulfoxide; LC-MS: liquid chromatography-mass spectrometry; NaCl: sodium chloride; MPLC: medium-pressure liquid chromatography preparation; EDCI: 1-ethyl-(3-dimethylaminopropyl)carbodiimide hydrochloride; Pd(OAc)2: palladium acetate; DMF: N,N-dimethylformamide; AIBN: azobisisobutyronitrile; NBS: N-bromosuccinimide; BPO: benzoyl peroxide.

[0076] 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 to 30°C. Unless otherwise specified in the examples, M is moles per liter.

[0077] The structures of the compounds were determined by nuclear magnetic resonance (NMR) and mass spectrometry (MS). NMR shifts (δ) are given in units of 10⁻⁶ (ppm). NMR determinations were performed using Bruker Avance III 400 and Bruker Avance 600 NMR spectrometers, 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 determinations were performed using a Shimadzu LC-MS 2020 (ESI) system. HPLC determinations were 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 uses Yantai Huanghai HSGF254 or Qingdao GF254 silica gel plates, with a thickness of 0.4 mm to 0.5 mm for product separation and purification. Column chromatography generally uses Yantai Huanghai 200-300 mesh silica gel as the carrier.

[0078] Example 1: Preparation of compound A1

[0079]

[0080] Step 1: Preparation of compound A-3

[0081]

[0082] Compound A-2 (50.00 mg, 211.65 μmol), DIPEA (132.25 mg, 0.80 mmol, 178.06 μL), HATU (250.96 mg, 0.66 mmol), and dichloromethane (2 mL) were added sequentially to a 50 mL reaction flask. After the reaction system temperature dropped to 0 °C, compound A-1 (86.29 mg, 1.06 mmol) was added. The reaction was stirred in an ice bath for 0.5 hours to quench the reaction (monitored by LC-MS). Extraction was completed with 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 solvent was removed by concentration under reduced pressure to obtain compound A-3 (crude product).

[0083] Step 2: Preparation of compound A-4

[0084]

[0085] Compound A-3 (crude product), sodium hydroxide (47.95 mg, 854.50 mmol), tetrahydrofuran (4 mL), and water (2 mL) were added sequentially to a 50 mL reaction flask. The reaction was quenched after stirring at room temperature for 8 hours (monitored by LC-MS). The pH of the system was adjusted to 6.0-7.0 with 1N hydrochloric acid solution. After concentrating the reaction solution, compound A-4 (crude product) was obtained and used directly in the next reaction.

[0086] Step 3: Synthesis of Compound A1

[0087]

[0088] Compound A-4 (8.70 mg, 34.90 μmol), EDCI (13.33 mg, 69.80 μmol), HOBT (9.43 mg, 69.80 μmol), and DIPEA (11.28 mg, 87.25 μmol, 15.20 μL) were added sequentially to a 50 mL reaction flask along with DMSO (1 mL). After the reaction was allowed to proceed at room temperature for 10 minutes, compound A-5 (4.89 mg, 34.90 μmol) was added. The reaction was stirred at room temperature for 1 hour, 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 A1 (2.30 mg, 6.20 μmol, 17.12% yield, 81.1% purity) was extracted by MPLC. LC-MS: C 18 H18 N3O4S, [M+H]+ 372.1; found 372.2. 1H NMR (400 MHz, Methanol-d4) δ7.98-7.95 (m, 2H), 7.54-7.49 (m, 2H), 4.44-4.05 (m, 4H), 3.21 (s, 3H), 3.13 (s,3H), 3.03-2.99 (m, 2H).

[0089] Referring to the synthesis method of compound A1, by replacing compound A-1 with raw material 1 from List 1 below, and replacing compound A-2 with raw material 2, while keeping other raw materials and operating methods unchanged, compounds A2-A11 can be obtained.

[0090]

[0091]

[0092]

[0093]

[0094]

[0095] Example 2, Preparation of compound A16

[0096]

[0097] Step 1: Synthesis of Compound A12

[0098]

[0099] Compounds A-16 (20.00 mg, 112.23 μmol), A-5 (15.73 mg, 112.23 μmol), EDCI (42.87 mg, 224.46 μmol), and pyridine (2 mL) were added sequentially to a 50 mL reaction flask. The reaction was stirred for 0.5 hours in an ice bath 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, purified by MPLC, and the solvent was evaporated to dryness. The solvent was then removed by concentration under reduced pressure to obtain compound A12 (10.00 mg, 33.30 μmol, 29.67% yield, 99.9% purity). LC-MS: C 15 H 13 N₂O₃S, [M+H] + 301.1; found 301.2.1 H NMR (600 MHz, DMSO-d6) δ 11.29 (s, 1H), 8.04 (d, J =7.8 Hz, 1H), 7.96 (d, J = 7.8 Hz, 1H), 7.86 (s, 1H), 7.51-7.44 (m, 2H), 4.71(d, J = 9.0 Hz, 1H), 4.61 (d, J = 9.0 Hz, 1H), 4.25 (m, 1H), 4.11 (m, 1H), 3.03 (s, 2H).

[0100] Following the synthetic method for compound A12, compounds A13 and A14 can be obtained by replacing compound A-16 with raw material 1 from List 2 below, while keeping other raw materials and procedures unchanged.

[0101]

[0102] Example 3: Preparation of compound A15

[0103]

[0104] Step 1: Synthesis of Compound A-20

[0105]

[0106] Compound A-19 (155.00 mg, 0.75 mmol), DIPEA (132.25 mg, 1.00 mmol, 178.06 μL), HATU (250.96 mg, 0.66 mmol), and dichloromethane (2 mL) were added sequentially to a 50 mL reaction flask. After the reaction system temperature dropped to 0 °C, compound A-1 (190.00 mg, 0.75 mmol) was added. The reaction was stirred for 0.5 hours under ice bath conditions to quench the reaction (monitored by LC-MS). Extraction was completed with 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 solvent was removed by concentration under reduced pressure to obtain compound A-20 (100.00 mg, crude product).

[0107] Step 2: Synthesis of Compound A-21

[0108]

[0109] Under nitrogen protection, A-20 (117.00 mg, 0.50 mmol), sodium carbonate (91.00 mg, 1.00 mmol), Mo(CO)6 (169.30 mg, 0.75 mmol), Pd(OAc)2 (9.6 mg, 0.05 mmol), and Bu3PBF4 (24.73 mg, 0.10 mmol) were dissolved in DMF / H2O (10 mL, 1:1). After heating at 85 °C for two hours, the solvent was evaporated, and the product was purified by MPLC to obtain A-21 (70.00 mg, 70.35%).

[0110] Step 3: Synthesis of Compound A15

[0111]

[0112] Compounds A-21 (20.00 mg, 100.39 μmol), A-5 (14.07 mg, 100.39 μmol), EDCI (19.17 mg, 100.39 μmol), and pyridine (2 mL) were added sequentially to a 50 mL reaction flask. The reaction was stirred in an ice bath for 0.5 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, purified by MPLC, and the solvent was evaporated to dryness. The solvent was then removed by concentration under reduced pressure to obtain compound A15 (5.00 mg, 15.56 μmol, 15.50% yield, 99.9% purity). LC-MS: C 14 H 16 N3O4S, [M+H] + 322.1; found 322.1. 1 H NMR (400 MHz, DMSO-d6) δ 11.23 (s, 1H), 7.72 (d, J =5.2 Hz, 1H), 7.21 (d, J = 5.2 Hz, 1H), 4.39-4.10 (m, 3H), 4.00 (m, 1H), 3.08-2.76 (m, 8H).

[0113] Example 4: Preparation of compound A16

[0114]

[0115] Step 1: Synthesis of Compound A-23

[0116]

[0117] Compounds A-22 (500.00 mg, 2.64 mmol), DMAP (322.83 mg, 2.64 mmol), and Boc2O (8.64 g, 39.64 mmol) were added sequentially to a 50 mL reaction flask. The reaction mixture was stirred overnight at room temperature. After evaporation of the solvent, the product was purified by MPLC to obtain A-23 (700.00 mg, 2.42 mmol, 91.56% yield).

[0118] Step 2: Synthesis of Compound A-24

[0119]

[0120] Compound A-23 (700.00 mg, 2.42 mmol), NBS (452.15 mg, 2.54 mmol), AIBN (79.46 mg, 483.88 μmol), and carbon tetrachloride (7 mL) were added sequentially to a 50 mL reaction flask. The reaction was refluxed overnight, and after evaporating the solvent, the product was purified by MPLC to obtain A-24 (700.00 mg, 1.90 mmol, 78.57% yield).

[0121] Step 3: Synthesis of Compound A-25

[0122]

[0123] Compounds A-1 (36.73 mg, 814.73 μmol), A-24 (100.00 mg, 271.58 μmol), potassium carbonate (187.39 mg, 1.36 mmol), and DMF (2 mL) were added sequentially to a 50 mL reaction flask. After stirring at 60°C for two hours, A-25 (50.00 mg, 150.42 μmol, 55.39% yield) was obtained by MPLC purification.

[0124] Step 4: Synthesis of Compound A-26

[0125]

[0126] Compound A-25 (50.00 mg, 150.42 μmol), sodium hydroxide (60.17 mg, 1.50 mmol), and MeOH / H2O (2 mL, 1:1) were added sequentially to a 50 mL reaction flask. The mixture was refluxed and stirred for two hours. The pH was adjusted with 1 M HCl aqueous solution, the solvent was evaporated to dryness, and MPLC was used to obtain A-26 (45.00 mg, 141.35 μmol, 93.97% yield).

[0127] Step 5: Synthesis of Compound A-27

[0128]

[0129] Compound A-26 (45.00 mg, 141.35 μmol) was dissolved in hydrochloric acid / dioxane (2 mL). After stirring at room temperature for two hours, the solvent was evaporated to give A-27 (30.00 mg, 137.46 μmol, 97.25% yield).

[0130] Step 6: Synthesis of Compound A16

[0131]

[0132] Compounds A-27 (45.00 mg, 206.18 μmol), A-5 (28.89 mg, 206.18 μmol), EDCI (78.97 mg, 412.37 μmol), and pyridine (2 mL) were added sequentially to a 25 mL reaction flask. The reaction was stirred in an ice bath for 0.5 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, purified by MPLC, and the solvent was evaporated to dryness. The solvent was then removed by concentration under reduced pressure to obtain compound A16 (8.60 mg, 25.27 μmol, 12.25% yield, 98.1% purity). LC-MS: C 18 H 21 N4O3, [M+H] + 341.2; found 341.1 1H NMR (400 MHz, DMSO-d6) δ 7.53 (m, 1H), 7.42 (dd, J = 6.6,2.4 Hz, 1H), 7.14 (m, 2H), 4.17 (d, J = 8.3 Hz, 2H), 4.01 (d, J = 9.2 Hz,2H), 3.78 (s, 2H), 2.97 (s, 2H), 2.22 (s, 6H).

[0133] Following the synthetic method for compound A16, by replacing compound A-22 with the starting materials listed in Table 3 below, while keeping other starting materials and procedures unchanged, compound A17 can be obtained.

[0134]

[0135] Example 5: Preparation of compound A18

[0136]

[0137] Step 1: Synthesis of Compound A-30

[0138]

[0139] Compounds A-29 (1.00 g, 5.20 mmol), A-1 (703.57 mg, 15.61 mmol), EDCI (2.99 g, 15.61 mmol), and pyridine (10 mL) were added sequentially to a 25 mL reaction flask. The reaction was stirred in an ice bath for 0.5 hours, and the reaction was quenched (monitored by LC-MS). Extraction was completed with saturated NaCl solution (10 mL) and ethyl acetate (3 × 20 mL). The organic phases were combined, dried over anhydrous sodium sulfate, purified by MPLC, and the solvent was evaporated to dryness. The solvent was removed by concentration under reduced pressure to obtain compound A-30 (1.13 g, 5.15 mmol, 99.05% yield).

[0140] Step 2: Synthesis of Compound A-31

[0141]

[0142] Compound A-30 (100.00 mg, 455.99 μmol), BPO (22.09 mg, 91.20 μmol), NBS (85.22 mg, 478.79 μmol), and DCM (5 mL) were added sequentially to a 25 mL reaction flask. After stirring overnight, the mixture was purified by MPLC to obtain A-31 (115.00 mg, 385.65 μmol, 84.57% yield).

[0143] Step 3: Synthesis of Compound A18

[0144]

[0145] Compounds A-31 (45.00 mg, 150.91 μmol), A-5 (21.15 mg, 150.91 μmol), sodium bicarbonate (63.38 mg, 754.53 μmol), and DMF (1 mL) were added sequentially to a 25 mL reaction flask. The mixture was stirred overnight at room temperature, and purified by MPLC to obtain A18 (17.85 mg, 49.94 μmol, 33.09% yield, 98.1% purity). LC-MS: C 18 H 21 N4O3, [M+H] + 341.2; found 341.1 1 H NMR (400 MHz, DMSO-d6) δ 7.53 (m, 1H), 7.42 (dd, J = 6.6, 2.4 Hz, 1H), 7.14 (m, 2H), 4.17 (d, J = 8.3 Hz, 2H), 4.01(d, J = 9.2 Hz, 2H), 3.78 (s, 2H), 2.97 (s, 2H), 2.22 (s, 6H).

[0146] Example 6: Preparation of compound A19

[0147]

[0148]

[0149] Step 1: Synthesis of Compound A-33

[0150]

[0151] Compound A-32 (500.00 mg, 2.85 mmol), HATU (1.08 g, 2.85 mmol), DIPEA (368.88 mg, 2.85 mmol, 497.14 μL), and dichloromethane (10 mL) were added sequentially to a 50 mL reaction flask. The mixture was stirred in an ice-water bath for 20 minutes, and then A-1 (128.67 mg, 2.85 mmol) was added. After stirring at room temperature for one hour, the solvent was evaporated, and the mixture was purified by MPLC to obtain A-33 (500.00 mg, 2.47 mmol, 86.62% yield).

[0152] Step 2: Synthesis of Compound A-34

[0153]

[0154] Compound A-33 (500.00 mg, 2.47 mmol), phosphorus oxychloride (1.13 g, 7.41 mmol), and DMF (15 mL) were added sequentially to a 50 mL reaction flask. The mixture was stirred at room temperature for two hours and purified by MPLC to obtain A-34 (430.00 mg, 1.88 mmol, 75.62%).

[0155] Step 3: Synthesis of Compound A-35

[0156]

[0157] Compound A-34 (430.00 mg, 1.88 mmol), sodium dihydrogen phosphate (381.26 mg, 2.44 mmol), hydrogen peroxide (76.70 mg, 2.26 mmol), and chlorite (238.00 mg, 2.63 mmol) were dissolved in acetonitrile / water (10 mL). The mixture was stirred overnight at room temperature. After evaporating the solvent, the solution was purified by MPLC to obtain A-35 (300.00 mg, 1.22 mmol, yield 64.89%).

[0158] Step 4: Synthesis of Compound A19

[0159]

[0160] Compounds A-35 (8.54 mg, 60.91 μmol), A-1 (15.00 mg, 60.91 μmol), EDCI (23.27 mg, 121.82 μmol), and pyridine (2 mL) were added sequentially to a 25 mL reaction flask. The reaction was stirred in an ice bath for 0.5 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, purified by MPLC, and the solvent was evaporated to dryness. The solvent was then removed by concentration under reduced pressure to obtain compound A19 (5.50 mg, 14.93 μmol, 24.51% yield, 98.4% purity). LC-MS: C 19 H 21 N4O4, [M+H] + 369.2; found 369.2 1 H NMR (600 MHz, DMSO-d6) δ 11.19 (s, 1H), 7.69 (d, J = 8.4Hz, 1H), 7.59 (d, J = 8.4 Hz, 1H), 7.31 (t, J = 7.8 Hz, 1H), 7.23 (t, J = 7.8Hz, 1H), 4.25-3.94 (m, 4H), 3.71 (s, 3H), 3.08 (s, 3H), 2.97 (s, 2H), 2.91(s, 3H).

[0161] Example 7: Preparation of compound A20

[0162]

[0163] Step 1: Synthesis of Compound A-37

[0164]

[0165] Compound A-36 (410.00 mg, 2.33 mmol), HATU (1.06 g, 2.79 mmol), DIPEA (902.37 mg, 6.98 mmol, 1.22 mL), and dichloromethane (10 mL) were added sequentially to a 50 mL reaction flask. The mixture was stirred in an ice-water bath for 20 minutes, and then A-1 (128.67 mg, 2.85 mmol) was added. After stirring at room temperature for one hour, the solvent was evaporated, and the mixture was purified by MPLC to obtain A-37 (397.00 mg, 1.95 mmol, 83.93% yield).

[0166] Step 2: Synthesis of Compound A-38

[0167]

[0168] Compound A-37 (560.00 mg, 2.76 mmol), NBS (637.54 mg, 3.58 mmol), AIBN (22.62 mg, 137.77 μmol), and chlorobenzene (10 mL) were added sequentially to a 50 mL reaction flask. The reaction was refluxed overnight, and after evaporating the solvent, A-38 (449.00 mg, 1.59 mmol, 57.76% yield) was purified by MPLC.

[0169] Step 3: Synthesis of Compound A-40

[0170]

[0171] Compounds A-38 (449.00 mg, 1.59 mmol), A-39 (478.13 mg, 6.37 mmol), and DMF (5 mL) were added sequentially to a 50 mL reaction flask. The mixture was stirred overnight at room temperature and purified by MPLC to obtain A-40 (187.00 mg, 860.88 μmol, 54.09% yield).

[0172] Step 4: Synthesis of Compound A-41

[0173]

[0174] Compound A-40 (187.00 mg, 860.88 μmol), sodium dihydrogen phosphate (381.26 mg, 2.44 mmol), hydrogen peroxide (35.14 mg, 1.03 mmol), and sodium chlorite (174.59 mg, 1.12 mmol) were dissolved in acetonitrile / water (10 mL). The mixture was stirred overnight at room temperature. After evaporating the solvent, the solution was purified by MPLC to obtain A-41 (140.00 mg, 600.29 μmol, 69.73% yield).

[0175] Step 5: Synthesis of Compound A20

[0176]

[0177] Compounds A-41 (30.00 mg, 128.63 μmol), A-5 (18.03 mg, 128.63 μmol), EDCI (25.47 mg, 128.63 μmol), and pyridine (3 mL) were added sequentially to a 25 mL reaction flask. The reaction was stirred in an ice bath for 0.5 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, purified by MPLC, and the solvent was evaporated to dryness. The solvent was then removed by concentration under reduced pressure to obtain compound A20 (8.00 mg, 22.51 μmol, 17.50% yield, 99.1% purity). LC-MS: C 18 H 18 N3O5, [M+H] + 355.1; found 355.2 1 H NMR (400 MHz, DMSO-d6) δ 11.23 (s, 1H), 7.78 (d, J = 7.6Hz, 1H), 7.72 (d, J = 8.4 Hz, 1H), 7.50 (t, J = 7.6 Hz, 1H), 7.42 (t, J = 7.6Hz, 1H), 4.35-3.93 (m, 4H), 3.07 (m, 6H), 2.96 (m, 2H).

[0178] The technical effects of this invention are illustrated by the following experimental examples:

[0179] Experimental Example 1: Detection of the inhibitory effect of compounds on CRBN / DDB1 activity (FRET)

[0180] 1. Experimental materials and reagents

[0181] 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).

[0182] 2. Experimental Methods

[0183] 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.

[0184] 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. o Pre-incubate at 250 rpm for 15 minutes at C. Dilute Anti-6His-Tb crypate Gold and FITC-labeled thalidomide analogue to twice the desired final concentration using 20 mM disodium hydrogen phosphate, 20 mM 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, yielding a mixture of Anti-6His-Tb crypate Gold and FITC-labeled thalidomide analogue. Add 5.0 μL of this mixture to a 384-well plate, centrifuge at 1000 rpm for 1 minute, and then incubate on a microplate shaker at 25°C. o Incubate at 250 rpm for 30 minutes at C. After the reaction, read the fluorescence signal values ​​in the 384-well plate using a microplate reader (Ex = 337 nm Em = 520 / 490 nm).

[0185] 3. Data Analysis

[0186] 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.

[0187] The formula for calculating the percentage of remaining activity at each concentration is as follows:

[0188] Remaining vitality (%) = 100% × (Flu)化合物组 -Flu 空白对照 ) / (Flu 阴性对照 -Flu 空白对照 )

[0189] Then, the IC was calculated by fitting the dose-effect curve using GraphPad 6.0. 50 value.

[0190]

[0191] Wherein, + represents 200 μM>IC50>100 μM, ++ represents 100 μM>IC50>10 μM, +++ represents 10 μM>IC50>1 μM, ++++ represents 1 μM>IC50>0.1 μM, and ++++ represents IC50<0.1 μM.

[0192] Experimental Example 2: Detection of the compound with CRBN / DDB1 using ITC (isothermal titration calorimetry).

[0193] 1. Experimental materials and reagents

[0194] Disodium hydrogen phosphate (Sigma), sodium dihydrogen phosphate (Sigma), Tween 20 (Sigma), dimethyl sulfoxide (Sigma), desalting column (Thermo Scientific, #89882), microplate reader (BMG PHERAstar FSX), CRBN / DDB1 protein (HitGen), MicroCal PEAQ-ITC (Malvern).

[0195] 2. Experimental Methods

[0196] Mix 6.1 mL of 200 mM disodium hydrogen phosphate aqueous solution with 3.9 mL of 200 mM sodium dihydrogen phosphate aqueous solution to obtain 200 mM PB buffer (pH 7.0). Take 600 μL of 200 mM PB buffer and 3 μL of 10% Tween 20 aqueous solution, add to 5397 μL of deionized water, and mix well to obtain a detection buffer with pH 7.0.

[0197] Using the detection buffer solution prepared above, press the desalting column Zeba TM Follow the instructions in the Spin Desalting Columns instructions (Thermo Scientific, #89882) to perform buffer replacement on the protein storage solution; after replacement, use a microplate reader (BMG PHERAstar FSX) to measure the OD value of the protein solution. 280nmThe UV absorbance was measured, and the concentration after displacement was calculated. Based on the measured concentration, the protein was diluted to 10 μM with detection buffer, and DMSO was added to a final concentration of 1%, for a total volume of 300 μL. After short-term centrifugation at room temperature, the protein was ready for use.

[0198] Dissolve the dry compound powder in DMSO, then dilute the compound to 100 μM with the above-mentioned detection buffer, and adjust the DMSO to a final concentration of 1%, for a total volume of 100 μL. Centrifuge the compound solution at 15,000 rpm for 5 minutes at room temperature, and then collect at least 75 μL of the supernatant for later use.

[0199] Clean the MicroCal PEAQ-ITC ITC instrument according to the instrument's procedure. After cleaning, perform a water droplet test to check the instrument's cleanliness and condition. During sample titration, add the protein sample to the sample cell and the compound solution to the titrator. Set the instrument temperature to 25°C, Reference Power (ucal / s) to 5.00, feedback mode to "High," and stirring speed to 750 rpm. Perform 19 drops in total. Discard the first drop and continue titrating the remaining 18 drops at 2 µL / 4 s intervals, equilibrating for 150 s between each drop. After titration, clean the instrument. Perform the water droplet test again to ensure the instrument is clean and in good condition. Then, perform the control titration. Prepare the compound as described above and add it to the titrator; separately prepare 300 µL of detection buffer containing 1% DMSO and add it to the sample cell. Set the instrument parameters as above and perform the titration.

[0200] 3. Data Analysis

[0201] Using the ITC instrument's built-in analysis software, the "one set of sites" mode was used for fitting. After subtracting the background from the control experiment, the reaction-related parameters, such as N, KD, δH, δG, and δS, were obtained.

[0202]

[0203] The above experiments show that compound A of the present invention has good CRBN binding ability and inhibitory effect, and can be effectively used for the treatment of diseases related to abnormal CRBN activity.

Claims

1. The compound represented by Formula I, or its stereoisomer, or its pharmaceutically acceptable salt: Formula I in, This indicates the presence of oxygen substitution; Ring A is selected from , , ; R A1 Selected from hydrogen, halogen, cyano, nitro, =O, =S, -C 1~3 Alkyl, halogen-substituted -C 1~3 alkyl; R 2 Independently selected , , , .

2. The compound according to claim 1, characterized in that: The compound is specifically: 、 A3、 A7、 A8、 A9、 A11、 、 。 3. Use of the compound of claim 1 or claim 2, or a stereoisomer thereof, or a pharmaceutically acceptable salt thereof, in the preparation of a medicament for treating diseases related to abnormal cell proliferation.

4. The use according to claim 3, characterized in that: The disease in question is cancer.

5. Use of the compound of claim 1 or claim 2, or its stereoisomer, or its pharmaceutically acceptable salt, in the preparation of a drug for targeting protein degradation.

6. Use of the compound of claim 1 or claim 2, or its stereoisomer, or its pharmaceutically acceptable salt, as an intermediate in the preparation of a drug targeting protein degradation.

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