Amide compound and use thereof

Abstract

The invention provides compounds shown as a formula (I) and a formula (II) and / or pharmaceutically acceptable salts, solvates, optical isomers, stereoisomers, polymorphs or isotope-enriched compounds thereof, a pharmaceutical composition containing the compounds, and application of the compounds and the pharmaceutical composition thereof in preparation of drugs.

Description

Technical Field

[0001] This invention belongs to the pharmaceutical field, specifically relating to amide compounds, pharmaceutical compositions comprising the same, and the use of the compounds and pharmaceutical compositions thereof. Background Technology

[0002] TRIM21, belonging to the TRIM family of E3 ligases, participates in the regulation of multiple cellular signaling pathways. Studies have shown that TRIM21 is associated with autoimmune diseases and inflammation, while other studies have indicated that it also participates in regulating tumor development and prognosis.

[0003] Therefore, developing TRIM21 small molecule ligands and using them to create targeted protein degradation compounds has many potential applications in the pharmaceutical field. Summary of the Invention

[0004] In a first aspect, the present invention provides a compound of formula (I), or a pharmaceutically acceptable salt, solvate, optical isomer, stereoisomer, polymorph, or isotope enrichment thereof:

[0005]

[0006] in,

[0007] Ring A is selected from 6-10 membered aromatic rings, 5-10 membered aromatic heterocycles, 3-10 membered cycloalkyl groups, and 3-10 membered heterocycles; ring A may optionally be substituted with one, two, three, or four independent R groups, wherein each R group is independently selected from hydrogen, halogen, cyano, nitro, substituted or unsubstituted C groups. 1-6 Alkyl, substituted or unsubstituted C 2-6 alkenyl, substituted or unsubstituted C 2-6 alkynyl group, -C 0-4 Alkylene-OR a -C 0-4 Alkylene-OC(=O)R a -C 0-4 Alkylene-SR a -C 0-4 Alkylene-S(=O)2R a -C 0-4 Alkylene-S(=O)R a -C 0-4 Alkylene-S(=O)2NR a R b -C 0-4 Alkylene-S(=O)NR a R b -C 0-4 Alkylene-C(=O)R a -C 0-4 Alkylene-C(=O)ORa -C 0-4 Alkylene-C(=O)NR a R b -C 0-4 Alkylene-NR a R b -C 0-4 Alkylene-NR a C(=O)R b -C 0-4 Alkylene-NR a S(=O)2R b -C 0-4 Alkylene-NR a S(=O)R b -C 0-4 Alkylene (3- to 10-membered cycloalkyl), -C 0-4 Alkylene (3- 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); wherein R a R b Each is independently selected from hydrogen and C. 1-6 Alkyl, C 2-6 alkenyl, C 2-6 Alkyne- and halogen-substituted C 1-6 Alkyl, halogen-substituted C 2-6 Alkenyl, halogen-substituted C 2-6 alkynyl group, -C 0-4 Alkylene (3- to 10-membered cycloalkyl), -C 0-4 Alkylene (3- 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);

[0008] X1, X2, and X3 are each independently selected from CH or N;

[0009] R1 is selected from -S(=O)2R 1a -P(=O)R 1a R 1b -C(=O)R 1a -S(=O)(=NH)R 1a -S(=O)(=NR) 1a )R 1b -N = S(=O)R 1a R 1b ;where R 1a and R 1b Each is independently selected from substituted or unsubstituted C 1-6 Alkyl, or R1a and R 1b Together with the atoms to which it is attached, it forms a 4- to 6-membered ring; the condition is that R1 is selected from -S(=O)2R 1a At that time, X1, X2, and X3 are not all CH at the same time;

[0010] R2 and R3 are each independently selected from hydrogen, -OR 2a C, substituted or unsubstituted 1-6 Alkyl, substituted or unsubstituted C 2-6 alkenyl, substituted or unsubstituted C 2-6 alkynyl group, -C 0-4 Alkylene-OR 2a -C 0-4 Alkylene-OC(=O)R 2a -C 0-4 Alkylene-SR 2a -C 0-4 Alkylene-S(=O)2R 2a -C 0-4 Alkylene-S(=O)R 2a -C 0-4 Alkylene-S(=O)2NR 2a R 2b -C 0-4 Alkylene-S(=O)NR 2a R 2b -C 0-4 Alkylene-C(=O)R 2a -C 0-4 Alkylene-C=(O)OR 2a -C 0-4 Alkylene-C(=O)NR 2a R 2b -C 0-4 Alkylene-NR 2a R 2b -C 0-4 Alkylene-NR 2a C(=O)R 2b -C 0-4 Alkylene-NR 2a S(=O)2R 2b -C 0-4 Alkylene-NR 2a S(=O)R 2b -C 0-4 Alkylene (3- to 10-membered cycloalkyl), -C 0-4 Alkylene (3- 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), wherein R 2a R2b Each is independently selected from hydrogen and C. 1-6 Alkyl, C 2-6 alkenyl, C 2-6 Alkyne- and halogen-substituted C 1-6 Alkyl, halogen-substituted C 2-6 Alkenyl, halogen-substituted C 2-6 Alkyne group.

[0011] In some embodiments, ring A is selected from 6- to 10-membered aromatic rings (such as phenyl, naphthyl), and ring A is optionally substituted with two independent Rs, wherein each R is independently selected from halogens (e.g., F, Cl, Br, or I), substituted or unsubstituted Cs. 1-6 Alkyl groups (e.g., methyl, ethyl, propyl, isopropyl, butyl), -C 0-4 Alkylene-OR a (e.g., -OH, -OCH3, -CH2OCH3, -CH2OCH2CH3, -OCH2CH3, -OCH2CH2CH3), where R a Independently selected from hydrogen and C 1-6 alkyl.

[0012] In some embodiments, ring A is selected from 5- to 10-membered aromatic heterocycles (e.g., pyrrole, thiophene, imidazole, oxazole, furan, thiazole, pyran ring, etc.), and ring A is optionally substituted by one or two independent Rs, wherein each R is independently selected from halogens (F, Cl, Br, or I), substituted or unsubstituted Cs. 1-6 Alkyl groups (e.g., methyl, ethyl, propyl, isopropyl, butyl), -C 0-4 Alkylene-OR a (e.g., -OH, -OCH3, -CH2OCH3, -CH2OCH2CH3, -OCH2CH3, -OCH2CH2CH3), where R a Independently selected from hydrogen and C 1-6 alkyl.

[0013] In some embodiments, ring A is selected from 3- to 10-membered cycloalkyl groups (e.g., cyclopropyl, cyclobutyric, cyclopentyl, cyclohexyl, etc.), and ring A is optionally substituted by one or two independent Rs, wherein each R is independently selected from a halogen (F, Cl, Br, or I), a substituted or unsubstituted C. 1-6 Alkyl groups (e.g., methyl, ethyl, propyl, isopropyl, butyl), -C 0-4 Alkylene-OR a (e.g., -OH, -OCH3, -CH2OCH3, -CH2OCH2CH3, -OCH2CH3, -OCH2CH2CH3), where R a Independently selected from hydrogen and C 1-6 alkyl.

[0014] In some embodiments, ring A is selected from 3- to 10-membered heterocycles (e.g., morpholine, pyrrolidine, tetrahydrothiophene, oxetane, etc.), and ring A is optionally substituted by one or two independent Rs, wherein each R is independently selected from a halogen (F, Cl, Br, or I), substituted or unsubstituted C. 1-6 Alkyl groups (e.g., methyl, ethyl, propyl, isopropyl, butyl), -C 0-4 Alkylene-OR a (e.g., -OH, -OCH3, -CH2OCH3, -CH2OCH2CH3, -OCH2CH3, -OCH2CH2CH3), where R a Independently selected from hydrogen and C 1-6 alkyl.

[0015] In some implementations, R2 and R3 are each independently selected from C. 1-6 Alkyl (e.g., methyl, ethyl, propyl, etc.) or -C 0-4 alkylene-(5-10 membered heteroaryl rings) (e.g.) ).

[0016] In some embodiments, the compound has the structure shown in formula (Ia):

[0017]

[0018] Wherein, R1 is selected from -P(=O)R 1a R 1b -C(=O)R 1a -S(=O)(=NH)R 1b -S(=O)(=NR) 1a )R 1b -N = S(=O)R 1a R 1b ;where R 1a and R 1b Each is independently selected from substituted or unsubstituted C 1-6 Alkyl, or R 1a and R 1b Together with the atoms to which they are attached, they form 4- to 6-membered rings. In some embodiments, the compound has the structure shown in formula (IIa):

[0019]

[0020] In this context, R is independently selected from: halogens, such as fluorine, chlorine, bromine, and iodine; C 1-6 Alkyl groups, such as methyl, ethyl, propyl, isopropyl; -OR a , where R a Independently selected from hydrogen, methyl, ethyl, and propyl;

[0021] R1 is selected from -S(=O)(=NH)R 1a -S(=O)(=NR) 1a )R 1b -N = S(=O)R 1a R 1b , where R 1a and R 1b C independently 1-6 Alkyl groups, such as methyl, ethyl, propyl, cyclopropyl, isopropyl, or R 1a and R 1b Together with the atoms to which it is attached, it forms a 4- to 6-membered ring; R1 is selected from, for example, -C(=O)CH3, -C(=O)CF3, -P(=O)CH3CH3, -S(=O)(=NH)CH3, -S(=O)(CH3)(CH3), -N=S(=O)(CH3)(CH2CH3), and

[0022] R2 and R3 are each independently selected from: C 1-6 Alkyl groups, such as methyl, ethyl, propyl, isopropyl; -C 1-4 alkylene rings (5- to 10-membered heteroaryl rings), for example

[0023] In some embodiments, the compound has the structure of formula (IIa1):

[0024]

[0025] In this context, R is independently selected from: halogens, such as fluorine, chlorine, bromine, and iodine; -OR a , where R a R1 is independently selected from hydrogen, methyl, ethyl, and propyl; R1 is selected from -N=S(=O)R 1a R 1b or -S(=O)(=NR) 1a )R 1b , where R 1a and R 1b C independently 1-6 Alkyl groups, such as methyl, ethyl, propyl, cyclopropyl, isopropyl, or R 1a and R 1b Together with the atoms to which it is attached, it forms a 4- to 6-membered ring; R3 is selected from C. 1-6 Alkyl groups, such as methyl, ethyl, propyl, and isopropyl.

[0026] In some embodiments, the compound has the structure shown in formula (Ib):

[0027]

[0028] Wherein, R1 is selected from -S(=O)2R 1a -C(=O)R 1a -P(=O)R 1a R 1b -S(=O)(=NH)R 1b -S(=O)(=NR) 1a )R 1b -N = S(=O)R 1a R 1b ;where R 1a and R 1b Each is independently selected from substituted or unsubstituted C 1-6 Alkyl, or R 1a and R 1b Together with the atoms attached to it, they form 4 to 6-membered rings.

[0029] In some embodiments, the compound has the structure shown in formula (IIb):

[0030]

[0031] In this context, R is independently selected from: halogens, such as fluorine, chlorine, bromine, and iodine; C 1-6 Alkyl groups, such as methyl, ethyl, propyl, cyclopropyl, isopropyl; -OR a , where R a Independently selected from hydrogen, methyl, ethyl, cyclopropyl, isopropyl, and propyl;

[0032] R1 is selected from -S(=O)2R 1a -P(=O)R 1a R 1b -S(=O)(=NR) 1a )R 1b -N = S(=O)R 1a R 1b ;That

[0033] Chinese R 1a and R 1b Each was independently selected from C 1-6 Alkyl groups, such as methyl, ethyl, propyl, isopropyl, or R 1a and R 1b Together with the atoms to which it is attached, it forms a 4- to 6-membered ring; R1 is selected from, for example, -S(=O)2CH3, -C(=O)CH3, -C(=O)CF3, -P(=O)CH3CH3, -S(=O)(=NH)CH3, -S(=O)(CH3)(CH3), -N=S(=O)(CH3)(CH2CH3), and

[0034] R2 and R3 are each independently selected from: C 1-6 Alkyl groups, such as methyl, ethyl, propyl, isopropyl; -C 1-4 alkylene rings (5- to 10-membered heteroaryl rings), for example

[0035] In some embodiments, the compound has the structure shown in formula (Ic):

[0036]

[0037] Wherein, R1 is selected from -S(=O)2R 1a -C(=O)R 1a -P(=O)R 1a R 1b -S(=O)(=NH)R 1b -S(=O)(=NR) 1a )R 1b -N = S(=O)R 1a R 1b ;where R 1a and R 1b Each is independently selected from substituted or unsubstituted C 1-6 Alkyl, or R 1a and R 1b Together with the atoms attached to it, they form 4 to 6-membered rings.

[0038] In some embodiments, the compound has the structure shown in formula (IIc):

[0039]

[0040] In this context, R is independently selected from: halogens, such as fluorine, chlorine, bromine, and iodine; C 1-6 Alkyl groups, such as methyl, ethyl, propyl, cyclopropyl, isopropyl; -OR a , where R a Independently selected from hydrogen, methyl, ethyl, and propyl;

[0041] R1 is selected from -S(=O)2R 1a -N = S(=O)R 1a R 1b ;where R 1a and R 1b Each was independently selected from C 1-6 Alkyl groups, such as methyl, ethyl, propyl, cyclopropyl, isopropyl, or R 1a and R 1bTogether with the atoms to which it is attached, it forms a 4- to 6-membered ring; R1 is selected from, for example, -S(=O)2CH3, -C(=O)CH3, -C(=O)CF3, -P(=O)CH3CH3, -S(=O)(=NH)CH3, -S(=O)(CH3)(CH3), -N=S(=O)(CH3)(CH2CH3),

[0042] R2 and R3 are each independently selected from: C 1-6 Alkyl groups, such as methyl, ethyl, cyclopropyl, propyl, isopropyl; -C 1-4 alkylene rings (5- to 10-membered heteroaryl rings), for example

[0043] In some embodiments, the compound has the structure shown in formula (Id):

[0044]

[0045] Wherein, R1 is selected from -S(=O)2R 1a -C(=O)R 1a -P(=O)R 1a R 1b -S(=O)(=NH)R 1b -S(=O)(=NR) 1a )R 1b -N = S(=O)R 1a R 1b ;where R 1a Each is independently selected from substituted or unsubstituted C 1-6 alkyl.

[0046] In some embodiments, the compound has the structure shown in formula (IId):

[0047]

[0048] In this context, R is independently selected from: halogens, such as fluorine, chlorine, bromine, and iodine; C 1-6 Alkyl groups, such as methyl, ethyl, propyl, isopropyl; -OR a , where R a Independently selected from hydrogen, methyl, ethyl, and propyl;

[0049] R1 is selected from -S(=O)2R 1a -C(=O)R 1a ;where R 1a Each was independently selected from C 1-6 Alkyl groups, such as methyl, ethyl, cyclopropyl, propyl, and isopropyl;

[0050] R2 and R3 are each independently selected from: C 1-6 Alkyl groups, such as methyl, ethyl, propyl, cyclopropyl, isopropyl; -C 1-4 alkylene rings (5- to 10-membered heteroaryl rings), for example

[0051] In some embodiments, compounds selected from the following, or their pharmaceutically acceptable salts, solvates, optical isomers, stereoisomers, polymorphs, or isotope enrichments, are used:

[0052]

[0053]

[0054]

[0055] In a second aspect, the present invention provides a compound of formula (II), or a pharmaceutically acceptable salt, solvate, optical isomer, stereoisomer, polymorph, or isotope enrichment thereof:

[0056]

[0057] Wherein, R1, R2, X1, X2, X3, and ring A can be applied to any of the limitations in the first aspect of the present invention;

[0058] L represents a linking group, typically a subunit containing heteroatoms such as nitrogen or oxygen, like PEG, saturated or unsaturated alkylene groups, or subunits containing aromatic rings, heteroaromatic rings, saturated or unsaturated cyclic alkylene groups, or saturated or unsaturated heterocyclic alkylene groups. The L structure selectively incorporates heteroatoms and cyclic structures to regulate the physicochemical properties and overall pharmacological efficacy of the molecule.

[0059] P represents the ligand group that targets and degrades the target protein. The target protein can be a pathogenic protein, including but not limited to proteins that cause diseases such as tumors, immune disorders, inflammation, and central nervous system disorders.

[0060] Among them, those skilled in the art can select or design L and P and their binding sites based on their specific understanding of the relevant disease protein targets.

[0061] Thirdly, the present invention provides a pharmaceutical composition comprising the compounds described above, and a pharmaceutically acceptable carrier.

[0062] Fourthly, the present invention provides the use of the compounds represented by Formula I or Formula II above, or pharmaceutically acceptable salts, solvates, optical isomers, stereoisomers, polymorphs or isotope enrichments thereof, in the preparation of pharmaceuticals, including but not limited to the use of the above compounds and derivatives, or the above pharmaceutical compositions, to induce intracellular TRIM21-dominated target protein degradation.

[0063] In some embodiments, the above-mentioned drugs are used to treat or prevent diseases or conditions related to TRIM21-dominant target proteins, including but not limited to tumors or cancers, inflammation, immune system diseases, neurodegenerative diseases, etc.

[0064] Fifthly, the present invention provides the application of the above-mentioned compounds, or their chemical synthetic routes, or their synthetic intermediates, in drug synthesis.

[0065] In a sixth aspect, the present invention provides a method for treating or preventing diseases or conditions related to TRIM21-dominant target proteins, comprising administering an effective amount of the compounds and / or pharmaceutical compositions described above to a subject, thereby achieving the effect of treating the related diseases.

[0066] In addition, this application also provides a kit comprising any of the compounds described above or pharmaceutically acceptable salts, solvates or stereoisomers, polymorphs, isotope enrichments or any of the compositions described above, which can be used to prepare a medicament for treating, inhibiting or preventing diseases or conditions related to the TRIM21-dominant target protein. Detailed Implementation

[0067] definition

[0068] To provide a clear and consistent understanding of the terminology used in this specification, some definitions are provided below. Furthermore, unless otherwise specified, all technical and scientific terms used in this invention have the same meaning as commonly understood by one of ordinary skill in the art to which this invention pertains.

[0069] When used in conjunction with the term "comprising" in the claims and / or description, the word "a" can mean "one / a type," but it also means "one / a type or more / a types," "at least one," and "one or more." Similarly, the word "another" can mean at least a second or more.

[0070] The terms “comprising,” “having,” “including,” and “containing” as used in this specification and claims are inclusive and open-ended and do not exclude additional unlisted elements or processing steps.

[0071] The term “about” is used to indicate that the value includes errors introduced by the instruments and methods used in determining the value.

[0072] As used herein, the term "substituted" or "having a substituent" means that a parent compound or part has at least one substituent group. The term "unsubstituted" or "not having a substituent" means that a parent compound or part does not have any other substituents except for an undetermined valence chemically saturated with hydrogen atoms. As described herein, "substituent" or "substituent group" means selected from halogens (F, Cl, Br, or I), hydroxyl, mercapto, amino, nitro, carbonyl, carboxyl, alkyl, alkoxy, alkylamino, aryl, aryloxy, arylamino, acyl, thionyl, sulfonyl, phosphonyl, or other organic moieties conventionally used and accepted in organic chemistry.

[0073] As used herein, the term "alkyl" refers to a saturated hydrocarbon having 1 to 6 carbon atoms, including straight-chain, branched, and cyclic alkyl groups. Examples of alkyl groups include, but are not limited to, methyl, ethyl, propyl, butyl, pentyl, hexyl, cyclopropyl, cyclopentyl, cyclohexyl, etc. The term alkyl includes both unsubstituted and substituted alkyl groups. The term "C1-C..." n Alkyl (where n is an integer from 2 to 6) means an alkyl group having 1 to the number of carbon atoms indicated by "n". Alkyl residues may be substituted or unsubstituted. In some embodiments, for example, the alkyl group may be substituted with groups such as hydroxyl, halogen (F, Cl, Br or I), amino, etc.

[0074] As used herein, the term "3- to 10-membered cycloalkyl" refers to a group having 3 to 10 carbon atoms (e.g., C36, C46, ​​C56, C6 ... 3-8 C 3-6 Or C 4-8 Cycloalkyl refers to monocyclic or polycyclic (e.g., fused ring, bridged ring, or spirocyclic) systems of saturated or unsaturated non-aromatic hydrocarbons. Examples of cycloalkyl groups include, but are not limited to, cyclopropyl, cyclobutyl, cyclopentyl, cyclohexyl, cyclopentenyl, and cyclohexenyl.

[0075] As used herein, the term "alkenyl" includes unsaturated aliphatic groups similar in length and possible substitution to the alkyl groups described above but containing at least one double bond. For example, the term "alkenyl" includes straight-chain alkenyl groups (e.g., vinyl, propenyl, butenyl, pentenyl, hexenyl) and branched alkenyl groups. In some embodiments, straight-chain or branched alkenyl groups have six or fewer carbon atoms in their backbone (e.g., C2-C6 for straight chains and C3-C6 for branched chains). The term "C 2-6 "Alkenyl" includes alkenyl groups containing 2 to 6 carbon atoms. The term "C" 3-6 "Alkenyl" includes alkenyl groups containing 3 to 6 carbon atoms.

[0076] As used herein, the term "alkynyl" includes unsaturated aliphatic groups similar in length and possible substitutions to the alkyl groups described above but containing at least one triple bond. For example, "alkynyl" includes straight-chain alkynyl groups (e.g., ethynyl, propynyl, butynyl, pentyynyl, hexynyl) and branched-chain alkynyl groups. In some embodiments, straight-chain or branched alkynyl groups have six or fewer carbon atoms in their main chain (e.g., C2-C6 for straight chains and C3-C6 for branched chains). A 6-carbon alkynyl group.

[0077] As used herein, the term "6- to 10-membered aromatic ring" refers to an aromatic group having "4n+2" (π) electrons and 6 to 10 ring atoms in a conjugated monocyclic or polycyclic system (fused or unfused), where n is an integer from 1 to 2. Aryl groups can be directly linked or linked via C1-C3 alkyl groups (also called arylalkyl or aralkyl groups). Examples of aryl groups include, but are not limited to, phenyl, benzyl, phenethyl, 1-phenylethyl, tolyl, naphthyl, etc.

[0078] As used herein, the term "aromatic heterocycle" refers to an aryl group as defined above, but with one to four heteroatoms (e.g., N, O, and S) in a ring structure. As used herein, the term "aromatic heterocycle" is intended to include stable 5-, 6-, or 7-membered monocyclic or 7-, 8-, 9-, 10-, 11-, or 12-membered bicyclic aromatic heterocycles composed of a carbon atom and one or more heteroatoms independently selected from the group consisting of nitrogen, oxygen, and sulfur (e.g., 1 or 1-2 or 1-3 or 1-4 heteroatoms). The nitrogen atom may be substituted or unsubstituted (i.e., N or NR, where R is H or other substituents as defined). The nitrogen and sulfur heteroatoms may optionally undergo oxidation (i.e., N → O and S(O)). p (where p = 1 or 2).

[0079] As used herein, the term "heterocycle" refers to any ring structure (saturated or unsaturated) containing at least one cyclic heteroatom (e.g., 1-4 heteroatoms selected from N, O, and S). Examples of heterocycles include, but are not limited to, morpholine, pyrrolidine, tetrahydrothiophene, piperidine, piperazine, oxobutane, pyran, tetrahydropyran, azabutane, and tetrahydrofuran.

[0080] As used in this article, “halogenated” or “halogen” refers to fluorine, chlorine, bromine and iodine, encompassing “partial halogenation” and “full halogenation”, where some or all of the hydrogen atoms in the group are replaced by halogen atoms.

[0081] As used in this article, the term "C" 0-4 "alkylene" indicates the absence of the alkylene group and C 1-4 The case of alkylene compounds (e.g., methylene, ethylene, propylene, butylene, etc.).

[0082] In this specification, for convenience, in some cases the structural formula of a compound represents a particular isomer, but the invention includes all isomers, such as geometric isomers, optical isomers based on asymmetric carbon, stereoisomers, tautomers, etc. Furthermore, compounds represented by general formulas can exist as crystalline polymorphs. It should be noted that any crystalline form, mixture of crystalline forms, or its anhydrous form or hydrate is included within the scope of this invention. The terms "crystalline polymorph," "polymorph," or "crystalline form" refer to crystalline structures in which the compound (or its salts or solvates) can crystallize in different crystalline stacks, all of which have the same elemental composition. Different crystalline forms typically have different X-ray diffraction patterns, infrared spectra, melting points, densities, hardness, crystal shapes, optical and electrical properties, stability, and solubility. Recrystallization solvents, crystallization rates, storage temperatures, and other factors can cause one crystalline form to dominate. Crystalline polymorphs of compounds can be prepared by crystallization under different conditions.

[0083] A “pharmaceutically acceptable salt” of a compound refers to a salt of a pharmaceutically acceptable compound. Ideally, a salt of a compound (basic, acidic, or charged functional group) should retain or improve the biological activity and properties of the parent compound as defined in this invention, and should not be biologically undesirable. Examples of pharmaceutically acceptable salts include those mentioned by Berge et al. in “Pharmaceutical Salts”, J. Pharm. Sci. 66, 1-19 (1977), including but not limited to:

[0084] (1) An acid addition salt formed by adding an acid to a basic or positively charged functional group, wherein inorganic acids such as hydrochloric acid, hydrobromic acid, hydroiodic acid, sulfuric acid, aminosulfonic acid, nitric acid, phosphoric acid, and carbonates may be added; or organic acids such as acetic acid, propionic acid, lactic acid, oxalic acid, glycolic acid, neopentanoic acid, tert-butylacetic acid, β-hydroxybutyric acid, valeric acid, hexanoic acid, cyclopentanepropionic acid, pyruvic acid, malonic acid, succinic acid, malic acid, fumaric acid, tartaric acid, citric acid, benzoic acid, 3-(4-hydroxybenzoyl)benzoic acid, cinnamic acid, and succinic acid may be added. Peach acid, mesylate, ethanesulfonic acid, 1,2-ethanedisulfonic acid, 2-hydroxyethanesulfonic acid, cyclohexylaminosulfonic acid, benzenesulfonic acid, sulfanilic acid, 4-chlorobenzenesulfonic acid, 2-naphthalenesulfonic acid, 4-toluenesulfonic acid, camphorsulfonic acid, 3-phenylpropionic acid, laurylsulfonic acid, laurylsulfonic acid, oleic acid, palmitic acid, stearic acid, lauric acid, primordic acid, pantothenic acid, lactobionic acid, alginic acid, galactobionic acid, galacturonic acid, gluconic acid, glucoheponic acid, glutamic acid, naphthoic acid, hydroxynaphthoic acid, salicylic acid, ascorbic acid, stearic acid, mucoconic acid, etc.

[0085] (2) When the parent compound contains an acidic proton or is replaced by a metal ion, an alkali addition salt is obtained by adding a base; wherein the metal ion includes basic metal ions (e.g., lithium, sodium, potassium), alkaline earth metal ions (magnesium, calcium, barium) or other metal ions such as aluminum, zinc, iron, etc.; or coordinated with an organic base, wherein the organic base is such as ammonia, ethylamine, diethylamine, N,N'-dibenzylethylenediamine, ethanolamine, diethanolamine, triethanolamine, aminobutanetriol, N-methylglucosamine, piperazine, chloroprocaine, procaine, choline, lysine, etc.

[0086] Pharmaceutically acceptable salts can be synthesized from parent compounds containing basic or acidic fragments using conventional chemical methods. Typically, such salts are prepared by reacting a compound (free acid or base) with an equistoichiometric base or acid in water, an organic solvent, or a mixture of both. Salts can be prepared in situ during the final separation or purification of the pharmaceutical preparation, or by reacting a purified compound of the invention in its free acid or base form separately with the desired corresponding base or acid and then separating the resulting salt. The term "pharmaceutically acceptable salt" also includes zwitterionic compounds containing cationic groups covalently bonded to anionic groups, which are referred to as "internal salts." It should be understood that all acidic, saltic, basic, and other ionic and nonionic forms of the compounds of the present invention are covered within the scope of the present invention. For example, if the compound of the present invention is an acid, the saltic form of that compound is also covered within the scope of the present invention. Similarly, if the compound of the present invention is a salt, the acidic and / or basic forms of that compound are also covered within the scope of the present invention.

[0087] As used in this invention, the term "isotope enrichment" refers to the enrichment or increase of one or more specific isotopes in a compound. Typically, in an isotope-enriched compound or derivative, a specific isotopic element at a specific position in the compound is enriched or increased. However, it should be understood that a compound may have two or more isotopic elements enriched or increased, including different isotopes of the same element and individual isotopes of different elements. Furthermore, isotope-enriched compounds can be a mixed form of isotope enrichment, i.e., containing multiple specific isotopes or elements, or both. Typically, the isotopes that can be used include... 1 H, D, T, 18 O、 17 O、 15 N and 13 C, etc.

[0088] "Pharmaceutical acceptable" means that the drug, pharmaceutical product, inert ingredient, etc., described by this term are suitable for contact with human and animal cells or tissues without adverse toxicity, incompatibility, instability, irritation, allergic reactions, etc., and are commensurate with a reasonable benefit / risk ratio. It typically refers to compounds or compositions approved or permitted by the government, or listed in recognized pharmacopoeias for use in animals, and more particularly for humans.

[0089] "Pharmaceutically acceptable carrier" refers to a diluent, adjuvant, excipient, carrier, or loading agent that is administered together with a compound.

[0090] "Pharmaceutical composition" means comprising compounds as described herein, and at least one component depending on the requirements of the route of administration and dosage form, including pharmaceutically acceptable carriers, diluents, adjuvants, excipients or loading agents, such as preservatives, fillers, disintegrants, wetting agents, emulsifiers, suspending agents, sweeteners, flavoring agents, aromatizers, antimicrobial agents, antifungal agents, lubricants and dispersants.

[0091] In some embodiments, the term "prevention" means at least a reduction in the likelihood of acquiring a disease or condition (or susceptibility) (i.e., causing at least one clinical symptom of a disease that has not yet developed in a patient, who may be exposed to or susceptible to the disease but has not yet experienced or exhibited symptoms of the disease). The term "treatment" means the relief of at least one disease or condition. In some embodiments, "treatment" means the relief of at least one bodily parameter, which may be identifiable or indistinguishable by the patient. In some embodiments, "treatment" means the suppression of a disease or condition physically (e.g., stabilization of identifiable symptoms) or physiologically (e.g., stabilization of bodily parameters) or both. In some embodiments, "treatment" means the improvement of quality of life or a side effect of a disease in a subject in need. "Effective amount" means an amount of compound administered to a subject for the treatment or prevention of a disease that is sufficient to achieve the effect of treating or preventing the disease. "Effective amount" will vary depending on the compound; the disease and its severity; the age, weight, etc., of the subject being treated or prevented from having the disease. As used herein, "effective amount" means a compound or composition sufficient to prevent, treat, inhibit, reduce, alleviate, or eliminate a disease, such as one or more causes, symptoms, or complications of cancer.

[0092] A “pharmaceutical composition” is a formulation containing a compound disclosed herein, in a form suitable for administration to a subject. In some embodiments, the pharmaceutical composition is in bulk or in unit dosage form. Unit dosage form is any of a variety of forms, including, for example, capsules, IV bags, tablets, single pumps or vials on aerosol inhalers. The amount of active ingredient (e.g., a formulation of the disclosed compound or its salts, hydrates, solvates or isomers) in a unit dose composition is an effective amount and varies depending on the specific treatment involved. Those skilled in the art will recognize that it is sometimes necessary to routinely vary the dosage according to the patient’s age and condition. The dosage will also depend on the route of administration, such as oral, pulmonary, rectal, parenteral, percutaneous, subcutaneous, intravenous, intramuscular, intraperitoneal, inhalation, buccal, sublingual, intrapleural, intrathecal, intranasal, etc. Dosage forms for topical or percutaneous administration of the compounds of the present invention include powders, sprays, ointments, pastes, creams, lotions, gels, solutions, patches, and inhalers.

[0093] Pharmaceutical compositions containing the compounds of this application can be prepared in conventionally known ways, such as by conventional mixing, dissolving, granulation, sugar coating, grinding, emulsification, encapsulation, embedding, or lyophilization processes. The pharmaceutical compositions can be formulated in a conventional manner using one or more pharmaceutically acceptable carriers (including excipients and / or adjuvants) that facilitate the processing of the active compound into a pharmaceutically usable formulation. Of course, the appropriate formulation depends on the chosen route of administration.

[0094] Suitable pharmaceutical compositions for injectable use comprise sterile aqueous solutions (in the water-soluble case) or dispersions, as well as sterile powders for the ad hoc preparation of sterile injectable solutions or dispersions. For intravenous administration, suitable carriers include physiological saline, antibacterial water, and Cremophor EL. TM(BASF) or phosphate-buffered saline (PBS). In all cases, the composition must be sterile and should have a fluidity sufficient for easy injection. It must be stable under the conditions of preparation and storage and must be preserved against contamination by microorganisms such as bacteria and fungi. The carrier can be a solvent or dispersion medium containing, for example, water, ethanol, polyols (e.g., glycerol, propylene glycol, and liquid polyethylene glycol, etc.) and suitable mixtures thereof. Appropriate fluidity can be maintained, for example, by using coatings such as lecithin, by maintaining the desired particle size in the case of dispersions, and by using surfactants. Protection against microorganisms can be achieved by various antibacterial and antifungal agents (e.g., parabens, chlorobutanol, phenol, ascorbic acid, thimerosal, etc.). In many cases, it is preferred to include isotonic agents (e.g., sugars, polyols such as mannitol and sorbitol, and sodium chloride) in the composition. Extended absorption of the injectable composition can be achieved by including agents that delay absorption (e.g., aluminum monostearate and gelatin) in the composition.

[0095] Sterile injectable solutions can be prepared by incorporating the desired amount of the active compound, as needed, with one or a combination of the ingredients listed above into a suitable solvent, followed by filtration and sterilization. Typically, dispersions are prepared by incorporating the active compound into a sterile carrier containing a base dispersion medium and other desired ingredients from those listed above. In the case of sterile powders used to prepare sterile injectable solutions, preparation methods include vacuum drying and freeze-drying, which produce powders of the active ingredient along with any other desired ingredients from its previously sterile filtered solution.

[0096] Oral compositions typically contain an inert diluent or an edible, pharmaceutically acceptable carrier. They can be encapsulated in gelatin capsules or compressed into tablets. For oral therapeutic administration, the active compound can be incorporated with excipients and administered in tablet, lozenge, or capsule form.

[0097] Pharmaceutically compatible binders and / or adjuvants may be included as part of the composition. Tablets, pills, capsules, lozenges, etc., may contain any of the following ingredients or compounds with similar properties: binders, such as microcrystalline cellulose, tragacanth gum, or gelatin; excipients, such as starch or lactose; disintegrants, such as alginate, Primogel, or corn starch; lubricants, such as magnesium stearate or sterotes; gliding agents, such as colloidal silica; sweeteners, such as sucrose or saccharin; or flavoring agents, such as peppermint, methyl salicylate, or orange flavoring.

[0098] For administration by inhalation, the compound is delivered as an aerosol spray from a pressurized container or dispenser (containing a suitable propellant, such as a gas, like carbon dioxide) or a nebulizer.

[0099] Systemic administration can also be achieved via mucosal or transdermal routes. For mucosal or transdermal administration, a penetrant suitable for the barrier to penetration is used in the formulation. Such penetrants are generally known in the art. Mucosal administration can be achieved by using nasal sprays or suppositories. For transdermal administration, as generally known in the art, the active compound is formulated as an ointment, cream, gel, or lotion.

[0100] In therapeutic applications, the dosage of the pharmaceutical composition used in this disclosure varies depending on the pharmaceutical agent, the age, weight and clinical condition of the patient receiving the treatment, the experience and judgment of the clinician or practicing physician administering the treatment, and other factors that influence the selected dosage.

[0101] The pharmaceutical composition may be included in a container, package, or dispenser along with the instructions for use.

[0102] The compounds of Formula I or Formula II provided in this application are effective for diseases or conditions related to TRIM21. TRIM21 can induce a series of tumor growth and spread (Alomari, M. Pharmaceutical Research 2021, 165, 105443) and induce the degradation of the pathogenic protein tau in brain nerve cells (Mukadam, ASet al). Science 2023, 379(6639,1336). As an intracellular antibody receptor or sensor, TRIM21 forms an innate immune defense against viral invasion. For viruses that have escaped the action of neutralizing antibodies extracellularly and are adsorbed by non-neutralizing antibodies, thus invading cells through the cell membrane, TRIM21 can exert antiviral effects by binding to the Fc fragment of antibodies in virus-antibody complexes, thereby inhibiting viral replication and inducing cellular immunity. TRIM21's dual effect of degrading viral particles and activating immune signaling is strictly regulated by its own ubiquitination and phosphorylation. Furthermore, TRIM21 can also synergize with the complement system. In summary, TRIM21 ligands and their derived target protein-degrading small molecules have broad potential applications in the treatment of many diseases, including tumors, neurodegenerative diseases, viral infections, and immune inflammation. Therefore, the compounds or compositions provided in this application can be used to treat many diseases or conditions, including tumors, neurodegenerative diseases, viral infections, and immune inflammation.

[0103] In some embodiments, the compounds and compositions of the present invention may be used in combination with one or more additional pharmaceutical agents. These additional agents may have TRIM21-regulating activity and / or may act through different mechanisms of action. In some embodiments, such agents comprise radiation (e.g., local or total radiotherapy) and / or other therapeutic forms of non-pharmacological nature. When used in combination therapy, the compounds and additional pharmaceutical agents of this application may be in the form of a single composition or multiple compositions, and the treatment may be administered simultaneously, sequentially, or through some other regimen. The combination therapy may have additive or synergistic effects.

[0104] Example

[0105] The invention will be more readily understood by referring to the following embodiments, which are used to illustrate the invention and should not be construed as limiting the scope of the invention in any way.

[0106] Unless otherwise defined or the context clearly requires, all technical and scientific terms used herein have the same meaning as commonly understood by one of ordinary skill in the art to which this invention pertains. It should be understood that any methods and materials similar to or equivalent to those described herein may be used in the practice or testing of this invention.

[0107] Example 1: Preparation of Compound 1

[0108]

[0109] Step 1: A mixture of compound 1-1 (200 mg, 0.587 mmol), dimethyl sulfinamide (54.64 mg, 0.587 mmol), Xantphos (33.94 mg, 0.059 mmol), Pd2(dba)3 (26.86 mg, 0.029 mmol), and Cs2CO3 (286.70 mg, 0.880 mmol) in dioxane (4 mL) was heated to 100 °C and stirred for 3 hours under nitrogen protection. After the reaction was complete, the reaction solution was poured into water (30 mL) and extracted with EtOAc (10 mL x 2). The combined extracts were washed with saturated brine (10 mL), dried over anhydrous Na2SO4, filtered, and the filtrate was concentrated to dryness to obtain compound 1-2 (170 mg, crude product), which was a yellow oil and was directly used in the next step of the reaction. MS-ESI(m / z):306.0 / 308.0[M+H] + .

[0110] Step 2: A mixture of dioxane (4 mL) and water (1 mL) containing compounds 1-2 (170 mg, 0.555 mmol), compounds 1-3 (141.55 mg, 0.833 mmol), Pd(dppf)Cl2 (40.63 mg, 0.056 mmol), and K2CO3 (191.84 mg, 1.388 mmol) was heated to 100 °C and stirred for 3 hours under nitrogen protection. After the reaction was complete, the reaction solution was concentrated under reduced pressure to remove most of the organic solvent, poured into water (30 mL), and extracted with EtOAc (10 mL x 2). The combined extracts were washed with saturated brine (10 mL), dried over anhydrous Na2SO4, filtered, and the filtrate was concentrated to dryness to obtain compound 1-4 (195 mg, crude product), which was a yellow oil and directly used in the next reaction. MS-ESI (m / z): 352.1 [M+H] + .

[0111] Step 3: Add NaOH (66.59 mg, 1.665 mmol) to a solution of compounds 1-4 (195 mg, 0.555 mmol) in MeOH (4 mL) and H₂O (2 mL). Heat the resulting mixture to reflux and stir for 1 hour. After the reaction is complete, dilute the reaction solution with H₂O (30 mL), adjust the pH to 6 with concentrated hydrochloric acid, extract with EtOAc (10 mL x 2), wash the combined extracts with saturated brine (10 mL), dry with anhydrous Na₂SO₄, filter, and concentrate the filtrate to obtain compounds 1-5 (180 mg, crude product), which is a yellowish-brown oil and can be used directly in the next step of the reaction.

[0112] MS-ESI (m / z): 338.1 [M+H] + .

[0113] Step 4: A DMF solution (1 mL) of compounds 1-5 (180 mg, 0.534 mmol), HATU (304.31 mg, 0.800 mmol), and DIEA (0.265 mL, 1.601 mmol) was stirred at 25°C for 5 minutes. Then, compounds 1-6 (73.72 mg, 0.534 mmol) were added to the above solution, and stirring was continued for 1 hour. After the reaction was complete, the reaction solution was poured into water (30 mL) and extracted with EtOAc (10 mL x 3). The combined extracts were washed with saturated brine (10 mL), dried over anhydrous Na2SO4, filtered, and the filtrate was concentrated to dryness to obtain compounds 1-7 (200 mg, crude product) as a yellow solid, which was directly used in the next reaction.

[0114] MS-ESI (m / z): 458.1 [M+H] + .

[0115] Step 5: Under nitrogen protection and at 25°C, NaH (69.94 mg, 1.749 mmol, 60% mineral oil) was added to a DMF (4 mL) solution of compounds 1-7 (200 mg, 0.437 mmol). The resulting reaction solution was stirred at this temperature for 10 minutes. Then, MeI (0.142 mL, 1.749 mmol) was added to the suspension, and the reaction was continued at this temperature for 1 hour. After the reaction was complete, H2O (1 mL) was slowly and carefully added dropwise to quench the reaction. The resulting solution was directly concentrated to dryness under reduced pressure to obtain compounds 1-8 (205 mg, crude product) as a yellow solid, which was directly used for the next reaction. MS-ESI (m / z): 472.2 [M+H] + .

[0116] Step 6: Add concentrated hydrochloric acid (1 mL) to a dioxane (2 mL) solution of compounds 1-8 (205 mg, 0.435 mmol). Heat the resulting reaction solution to 100 °C and stir for 6 hours. After the reaction is complete, adjust the pH of the reaction solution to 8 with saturated NaHCO3, extract with EtOAc (10 mL x 3), wash the combined extracts with saturated brine (10 mL), dry with anhydrous Na2SO4, filter, and concentrate the filtrate to dryness under reduced pressure. Purify the residue by preparative thin-layer chromatography (DCM / MeOH = 10 / 1) to obtain compound 1 (15 mg, total yield of six steps: 5.6%) as a yellow solid.

[0117] 1 H-NMR(400MHz,DMSO-d6)δ11.48(1H,brs),7.34-7.40(2H,m),7.21-7.30(1H,m),7.15-7.20(1H,m),6.97-7.09(2H,m),6.85-6 .92(1H,m),6.23-6.52(1H,m),6.04-6.21(1H,m),3.78-3.85(3H,m),3.34-3.40(5H,m),3.22-3.31(3H,m),2.82-2.90(3H,m).

[0118] MS-ESI (m / z): 458.1 [M+H] + .

[0119] Example 2: Preparation of Compound 2

[0120]

[0121] Step 1: Triethylamine (1.871 mL, 13.462 mmol) and di-tert-butyl dicarbonate (0.773 mL, 3.365 mmol) were added to a solution of compounds 1-6 (465 mg, 3.365 mmol) in dichloromethane (5 mL). The mixture was stirred at room temperature for 2 hours, then concentrated to dryness under reduced pressure. The residue was purified by silica gel column chromatography (PE / EtOAc = 5 / 1) to give compound 2-1 (648 mg, yield: 80.8%) as a yellow oil. MS-ESI (m / z): 239.1 [M+H] + .

[0122] Step 2: In an ice bath, NaH (261.06 mg, 10.878 mmol, 60% mineral oil mixture) was added dropwise over 10 minutes to a tetrahydrofuran (12 mL) solution of compound 2-1 (648 mg, 2.719 mmol). Iodomethane (0.331 mL, 4.079 mmol) was then added dropwise. The mixture was stirred at room temperature for 3 hours. The mixture was purified by silica gel column chromatography (PE / EtOAc = 5 / 1) to give compound 2-2 (539 mg, yield: 78.6%) as a colorless oil. MS-ESI (m / z): 253.1 [M+H] + .

[0123] Step 3: Add concentrated hydrochloric acid (1 mL) to a 2 mL solution of dioxane (360 mg, 1.427 mmol) of compound 2-2. Heat the resulting reaction solution to 100 °C and stir for 6 hours. After the reaction is complete, concentrate the reaction solution directly under reduced pressure to dryness to obtain compound 2-3 (197 mg, crude product) as a white solid.

[0124] MS-ESI (m / z): 139.2 [M+H] + .

[0125] Step 4: A mixture of compound 1-1 (400 mg, 0.587 mmol), dimethylphosphine oxide (99.42 mg, 1.291 mmol), Xantphos (135.77 mg, 0.253 mmol), Pd2(dba)3 (107.44 mg, 0.117 mmol), and K3PO4 (540.34 mg, 2 mmol) in dioxane (8 mL) was heated to 100 °C and stirred for 8 hours under nitrogen protection. After the reaction was complete, the reaction solution was poured into water (60 mL) and extracted with EtOAc (20 mL x 2). The combined extracts were washed with saturated brine (10 mL), dried over anhydrous Na2SO4, filtered, and concentrated to dryness. The residue was purified by silica gel column chromatography (PE:EtOAc = 10:1 to pure EtOAc) to obtain compound 2-4 (177 mg, yield 48.05%) as a yellow oil.

[0126] MS-ESI(m / z):290.9 / 293.0[M+H] + .

[0127] Step 5: A mixture of dioxane (4 mL) and water (1 mL) containing compounds 2-4 (177 mg, 0.555 mmol), compounds 1-3 (103.34 mg, 0.608 mmol), Pd(dppf)Cl2 (44.49 mg, 0.061 mmol), and K2CO3 (252.11 mg, 1.824 mmol) was heated to 100 °C and stirred for 3 hours under nitrogen protection. After the reaction was complete, the reaction solution was concentrated under reduced pressure to remove most of the organic solvent, poured into water (30 mL), and extracted with EtOAc (10 mL x 2). The combined extracts were washed with saturated brine (10 mL), dried over anhydrous Na2SO4, filtered, and the filtrate was concentrated to dryness to obtain compound 2-5 (116 mg, crude product), which was a brown oil and directly used in the next reaction. MS-ESI (m / z): 337.1 [M+H] + .

[0128] Step 6: Add NaOH (41.39 mg, 1.035 mmol) to a solution of compound 2-5 (116 mg, 0.555 mmol) in MeOH (4 mL) and H₂O (2 mL). Heat the resulting mixture to reflux and stir for 1 hour. After the reaction is complete, dilute the reaction solution with H₂O (20 mL), adjust the pH to 6 with concentrated hydrochloric acid, extract with EtOAc (10 mL x 2), wash the combined extracts with saturated brine (10 mL), dry with anhydrous Na₂SO₄, filter, and concentrate the filtrate to obtain compound 2-6 (110 mg, crude product) as a brown solid, which can be used directly in the next step of the reaction. MS-ESI (m / z): 323.1 [M+H]+.

[0129] Step 7: A DMF solution (1 mL) of compounds 2-6 (60 mg, 0.186 mmol), HATU (77.87 mg, 0.205 mmol), and DIEA (0.092 mL, 0.559 mmol) was stirred at 25 °C for 5 minutes. Then, compound 2-3 (73.72 mg, 0.534 mmol) was added to the above solution, and stirring was continued for 1 hour. After the reaction was complete, the reaction solution was poured into water (10 mL) and extracted with EtOAc (5 mL x 3). The combined extracts were washed with saturated brine (5 mL), dried over anhydrous Na2SO4, filtered, and the filtrate was concentrated to dryness under reduced pressure. The residue was purified by preparative thin-layer chromatography (DCM / MeOH = 10 / 1) to obtain compound 2 (3.96 mg, yield: 5.6%) as a white solid.

[0130] 1 H-NMR(400MHz,DMSO-d6)δ11.45(brs,1H),7.82-7.87(m,1H),7.62-7.72(m,1H),7.29-7.47(m,3H),7.02-7.10(m,1H),6 .84-6.95(m,1H),6.17-6.39(m,2H),4.15-4.49(m,2H),3.75-3.85(m,3H),2.77-2.88(m,3H),1.74(s,3H),1.70(s,3H).

[0131] MS-ESI (m / z): 433.1 [M+H] + .

[0132] Example 3: Preparation of Compound 3

[0133]

[0134] Step 1: A mixture of compound 1-1 (100 mg, 0.293 mmol), compound 3-1 (39.07 mg, 0.293 mmol), Xantphos (16.97 mg, 0.029 mmol), Pd2(dba)3 (11.87 mg, 0.015 mmol), and K3PO4 (124.51 mg, 0.587 mmol) in dioxane (4 mL) was heated to 100 °C and stirred for 8 hours under nitrogen protection. After the reaction was complete, the reaction solution was poured into water (20 mL) and extracted with EtOAc (10 mL x 2). The combined extracts were washed with saturated brine (10 mL), dried over anhydrous Na2SO4, filtered, and the filtrate was concentrated to dryness to obtain compound 3-2 (68 mg, crude product), which was a yellow oil and directly used in the next reaction. MS-ESI(m / z):347.90 / 350.00[M+H] + .

[0135] Step 2: A mixture of compound 3-2 (68 mg, 0.196 mmol), compound 1-3 (33.38 mg, 0.196 mmol), Pd(dppf)Cl2 (14.37 mg, 0.020 mmol), and K2CO3 (81.43 mg, 0.589 mmol) in dioxane (4 mL) and water (1 mL) was heated to 100 °C and stirred for 3 hours under nitrogen protection. After the reaction was complete, the reaction solution was concentrated under reduced pressure to remove most of the organic solvent, poured into water (20 mL), and extracted with EtOAc (10 mL x 2). The combined extracts were washed with saturated brine (10 mL), dried over anhydrous Na2SO4, filtered, and the filtrate was concentrated to dryness to obtain compound 3-3 (70 mg, crude product), which was a brown oil and directly used in the next reaction. MS-ESI (m / z): 392.10 [M+H] + .

[0136] Step 3: Add NaOH (21.46 mg, 0.536 mmol) to a solution of compound 3-3 (70 mg, 0.179 mmol) in MeOH (4 mL) and H₂O (2 mL). Heat the resulting mixture to reflux and stir for 1 hour. After the reaction is complete, dilute the reaction solution with H₂O (10 mL), adjust the pH to 6 with concentrated hydrochloric acid, extract with EtOAc (5 mL x 2), wash the combined extracts with saturated brine (5 mL), dry with anhydrous Na₂SO₄, filter, and concentrate the filtrate to obtain compound 3-4 (65 mg, crude product), which is a brown oil and can be used directly in the next reaction. MS-ESI (m / z): 378.00 [M+H] + .

[0137] Step 4: A DMF solution (1 mL) of compounds 3-4 (65 mg, 0.172 mmol), HATU (130.97 mg, 0.344 mmol), and DIEA (0.085 mL, 0.517 mmol) was stirred at 25°C for 5 minutes. Then, compounds 1-6 (28.55 mg, 0.207 mmol) were added to the above solution, and stirring was continued for 1 hour. After the reaction was complete, the reaction solution was poured into water (20 mL) and extracted with EtOAc (10 mL x 3). The combined extracts were washed with saturated brine (10 mL), dried over anhydrous Na2SO4, filtered, and the filtrate was concentrated to dryness to obtain compound 3-5 (100 mg, crude product), which was a brown oil and could be used directly in the next step of the reaction.

[0138] MS-ESI (m / z): 498.10 [M+H] + .

[0139] Step 5: Under nitrogen protection and at 25°C, NaH (24.12 mg, 0.603 mmol, 60% mineral oil mixture) was added to a THF (4 mL) solution of compound 3-5 (100 mg, 0.201 mmol). The resulting reaction solution was stirred at this temperature for 10 minutes. Then, MeI (0.049 mL, 0.603 mmol) was added to the suspension, and the reaction was carried out at 50°C for 1 hour. After the reaction was complete, H2O (1 mL) was slowly and carefully added dropwise to quench the reaction. The resulting solution was directly concentrated to dryness under reduced pressure to obtain compound 3-6 (100 mg, crude product) as a brown solid, which was directly used for the next reaction. MS-ESI (m / z): 512.10 [M+H] + .

[0140] Step 6: Synthesis of Compound 1

[0141] Concentrated hydrochloric acid (1 mL) was added to a 2 mL solution of dioxane (100 mg, 0.435 mmol) of compounds 3-6. The resulting reaction solution was heated to 100 °C and stirred for 6 hours. After the reaction was complete, the pH of the reaction solution was adjusted to 8 with saturated NaHCO3 aqueous solution, and extracted with EtOAc (10 mL x 3). The combined extracts were washed with saturated brine (10 mL), dried over anhydrous Na2SO4, filtered, and the filtrate was concentrated to dryness under reduced pressure. The residue was purified by preparative thin-layer chromatography (DCM / MeOH = 10 / 1) to give compound 3 (7.51 mg, total yield of six steps: 5.1%) as a white solid.

[0142] 1 H-NMR(400MHz,DMSO-d6)δ11.45(brs,1H),7.29-7.40(m,2H),7.19-7.34(m ,1H),7.15(t,J=6.8Hz,1H),6.94-7.06(m,2H),6.81-6.88(m,1H),6.25-6. 51(m,1H),6.07-6.17(m,1H),4.04-4.96(m,2H),3.78(s,3H),3.44-3.64(m ,2H),3.23-3.29(m,2H),2.89(s,3H),1.71-2.05(m,4H),1.46-1.67(m,2H).

[0143] MS-ESI (m / z): 498.20 [M+H] + .

[0144] Example 4: Preparation of Compound 4

[0145]

[0146] Step 1: A mixture of compound 1-1 (150 mg, 0.440 mmol), compound 4-1 (52.43 mg, 0.440 mmol), Xantphos (25.46 mg, 0.044 mmol), Pd2(dba)3 (20.14 mg, 0.022 mmol), and Cs2CO3 (93.39 mg, 0.440 mmol) in dioxane (2 mL) was heated to 110 °C and stirred for 18 hours under nitrogen protection. After the reaction was complete, the reaction mixture was poured into water (20 mL) and extracted with EtOAc (10 mL x 2). The combined extracts were washed with saturated brine (10 mL), dried over anhydrous Na2SO4, filtered, and the filtrate was concentrated to dryness. The residue was purified by preparative thin-layer chromatography (PE / EtOAc = 1 / 1) to give compound 4-2 (110 mg, yield: 75.2%) as a yellow solid.

[0147] MS-ESI(m / z):332.0 / 334.0[M+H] + .

[0148] Step 2: A mixture of dioxane (1 mL) and water (1 mL) of compound 4-2 (110 mg, 0.331 mmol), compound 1-3 (56.27 mg, 0.331 mmol), Pd(dppf)Cl2 (27.04 mg, 0.033 mmol), and K2CO3 (137.28 mg, 0.993 mmol) was heated to 110 °C and stirred for 2 hours under nitrogen protection. After the reaction was complete, the reaction solution was concentrated to dryness under reduced pressure. The residue was purified by silica gel column chromatography (DCM / MeOH = 20 / 1) to give compound 4-3 (71 mg, yield: 56.8%) as a white solid.

[0149] MS-ESI(m / z):378.1 / 380.1[M+H] + .

[0150] Step 3: Add a LiOH (23.68 mg, 0.564 mmol) H₂O (0.5 mL) solution to a MeOH (1 mL) and THF (1 mL) solution of compound 4-3 (71 mg, 0.188 mmol). Heat the resulting mixture to reflux and stir for 20 minutes. After the reaction is complete, adjust the pH of the reaction solution to 7-8 with concentrated hydrochloric acid, and concentrate to dryness to obtain compound 4-4 (68 mg, crude product) as a white solid, which can be used directly in the next step of the reaction.

[0151] MS-ESI (m / z): 364.1 [M+H] + .

[0152] Step 4: Compound 1-6 (25.85 mg, 0.187 mmol) was added to a DMF (2 mL) solution of compound 4-4 (68 mg, 0.187 mmol), HATU (85.38 mg, 0.225 mmol), and DIEA (0.093 mL, 0.561 mmol), and the mixture was stirred for 1 hour. After the reaction was complete, the reaction solution was poured into water (30 mL) and extracted with EtOAc (10 mL x 3). The combined extracts were washed with saturated brine (10 mL), dried over anhydrous Na2SO4, filtered, and the filtrate was concentrated to dryness. The residue was purified by silica gel column chromatography (DCM / MeOH = 20 / 1) to give compound 4-5 (90 mg, yield: 99.4%) as a yellow oil.

[0153] MS-ESI (m / z): 484.1 [M+H] + .

[0154] Step 5: Under ice bath conditions, NaH (13.40 mg, 0.558 mmol) was added to a DMF (4 mL) solution of compound 4-5 (90 mg, 0.186 mmol). The resulting reaction solution was stirred at this temperature for 10 minutes. Then, MeI (0.045 mL, 0.558 mmol) was added, and the reaction was carried out at 40 °C for 6 hours. After the reaction was complete, H2O (0.5 mL) was slowly and carefully added dropwise to quench the reaction, and the pH was adjusted to 7-8 with dilute hydrochloric acid. The resulting solution was directly concentrated to dryness under reduced pressure to obtain compound 4-6 (92 mg, crude product) as a brown solid, which was directly used for the next reaction. MS-ESI (m / z): 498.1 [M+H] + .

[0155] Step 6: Compound 4-6 (92 mg, 0.185 mmol) was added to dioxane hydrochloride (4 M, 2 mL), and the resulting reaction solution was heated to 80 °C and stirred for 8 hours. After the reaction was complete, the reaction solution was concentrated to dryness under reduced pressure. The residue was purified by preparative thin-layer chromatography (DCM / MeOH = 20 / 1) to obtain the crude product, which was then separated by reversed-phase chromatography to obtain compound 4 (3.13 mg, total yield of six steps: 1.5%) as a yellow solid.

[0156] 1H-NMR(400MHz,MeOD)δ7.30-7.40(m,1H),7.25-7.30(m,1H),7.17-7.24(m,1 H),7.08-7.16(m,1H),6.94-7.05(m,1H),6.72-6.80(m,1H),6.63-6.71(m,1H ),6.57-6.63(m,1H),6.21-6.36(m,1H),4.04-4.72(m,2H),3.68-3.74(m,3H) ,3.34-3.59(m,2H),3.02-3.17(m,2H),2.86-2.98(m,3H),2.06-2.24(m,4H).

[0157] MS-ESI (m / z): 484.1 [M+H] + .

[0158] Example 5: Preparation of Compound 5

[0159]

[0160] Step 1: Sodium methanethiol (1.11 g, 16 mmol) was added to a THF (80 mL) solution of compound 5-1 (4.0 g, 16 mmol) at 0 °C, and stirring was continued at 0 °C for 1 hour. The temperature was then increased to 25 °C and stirred for 20 hours. After the reaction was complete, the reaction mixture was poured into water (80 mL) and extracted with DCM (30 mL x 3). The combined extracts were washed with saturated brine (10 mL), dried over anhydrous Na₂SO₄, filtered, and the filtrate was concentrated to dryness under reduced pressure. The residue was subjected to silica gel column chromatography (PE / EtOAc = 10 / 1) to give compound 5-2 (1.32 g, yield: 23%) as a white solid.

[0161] 1 H-NMR (400MHz, CDCl3) δ8.48(d,J=2.0Hz,1H),7.75(d,J=2.0Hz,1H),4.00(s,3H),2.46(s,3H).

[0162] MS-ESI(m / z):262.0 / 264.0[M+H] + .

[0163] Step 2: At 0°C, m-CPBA (1.45 g, 8.4 mmol, 85% purity) was added in portions to a DCM (10 mL) solution of compound 5-2 (1.1 g, 4.2 mmol). After the addition was complete, the reaction mixture was heated to 25°C and stirred for 18 hours. After the reaction was complete, the reaction solution was diluted with water (100 mL), and the resulting mixture was extracted with DCM (30 mL x 3). The combined extracts were washed with saturated Na₂S₂O₄ aqueous solution (20 mL), and the organic phase was further washed with saturated brine (10 mL). Finally, the mixture was dried over anhydrous Na₂SO₄, filtered, and the filtrate was concentrated to dryness under reduced pressure. The residue was purified by silica gel column chromatography (PE / DCM = 5 / 1 to 1 / 1) to obtain compound 5-3 (630 mg, yield 73%) as a white solid.

[0164] 1 H-NMR (400MHz, CDCl3) δ8.90(d,J=2.0Hz,1H),8.56(d,J=2.0Hz,1H),4.04(s,3H),3.40(s,3H).

[0165] MS-ESI (m / z): 294.0 [M+H] + .

[0166] Step 3: A solution of dioxane (3 mL) and water (1 mL) of compounds 5-3 (30 mg, 0.102 mmol), 1-6 (19.07 mg, 0.112 mmol), Pd(dppf)Cl2 (7.46 mg, 0.010 mmol), and K2CO3 (42.29 mg, 0.306 mmol) was heated to 100 °C and stirred for 3 hours under nitrogen protection. After the reaction was complete, the solution was concentrated under reduced pressure to remove most of the organic solvent, diluted with water (6 mL), and the pH was adjusted to 6 with concentrated hydrochloric acid. The precipitated solid was filtered, the filter cake was washed with water (1 mL x 3), and finally dried under high vacuum to obtain compound 5-4 (31 mg, yield: 93.4%) as a white solid.

[0167] MS-ESI (m / z): 326.0 [M+H] + .

[0168] Step 4: A DMF (1 mL) solution of compounds 5-4 (20.0 mg, 0.061 mmol), compounds 2-3 (7.63 mg, 0.061 mmol), HATU (28.05 mg, 0.074 mmol), and DIEA (0.030 mL, 0.184 mmol) was stirred at 25 °C for half an hour. After the reaction was complete, the reaction solution was concentrated to dryness under reduced pressure. The residue was purified by preparative thin-layer chromatography (DCM / MeOH = 10 / 1) to obtain compound 5 (15 mg, yield: 54.8%) as a yellow solid.

[0169] 1 H-NMR(400MHz,MeOD)δ8.78-8.95(m,1H),8.40-8.50(m,1H),7.25-7.50(m,2H),6.80-6.90(m,1H),6.70-7 .78(m,1H),6.40-6.60(m,2H),4.10-4.60(m,2H),3.68-3.78(m,3H),3.23-3.28(m,3H),2.75-3.00(m,3H).

[0170] MS-ESI (m / z): 446.1 [M+H] + .

[0171] Example 6: Preparation of Compound 6

[0172]

[0173] Step 1: A mixture of compound 1-1 (70 mg, 0.205 mmol), compound 6-1 (21.59 mg, 0.205 mmol), Xantphos (11.88 mg, 0.021 mmol), Pd2(dba)3 (9.40 mg, 0.010 mmol), and Cs2CO3 (43.58 mg, 0.205 mmol) in dioxane (1 mL) was heated to 110 °C and stirred for 18 hours under nitrogen protection. After the reaction was complete, the reaction mixture was poured into water (20 mL) and extracted with EtOAc (10 mL x 2). The combined extracts were washed with saturated brine (10 mL), dried over anhydrous Na2SO4, filtered, and the filtrate was concentrated to dryness. The residue was purified by preparative thin-layer chromatography (PE / EtOAc = 1 / 1) to give compound 6-2 (47 mg, yield: 71.9%) as a yellow solid.

[0174] MS-ESI(m / z):319.2 / 321.2[M+H] + .

[0175] Step 2: A mixture of dioxane (2 mL) and water (25.10 mg, 0.148 mmol) of compound 6-2 (47 mg, 0.148 mmol), compound 1-3 (25.10 mg, 0.148 mmol), Pd(dppf)Cl2 (10.81 mg, 0.015 mmol), and K2CO3 (61.24 mg, 0.443 mmol) was heated to 110 °C and stirred for 2 hours under nitrogen protection. After the reaction was complete, the reaction solution was concentrated to dryness under reduced pressure. The residue was purified by silica gel column chromatography (DCM / MeOH = 20 / 1) to give compound 6-3 (41 mg, yield: 76.3%) as a white solid.

[0176] MS-ESI(m / z):364.1 / 366.1[M+H] + .

[0177] Step 3: To a solution of compound 6-3 (41 mg, 0.113 mmol) in MeOH (1 mL) and THF (1 mL), add a solution of LiOH (14.20 mg, 0.338 mmol) in H₂O (0.5 mL). Heat the resulting mixture to reflux and stir for 20 minutes. After the reaction is complete, adjust the pH of the reaction solution to 7-8 with concentrated hydrochloric acid and concentrate to dryness under reduced pressure. Purify the residue by silica gel column chromatography (DCM / MeOH = 20 / 1) to give compound 6-4 (26 mg, yield: 65.9%) as a white solid. MS-ESI (m / z): 350.0 [M+H] + .

[0178] Step 4: Compound 2-3 (26 mg, 0.074 mmol) was added to a DMF (1 mL) solution of compound 6-4 (10.28 mg, 0.074 mmol), HATU (42.44 mg, 0.112 mmol), and DIEA (0.061 mL, 0.372 mmol), and the mixture was stirred for 1 hour. After the reaction was complete, the reaction solution was poured into water (30 mL) and extracted with EtOAc (10 mL x 3). The combined extracts were washed with saturated brine (10 mL), dried over anhydrous Na2SO4, filtered, and the filtrate was concentrated to dryness. The residue was purified by preparative thin-layer chromatography (DCM / MeOH = 20 / 1) to obtain the crude product, which was then separated by reversed-phase chromatography to obtain compound 6 (2.38 mg, total yield of four steps: 2.4%) as a yellow solid.

[0179] 1H-NMR(400MHz,MeOD)δ7.27-7.35(m,1H),7.18-7.26(m,1H),7.16-7.18(m,1H),7.10-7.15(m,1H),6.96-7.07(m,1H),6.72-6.81(m,1H) ),6.66-6.71(m,1H),6.59-6.66(m,1H),6.20-6.37(m,1H),3.97-4.53(m,6H),3.67-3.74(m,3H),2.85-2.99(m,3H),2.10-2.34(m,2H).

[0180] MS-ESI (m / z): 470.1 [M+H] + .

[0181] Example 7: Preparation of Compound 7

[0182]

[0183] Step 1: A mixture of compound 1-1 (100 mg, 0.293 mmol), compound 7-1 (39.07 mg, 0.293 mmol), Xantphos (16.97 mg, 0.029 mmol), Pd2(dba)3 (11.87 mg, 0.015 mmol), and K3PO4 (124.51 mg, 0.587 mmol) in dioxane (4 mL) was heated to 100 °C and stirred for 8 hours under nitrogen protection. After the reaction was complete, the reaction solution was poured into water (20 mL) and extracted with EtOAc (10 mL x 2). The combined extracts were washed with saturated brine (10 mL), dried over anhydrous Na2SO4, filtered, and the filtrate was concentrated to dryness to obtain compound 7-2 (68 mg, crude product), which was a yellow oil and directly used in the next step of the reaction. MS-ESI(m / z):349.90 / 351.00[M+H] + .

[0184] Step 2: A mixture of dioxane (4 mL) and water (1 mL) of compound 7-2 (68 mg, 0.195 mmol), compound 1-3 (33.19 mg, 0.195 mmol), Pd(dppf)Cl2 (14.29 mg, 0.020 mmol), and K2CO3 (80.96 mg, 0.586 mmol) was heated to 100 °C and stirred for 3 hours under nitrogen protection. After the reaction was complete, the reaction solution was concentrated under reduced pressure to remove most of the organic solvent, poured into water (20 mL), and extracted with EtOAc (10 mL x 2). The combined extracts were washed with saturated brine (10 mL), dried over anhydrous Na2SO4, filtered, and the filtrate was concentrated to dryness to obtain compound 7-3 (72 mg, crude product), which was a brown oil and directly used in the next reaction. MS-ESI (m / z): 394.00 [M+H] + .

[0185] Step 3: Add NaOH (21.96 mg, 0.549 mmol) to a solution of compound 7-3 (72 mg, 0.179 mmol) in MeOH (4 mL) and H₂O (2 mL). Heat the resulting mixture to reflux and stir for 1 hour. After the reaction is complete, dilute the reaction solution with H₂O (10 mL), adjust the pH to 6 with concentrated hydrochloric acid, extract with EtOAc (5 mL x 2), wash the combined extracts with saturated brine (5 mL), dry with anhydrous Na₂SO₄, filter, and concentrate the filtrate to obtain compound 7-4 (69 mg, crude product), which is a brown oil and can be used directly in the next step. MS-ESI (m / z): 380.00 [M+H] + .

[0186] Step 4: A DMF solution (1 mL) of compound 7-4 (69 mg, 0.182 mmol), HATU (138.17 mg, 0.364 mmol), and DIEA (0.120 mL, 0.727 mmol) was stirred at 25°C for 5 minutes. Then, compound 1-6 (25.13 mg, 0.182 mmol) was added to the above solution, and stirring was continued for 1 hour. After the reaction was complete, the reaction solution was poured into water (20 mL) and extracted with EtOAc (10 mL x 3). The combined extracts were washed with saturated brine (10 mL), dried over anhydrous Na2SO4, filtered, and the filtrate was concentrated to dryness to obtain compound 7-5 (90 mg, crude product), which was a brown oil and could be used directly in the next step of the reaction.

[0187] MS-ESI (m / z): 500.10 [M+H] + .

[0188] Step 5: Under nitrogen protection and at 25°C, NaH (43.24 mg, 1.801 mmol, 60% mineral oil mixture) was added to a THF (4 mL) solution of compound 7-5 (90 mg, 0.180 mmol). The resulting reaction solution was stirred at this temperature for 10 minutes. Then, MeI (0.088 mL, 1.801 mmol) was added to the suspension, and the reaction was carried out at 50°C for 1 hour. After the reaction was complete, H2O (1 mL) was slowly and carefully added dropwise to quench the reaction. The resulting solution was directly concentrated to dryness under reduced pressure to obtain compound 7-6 (90 mg, crude product) as a brown solid, which was directly used for the next reaction. MS-ESI (m / z): 514.10 [M+H] + .

[0189] Step 6: Add concentrated hydrochloric acid (1 mL) to a dioxane (2 mL) solution of compound 7-6 (90 mg, 0.175 mmol). Heat the resulting reaction solution to 100 °C and stir for 6 hours. After the reaction is complete, adjust the pH of the reaction solution to 8 with saturated NaHCO3 aqueous solution, extract with EtOAc (10 mL x 3), wash the combined extracts with saturated brine (10 mL), dry with anhydrous Na2SO4, filter, and concentrate the filtrate under reduced pressure to dryness. Purify the residue by preparative thin-layer chromatography (DCM / MeOH = 10 / 1) to obtain compound 7 (14.92 mg, total yield of six steps: 10.2%) as a white solid.

[0190] 1 H-NMR(400MHz,MeOD)δ7.37-7.44(m,2H),7.27-7.35(m,1H),7.20-7.25(m,1H),7.06-7.14(m,1H),6.85-6.90(m,1H),6.83-6.45(m,2H),6.44-6 .30(m,1H),5.05-5.11(m,1H),4.21-.34(m,1H),3.99-4.11(m,4H),3.6 0-3.83(m,3H),3.57-3.78(m,2H),3.35-3.50(m,2H),2.99-3.09(m,3H).

[0191] MS-ESI (m / z): 500.10 [M+H] + .

[0192] Example 8: Preparation of Compound 8

[0193]

[0194] Step 1: Compound 2-2 (263 mg, 1.042 mmol) was added to a solution of dioxane hydrochloride (4 M, 3 mL), and stirred at room temperature for 1 hour. The mixture was concentrated to dryness under reduced pressure to obtain compound 8-1 (158 mg, crude product) as a white solid. MS-ESI (m / z): 153.2 [M+H] + .

[0195] Step 2: Compound 8-1 (27.07 mg, 0.178 mmol) was added to a DMF (2 mL) solution of compounds 1-5 (50 mg, 0.148 mmol), HATU (67.62 mg, 0.178 mmol), and DIEA (0.122 mL, 0.741 mmol), and the mixture was stirred for 1 hour. After the reaction was complete, the reaction solution was poured into water (30 mL) and extracted with EtOAc (10 mL x 3). The combined extracts were washed with saturated brine (10 mL), dried over anhydrous Na2SO4, filtered, and the filtrate was concentrated to dryness. The residue was separated by reversed-phase chromatography to obtain compound 8 (30 mg, overall yield of two steps: 42.5%) as a white solid.

[0196] 1 H-NMR(400MHz,DMSO-d6)δ8.08-8.19(m,1H),7.30-7.39(m,1H),7.20-7.30(m ,1H),7.12-7.20(m,1H),7.03-7.08(m,1H),7.02-7.03(m,1H),6.95-7.02(m,1 H),6.87-6.90(m,1H),6.69-6.87(m,1H),4.14-5.21(m,2H),3.83-3.89(m,3H ),3.77-3.83(m,3H),3.25-3.30(m,3H),3.12-3.25(m,3H),2.76-2.90(m,3H).

[0197] MS-ESI (m / z): 472.2 [M+H] + .

[0198] Example 9: Preparation of Compound 9

[0199]

[0200] Step 1: A mixture of compound 1-1 (150 mg, 0.440 mmol), diethylphosphine oxide (50.86 mg, 0.484 mmol), Xantphos (50.91 mg, 0.088 mmol), Pd2(dba)3 (40.29 mg, 0.044 mmol), and K3PO4 (202.63 mg, 0.880 mmol) in dioxane (4 mL) was heated to 100 °C and stirred for 8 hours under nitrogen protection. After the reaction was complete, the reaction mixture was poured into water (30 mL) and extracted with EtOAc (10 mL x 2). The combined extracts were washed with saturated brine (5 mL), dried over anhydrous Na2SO4, filtered, and concentrated to dryness. The residue was purified by silica gel column chromatography (PE / EtOAc = 10:1 to pure EtOAc) to give compound 9-1 (55 mg, yield: 39.2%) as a yellow oil.

[0201] MS-ESI(m / z):319.00 / 321.90[M+H] + .

[0202] Step 2: A mixture of compound 9-1 (55 mg, 0.172 mmol), compound 1-3 (32.22 mg, 0.190 mmol), Pd(dppf)Cl2 (12.61 mg, 0.017 mmol), and K2CO3 (71.45 mg, 0.517 mmol) in dioxane (2 mL) and water (0.5 mL) was heated to 100 °C and stirred for 3 hours under nitrogen protection. After the reaction was complete, the reaction solution was concentrated under reduced pressure to remove most of the organic solvent, poured into water (10 mL), and extracted with EtOAc (5 mL x 2). The combined extracts were washed with saturated brine (5 mL), dried over anhydrous Na2SO4, filtered, and the filtrate was concentrated to dryness to obtain compound 9-2 (60 mg, crude product), which was a brown oil and directly used in the next reaction. MS-ESI (m / z): 365.1 [M+H] + .

[0203] Step 3: Add NaOH (19.76 mg, 0.494 mmol) to a solution of compound 9-2 (60 mg, 0.165 mmol) in MeOH (2 mL) and H₂O (0.5 mL). Heat the resulting mixture to 50 °C and stir for 1 hour. After the reaction is complete, dilute the reaction solution with H₂O (10 mL), adjust the pH to 6 with concentrated hydrochloric acid, extract with EtOAc (5 mL x 2), wash the combined extracts with saturated brine (5 mL), dry with anhydrous Na₂SO₄, filter, and concentrate the filtrate to obtain compound 9-3 (42 mg, crude product) as a brown solid, which can be used directly in the next step of the reaction. MS-ESI (m / z): 351.1 [M+H] + .

[0204] Step 4: A DMF solution (1 mL) of compound 9-3 (42 mg, 0.120 mmol), HATU (68.38 mg, 0.180 mmol), and DIEA (0.059 mL, 0.360 mmol) was stirred at 25 °C for 5 minutes. Then, compound 2-3 (16.56 mg, 0.120 mmol) was added to the above solution, and stirring was continued for 1 hour. After the reaction was complete, the reaction solution was poured into water (10 mL) and extracted with EtOAc (5 mL x 3). The combined extracts were washed with saturated brine (5 mL), dried over anhydrous Na2SO4, filtered, and the filtrate was concentrated to dryness under reduced pressure. The residue was purified by preparative thin-layer chromatography (DCM / MeOH = 10 / 1) to obtain compound 9 (8.89 mg, yield: 15.6%) as a white solid.

[0205] 1 H-NMR(400MHz,DMSO-d6)δ11.45(brs,1H),7.77-7.85(m,1H),7.61-7.71(m,1H),7.27-7.46(m,3H),7.02-7.10(m,1H),6.85-6 .95(m,1H),6.19-6.39(m,2H),4.13-4.47(m,2H),3.78-3.80(m,3H),2.77-2.87(m,3H),1.89-2.06(m,4H),0.93-1.08(m,6H).

[0206] MS-ESI (m / z): 471.20 [M+H] + .

[0207] Example 10: Preparation of Compound 10

[0208]

[0209] Step 1: Compound 10⁻¹ (165 mg, 0.629 mmol), compound 1-3 (117.68 mg, 0.692 mmol), Pd(dppf)Cl₂ (51.41 mg, 0.063 mmol), and K₂CO₃ (260.98 mg, 1.888 mmol) were dissolved in a mixed solution of dioxane (2.5 mL) and water (2.5 mL). The solution was heated to 80 °C and stirred for 2 hours under nitrogen protection. After the reaction was complete, the reaction solution was concentrated to dryness under reduced pressure. The residue was subjected to silica gel column chromatography (EtOAc / PE = 50%-100%) to give compound 10⁻² (166 mg, yield: 85.8%) as a white solid. MS-ESI (m / z): 308.1 [M+H] + .

[0210] Step 2: Compound 10⁻² (166 mg, 0.540 mmol) and NaOH (108.02 mg, 2.701 mmol) were added to a mixed solution of methanol (2 mL) and water (2 mL). The reaction solution was stirred at 25 °C for 1 hour. After the reaction was complete, the pH was adjusted to 6-7 with 1 M hydrochloric acid solution, and DCM (10 mL x 3) was added for extraction. The combined extracts were dried over anhydrous Na₂SO₄, filtered, and the filtrate was concentrated under reduced pressure to obtain compound 10⁻³ (150 mg, crude product) as a white solid, which was directly used in the next step of the reaction.

[0211] MS-ESI (m / z): 292.1 [MH] + .

[0212] Step 3: Compound 10⁻³ (150 mg, 0.511 mmol), compound 2⁻³ (141.32 mg, 1.023 mmol), HATU (213.90 mg, 0.563 mmol), and DIEA (0.423 mL, 2.557 mmol) were dissolved in DMF (3 mL). The reaction solution was stirred at 25 °C for 1 hour. After the reaction was complete, water (20 mL) was added to the reaction solution, and the mixture was extracted with EtOAc (20 mL x 3). The combined extracts were dried over anhydrous Na₂SO₄, filtered, and the filtrate was concentrated to dryness under reduced pressure. The residue was subjected to silica gel column chromatography (MeOH / DCM = 0%-10%) to obtain compound 10⁻⁴ (160 mg, total yield of two steps 71.7%) as a white solid.

[0213] MS-ESI (m / z): 414.1 [M+H] + .

[0214] Step 4: Compound 10-4 (160 mg, 0.387 mmol) was dissolved in methanol (5 mL). Ammonium carbonate (55.78 mg, 0.580 mmol) and iodophenyldiacetic acid (288.47 mg, 0.890 mmol) were added to the solution, and the mixture was stirred at 25 °C for 2 hours. After the reaction was completed, the reaction solution was concentrated under reduced pressure. The residue was subjected to silica gel column chromatography (MeOH / DCM = 0%-15%) to give compound 10 (150 mg, yield: 87.2%) as a white solid.

[0215] 1H-NMR(400MHz,DMSO-d6)δ11.52(brs,1H),8.82-9.00(m,1H),8.37-8.47(m,1H),7.52-7.61(m,1H),7.29-7.40(m,1H),7.09-7.19(m,1H ),6.91-7.03(m,1H),6.18-6.42(m,2H),4.49-4.61(m,2H),4.01-4.15(m,1H),3.80-3.91(m,3H),3.13-3.27(m,3H),2.72-2.92(m,3H).

[0216] MS-ESI (m / z): 445.1 [M+H] + .

[0217] Example 11: Preparation of Compound 11

[0218]

[0219] Step 1: Sodium methanethiol (180.43 mg, 2.575 mmol) was added to a solution of compound 11-1 (600 mg, 2.575 mmol) in N,N-dimethylacetamide (5 mL). The mixture was heated to 50 °C and stirred for 3 hours. After the reaction was complete, the reaction solution was poured into water (50 mL) and extracted with EtOAc (10 mL x 2). The combined extracts were washed with saturated brine (10 mL), dried over anhydrous Na₂SO₄, filtered, and the filtrate was concentrated to dryness. The residue was purified by silica gel column chromatography (PE / EtOAc = 10 / 1) to give compound 11-2 (414 mg, yield: 65.1%) as a yellow oil. MS-ESI (m / z): 228.9 / 230.9 [M-OMe] + .

[0220] Step 2: A mixture of dioxane (10 mL) and water (2 mL) of compound 11-2 (414 mg, 1.675 mmol), compound 1-3 (284.73 mg, 1.675 mmol), Pd(dppf)Cl2 (122.59 mg, 0.168 mmol), and K2CO3 (694.61 mg, 5.026 mmol) was heated to 80 °C and stirred for 1 hour under nitrogen protection. After the reaction was complete, the reaction solution was concentrated to dryness under reduced pressure. The residue was purified by silica gel column chromatography (PE / EtOAc = 10 / 1) to give compound 11-3 (260 mg, yield: 50.6%) as a yellow solid.

[0221] MS-ESI (m / z): 275.0 [M-OMe] + .

[0222] Step 3: Add a solution of NaOH (66.59 mg, 1.665 mmol) in H₂O (0.4 mL) to a solution of compound 11-3 (260 mg, 0.849 mmol) in MeOH (2 mL) and tetrahydrofuran (2 mL). Heat the resulting mixture to 65 °C and stir for 2 hours. After the reaction is complete, adjust the pH of the reaction solution to 7-8 with concentrated hydrochloric acid, and concentrate to dryness to obtain compound 11-4 (248 mg, crude product) as a white solid, which can be used directly in the next step of the reaction. MS-ESI (m / z): 291.1 [MH] + .

[0223] Step 4: Compound 2-3 (140.67 mg, 1.018 mmol) was added to a DMF (3 mL) solution of compound 11-4 (248 mg, 0.848 mmol), HATU (387.11 mg, 1.018 mmol), and DIEA (0.701 mL, 4.242 mmol), and the mixture was stirred for another 20 minutes. After the reaction was complete, the reaction mixture was poured into water (30 mL) and extracted with EtOAc (10 mL x 3). The combined extracts were washed with saturated brine (10 mL), dried over anhydrous Na2SO4, filtered, and the filtrate was concentrated to dryness. The residue was purified by silica gel column chromatography (DCM / MeOH = 10 / 1) to give compound 11-5 (160 mg, yield: 45.7%) as a yellow solid.

[0224] MS-ESI (m / z): 413.1 [M+H] + .

[0225] Step 5: Add ammonium carbonate (55.91 mg, 0.582 mmol) to a methanol (4 mL) solution of compound 11-5 (160 mg, 0.388 mmol), and stir the mixture at room temperature for 5 minutes. Then add iodophenyldiacetic acid (289.16 mg, 0.892 mmol) and continue the reaction for 30 minutes. After the reaction is complete, concentrate the mixture under reduced pressure to dryness, and purify the residue by silica gel column chromatography (DCM / MeOH = 10 / 1) to give compound 11 (80 mg, overall yield of five steps: 7.0%) as a yellow solid.

[0226] 1H-NMR(400MHz,DMSO-d6)δ11.46(brs,1H),7.98-8.22(m,1H),7.65-7.92(m,1H),7.29-7.53(m,3H),7.03-7.13(m,1H ),6.86-7.01(m,1H),6.06-6.41(m,2H),3.89-4.88(m,3H),3.78-3.86(m,3H),3.09-3.29(m,3H),2.66-2.94(m,3H).

[0227] MS-ESI (m / z): 444.1 [M+H] + .

[0228] Example 12: Preparation of Compound 12

[0229]

[0230] Step 1: Compound 12-1 (0.204 mL, 2.360 mmol) and potassium carbonate (385.47 mg, 2.789 mmol) were added to a DMF (3 mL) solution of compound 11-1 (500 mg, 2.146 mmol). The mixture was heated to 100 °C and stirred for 18 hours. After the reaction was complete, the reaction solution was poured into water (30 mL) and extracted with EtOAc (10 mL x 3). The combined extracts were washed with saturated brine (10 mL), dried over anhydrous Na2SO4, filtered, and the filtrate was concentrated to dryness. The residue was purified by silica gel column chromatography (DCM / MeOH = 10 / 1) to give compound 12-2 (548 mg, yield: 83.6%) as a white oil.

[0231] MS-ESI(m / z):328.9 / 330.9[M+Na] + .

[0232] Step 2: A mixture of compound 12-2 (360 mg, 1.180 mmol), compound 1-3 (200.47 mg, 1.180 mmol), Pd(dppf)Cl2 (86.31 mg, 0.118 mmol), and K2CO3 (489.06 mg, 3.539 mmol) in dioxane (10 mL) and water (2 mL) was heated to 80 °C and stirred for 1 hour under nitrogen protection. After the reaction was complete, the reaction solution was concentrated to dryness under reduced pressure. The residue was purified by silica gel column chromatography (PE / EtOAc = 10 / 1) to give compound 12-3 (375 mg, yield: 99.8%) as a yellow solid.

[0233] MS-ESI (m / z): 319.0 [M+H] + .

[0234] Step 3: Add a solution of NaOH (238.10 mg, 5.952 mmol) in H₂O (2 mL) to a solution of compound 12-3 (379 mg, 1.190 mmol) in MeOH (5 mL) and tetrahydrofuran (5 mL). Heat the resulting mixture to 65 °C and stir for 2 hours. After the reaction is complete, adjust the pH of the reaction solution to 7-8 with concentrated hydrochloric acid, and concentrate to dryness to obtain compound 12-4 (400 mg, crude product) as a black solid, which can be used directly in the next step of the reaction. MS-ESI (m / z): 335.2 [MH] - .

[0235] Step 4: Under ice bath conditions, thionyl chloride (0.345 mL, 4.757 mmol) was slowly added dropwise to a DCE (3 mL) solution of compound 12-4 (400 mg, 1.189 mmol). After stirring the mixture at room temperature for 1 hour, it was concentrated under reduced pressure to obtain compound 12-5 (443 mg, crude product) as a black solid, which was directly used in the next reaction. MS-ESI (m / z): 337.1 / 339.0 [M+H-Cl] + .

[0236] Step 5: Compound 12-5 (64.85 mg, 0.469 mmol) was added to an 8 mL solution of compound 2-3 (64.85 mg, 0.469 mmol) and triethylamine (0.701 mL, 4.242 mmol) in DCM. The mixture was stirred at room temperature for 1 hour. After the reaction was complete, the reaction solution was concentrated to dryness under reduced pressure. The residue was purified by silica gel column chromatography (DCM / MeOH = 10 / 1) to give compound 12-6 (70 mg, yield: 37.6%) as a yellow solid.

[0237] MS-ESI (m / z): 475.1 [M+H] + .

[0238] Step 6: Ammonium carbonate (21.24 mg, 0.221 mmol) was added to a methanol (1 mL) solution of compound 12-6 (70 mg, 0.147 mmol), and the mixture was stirred at room temperature for 5 minutes. Then, iodophenyldiacetic acid (109.86 mg, 0.339 mmol) was added, and the reaction was continued for 30 minutes. After the reaction was complete, the mixture was concentrated to dryness under reduced pressure, and the residue was purified by silica gel column chromatography (DCM / MeOH = 10 / 1) to give compound 12-7 (30 mg, yield: 43.3%) as a yellow solid. MS-ESI (m / z): 506.1 [M+H] + .

[0239] Step 7: Compound 12-7 (30 mg, 0.059 mmol) was added to a 37% ammonia solution (30 mL), and a few drops of MeOH were added dropwise. The mixture was then stirred at 80 °C for 2 hours. After concentrating under reduced pressure to dryness, the residue was purified by reversed-phase column chromatography to obtain the crude product. The crude product was purified by silica gel column chromatography (DCM / MeOH = 10 / 1) to obtain compound 12-8 (13 mg, total yield of seven steps: 6.3%) as a white solid.

[0240] 1 H-NMR(400MHz,DMSO-d6)δ11.50(brs,1H),7.69-8.16(m,2H),7.40-7.59(m,2H),7.28-7.38(m,1H),7.04-7.13(m,1H ),6.83-6.98(m,1H),6.14-6.47(m,2H),3.77-4.21(m,5H),3.35-3.66(m,4H),2.75-2.93(m,3H),2.10-2.38(m,2H).

[0241] MS-ESI (m / z): 470.1 [M+H] + .

[0242] Example 13: Preparation of Compound 13

[0243]

[0244] Step 1: Compound 10 (40 mg, 0.090 mmol), formaldehyde aqueous solution (0.007 mL, 0.099 mmol), triethylsilane (31.39 mg, 0.270 mmol), and InCl3 (1.00 mg, 0.004 mmol) were dissolved in 1,2-dichloroethane (1 mL). The reaction solution was stirred at 70 °C for 6 hours. After the reaction was completed, the reaction solution was concentrated to dryness under reduced pressure. The residue was purified by silica gel column chromatography (MeOH / DCM = 0%-10%) to obtain compound 13 (3.77 mg, 9.1%) as a white solid.

[0245] 1H-NMR(400MHz,DMSO-d6)δ11.50(brs,1H),8.87-8.97(m,1H),8.25-8.33(m,1H),7.53-7.62(m,1H),7.29-7.46(m,2H),7.11-7.17(m,1H) ),6.93-7.01(m,1H),6.29-6.41(m,1H),4.43-4.69(m,2H),3.82-3.87(m,3H),3.25-3.29(m,3H),2.77-2.89(m,3H),2.58-2.63(m,3H).

[0246] MS-ESI (m / z): 459.1 [M+H] + .

[0247] Comparative Example 1: Preparation of the Comparative Compound

[0248] Compound 57 was prepared according to the description in WO2024179572.

[0249]

[0250] Compare with compound 57 in WO2024179572.

[0251] Test case

[0252] 1. ITC Experiment

[0253] Instrument: MicroCal PEAQ-ITC

[0254] Consumables: 1ml, 200ul and 10ul pipettes and tips, 1.5ml EP tubes, buffer solutions, proteins, and compounds.

[0255] Preparation: Use the website https: / / web.expasy.org / cgi-bin / protparam / protparam to calculate the protein molecular weight and absorbance.

[0256] 1. PRYSPRY protein was dissolved in buffer A: 150 mM NaCl, 20 mM Hepes, pH = 8.0

[0257] 2. Compound: 20 mM stock solution in DMSO

[0258] 3. The compound was diluted 100-fold with buffer A to obtain a 1% DMSO solution.

[0259] 4. Protein buffer B: 150mM NaCl, 20mM Hepes, pH=8.0 + 1% DMSO replacement (change the buffer 4 times), concentrate and then measure the concentration.

[0260] 5. Final concentration: 200 μM for compound, ≥20 μM for protein (increase concentration if signal is insufficient), in buffer B.

[0261] Experimental procedure:

[0262] 1. Check the washing solution (more than two-thirds, DECAN90 to pure water volume ratio is 14%) and waste liquid (less than one-third).

[0263] 2. Turn on the computer and other machines.

[0264] 3. Open the ITC software on your computer and wait a while. You can control the temperature and the cleaning status of the sample cell and titration cell below.

[0265] 4. Cleaning:

[0266] Place the sample cleaner into the sample cell and press firmly; remove the titration cell, insert the FPA tube, place it in the wash position, and press and lock the buckle; select Cell clean:wash (methanol cleaning) and Syringe clean:wash in the software.

[0267] After the instrument is cleaned, use the sampling needle on the table to draw out the residual liquid in the sample cell (pick it onto a paper towel, wipe the syringe with the paper towel until no more liquid comes out and there is no liquid film in the sample cell, about 20 times). Then, suspend the sample cell and titration cell in the air and let them air dry for another 5 minutes.

[0268] 5. Sample loading: First, turn on the small light.

[0269] Sample cell: Wash the sample needle twice with water and twice with buffer solution to remove air bubbles. Use the sample needle to draw 300 μL of protein sample, insert it to the bottom of the sample cell, and add the sample very slowly. Once liquid overflows from the sample well, slowly pull it up while continuing to add the sample slowly. After completely pulling it out, aspirate the excess liquid. The excess volume should be about 20 μL, and the sample cell should contain 280 μL.

[0270] Titration cell: Add 60 μL of the compound to the small tube, place it in the load position, keep the FPA inserted in the titration cell, place it in the small tube, click load on the software, and observe that there are no air bubbles in the titration cell after completion. Turn off the indicator light.

[0271] 6. Start running:

[0272] Remove the FPA from the titration cell, slowly place the titration cell into the sample cell, select the injection 19 method in the software, input the concentrations of the compound (syringe: 200e-6) and protein (cell: 20e-6), save the running file, and click start.

[0273] The baseline height is set to 10 ucal / s, and after equilibration it should be 10 plus or minus 0.5. If the difference is large, the sample should be loaded again; each drop is spaced 90s apart.

[0274] 7. Analyze the data:

[0275] In the ITC software, directly click "analyze" to open the analysis software, then go to "open-presentation-finalfigure," select "subtract baseline," "show result," and export the data and image.

[0276] 8. Clean up the equipment after the experiment:

[0277] Wash the sample cell and titration cell, and return the sample cell washer to its original position. Fill the sample cell with water using a syringe. Return the titration cell to the clean position, keeping it in the pop-out state. Clean the sample loading needle and empty the waste liquid.

[0278] 9. Experimental Results:

[0279]

[0280]

[0281]

[0282] 2. BLI Experiment

[0283] Instrument: BLI-R8 Molecular Interaction Analyzer

[0284] Consumables: 1ml, 200ul and 10ul pipettes and tips, 1.5ml EP tubes, buffer solutions, proteins, compounds, SSA sensors

[0285] Preparation: Use the website https: / / web.expasy.org / cgi-bin / protparam / protparam to calculate the protein molecular weight and absorbance.

[0286] 1. PRYSPRY protein was dissolved in buffer C: PBS + 0.02% Tween-20, pH = 6.95

[0287] 2. Compound: 20 mM stock solution in DMSO

[0288] 3. The compound was diluted 100 times with buffer C to obtain a 1% DMSO solution.

[0289] 4. Protein Buffer D: PBS + 0.02% Tween-20 + 1% DMSO, pH = 6.95. Replace the buffer 4 times, concentrate, and then measure the concentration.

[0290] 5. Final concentration: Compound 200 μM, protein, in buffer D

[0291] Experimental procedure:

[0292] 1. Biotinylate the protein using a biotinylation kit. Incubate at room temperature for 1 hour, then elute the biotinylated protein using a desalting column. Measure the protein concentration.

[0293] 2. First turn on the instrument (the power switch is in the lower right corner at the back), then turn on the computer. Open the Octet BLIdiscovery software.

[0294] 3. Take 2N+2 (N is the number of compounds to be measured at the same time; use 4 sensors when N=1) sensors, put them in an empty box, place a pre-humidified plate underneath, and add 200ul of protein buffer under each sensor.

[0295] Pre-wet for 10 minutes.

[0296] 4. Solidified protein:

[0297] Wait for the system to perform a self-check. Once it's ready, select kinetics and then blank experiment.

[0298] 1) Plate definition (Set sample plate)

[0299] In the first column, select "buffer" to add buffer. In the second column, select "load" to add proteins (select from top to bottom, the quantity is the number of compounds + 1).

[0300] 2) Assay definition (setting the experimental method)

[0301] Add baseline & loading & baseline

[0302] Adjusted times: baseline1 (buffer wells): 60s, loading (protein wells): 600s, baseline2 (buffer wells): 120s

[0303] Select sensor type: SSA

[0304] 3) Sensor assignment (setting up the sensor and methods)

[0305] Check the box to replace sensor

[0306] Delete the empty columns at the beginning; the next column will be the default sensor position (same as step 1). Change all sensor types to SSA.

[0307] PS: Select the sensors in the left column for loading, and the sensors in the right column are the protein-free control.

[0308] 4) Review experiment (simulation experiment)

[0309] To check for errors, click → Simulate Run.

[0310] 5) Run the experiment

[0311] Select the folder to save the data, rename it, and delete the delay time and temperature control settings.

[0312] 5. Determination of small molecule affinity:

[0313] After loading is complete, rebuild the experiment method:

[0314] kinetics, blank experiment

[0315] 1) Plate definition (Set sample plate)

[0316] In the first column, select "buffer" to add buffer. In the second column, select "load" to add proteins (select from top to bottom, the quantity is the number of compounds + 1).

[0317] 2) Assay definition (setting the experimental method)

[0318] Add association & dissociation

[0319] Adjusted time: baseline (buffer well): 60s, association (protein well): 90s, dissociation (buffer well): 90s

[0320] Select sensor type: SSA

[0321] 3) Sensor assignment (setting up the sensor and methods)

[0322] Check the box to replace sensor

[0323] Delete the empty columns at the beginning; the next column will be the default sensor position (same as step 1). Change all sensor types to SSA.

[0324] PS: Select the sensors in the left column to load.

[0325] 4) Review experiment (simulation experiment)

[0326] To check for errors, click → Simulate Run.

[0327] 5) Run the experiment

[0328] Select the folder to save the data, rename it, and delete the delay time and temperature control settings.

[0329] Single-concentration experiments only test 200 μM compounds. Multi-concentration experiments: samples are processed from low to high concentrations (concentrations are entered in advance).

[0330] 6. Results Analysis:

[0331] Open Octet Analysis Studio 13.0 software.

[0332] 1. Open the file

[0333] 2. In the processed data, the reference sensor and reference sample are subtracted from each other.

[0334] For data correction, select "average of baseline step" and check the box for "savitzky-Golayfiltering".

[0335] 3. Kinetic analysis

[0336] Choose association and dissociation, type: global (select global for multi-concentration experiments, select single for single-concentration experiments), Rmax: sensor. Finally, select response.

[0337] The curve fit shown on the right can remove concentration points with no signal.

[0338] In the graph settings above, you can disable "show baseline". Click the last group graph options, add a sample ID and KD, and check "steady-state" in the lower left corner.

[0339] 7. View the analysis results:

[0340] 1. If the signal height is less than 0.01, the presence of compound binding is not considered.

[0341] 2. If it is a negative signal, and the absolute value of the signal from the empty sensor is greater than that from the protein sensor, then the binding of the compound is not considered.

[0342] 3. For multi-concentration experiments, the steady-state fitted KD is used as the standard.

[0343] Note: 1. Double subtraction method experimental design is used for small molecules. Double subtraction method means that the last row contains only protein and no small molecule (reference well), and the second column contains only sensor and no protein (reference sensor).

[0344] 3. Add 200ul of buffer or sample to all wells. The protein concentration is generally 50ug / ml, but can be higher.

[0345] 4. The loading (curing) time can be extended (originally 600s) to ensure a binding height > 4nm.

[0346] 5. The left side shows the sensor and pre-wetted plate, and the right side shows the sample plate.

[0347] 3. Liver microsomal stability

[0348] Experimental Methods: 30 μL of a mixture of the test compound and liver microsomes was added to each well of a 96-well plate to prepare two parallel samples. After pre-incubation at 37°C for 10 min, 15 μL of NADPH solution (6 mM) was added at time points. The final concentration of the test compound was 1 μM, the concentration of liver microsomes was 0.5 mg / mL, and the final concentration of NADPH was 2 mM. Incubation was performed for 0, 15, 30, and 60 min, respectively. After incubation, 150 mL of acetonitrile (containing internal standard) was added to the mixture. The acetonitrile-diluted sample was centrifuged at 4000 rpm for 5 min, and 150 μL of the supernatant was analyzed by LC-MS / MS. The half-life (T5) of the compound in liver microsome solutions of different species was obtained. 1 / 2 ) and the corresponding CL int(liver) Parameters such as values ​​were analyzed. The metabolic rate of the compounds of this invention in liver microsomes of different species was analyzed.

[0349] 4. In vivo pharmacokinetic properties

[0350] Experimental objective: To determine the pharmacokinetic properties of the compound in balb / c mice.

[0351] Experimental materials: male Balb / c mice

[0352] Experimental methods:

[0353] 1. Six male mice were used. They were weighed before administration, and the dosage was calculated based on their body weight. The mice were then divided into two groups. One group of three mice was administered the drug via intravenous injection, and the other group of three mice was administered the drug orally.

[0354] 2. Collect plasma samples at 0.083, 0.25, 0.5, 1, 2, 4, 8, and 24 hours after drug administration. Collect approximately 0.05 mL of each sample, anticoagulated with sodium heparin, and place on moist ice after collection.

[0355] 3. After blood sample collection, place it on ice and centrifuge to separate the plasma within 1 hour. Plasma samples should be stored at -80°C before analysis.

[0356] 4. Perform LC-MS / MS analysis on the collected samples and acquire data. Use PhoenixWinNonlin 8.2.0 software to calculate relevant pharmacokinetic parameters, such as peak concentration (Cmax), clearance (Cl), half-life (T1 / 2), area under the curve (AUC), and bioavailability.

[0357] The pharmacokinetic properties of the compounds of this invention in mice were analyzed based on the above data.

[0358] Although the invention has been described in detail with reference to embodiments thereof, these embodiments are provided for illustration and not limitation. Other embodiments that can be obtained according to the principles of the invention fall within the scope defined by the claims of the invention.

[0359] The contents of all documents and references listed in this article are incorporated herein by reference in their entirety.

Claims

1. A compound represented by Formula (I), or a pharmaceutically acceptable salt, solvate, optical isomer, stereoisomer, polymorph, or isotopically enriched compound thereof: wherein, X1, X2, X3are each independently selected from CH or N; R1, R2, and A ring are defined according to any one of claims 1 to 11; L represents a linker group, P represents a ligand group targeting degradation of a target protein.

2. The compound of claim 1, wherein the compound has a structure represented by Formula (Ia): A ring is selected from a 6-10 membered aromatic ring, a 5-10 membered aromatic heterocyclic ring, a 3-10 membered cycloalkyl ring, a 3-10 membered heterocyclic ring; said A ring is optionally substituted with one, two, three, or four independent R, wherein each R is independently selected from hydrogen, halogen, cyano, nitro, substituted or unsubstituted C 1-6 alkyl, substituted or unsubstituted C 2-6 alkenyl, substituted or unsubstituted C 2-6 alkynyl, -C 0-4 alkylene-OR a , -C 0-4 alkylene-OC(=O)R a , -C 0-4 alkylene-SR a , -C 0-4 alkylene-S(=O)2R a , -C 0-4 alkylene-S(=O)R a , -C 0-4 alkylene-S(=O)2NR a R b , -C 0-4 alkylene-S(=O)NR a R b , -C 0-4 alkylene-C(=O)R a , -C 0-4 alkylene-C(=O)OR a , -C 0-4 alkylene-C(=O)NR a R b , -C 0-4 alkylene-NR a R b , -C 0-4 alkylene-NR a C(=O)R b , -C 0-4 alkylene-NR a S(=O)2R b , -C 0-4 alkylene-NR a S(=O)R b , -C 0-4 alkylene-(3-10 membered cycloalkyl), -C 0-4 alkylene-(3-10 membered heterocycloalkyl), -C 0-4 alkylene-(6-10 membered aromatic ring), -C 0-4 alkylene-(5-10 membered heteroaromatic ring); wherein each R a , R b is independently selected from hydrogen, C 1-6 alkyl, C 2-6 alkenyl, C 2-6 alkynyl, halogen-substituted C 1-6 alkyl, halogen-substituted C 2-6 alkenyl, halogen-substituted C 2-6 alkynyl, -C 0-4 alkylene-(3-10 membered cycloalkyl), -C 0-4 alkylene-(3-10 membered heterocycloalkyl), -C 0-4 alkylene-(6-10 membered aryl), -C 0-4 alkylene-(5-10 membered heteroaryl); 3. The compound of claim 1, wherein the compound has a structure represented by Formula (IIa): R1is selected from -S(=0)2R 1a , -P(=0)R 1a R 1b , -C(=0)R 1a , -S(=0)(=NH)R 1a , -S(=O)(=NR 1a )R 1b ; -N=S(=O)R 1a R 1b ; wherein R 1a and R 1b are each independently selected from substituted or unsubstituted C 1-6 1-6 alkyl, or R 1a and R 1b together with the atoms to which they are attached form a 4- to 6-membered ring; provided that when R1is selected from -S(=O)2R 1a , X1, X2, X3are not simultaneously CH; R2and R3are each independently selected from the group consisting of hydrogen, -OR 2a , substituted or unsubstituted C 1-6 alkyl, substituted or unsubstituted C 2-6 alkenyl, substituted or unsubstituted C 2-6 alkynyl, -C 0-4 alkylene-OR 2a , -C 0-4 alkylene-OC(=O)R 2a , -C 0-4 alkylene-SR 2a , -C 0-4 alkylene-S(=O)2R 2a , -C 0-4 alkylene-S(=O)R 2a , -C 0-4 alkylene-S(=O)2NR 2a R 2b , -C 0-4 alkylene-S(=O)NR 2a R 2b , -C 0-4 alkylene-C(=O)R 2a , -C 0-4 alkylene-C(=O)OR 2a , -C 0-4 alkylene-C(=O)NR 2a R 2b , -C 0-4 alkylene-NR 2a R 2b , -C 0-4 alkylene-NR 2a C(=O)R 2b , -C 0-4 alkylene-NR 2a S(=O)2R 2b , -C 0-4 alkylene-NR 2a S(=O)R 2b , -C 0-4 alkylene-(3-10 membered cycloalkyl), -C 0-4 alkylene-(3-10 membered heterocycloalkyl), -C 0-4 alkylene-(6-10 membered aryl), -C 0-4 alkylene-(5-10 membered heteroaryl), wherein R 2a , R 2b are each independently selected from the group consisting of hydrogen, C 1-6 alkyl, C 2-6 alkenyl, C 2-6 alkynyl, halogen-substituted C 1-6 alkyl, halogen-substituted C 2-6 alkenyl, halo-substituted C 2-6 alkynyl.

2. The compound of claim 1, wherein, The A ring is selected from a 6-10 membered aromatic ring, and the A ring is optionally substituted with two independent R, wherein each R is independently selected from halogen, substituted or unsubstituted C 1-6 alkyl, -C 0-4 alkylene-OR a wherein R a is independently selected from hydrogen, C 1-6 alkyl.

3. The compound of claim 1 or 2, wherein, R2and R3are each independently selected from C 1-6 alkyl or -C 0-4 alkylene-(5- to 10-membered heteroaryl ring).

4. The compound according to any one of claims 1 to 3, wherein, 4. The compound of claim 1, wherein the compound has a structure represented by Formula (Ib): wherein R1is selected from -P(=O)R 1a R 1b ; -C(=O)R 1a ; -S(=O)(=NH)R 1b ; -S(=O)(=NR 1a )R 1b ; -N=S(=O)R 1a R 1b ; wherein R 1a and R 1b are each independently selected from substituted or unsubstituted C 1-6 1-6 alkyl, or R 1a and R 1b together with the atoms to which they are attached form a 4- to 6-membered ring.

5. The compound of claim 4, wherein, 5. The compound of claim 1, wherein the compound has a structure represented by Formula (IIb): wherein each R is independently selected from the group consisting of: halogen, such as fluorine, chlorine, bromine and iodine; C 1-6 alkyl, such as methyl, ethyl, propyl, isopropyl; -OR a wherein R a is independently selected from the group consisting of hydrogen, methyl, ethyl, propyl, cyclopropyl, isopropyl; R1is selected from -S(=0)(=NH)R 1a , -S(=0)(=NR 1a )R 1b , -N=S(=0)R 1a R 1b wherein R 1a and R 1b are each independently C 1-6 1-6 alkyl, for example methyl, ethyl, propyl, cyclopropyl, isopropyl, or R 1a and R 1b together with the atom to which they are attached form a 4- to 6-membered ring; and R2and R3are each independently selected from the group consisting of: C 1-6 alkyl, such as methyl, ethyl, cyclopropyl, propyl, isopropyl; -C 1-4 alkylene-(5- to 10-membered heteroaromatic ring), such as 6. The compound according to any one of claims 1 to 3, wherein, 6. The compound of claim 1, wherein the compound has a structure represented by Formula (Ic): wherein R1is selected from -S(=O)2R 1a , -C(=O)R 1a , -P(=O)R 1a R 1b , -S(=O)(=NH)R 1b , -S(=O)(=NR 1a )R 1b , -N=S(=O)R 1a R 1b ; wherein R 1a and R 1b are each independently selected from substituted or unsubstituted C 1-6 alkyl, or R 1a and R 1b together with the atom to which they are attached form a 4- to 6-membered ring.

7. The compound of claim 6, wherein, 7. The compound of claim 1, wherein the compound has a structure represented by Formula (IIc): wherein each R is independently selected from the group consisting of: halogen, such as fluorine, chlorine, bromine and iodine; C 1-6 alkyl, such as methyl, ethyl, propyl, cyclopropyl, isopropyl; -OR a wherein R a is independently selected from the group consisting of hydrogen, methyl, ethyl, cyclopropyl, isopropyl, propyl; R1is selected from -S(=0)2R 1a , -P(=0)R 1a R 1b , -S(=0)(=NR 1a )R 1b , -N=S(=0)R 1a R 1b ; R 1a R 1b each independently is selected from C 1-6 alkyl, for example methyl, ethyl, propyl, isopropyl, or R 1a R 1b together with the atom to which they are attached form a 4- to 6-membered ring; and R2and R3are each independently selected from the group consisting of: C 1-6 alkyl, such as methyl, ethyl, cyclopropyl, propyl, isopropyl; -C 1-4 alkylene-(5- to 10-membered heteroaromatic ring), such as 8. The compound according to any one of claims 1 to 3, wherein, 8. The compound of claim 1, wherein the compound has a structure represented by Formula (Id): wherein R1is selected from -S(=O)2R 1a , -C(=O)R 1a , -P(=O)R 1a R 1b , -S(=O)(=NH)R 1b , -S(=O)(=NR 1a )R 1b , -N=S(=O)R 1a R 1b ; wherein R 1a and R 1b are each independently selected from substituted or unsubstituted C 1-6 alkyl, or R 1a and R 1b together with the atoms to which they are attached form a 4- to 6-membered ring.

9. The compound of claim 8, wherein, 9. The compound of claim 1, wherein the compound has a structure represented by Formula (IId): wherein each R is independently selected from the group consisting of: halogen, such as fluorine, chlorine, bromine and iodine; C 1-6 alkyl, such as methyl, ethyl, propyl, cyclopropyl, isopropyl; -OR a wherein R a is independently selected from the group consisting of hydrogen, methyl, ethyl, propyl; R1is selected from -S(=0)2R 1a , -N=S(=0)R 1a R 1b ; wherein R 1a and R 1b are each independently selected from C 1-6 1-6 alkyl, for example methyl, ethyl, propyl, cyclopropyl, isopropyl, or R 1a and R 1b together with the atoms to which they are attached form a 4- to 6-membered ring; R2and R3are each independently selected from the group consisting of: C 1-6 alkyl, such as methyl, ethyl, cyclopropyl, propyl, isopropyl; -C 1-4 alkylene-(5- to 10-membered heteroaromatic ring), such as 10. The compound according to any one of claims 1 to 3, wherein, 12. A compound selected from the following, or a pharmaceutically acceptable salt, solvate, optical isomer, stereoisomer, polymorph, or isotopically enriched compound thereof: wherein R1is selected from -S(=O)2R 1a , -C(=O)R 1a , -P(=O)R 1a R 1b , -S(=O)(=NH)R 1b , -S(=O)(=NR 1a )R 1b , -N=S(=O)R 1a R 1b ; wherein each R 1a is independently selected from substituted or unsubstituted C 1-6 alkyl.

11. The compound of claim 10, wherein, 13. A compound represented by Formula (II), or a pharmaceutically acceptable salt, solvate, optical isomer, stereoisomer, polymorph, or isotopically enriched compound thereof: wherein, R1, R2, X1, X2, X3, and A ring are defined according to any one of claims 1 to 11; L represents a linker group, P represents a ligand group targeting degradation of a target protein. wherein each R is independently selected from the group consisting of: halogen, such as fluorine, chlorine, bromine and iodine; C 1-6 alkyl, such as methyl, ethyl, propyl, isopropyl; -OR a wherein R a is independently selected from the group consisting of hydrogen, methyl, ethyl, propyl; R1is selected from -S(=0)2R 1a , -C(=0)R 1a ; wherein R 1a each independently is selected from C 1-6 alkyl, such as methyl, ethyl, cyclopropyl, propyl, isopropyl; R2and R3are each independently selected from the group consisting of: C 1-6 alkyl, such as methyl, ethyl, cyclopropyl, propyl, isopropyl; -C 1-4 alkylene-(5- to 10-membered heteroaromatic ring), such as 14. A pharmaceutical composition comprising a compound according to any one of claims 1 to 13, or a pharmaceutically acceptable salt, solvate, optical isomer, stereoisomer, polymorph, or isotopically enriched compound thereof, and a pharmaceutically acceptable carrier.

15. Use of a compound according to any one of claims 1 to 13, or an optical isomer, stereoisomer, polymorph, or isotopically enriched compound thereof, or a pharmaceutical composition of claim 14, in the manufacture of a medicament, including but not limited to inducing TRIM21 -dominated targeted protein degradation in cells using a compound and derivatives according to any one of claims 1 to 13, or a pharmaceutical composition of claim 14. wherein 16. Use of a compound according to any one of claims 1 to 13, or a chemical synthesis route thereof, or a synthetic intermediate thereof, in the synthesis of a medicament. ​ ​ ​ ​ ​

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