CN111303133A - Small-molecule compounds that degrade EZH2 protein - Google Patents

Abstract

The invention provides a compound which is a compound shown as a formula I or a stereoisomer, a geometric isomer, a tautomer, a nitrogen oxide, a hydrate, a solvate, a metabolite, a pharmaceutically acceptable salt or a prodrug thereof: wherein X represents a ligand of EZH2 protein, Z represents a ligand of E3 ligase, and Y represents a chain connecting X and Z. The compound provided by the invention is a PROTAC molecule capable of degrading EZH2, and compared with the existing EZH2 inhibitor, the small molecule compound provided by the invention has the capability of rapidly inhibiting tumor proliferation, and has the potential to become an effective treatment mode for treating malignant tumors. X-Y-Z is formula I.

Description

Small-molecule compounds that degrade EZH2 protein

technical field

The present invention relates to the field of medicine, in particular, the present invention relates to a small molecule compound that degrades EZH2 protein.

Background technique

The enhancer of zeste homolog 2 (EZH2) of the Drosophila zeste gene enhancer is the catalytic subunit of the polycomb repressive complex 2 (PRC2), which trimethylates histone H3 Lysine at position 27 inhibits the transcription of target genes and participates in the regulation of physiological or pathological processes such as cell cycle, cell senescence, and cell differentiation. Studies have found that EZH2 overexpression and abnormal regulation can be detected in various cancer cells. During tumorigenesis, the expression level of EZH2 is continuously and steadily increased, and it is often directly related to the high aggressiveness and poor prognosis of cancer. Therefore, inhibition of EZH2 protein activity can be used to treat related tumors.

EZH2 inhibitors can inhibit the methyl transfer activity of the protein in some tumor cells, including diffuse large B-cell lymphoma (DLBCL) and malignant rhabdoid tumor (MRT) cells and The ability to inhibit proliferation is evident in animal models. Although abnormal mutations in EZH2 activity also exist in other types of lymphomas and solid tumors, they are resistant to currently known EZH2 inhibitors. To expand the application of EZH2 inhibitors in other tumor types, it is of great significance to develop EZH2 protein-targeting molecules with novel mechanisms of action. Meanwhile, in addition to its role in transcriptional silencing, some studies have found some functions of EZH2 in transcriptional activation that are independent of the PRC2 complex. For example, EZH2 is involved in transcriptional activation of NOTCH1 and Wnt signaling pathways in breast cancer. In addition, EZH2 was found to bind to the β-catenin transcriptional complex in colon cancer to specifically increase the transcription level of Wnt target genes, independent of its methyltransferase function. EZH2 has a transcriptional co-activation effect on AR expression in castration-resistant prostate cancer (CRPC). Therefore, it is difficult for traditional inhibitors to target methyltransferase pockets to play a role in these aspects that are not dependent on the function of methyltransferases.

Proteolysis Targeting Chimera (PROTAC) utilizes the inherent ubiquitin-proteasome system of cells to regulate the degradation of targeted proteins, and has become a highly potential disease treatment method. Compared with traditional small molecules, PROTAC molecules have high selectivity, anti-mutation, and can inhibit the non-enzymatic function of proteins.

SUMMARY OF THE INVENTION

This discovery involves a PROTAC molecule that can degrade EZH2. The structure is shown in Figure 1. One end of this molecule is targeted to bind to E3 ligase, and the other end of the molecule is targeted to bind to the target protein (EZH2 protein) to be degraded. The structures are connected by chains (linkers) to form a complete compound molecule. Compared with the existing EZH2 inhibitors, the small molecule compounds involved in the present invention have the ability to rapidly inhibit tumor proliferation, and have the potential to become an effective treatment method for treating malignant tumors.

In the first aspect of the present invention, the present invention provides a compound, which is a compound represented by formula I or its stereoisomer, geometric isomer, tautomer, nitrogen oxide, hydrate, solvate , metabolites, pharmaceutically acceptable salts or prodrugs:

X-Y-Z

Formula I.

Among them, X represents the ligand of EZH2 protein, Z represents the ligand of E3 ligase, and Y represents the chain connecting X and Z. The compound proposed in the present invention is a PROTAC molecule that can degrade EZH2. Compared with the existing EZH2 inhibitors, the small molecule compound involved in the present invention has the ability to rapidly inhibit tumor proliferation and has the potential to become an effective treatment for malignant tumors. Way.

According to an embodiment of the present invention, the above-mentioned compound may further include at least one of the following additional technical features:

According to an embodiment of the present invention, the X is an optionally substituted structure shown below:

According to an embodiment of the present invention, the Z is an optionally substituted structure shown below:

According to an embodiment of the present invention, the Y is an optionally substituted structure shown below:

Here, n1 is an integer of 0-3, m is an integer of 1-5, and n2 is an integer of 1-12.

According to an embodiment of the present invention, the X, Y, Z are independently optionally selected by F, Cl, Br, CN, NO ₂ , OR ^b , -NR ^c R ^d , R ^b OC _1-4 alkylene, R ^d R ^c NC _1-4 alkylene, -C(=O)R ^a , -C(=O)OR ^b , -C(=O)NR ^c R ^d , C _1-12 alkyl, C _{1- 6} haloalkyl, C _3-6 cycloalkyl, C _3-6 cycloalkyl-C _1-4 alkylene, heterocyclyl composed of 3-8 atoms, (heterocyclyl composed of 3-8 atoms) )-C _1-4 alkylene, C _6-10 aryl, C _6-10 aryl-C _1-4 alkylene, heteroaryl consisting of 5-8 atoms or (5-8 atoms) Heteroaryl)-C _1-4 alkylene substituted,

Wherein, R ^a , R ^b , R ^c , and R ^d are each independently H, D, C _1-6 haloalkyl, C _1-6 alkyl, C _2-6 alkenyl, C _2-6 alkynyl, C _3-6 carbocyclyl, C _3-6 carbocyclyl-C _1-4 alkylene, heterocyclyl composed of 3-12 atoms, (heterocyclyl composed of 3-12 atoms)-C _{1- 4} alkylene group, C _6-10 aryl group, C _6-10 aryl group-C _1-4 alkylene group, heteroaryl group composed of 5-10 atoms, (heteroaryl group composed of 5-10 atoms) -C _1-4 alkylene, or R ^c , R ^d and the nitrogen atom to which they are attached together form a 3-8 atom heterocyclyl group or a 5-8 atom heteroaryl group, wherein the C _1-6 alkyl, C _2-6 alkenyl, C _2-6 alkynyl, C _3-6 carbocyclyl, C _3-6 carbocyclyl-C _1-4 alkylene, 3-12 atoms Heterocyclic group composed of, (heterocyclic group composed of 3-12 atoms)-C _1-4 alkylene group, C _6-10 aryl group, C _6-10 aryl group-C _1-4 alkylene group, 5 -Heteroaryl consisting of 10 atoms, (heteroaryl consisting of 5-10 atoms)-C _1-4 alkylene, heterocyclyl consisting of 3-8 atoms and heterocyclic group consisting of 5-8 atoms Aryl groups are each independently unsubstituted or substituted with 1, 2, 3 or 4 substituents independently selected from D, F, Cl, CN, OH, _NH2 , _C1-6 alkyl , C _1-6 haloalkyl, C _1-6 alkoxy or C _1-6 alkylamino.

According to an embodiment of the present invention, the R ^a , R ^b , R ^c , and R ^d are each independently trifluoromethyl, methyl, ethyl, isopropyl, n-propyl, n-butyl, tert-butyl , C _3-6 carbocyclic group, heterocyclic group composed of 5-6 atoms, phenyl, heteroaryl group composed of 5-6 atoms, (heteroaryl group composed of 5-6 atoms) -C _{1- 4} Alkylene, or R ^c , R ^d and the nitrogen atom to which they are attached together form a heterocyclic group of 5-6 atoms or a heteroaryl group of 5-6 atoms, wherein the methyl, ethyl base, isopropyl, n-propyl, n-butyl, tert-butyl, C _3-6 carbocyclyl, heterocyclyl of 5-6 atoms, phenyl, heteroaryl of 5-6 atoms and (5-6 atoms of heteroaryl)-C _1-4 alkylene are each independently unsubstituted or substituted with 1, 2, 3 or 4 substituents independently selected from D, F, Cl, CN, OH, _NH2 , _C1-3 alkyl, _C1-3 haloalkyl or methoxy.

In the second aspect of the present invention, the present invention provides a compound, which is the compound shown below or its stereoisomers, geometric isomers, tautomers, nitrogen oxides, hydrates, solvates, metabolites, pharmaceutically acceptable salts or prodrugs,

In the third aspect of the present invention, the present invention provides a pharmaceutical composition. According to an embodiment of the present invention, the pharmaceutical composition comprises the aforementioned compound.

According to an embodiment of the present invention, the pharmaceutical composition further comprises a pharmaceutically acceptable carrier, excipient, diluent, adjuvant, vehicle or a combination thereof.

According to an embodiment of the present invention, the pharmaceutical composition further includes other drugs for treating or preventing tumors.

In the fourth aspect of the present invention, the present invention proposes the use of the aforementioned compound or the aforementioned pharmaceutical composition in the preparation of a medicament for degrading EZH2 or inhibiting EZH2.

In the fifth aspect of the present invention, the present invention proposes the use of the aforementioned compound or the aforementioned pharmaceutical composition in degrading EZH2.

In the sixth aspect of the present invention, the present invention proposes the use of the aforementioned compound or the aforementioned pharmaceutical composition in preparing a medicament for treating or preventing EZH2-related diseases.

According to an embodiment of the present invention, the EZH2-related disease is lymphoma or malignant rhabdoid tumor, optionally, the lymphoma comprises Hodgkin's lymphoma, non-Hodgkin's lymphoma (NHL), diffuse large At least one of B-cell lymphoma (DLBCL), breast cancer and prostate cancer.

In the seventh aspect of the present invention, the present invention proposes the use of the aforementioned compound or the aforementioned pharmaceutical composition in anti-tumor.

According to an embodiment of the present invention, the tumor is lymphoma or malignant rhabdoid tumor.

According to an embodiment of the present invention, the lymphoma comprises at least one selected from the group consisting of Hodgkin's lymphoma, non-Hodgkin's lymphoma (NHL), diffuse large B-cell lymphoma (DLBCL), breast cancer and prostate cancer.

In yet another aspect of the present invention, the present invention presents general synthetic routes for compounds of formula I. According to an embodiment of the present invention, the compound represented by formula I can be connected by a click reaction or an amide condensation reaction between Pomalidomide or Lenanidomide or RG-7112 terminal derivative and X part derivative, as shown in FIG. 2 . Among them, for the preparation method of Pomalidomide terminal derivatives, please refer to the literature Chemistry & Biology 22, 755-763 (2015). The preparation method of 7112-terminal derivatives can be referred to in the literature Bioorg.Med.Chem.Lett.18, 5904-5908 (2008).ACSMed.Chem.Lett.4,466-469 (2013). Synthesis of the parent nucleus of part X reference J.Med.Chem (DOI: 10.1021/acs.jmedchem.5b01501)

Description of drawings

Fig. 1 is the basic technical route of PROTACs according to an embodiment of the present invention;

Fig. 2 is according to the embodiment of the present invention by click reaction and amide condensation reaction to construct the schematic diagram shown in formula I;

Fig. 3-Fig. 5 is the degradation effect of the molecule according to the embodiment of the present invention on the EZH2 protein on some cell lines;

6-8 are the in vitro inhibitory effects of Formula 10 and Formula 12 on various DLBCL cells according to an embodiment of the present invention;

Figures 9-10 show the inhibitory effect of the mouse xenograft of formula 10 according to the embodiment of the present invention.

Detailed ways

The following describes in detail the embodiments of the present invention, examples of which are illustrated in the accompanying drawings. The embodiments described below with reference to the accompanying drawings are exemplary, and are intended to explain the present invention and should not be construed as limiting the present invention.

As used herein, the term "administering to a patient a compound as previously described or a stereoisomer, geometric isomer, tautomer, nitroxide, hydrate, solvate, metabolite, pharmaceutically acceptable The term "salts or prodrugs or the aforementioned pharmaceutical compositions" refers to the introduction of a predetermined amount of a substance into a patient by some suitable means. The compounds of formula I of the present invention or their stereoisomers, geometric isomers, tautomers, nitrogen oxides, hydrates, solvates, metabolites, pharmaceutically acceptable salts or prodrugs or drugs The composition can be administered by any common route so long as it can reach the intended tissue. Various modes of administration are contemplated, including peritoneal, intravenous, intramuscular, subcutaneous, cortical, oral, topical, nasal, pulmonary and rectal, but the invention is not limited to these exemplified modes of administration. However, due to oral administration, the active ingredient of an orally administered composition should be coated or formulated to prevent its degradation in the stomach. In addition, the compounds of formula I or the pharmaceutical compositions of the present invention may be administered using specific devices that deliver the active ingredient to target cells.

The frequency and dosage of administration of the pharmaceutical compositions of the present invention can be determined by a number of relevant factors, including the type of disease to be treated, the route of administration, the age, sex, weight, and severity of the disease of the patient as the active ingredient type of drug.

The term "therapeutically effective amount" refers to an amount of a compound sufficient to significantly ameliorate certain symptoms associated with a disease or disorder, ie, an amount to provide a therapeutic effect for a given disorder and dosage regimen. A therapeutically effective amount of a drug or compound is not required to cure the disease or disorder, but will provide a treatment for the disease or disorder such that the onset of the disease or disorder in an individual is delayed, prevented, or prevented, or the symptoms of the disease or disorder are alleviated, or the disease or disorder is The duration of the condition is altered, or, for example, the disease or condition is less severe, or recovery is accelerated.

The term "treating" is used to refer to obtaining a desired pharmacological and/or physiological effect. The effect may be prophylactic in terms of complete or partial prevention of the disease or symptoms thereof, and/or therapeutic in terms of partial or complete cure of the disease and/or adverse effects caused by the disease. "Treatment" as used herein encompasses the treatment of a disease in a mammal, particularly a human, including: (a) preventing the occurrence of a disease or disorder in individuals susceptible to but not yet diagnosed with the disease; (b) inhibiting the disease; or (c) Alleviate disease, such as reducing symptoms associated with disease. "Treatment" as used herein encompasses any administration of a drug or compound to an individual to treat, cure, alleviate, ameliorate, alleviate or inhibit a disease in the individual, including, but not limited to, administering a compound or drug combination of Formula I or Formula II described herein The giving of things to individuals in need.

According to an embodiment of the present invention, the adjuvants include pharmaceutically acceptable excipients, lubricants, fillers, diluents, disintegrants, stabilizers, preservatives, emulsifiers, cosolvents, and colorants that are well known in the formulation field. , sweetener, made into tablets, pills, capsules, injections and other different dosage forms.

Certain embodiments of the invention will now be described in detail, examples of which are illustrated by the accompanying structural and chemical formulae. The present invention is intended to cover all alternatives, modifications and equivalents, which are included within the scope of the present invention as defined by the claims. One skilled in the art will recognize that many methods and materials similar or equivalent to those described herein could be used in the practice of the present invention. The present invention is in no way limited to the methods and materials described herein. In the event that one or more of the incorporated literature, patents, and similar materials differs from or contradicts this application (including, but not limited to, terms defined, uses of terms, techniques described, etc.), this Application shall prevail.

It should further be appreciated that certain features of the invention, which are, for clarity, described in the context of multiple separate embodiments, can also be provided in combination in a single embodiment. Conversely, various features of the invention, which are, for brevity, described in the context of a single embodiment, can also be provided separately or in any suitable subcombination.

Unless otherwise defined, 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 belongs. All patents and publications referred to herein are incorporated by reference in their entirety.

Unless otherwise stated, the following definitions as used herein shall apply. For the purposes of the present invention, chemical elements are in accordance with the Periodic Table of the Elements, CAS Edition, and Handbook of Chemistry and Physics, 75th Edition, 1994. In addition, general principles of organic chemistry may be referred to as described in "Organic Chemistry", Thomas Sorrell, University Science Books, Sausalito: 1999, and "March's Advanced Organic Chemistry" by Michael B. Smith and Jerry March, John Wiley & Sons, New York: 2007, Its entire contents are incorporated herein by reference.

As used herein, the articles "a", "an" and "the" are intended to include "at least one" or "one or more" unless stated otherwise or otherwise clearly contradicted by context. Thus, as used herein, these articles refer to one or more than one (ie, at least one) object of the article. For example, "a component" refers to one or more components, ie, there may be more than one component contemplated for use or use in the implementation of the described embodiments.

The term "comprising" is an open-ended expression, that is, it includes the contents specified in the present invention, but does not exclude other aspects.

"Stereoisomers" refer to compounds that have the same chemical structure, but differ in the arrangement of atoms or groups in space. Stereoisomers include enantiomers, diastereomers, conformational isomers (rotamers), geometric isomers (cis/trans), atropisomers, etc. .

"Chiral" is a molecule that has the property of being non-superimposable with its mirror image; while "achiral" refers to a molecule that is superimposable with its mirror image.

"Enantiomer" refers to two nonsuperimposable, but mirror-image isomers of a compound.

"Diastereomer" refers to a stereoisomer having two or more centers of chirality and whose molecules are not mirror images of each other. Diastereomers have different physical properties such as melting point, boiling point, spectral properties and reactivity. Diastereomeric mixtures can be separated by high resolution analytical procedures such as electrophoresis and chromatography, eg HPLC.

Stereochemical definitions and rules used in the present invention generally follow S.P. Parker, Ed., McGraw-Hill Dictionary of Chemical Terms (1984) McGraw-Hill Book Company, New York; and Eliel, E. and Wilen, S., "Stereochemistry of Organic Compounds", John Wiley & Sons, Inc., New York, 1994.

Many organic compounds exist in optically active forms, that is, they have the ability to rotate the plane of plane-polarized light. In describing optically active compounds, the prefixes D and L or R and S are used to denote the absolute configuration of the molecule about one or more of its chiral centers. The prefixes d and 1 or (+) and (-) are symbols used to designate the rotation of plane polarized light by the compound, where (-) or 1 indicates that the compound is levorotatory. Compounds prefixed with (+) or d are dextrorotatory. A specific stereoisomer is an enantiomer, and a mixture of such isomers is called an enantiomeric mixture. A 50:50 mixture of enantiomers is called a racemic mixture or racemate, which can occur when there is no stereoselectivity or stereospecificity in a chemical reaction or process.

Any asymmetric atom (eg, carbon, etc.) of the compounds disclosed herein may exist in racemic or enantiomerically enriched forms, such as (R)-, (S)- or (R,S)-configurations exist. In certain embodiments, each asymmetric atom has at least 50% enantiomeric excess, at least 60% enantiomeric excess, at least 70% enantiomeric excess, at least 70% enantiomeric excess in the (R)- or (S)-configuration 80% enantiomeric excess, at least 90% enantiomeric excess, at least 95% enantiomeric excess, or at least 99% enantiomeric excess.

Depending on the choice of starting materials and methods, the compounds of the present invention may be present as one of the possible isomers or as mixtures thereof, such as racemates and diastereoisomeric mixtures (depending on the number of asymmetric carbon atoms) ) in the form of. Optically active (R)- or (S)-isomers can be prepared using chiral synthons or chiral reagents, or resolved using conventional techniques. If the compound contains a double bond, the substituent may be in the E or Z configuration; if the compound contains a disubstituted cycloalkyl, the cycloalkyl substituent may have the cis or trans configuration.

The resulting mixtures of any stereoisomers may be separated into pure or substantially pure geometric isomers, enantiomers, diastereomers on the basis of differences in the physicochemical properties of the components, for example, by chromatography and/or fractional crystallization.

Any resulting racemate of the final product or intermediate can be resolved into the optical enantiomers by known methods by methods familiar to those skilled in the art, eg, by performing diastereomeric salts thereof obtained. separation. Racemic products can also be separated by chiral chromatography, eg, high performance liquid chromatography (HPLC) using chiral adsorbents. In particular, enantiomers can be prepared by asymmetric synthesis, for example, see Jacques, et al., Enantiomers, Racemates and Resolutions (Wiley Interscience, New York, 1981); Principles of Asymmetric Synthesis (2 ^nd Ed. Robert E. Gawley, Jeffrey Aubé, Elsevier, Oxford, UK, 2012); Eliel, EL Stereochemistry of Carbon Compounds (McGraw-Hill, NY, 1962); Wilen, SH Tables of Resolving Agents and Optical Resolutions p.268 (ELEliel, Ed., Univ. of NotreDame Press, Notre Dame, IN 1972); Chiral Separation Techniques: A Practical Approach (Subramanian, G. Ed., Wiley-VCH Verlag GmbH & Co. KGaA, Weinheim, Germany, 2007).

The term "tautomer" or "tautomeric form" refers to structural isomers of different energies that are interconvertible through a low energy barrier. A chemical equilibrium of tautomers can be achieved if tautomerism is possible (eg, in solution). For example, protontautomers (also known as prototropic tautomers) include interconversions by migration of protons, such as keto-enol isomerization and imine-ene Amine isomerization. Valence tautomers include interconversions by recombination of some of the bonding electrons. A specific example of keto-enol tautomerism is the interconversion of pentane-2,4-dione and 4-hydroxypent-3-en-2-one tautomers. Another example of tautomerism is phenol-ketone tautomerism. A specific example of phenol-ketone tautomerism is the interconversion of pyridin-4-ol and pyridin-4(lH)-one tautomers. Unless otherwise indicated, all tautomeric forms of the compounds of the present invention are within the scope of the present invention.

As described herein, the compounds of the present invention may be optionally substituted with one or more substituents, such as the compounds of the general formula above, or as in the Examples, subclasses, and subclasses encompassed by the present invention. a class of compounds. It is to be understood that the term "optionally substituted" is used interchangeably with the term "substituted or unsubstituted". In general, the term "substituted" means that one or more hydrogen atoms in a given structure have been replaced with a specified substituent. Unless otherwise indicated, an optional substituent group may be substituted at each substitutable position of the group. When more than one position in a given formula can be substituted with one or more substituents selected from a particular group, the substituents can be substituted identically or differently at each position.

In addition, it should be noted that, unless clearly stated otherwise, the description modes "each independently" and "...independently" and "...independently" used in the present invention can be interchanged, and both are interchangeable. It should be understood in a broad sense. It can either mean that in different groups, the specific options expressed between the same symbols do not affect each other, or it can mean that in the same group, the specific options expressed between the same symbols do not affect each other.

In various parts of this specification, the substituents of the compounds disclosed in the present invention are disclosed in terms of group type or scope. Specifically, the present invention includes each and every independent subcombination of each member of these group species and ranges. For example, the term "C1-6 alkyl" specifically refers to the independently disclosed methyl, ethyl, C3 alkyl, C4 alkyl, C5 alkyl and C6 alkyl groups.

In various parts of the present invention, linking substituents are described. When the structure clearly requires a linking group, the Markush variables listed for that group should be understood to be the linking group. For example, if the structure requires a linking group and the Markush group definition for that variable recites "alkyl" or "aryl", it should be understood that the "alkyl" or "aryl" respectively represents the linking An alkylene group or an arylene group.

The term "alkyl" or "alkyl group" used in the present invention refers to a saturated linear or branched monovalent hydrocarbon group containing 1 to 20 carbon atoms, wherein the alkyl group may optionally be is substituted with one or more substituents described herein. Unless otherwise specified, alkyl groups contain 1-20 carbon atoms. In one embodiment, the alkyl group contains 1-12 carbon atoms; in another embodiment, the alkyl group contains 1-6 carbon atoms; in yet another embodiment, the alkyl group contains 1 -4 carbon atoms; in yet another embodiment, the alkyl group contains 1-3 carbon atoms.

Examples of alkyl groups include, but are not limited to, methyl (Me, -CH3), ethyl (Et, -CH2CH3), n-propyl (n-Pr, -CH2CH2CH3), isopropyl (i-Pr) , -CH(CH3)2), n-butyl (n-Bu, -CH2CH2CH2CH3), isobutyl (i-Bu, -CH2CH(CH3)2), sec-butyl (s-Bu, -CH(CH3) CH2CH3), tert-butyl (t-Bu, -C(CH3)3), n-pentyl (-CH2CH2CH2CH2CH3), 2-pentyl (-CH(CH3)CH2CH2CH3), 3-pentyl (-CH(CH2CH3) 2), 2-methyl-2-butyl (-C(CH3)2CH2CH3), 3-methyl-2-butyl (-CH(CH3)CH(CH3)2), 3-methyl-1- Butyl (-CH2CH2CH(CH3)2), 2-methyl-1-butyl (-CH2CH(CH3)CH2CH3), n-hexyl (-CH2CH2CH2CH2CH2CH3), 2-hexyl (-CH(CH3)CH2CH2CH2CH3), 3- Hexyl (-CH(CH2CH3)(CH2CH2CH3)), 2-methyl-2-pentyl (-C(CH3)2CH2CH2CH3), 3-methyl-2-pentyl (-CH(CH3)CH(CH3)CH2CH3 ), 4-methyl-2-pentyl (-CH(CH3)CH2CH(CH3)2), 3-methyl-3-pentyl (-C(CH3)(CH2CH3)2), 2-methyl- 3-pentyl (-CH(CH2CH3)CH(CH3)2), 2,3-dimethyl-2-butyl (-C(CH3)2CH(CH3)2), 3,3-dimethyl- 2-Butyl (-CH(CH3)C(CH3)3), n-heptyl, n-octyl, etc.

The term "alkylene" refers to a saturated divalent hydrocarbyl group obtained by removing two hydrogen atoms from a saturated straight or branched chain hydrocarbyl group. Unless otherwise specified, alkylene groups contain 1-12 carbon atoms. In one embodiment, the alkylene group contains 1-6 carbon atoms; in another embodiment, the alkylene group contains 1-4 carbon atoms; in yet another embodiment, the alkylene group A group contains 1-3 carbon atoms; in yet another embodiment, an alkylene group contains 1-2 carbon atoms. Examples of such include methylene (-CH2-), ethylene (-CH2CH2-), isopropylidene (-CH(CH3)CH2-), and the like.

The term "alkenyl" refers to a linear or branched monovalent hydrocarbon group containing 2 to 12 carbon atoms, wherein there is at least one site of unsaturation, i.e., a carbon-carbon sp2 double bond, wherein the alkenyl group Can be optionally substituted with one or more of the substituents described herein, including the "cis" and "tans" positions, or the "E" and "Z" positions. In one embodiment, the alkenyl group contains 2-8 carbon atoms; in another embodiment, the alkenyl group contains 2-6 carbon atoms; in yet another embodiment, the alkenyl group contains 2 -4 carbon atoms. Examples of alkenyl groups include, but are not limited to, vinyl (-CH=CH2), allyl (-CH2CH=CH2), and the like.

The term "alkynyl" refers to a linear or branched monovalent hydrocarbon group containing 2-12 carbon atoms, wherein there is at least one site of unsaturation, i.e., a carbon-carbon sp triple bond, wherein the alkynyl group Can be optionally substituted with one or more of the substituents described herein. In one embodiment, the alkynyl group contains 2-8 carbon atoms; in another embodiment, the alkynyl group contains 2-6 carbon atoms; in yet another embodiment, the alkynyl group contains 2 -4 carbon atoms. Examples of alkynyl groups include, but are not limited to, ethynyl (-C≡CH), propargyl (-CH2C≡CH), 1-propynyl (-C≡C-CH3), and the like.

The term "heteroalkyl" denotes the insertion of one or more heteroatoms into the alkyl chain, wherein the alkyl group and the heteroatom have the meanings as described herein. Unless otherwise specified, heteroalkyl groups contain 2-10 carbon atoms, in other embodiments, heteroalkyl groups contain 2-8 carbon atoms, and in other embodiments, heteroalkyl groups contain 2 -6 carbon atoms, in other embodiments, the heteroalkyl group contains 2-4 carbon atoms, and in other embodiments, the heteroalkyl group contains 2-3 carbon atoms. Such examples include, but are not limited to, CH3OCH2-, CH3CH2OCH2-, CH3SCH2-, (CH3)2NCH2-, (CH3)2CH2OCH2-, CH3OCH2CH2-, CH3CH2OCH2CH2-, and the like.

The term "alkenylene" refers to an alkenyl group obtained by removing two hydrogen atoms from a linear or branched alkene. And the alkenylene can be substituted or unsubstituted, wherein the substituent can be, but not limited to, deuterium, hydroxyl, amino, halogen, cyano, aryl, heteroaryl, alkoxy, alkyl, Alkenyl, alkynyl, heterocyclyl, mercapto, nitro or aryloxy. Such examples include, but are not limited to, vinylidene (-CH=CH-), isopropenylene (-C(CH3)=CH-), 3-methoxypropene-1,1-diyl, 2-methylbutene-1,1-diyl and the like.

The term "carbocyclylene" ("cycloalkylene") refers to a saturated divalent hydrocarbon ring obtained by removing two hydrogen atoms from a monocyclic ring of 3-12 carbon atoms or a bicyclic ring of 7-12 carbon atoms, Where carbocyclyl or cycloalkyl has the meaning as described herein, such examples include, but are not limited to, cyclopropylene, cyclobutylene, cyclopentylene, 1-cyclopent-1-ene Alkenyl, 1-cyclopent-2-alkenylene, etc.

The term "heterocyclylene" refers to a monocyclic, bicyclic or tricyclic ring system in which one or more atoms in the ring are independently selected from heteroatoms, and may be fully saturated or contain one or more unsaturations, but not Belongs to the aromatic class, having two points of attachment to the rest of the molecule, where the heterocyclyl group has the meaning as described herein. Such examples include, but are not limited to, piperidine-1,4-diyl, piperazine-1,4-diyl, tetrahydrofuran-2,4-diyl, tetrahydrofuran-3,4-diyl, aza Cyclobutane-1,3-diyl, pyrrolidine-1,3-diyl, etc.

The term "alkoxy" means that an alkyl group is attached to the remainder of the molecule through an oxygen atom, wherein the alkyl group has the meaning as described herein. Unless otherwise specified, the alkoxy groups contain 1-12 carbon atoms. In one embodiment, the alkoxy group contains 1-6 carbon atoms; in another embodiment, the alkoxy group contains 1-4 carbon atoms; in yet another embodiment, the alkoxy group The group contains 1-3 carbon atoms. The alkoxy groups may be optionally substituted with one or more substituents described herein.

Examples of alkoxy groups include, but are not limited to, methoxy (MeO, -OCH3), ethoxy (EtO, -OCH2CH3), 1-propoxy (n-PrO, n-propoxy, -OCH2CH2CH3), 2-propoxy (i-PrO, i-propoxy, -OCH(CH3)2), 1-butoxy (n-BuO, n-butoxy, -OCH2CH2CH2CH3), 2- Methyl-l-propoxy (i-BuO, i-butoxy, -OCH2CH(CH3)2), 2-butoxy (s-BuO, s-butoxy, -OCH(CH3)CH2CH3) , 2-methyl-2-propoxy (t-BuO, t-butoxy, -OC(CH3)3), 1-pentyloxy (n-pentyloxy, -OCH2CH2CH2CH2CH3), 2-pentyloxy base (-OCH(CH3)CH2CH2CH3), 3-pentyloxy (-OCH(CH2CH3)2), 2-methyl-2-butoxy (-OC(CH3)2CH2CH3), 3-methyl-2- Butoxy(-OCH(CH3)CH(CH3)2), 3-methyl-l-butoxy(-OCH2CH2CH(CH3)2), 2-methyl-l-butoxy(-OCH2CH(CH3 )CH2CH3), etc.

The terms "haloalkyl", "haloalkenyl" or "haloalkoxy" mean an alkyl, alkenyl or alkoxy group substituted with one or more halogen atoms, examples of which include, but are not limited to, Trifluoromethyl, trifluoromethoxy, etc.

The terms "hydroxyalkyl" and "hydroxy-substituted alkyl" mean that an alkyl group is substituted with one or more hydroxy groups, wherein the alkyl group has the meaning set forth herein. Such examples include, but are not limited to, hydroxymethyl, hydroxyethyl, 1,2-dihydroxyethyl, and the like.

The term "carbocyclyl" or "carbocycle" refers to a monovalent or polyvalent non-aromatic saturated or partially unsaturated monocyclic, bicyclic or tricyclic ring system containing from 3 to 12 carbon atoms. Carbobicyclyl groups include spirocarbobicyclyl groups and fused carbobicyclyl groups, and suitable carbocyclyl groups include, but are not limited to, cycloalkyl, cycloalkenyl, and cycloalkynyl. Examples of carbocyclyl groups further include, cyclopropyl, cyclobutyl, cyclopentyl, 1-cyclopentyl-1-enyl, 1-cyclopentyl-2-enyl, 1-cyclopentyl- 3-alkenyl, cyclohexyl, 1-cyclohexyl-1-enyl, 1-cyclohexyl-2-enyl, 1-cyclohexyl-3-enyl, cyclohexadienyl, cycloheptyl, cyclooctyl cyclononyl, cyclodecyl, cycloundecyl, cyclododecyl, and the like.

The term "cycloalkyl" denotes a monovalent or polyvalent saturated monocyclic, bicyclic or tricyclic ring system containing 3 to 12 carbon atoms. In one embodiment, the cycloalkyl group contains 3-12 carbon atoms; in another embodiment, the cycloalkyl group contains 3-8 carbon atoms; in yet another embodiment, the cycloalkyl group contains 3-6 carbon atoms carbon atom. The cycloalkyl groups may independently be unsubstituted or substituted with one or more substituents described herein.

基，二氮杂

基，硫氮杂

基，吲哚啉基，1,2,3,4-四氢异喹啉基、1,3-苯并二噁茂基、2-氧杂-5-氮杂双环[2.2.1]庚-5-基。杂环基中-CH2-基团被-C(O)-取代的实例包括，但不限于，2-氧代吡咯烷基、氧代-1,3-噻唑烷基、2-哌啶酮基、3,5-二氧代哌啶基和嘧啶二酮基。杂环基中硫原子被氧化的实例包括，但不限于，环丁砜基、1,1-二氧代硫代吗啉基。所述的杂环基基团可以任选地被一个或多个本发明所描述的取代基所取代。The terms "heterocyclyl" and "heterocycle" are used interchangeably herein, and both refer to saturated or partially unsaturated monocyclic, bicyclic or tricyclic rings containing 3 to 12 ring atoms, at least one of which is selected from the group consisting of Nitrogen, sulfur and oxygen atoms. Unless otherwise specified, a heterocyclyl group can be carbon or nitrogen, and the -CH2- group can be optionally replaced by -C(O)-. Ring sulfur atoms can optionally be oxidized to S-oxides. The nitrogen atoms of the rings can be optionally oxidized to N-oxygen compounds. Examples of heterocyclyl groups include, but are not limited to: oxiranyl, azetidinyl, oxetanyl, thietanyl, pyrrolidinyl, 2-pyrrolinyl, 3-pyrrolinyl , pyrazolinyl, pyrazolidine, imidazolinyl, imidazolidinyl, tetrahydrofuranyl, dihydrofuranyl, tetrahydrothienyl, dihydrothienyl, 1,3-dioxocyclopentyl, disulfide Amyl, tetrahydropyranyl, dihydropyranyl, 2H-pyranyl, 4H-pyranyl, tetrahydrothiopyranyl, piperidinyl, morpholinyl, thiomorpholinyl, piperazinyl , Dioxanyl, Dithianyl, Thioxanyl, Homopiperazinyl, Homopiperidinyl, Oxepanyl, Thiepanyl, Oxazepine

base, diazepine

base, thiazepine

base, indolinyl, 1,2,3,4-tetrahydroisoquinolinyl, 1,3-benzodioxinyl, 2-oxa-5-azabicyclo[2.2.1]hept-5 -base. Examples of heterocyclyls where the -CH2-group is substituted with -C(O)- include, but are not limited to, 2-oxopyrrolidinyl, oxo-1,3-thiazolidinyl, 2-piperidinyl , 3,5-dioxopiperidyl and pyrimidinedione. Examples of the oxidized sulfur atom in the heterocyclyl group include, but are not limited to, sulfolane, 1,1-dioxothiomorpholinyl. The heterocyclyl group may be optionally substituted with one or more substituents described herein.

基，二氮杂

基，硫氮杂

基。杂环基中-CH2-基团被-C(O)-取代的实例包括，但不限于，2-氧代吡咯烷基、氧代-1,3-噻唑烷基、2-哌啶酮基、3,5-二氧代哌啶基和嘧啶二酮基。杂环基中硫原子被氧化的实例包括，但不限于，环丁砜基、1,1-二氧代硫代吗啉基。所述的4-7个原子组成的杂环基基团可以任选地被一个或多个本发明所描述的取代基所取代。In one embodiment, the heterocyclyl group is a heterocyclyl group consisting of 4-7 atoms, which refers to a saturated or partially unsaturated monocyclic ring containing 4-7 ring atoms, wherein at least one ring atom is selected from nitrogen, sulfur and oxygen atoms. Unless otherwise specified, heterocyclyl groups of 4-7 atoms may be carbon or nitrogen, and -CH2- groups may be optionally replaced by -C(O)-. Ring sulfur atoms can optionally be oxidized to S-oxides. The nitrogen atoms of the rings can be optionally oxidized to N-oxygen compounds. Examples of heterocyclyl groups of 4-7 atoms include, but are not limited to: azetidine, oxetanyl, thietanyl, pyrrolidinyl, 2-pyrrolinyl, 3-pyrroline base, pyrazolinyl, pyrazolidine, imidazolinyl, imidazolidinyl, tetrahydrofuranyl, dihydrofuranyl, tetrahydrothienyl, dihydrothienyl, 1,3-dioxocyclopentyl, disulfide Cyclopentyl, tetrahydropyranyl, dihydropyranyl, 2H-pyranyl, 4H-pyranyl, tetrahydrothiopyranyl, piperidinyl, morpholinyl, thiomorpholinyl, piperazine base, dioxanyl, dithianyl, thioxanyl, homopiperazinyl, homopiperidinyl, oxepanyl, thiepanyl, oxazepine

base, diazepine

base, thiazepine

base. Examples of heterocyclyl groups where the -CH2-group is substituted with -C(O)- include, but are not limited to, 2-oxopyrrolidinyl, oxo-1,3-thiazolidinyl, 2-piperidinone , 3,5-dioxopiperidyl and pyrimidinedione. Examples of the oxidized sulfur atom in the heterocyclyl group include, but are not limited to, sulfolanyl, 1,1-dioxothiomorpholinyl. Said heterocyclyl group consisting of 4-7 atoms can be optionally substituted by one or more substituents described in the present invention.

In another embodiment, heterocyclyl is a 4-atom heterocyclyl, which refers to a saturated or partially unsaturated monocyclic ring containing 4 ring atoms, wherein at least one ring atom is selected from nitrogen, sulfur and oxygen atoms replaced. Unless otherwise specified, a 4-atom heterocyclyl group may be carbon or nitrogen, and the -CH2- group may be optionally replaced by -C(O)-. Ring sulfur atoms can optionally be oxidized to S-oxides. The nitrogen atoms of the rings can be optionally oxidized to N-oxygen compounds. Examples of 4-atom heterocyclyl groups include, but are not limited to: azetidine, oxetanyl, thietanyl. The 4-atom heterocyclyl group may be optionally substituted with one or more substituents described herein.

In another embodiment, heterocyclyl is a 5-atom heterocyclyl, which refers to a saturated or partially unsaturated monocyclic ring containing 5 ring atoms, wherein at least one ring atom is selected from nitrogen, sulfur and oxygen atoms . Unless otherwise specified, a 5-atom heterocyclyl group may be carbon or nitrogen, and the -CH2- group may be optionally replaced by -C(O)-. Ring sulfur atoms can optionally be oxidized to S-oxides. The nitrogen atoms of the rings can be optionally oxidized to N-oxygen compounds. Examples of 5-atom heterocyclic groups include, but are not limited to: pyrrolidinyl, 2-pyrrolinyl, 3-pyrrolinyl, pyrazolinyl, pyrazolidinyl, imidazolinyl, imidazolidinyl, Tetrahydrofuranyl, dihydrofuranyl, tetrahydrothienyl, dihydrothienyl, 1,3-dioxocyclopentyl, dithiocyclopentyl. Examples of the -CH2- group in the heterocyclyl group substituted with -C(O)- include, but are not limited to, 2-oxopyrrolidinyl, oxo-1,3-thiazolidinyl. Examples of oxidized sulfur atoms in heterocyclyl groups include, but are not limited to, sulfolane groups. The 5-atom heterocyclyl group may be optionally substituted with one or more of the substituents described herein.

In another embodiment, heterocyclyl is a 6-atom heterocyclyl, which refers to a saturated or partially unsaturated monocyclic ring containing 6 ring atoms, wherein at least one ring atom is selected from nitrogen, sulfur and oxygen atoms . Unless otherwise specified, a 6-atom heterocyclyl group may be carbon or nitrogen, and the -CH2- group may be optionally replaced by -C(O)-. Ring sulfur atoms can optionally be oxidized to S-oxides. The nitrogen atoms of the rings can be optionally oxidized to N-oxygen compounds. Examples of 6-atom heterocyclic groups include, but are not limited to: tetrahydropyranyl, dihydropyranyl, 2H-pyranyl, 4H-pyranyl, tetrahydrothiopyranyl, piperidinyl, Morpholinyl, thiomorpholinyl, piperazinyl, dioxanyl, dithianyl, thioxanyl. Examples of heterocyclyl groups where the -CH2-group is substituted with -C(O)- include, but are not limited to, 2-piperidinyl, 3,5-dioxopiperidinyl, and pyrimidinedione. Examples of oxidized sulfur atoms in a heterocyclyl group include, but are not limited to, 1,1-dioxothiomorpholinyl. The 6-atom heterocyclyl group may be optionally substituted with one or more substituents described herein.

In another embodiment, the heterocyclyl group is a heterocyclyl group consisting of 7-12 atoms, and refers to a saturated or partially unsaturated spirobicyclic or fused bicyclic ring containing 7-12 ring atoms, wherein at least one ring atom is selected from nitrogen, sulfur and oxygen atoms. Unless otherwise specified, heterocyclyl groups of 7-12 atoms may be carbon or nitrogen, and -CH2- groups may be optionally replaced by -C(O)-. Ring sulfur atoms can optionally be oxidized to S-oxides. The nitrogen atoms of the rings can be optionally oxidized to N-oxygen compounds. Examples of heterocyclyl groups of 7-12 atoms include, but are not limited to: indolinyl, 1,2,3,4-tetrahydroisoquinolinyl, 1,3-benzodioxinyl, 2- Oxa-5-azabicyclo[2.2.1]heptan-5-yl. The heterocyclyl group consisting of 7-12 atoms can be optionally substituted by one or more substituents described in the present invention.

The terms "fused bicyclic", "fused ring", "fused bicyclic group" and "fused ring group" are used interchangeably herein and all refer to a monovalent or polyvalent saturated or partially unsaturated bridged ring system, so The bridged ring system refers to a non-aromatic bicyclic ring system. Such systems may contain independent or conjugated unsaturated systems, but whose core structure does not contain aromatic rings or aromatic heterocycles (although aromatic groups may serve as substituents thereon).

The terms "spirocyclyl", "spirocycle", "spirobicyclyl" or "spirobicyclyl" are used interchangeably herein to refer to a monovalent or polyvalent saturated or partially unsaturated ring system in which one ring originates from the other A specific ring carbon atom in a ring. For example, as described below, a saturated bridged ring system (Rings B and B') is referred to as a "fused bicyclic", while Rings A and B share a carbon atom in the two saturated ring systems and are referred to by Known as "spiro" or "spirobicycle". Each ring in the fused bicyclyl and spirobicyclyl groups can be carbocyclyl or heterocyclyl, and each ring is optionally substituted with one or more substituents described herein.

The term "heterocycloalkyl" refers to a monovalent or polyvalent saturated monocyclic, bicyclic or tricyclic ring system containing from 3 to 12 ring atoms, wherein at least one ring atom is selected from nitrogen, sulfur or oxygen atoms.

The term "consisting of n atoms", where n is an integer, typically describes the number of ring-forming atoms in a molecule where the number of ring-forming atoms is n. For example, piperidinyl is a 6-atom heterocycloalkyl group, while 1,2,3,4-tetrahydronaphthalene is a 10-atom cycloalkyl group.

The term "unsaturated" as used herein means that the group contains one or more degrees of unsaturation.

The term "heteroatom" refers to O, S, N, P, and Si, including N, S, and P in any oxidation state; in the form of primary, secondary, tertiary amines, and quaternary ammonium salts; or on a nitrogen atom in a heterocyclic ring. Hydrogen substituted form, for example, N (like N in 3,4-dihydro-2H-pyrrolidinyl), NH (like NH in pyrrolidinyl) or NR (like in N-substituted pyrrolidinyl) NR).

The term "halogen" refers to fluorine (F), chlorine (Cl), bromine (Br) or iodine (I).

The term "aryl" refers to monocyclic, bicyclic and tricyclic carbocyclic ring systems containing 6-14 ring atoms, or 6-12 ring atoms, or 6-10 ring atoms, wherein at least one ring system is aromatic A family of rings in which each ring system contains a ring of 3-7 atoms with one or more points of attachment to the rest of the molecule. The term "aryl" is used interchangeably with the term "aromatic ring". Examples of aryl groups may include phenyl, naphthyl, and anthracene. The aryl groups can be independently optionally substituted with one or more substituents described herein.

The term "heteroaryl" refers to monocyclic, bicyclic and tricyclic ring systems containing 5-12 ring atoms, or 5-10 ring atoms, or 5-6 ring atoms, wherein at least one ring system is aromatic, And at least one ring system contains one or more heteroatoms, wherein each ring system contains a ring of 5-7 atoms and has one or more points of attachment to the rest of the molecule. The term "heteroaryl" may be used interchangeably with the terms "heteroaromatic ring" or "heteroaromatic". The heteroaryl group is optionally substituted with one or more substituents described herein. In one embodiment, a heteroaryl group of 5-10 atoms contains 1, 2, 3 or 4 heteroatoms independently selected from O, S and N.

Examples of heteroaryl groups include, but are not limited to, 2-furyl, 3-furyl, N-imidazolyl, 2-imidazolyl, 4-imidazolyl, 5-imidazolyl, 3-isoxazolyl , 4-isoxazolyl, 5-isoxazolyl, 2-oxazolyl, 4-oxazolyl, 5-oxazolyl, N-pyrrolyl, 2-pyrrolyl, 3-pyrrolyl, 2- Pyridyl, 3-pyridyl, 4-pyridyl, 2-pyrimidinyl, 4-pyrimidinyl, 5-pyrimidinyl, pyridazinyl (such as 3-pyridazinyl), 2-thiazolyl, 4-thiazolyl, 5-thiazolyl, tetrazolyl (such as 5-tetrazolyl), triazolyl (such as 2-triazolyl and 5-triazolyl), 2-thienyl, 3-thienyl, pyrazolyl (such as 2-pyrazolyl), isothiazolyl, 1,2,3-oxadiazolyl, 1,2,5-oxadiazolyl, 1,2,4-oxadiazolyl, 1,2,3-oxadiazolyl Triazolyl, 1,2,3-thiodiazolyl, 1,3,4-thiodiazolyl, 1,2,5-thiodiazolyl, pyrazinyl, 1,3,5- Triazinyl; also including, but in no way limited to, the following bicyclic rings: benzimidazolyl, benzofuranyl, benzothienyl, indolyl (eg 2-indolyl), purinyl, quinolinyl (such as 2-quinolinyl, 3-quinolinyl, 4-quinolinyl), isoquinolinyl (such as 1-isoquinolinyl, 3-isoquinolinyl or 4-isoquinolinyl), imidazole [1,2-a]pyridyl, pyrazolo[1,5-a]pyridyl, pyrazolo[1,5-a]pyrimidinyl, imidazo[1,2-b]pyridazinyl, [1,2,4]Triazolo[4,3-b]pyridazinyl, [1,2,4]triazolo[1,5-a]pyrimidinyl, [1,2,4]triazole and [1,5-a]pyridyl, etc.

The term "carboxy", either alone or in combination with other terms, such as "carboxyalkyl", means _-CO2H ; the term "carbonyl", either alone or in combination with other terms, such as "aminocarbonyl" or "Acyloxy" means -(C=O)-.

The term "alkylamino" includes "N-alkylamino" and "N,N-dialkylamino" wherein the amino group is independently substituted with one or two alkyl groups, respectively. In some embodiments, alkylamino is a lower alkylamino group with one or two _C1-6 alkyl groups attached to a nitrogen atom. In other embodiments, the alkylamino group is a _C1-3 lower alkylamino group. Suitable alkylamino groups may be monoalkylamino or dialkylamino, examples of which include, but are not limited to, N-methylamino, N-ethylamino, N,N-dimethylamino, N,N -Diethylamino, etc.

The term "arylamino" means that an amino group is substituted with one or two aryl groups, examples of which include, but are not limited to, N-phenylamino. In some embodiments, the aromatic ring on the arylamino group may be further substituted.

The term "aminoalkyl" includes a C _1-10 straight or branched chain alkyl group substituted with one or more amino groups. In some embodiments, aminoalkyl is a C _1-6 "lower aminoalkyl" substituted with one or more amino groups, such examples include, but are not limited to, aminomethyl, amino Ethyl, aminopropyl, aminobutyl and aminohexyl.

The term "prodrug" as used in the present invention refers to the conversion of a compound into a compound of formula (I) in vivo. Such conversion is effected by hydrolysis of the prodrug in blood or enzymatic conversion to the parent structure in blood or tissue. The prodrug compounds of the present invention can be esters. In the existing invention, esters that can be used as prodrugs include phenyl esters, aliphatic (C _1-24 ) esters, acyloxymethyl esters, carbonate esters , carbamates and amino acid esters. For example, a compound of the present invention contains a hydroxyl group, which can be acylated to give the compound in prodrug form. Other prodrug forms include phosphates, such as these phosphates are phosphorylated by the hydroxyl group on the parent. A complete discussion of prodrugs can be found in the following literature: T. Higuchi and V. Stella, Pro-drugs as Novel Delivery Systems, Vol. 14 of the ACSSymposium Series, Edward B. Roche, ed., Bioreversible Carriers in Drug Design, American Pharmaceutical Association and Pergamon Press, 1987, J. Rautio et al., Prodrugs: Design and Clinical Applications, Nature Review Drug Discovery, 2008, 7, 255-270, and SJ Hecker et al., Prodrugs of Phosphates and Phosphonates, Journal of Medicinal Chemistry, 2008, 51, 2328 -2345.

"Metabolite" refers to a product obtained by metabolism of a specific compound or salt thereof in vivo. Metabolites of a compound can be identified by techniques well known in the art, and their activity can be characterized using assays as described herein. Such products may be obtained by subjecting the administered compound to oxidation, reduction, hydrolysis, amidation, deamidation, esterification, delipidation, enzymatic cleavage, and the like. Accordingly, the present invention includes metabolites of compounds, including metabolites produced by contacting a compound of the present invention with a mammal for a sufficient period of time.

As used herein, "pharmaceutically acceptable salts" refer to organic and inorganic salts of the compounds of the present invention. Pharmaceutically acceptable salts are well known in the art, as described in the literature: SM Berge et al., describe pharmaceutically acceptable salts in detail in J. Pharmaceutical Sciences, 1977, 66: 1-19. Pharmaceutically acceptable non-toxic acid salts include, but are not limited to, inorganic acid salts formed by reaction with amino groups including hydrochloride, hydrobromide, phosphate, sulfate, perchlorate, And organic acid salts such as acetate, oxalate, maleate, tartrate, citrate, succinate, malonate, or obtained by other methods such as ion exchange method described in books and literature these salts. Other pharmaceutically acceptable salts include adipate, alginate, ascorbate, aspartate, besylate, benzoate, bisulfate, borate, butyrate, camphoric acid Salt, camphorsulfonate, cyclopentylpropionate, digluconate, lauryl sulfate, ethanesulfonate, formate, fumarate, glucoheptonate, glycerophosphate Salt, Gluconate, Hemisulfate, Heptanoate, Caproate, Hydroiodide, 2-Hydroxy-ethanesulfonate, Lacturonate, Lactate, Laurate, Lauryl Sulfate, Malonate, Malonate, Mesylate, 2-Naphthalenesulfonate, Nicotinate, Nitrate, Oleate, Palmitate, Pamoate, Pectate, Persulfate, 3 - Phenylpropionate, picrate, pivalate, propionate, stearate, thiocyanate, p-toluenesulfonate, undecanoate, valerate, etc. Salts obtained with appropriate bases include alkali metal, alkaline earth metal, ammonium and N ⁺ (C _1-4 alkyl) ₄ salts. The present invention also contemplates quaternary ammonium salts formed from any compound containing an N-group. Water- or oil-soluble or dispersible products can be obtained by quaternization. Alkali metal or alkaline earth metal salts include sodium, lithium, potassium, calcium, magnesium, and the like. Pharmaceutically acceptable salts further include appropriate, non-toxic ammonium, quaternary ammonium salts and amine cations that resist the formation of counterions, such as halides, hydroxides, carboxylates, sulfates, phosphates, nitrates, _{C1 -8} Sulfonates and aromatic sulfonates.

A "solvate" of the present invention refers to an association of one or more solvent molecules with a compound of the present invention. Solvate-forming solvents include, but are not limited to, water, isopropanol, ethanol, methanol, dimethyl sulfoxide, ethyl acetate, acetic acid, and aminoethanol. The term "hydrate" refers to an association in which the solvent molecule is water.

The term "treating" any disease or disorder, as used herein, in some embodiments refers to ameliorating the disease or disorder (ie, slowing or arresting or alleviating the development of the disease or at least one clinical symptom thereof). In other embodiments, "treating" refers to alleviating or improving at least one physical parameter, including a physical parameter that may not be perceived by a patient. In other embodiments, "treating" refers to modulating a disease or disorder physically (eg, stabilizing an observable symptom) or physiologically (eg, stabilizing a physical parameter), or both. In other embodiments, "treating" refers to preventing or delaying the onset, occurrence or worsening of a disease or disorder.

Pharmaceutically acceptable acid addition salts can be formed with inorganic and organic acids such as acetates, aspartates, benzoates, benzenesulfonates, bromides/hydrobromides, bicarbonates/ Carbonate, bisulfate/sulfate, camphorsulfonate, chloride/hydrochloride, chlorophylline, citrate, ethanedisulfonate, fumarate, glucoheptonate, gluconate Sugar, glucuronate, hippurate, hydroiodate/iodide, isethionate, lactate, lacturonate, lauryl sulfate, malate, maleate acid salt, malonate, mandelate, mesylate, methosulfate, naphthoate, naphthalenesulfonate, nicotinate, nitrate, octadecanoate, oleate, oxalic acid Salt, Palmitate, Pamoate, Phosphate/Hydrogen Phosphate/Dihydrogen Phosphate, Polygalactonate, Propionate, Stearate, Succinate, Sulfosalicylates, Tartrate , tosylate and trifluoroacetate.

Inorganic acids from which salts can be derived include, for example, hydrochloric acid, hydrobromic acid, sulfuric acid, nitric acid, phosphoric acid, and the like.

Organic acids from which salts can be derived include, for example, acetic acid, propionic acid, glycolic acid, oxalic acid, maleic acid, malonic acid, succinic acid, fumaric acid, tartaric acid, citric acid, benzoic acid, mandelic acid, methanesulfonic acid , ethanesulfonic acid, p-toluenesulfonic acid, sulfosalicylic acid, etc.

Pharmaceutically acceptable base addition salts can be formed with inorganic and organic bases.

Inorganic bases from which salts can be derived include, for example, ammonium salts and metals from Groups I to XII of the Periodic Table. In certain embodiments, the salts are derived from sodium, potassium, ammonium, calcium, magnesium, iron, silver, zinc and copper; particularly suitable salts include ammonium, potassium, sodium, calcium and magnesium salts.

Organic bases from which salts can be derived include primary, secondary, and tertiary amines, and substituted amines include naturally occurring substituted amines, cyclic amines, basic ion exchange resins, and the like. Certain organic amines include, for example, isopropylamine, benzathine, choline, diethanolamine, diethylamine, lysine, meglumine, piperazine, and tromethamine .

The pharmaceutically acceptable salts of the present invention can be synthesized from the parent compound, basic or acidic moieties, using conventional chemical methods. In general, such salts can be prepared by reacting the free acid forms of these compounds with a stoichiometric amount of a suitable base (eg, hydroxide, carbonate, bicarbonate, etc. of Na, Ca, Mg, or K), or by The preparations are carried out by reacting the free base forms of these compounds with a stoichiometric amount of the appropriate acid. Such reactions are usually carried out in water or an organic solvent or a mixture of the two. Generally, where appropriate, the use of non-aqueous media such as diethyl ether, ethyl acetate, ethanol, isopropanol or acetonitrile is required. In, for example, "Remington's Pharmaceutical Sciences", 20th Edition, Mack Publishing Company, Easton, Pa., (1985); and "Handbook of Pharmaceutical Salts: Properties, Selection, and Use )", Stahl and Wermuth (Wiley-VCH, Weinheim, Germany, 2002) may find additional lists of suitable salts.

In addition, the compounds disclosed herein, including their salts, can also be obtained in their hydrate form or in a form containing a solvent (eg, ethanol, DMSO, etc.) for their crystallization. Compounds of the present disclosure may form solvates, inherently or by design, with pharmaceutically acceptable solvents, including water; thus, the present invention is intended to include both solvated and unsolvated forms.

Any structural formula given herein is also intended to represent both isotopically unenriched as well as isotopically enriched forms of these compounds. Isotopically enriched compounds have the structures depicted by the general formulae given herein, except that one or more atoms are replaced by atoms having a selected atomic weight or mass number. Exemplary isotopes that can be incorporated into the compounds of the present invention include isotopes of ^hydrogen , carbon, nitrogen, oxygen, phosphorus, sulfur, fluorine, and chlorine, such as 2H, ^3H , ^11C , ^13C , ^14C , ^15N , ^17O , ¹⁸ O, ¹⁸ F, ³¹ P, ³² P, ³⁵ S, ³⁶ Cl and ¹²⁵ I.

In another aspect, the compounds of the present invention include isotopically enriched compounds as defined by the present invention, for example, those in which radioactive isotopes, such as ³ H, ¹⁴ C and ¹⁸ F are present, or in which non-radioactive isotopes, such as ² H and ¹³ C. Such isotopically enriched compounds can be used in metabolic studies (using ¹⁴ C), reaction kinetic studies (using eg ² H or ³ H), detection or imaging techniques such as positron emission tomography (PET) or including drugs or Single photon emission computed tomography (SPECT) for substrate tissue distribution measurements, or may be used in radiation therapy of patients. ¹⁸ F-enriched compounds are particularly ideal for PET or SPECT studies. Isotopically enriched compounds of formula I or formula II can be prepared by using a suitable isotope-labeled reagent to replace the unlabeled reagent originally used by conventional techniques familiar to those skilled in the art or as described in the examples and preparation process of the present invention. .

In addition, substitution of heavier isotopes, particularly deuterium (ie, ² H or D), may provide certain therapeutic advantages that result from greater metabolic stability. For example, increased in vivo half-life or reduced dosage requirements or improved therapeutic index. The concentration of such heavier isotopes, especially deuterium, can be defined by an isotopic enrichment factor. If a substituent of a compound of the present invention is designated as deuterium, the compound has for each designated deuterium atom at least 3500 (52.5% deuterium incorporation at each designated deuterium atom), at least 4000 (60% deuterium incorporation), At least 4500 (67.5% deuterium incorporated), at least 5000 (75% deuterium incorporated), at least 5500 (82.5% deuterium incorporated), at least 6000 (90% deuterium incorporated), at least 6333.3 (95% deuterium incorporated) deuterium incorporation), an isotopic enrichment factor of at least 6466.7 (97% deuterium incorporation), at least 6600 (99% deuterium incorporation), or at least 6633.3 (99.5% deuterium incorporation). Pharmaceutically acceptable solvates of the present invention include those in which the crystallization solvent may be isotopically substituted, eg, _D2O , acetone _- d6, _DMSO -d6.

In another aspect, the present invention relates to intermediates for the preparation of compounds encompassed by formula I.

In another aspect, the present invention relates to methods for the preparation, isolation and purification of compounds encompassed by formula I.

In another aspect, the present invention provides a pharmaceutical composition comprising a compound of the present invention, a pharmaceutically acceptable carrier, excipient, diluent, adjuvant, vehicle, or a combination thereof. In some embodiments, the pharmaceutical composition may be in liquid, solid, semi-solid, gel or spray form.

"Combination" means a fixed combination in a single dosage unit form or a kit of parts for combined administration, wherein a compound disclosed herein and a combination partner may be administered independently at the same time or may be administered separately at intervals, In particular, the joint partners are made to exhibit cooperation, eg synergy. The terms "co-administration" or "co-administration" and the like are intended to encompass the administration of a selected combination partner to a single individual (eg, a patient) in need thereof, and are intended to encompass treatment regimens in which the substances are not necessarily administered by the same route of administration or at the same time . The term "pharmaceutical combination" as used herein refers to a product resulting from mixing or combining more than one active ingredient, and includes both fixed and non-fixed combinations of active ingredients. The term "fixed combination" means that the active ingredients, such as the compounds disclosed herein, and the combination partner are administered to a patient simultaneously in the form of a single entity or dose. The term "non-fixed combination" means that the active ingredients, such as the disclosed compounds and the combination partner, are administered to a patient simultaneously, jointly or sequentially without a specific time limit as separate entities, wherein the administration provides therapeutically effective levels of both compounds in the patient. .

The experimental methods used in the following examples are conventional methods unless otherwise specified.

The materials, reagents, etc. used in the following examples can be obtained from commercial sources unless otherwise specified.

The Pomalidomide terminal derivatives used in the following examples were prepared according to the method disclosed in the literature Chemistry & Biology 22, 755-763 (2015). The Lenalidomide terminal derivative was prepared according to the method disclosed in the document J.Med.Chem (DOI: 10.1021/acs.jmedchem.6b01816). The RG-7112 terminal carboxylic acid derivative was prepared according to the document Bioorg.Med.Chem.Lett.18,5904 -5908 (2008). and the methods disclosed in ACS Med. Chem. Lett. 4, 466-469 (2013). For the synthesis of the parent nucleus of part X, reference is made to J. Med. Chem (DOI: 10.1021/acs.jmedchem.5b01501). (See Figure 2).

1.X Partial Nucleosynthesis

Reference J.Med.Chem (DOI:10.1021/acs.jmedchem.5b01501)

2. Synthesis of some intermediates

In a round bottom flask add B6,tert-butyl4-(4-(4,4,5,5-tetramethyl-1,3,2-dioxaborolan-2-yl)benzyl)piperazine-1-carboxylate(1.5eq), Na2CO3 (3.0eq), Pd(PPh3)4 (0.1eq), 1,4 dioxane: water = 5:1 as solvent, protected by argon, and reacted at 80°C overnight. After the reaction was completed, 1,4 dioxane was suspended, the mixture was extracted three times with 50 mL×3 dichloromethane, the organic phases were combined, dried over anhydrous sodium sulfate, the solvent was spin-dried, and a 200-300 mesh silica gel column was used Separation and purification. Then at room temperature, an equal volume of mixed solvent of DCM/TFA was added to react. After the reaction was completed, it was basified with saturated NaHCO3 solution and extracted with DCM to obtain intermediate A1.

Add B6,4,4,5,5-tetramethyl-2-(4-nitrophenyl)-1,3,2-dioxaborolane(1.5eq),Na2CO3(3.0eq),Pd(PPh3)4( 0.1eq), 1,4 dioxane: water = 5: 1 as solvent, protected with argon, and reacted at 80 °C overnight. After the reaction was completed, 1,4 dioxane was suspended, the mixture was extracted three times with 50 mL×3 dichloromethane, the organic phases were combined, dried over anhydrous sodium sulfate, the solvent was spin-dried, and a 200-300 mesh silica gel column was used Separation and purification. The purified intermediate is under Fe (4eq), ammonium chloride (2eq) conditions, water and ethanol are used as solvents, the volume ratio of water and ethanol is 1:3, and reflux. After completion, filter while hot, cool, spin dry, and extract. The solvent was then spun dry, and the intermediate A2 was obtained by separation and purification using a 200-300 mesh silica gel column.

Add B6,tert-butyl(4-(4,4,5,5-tetramethyl-1,3,2-dioxaborolan-2-yl)benzyl)carbamate(1.5eq),Na2CO3(3.0eq) to a round bottom flask , Pd (PPh3) 4 (0.1eq), 1,4 dioxane: water = 5: 1 as solvent, protected with argon, 80 ℃ reaction overnight. After the reaction was completed, 1,4 dioxane was suspended, the mixture was extracted three times with 50 mL×3 dichloromethane, the organic phases were combined, dried over anhydrous sodium sulfate, the solvent was spin-dried, and a 200-300 mesh silica gel column was used Separation and purification. Then at room temperature, an equal volume of mixed solvent of DCM/TFA was added to react. After the reaction was completed, it was basified with saturated NaHCO3 solution and extracted with DCM to obtain intermediate A3.

Add B6, but-3-yn-1-ol (1.5eq), Na2CO3 (3.0eq), CuI (0.2eq), Pd(PPh3)2Cl2 (0.1eq) to a round bottom flask, DMF as solvent, with argon Under gas protection, the reaction was carried out at 80°C overnight. After the reaction was completed, the mixture was extracted three times with ethyl acetate and water, the organic phases were combined, dried over anhydrous sodium sulfate, the solvent was spin-dried, and separated and purified using a 200-300 mesh silica gel column to obtain Intermediate A4.

In a 100 mL round bottom flask, add 1.6 g of sodium azide and 25 mL of water, and add 4.9 g of 2-(2-hydroxyethoxy)ethyl-4-methylbenzenesulfonate. The reaction solution was stirred at 90 degrees Celsius for 24 hours. 50 mL of saturated aqueous sodium bicarbonate solution was added to quench the reaction, the mixture was extracted three times with 40 mL × 3 dichloromethane, the organic phases were combined, dried over anhydrous sodium sulfate, and the solvent was spin-dried to obtain an intermediate with a yield of 46%. In a 50mL round-bottomed flask, 787mg of the obtained product, 8mL of anhydrous dichloromethane and 1.67mL of triethylamine were added, and 1.72g of TsCl in anhydrous dichloromethane solution was slowly added to the above reaction solution under stirring conditions. Stir for 24 hours. The reaction was quenched by adding 100 mL of saturated aqueous sodium bicarbonate solution, the mixture was extracted three times with 50 mL×3 dichloromethane, the organic phases were combined, dried over anhydrous sodium sulfate, the solvent was spin-dried, and a 200-300 mesh silica gel column was used for separation After purification, the mobile phase is petroleum ether:ethyl acetate=5:1, and intermediate A5 is obtained with a yield of 67%.

A 100 mL round bottom flask was charged with 12.6 g of 2,2'-(ethane-1,2-diylbis(oxy))bis(ethan-1-ol), 50 mL of anhydrous THF and 5 g of potassium tert-butoxide. The reaction solution was stirred for 15 minutes under argon protection at room temperature, and 3.5 mL of 3-bromopropyne was added dropwise. The reaction solution was kept stirring for 12 hours under argon protection at room temperature. The solvent was spin-dried, 30 mL of saturated aqueous sodium chloride solution was added to quench the reaction, the pH was adjusted to 3-5 with 1M HCl, the mixture was extracted three times with 30 mL×3 dichloromethane, the organic phases were combined, and anhydrous sodium sulfate was used. Dry, spin to dry the solvent, and use a 200-300 mesh silica gel column for separation and purification. The mobile phase is petroleum ether:ethyl acetate=3:1, and intermediate A6 is obtained with a yield of 77%.

In a 100 mL round bottom flask, 4.4 mL of pentane-1,5-diol, 30 mL of anhydrous THF, and 2.5 g of potassium tert-butoxide were added. The reaction solution was kept stirring for 15 minutes under argon protection at room temperature, and 1.73 mL of 3-bromopropyne was added dropwise. The reaction solution was kept stirring for 12 hours under argon protection at room temperature. The solvent was spin-dried, 30 mL of saturated aqueous sodium chloride solution was added to quench the reaction, the pH was adjusted to 3-5 with 1M HCl, the mixture was extracted three times with 30 mL×3 dichloromethane, the organic phases were combined, and anhydrous sodium sulfate was used. Dry, spin to dry the solvent, and use a 200-300 mesh silica gel column for separation and purification. The mobile phase is petroleum ether:ethyl acetate=3:1, and intermediate A7 is obtained with a yield of 74%.

In a 5 mL round bottom flask was added 12 mg 3-aminopropanoic acid, 70 μL DIEA, 0.3 mL DMF, and 35 mg 2-(2,6-dioxopiperidin-3-yl)-4-fluoroisoindoline-1,3-dione. The reaction was carried out at 80°C for 2 hours. At the end of the reaction, the mixture was extracted three times with 5 mL×3 ethyl acetate, the organic phases were combined, dried over anhydrous sodium sulfate, the solvent was spin-dried, and the separation and purification were carried out using a 200-300 mesh silica gel column. The mobile phase was dichloromethane: Methanol=40:1 to obtain intermediate A8.

In a 25mL round bottom flask, add 3-(4-bromo-1-oxoisoindolin-2-yl)piperidine-2,6-dione, 1-azido-4-(prop-2-yn-1-yloxy)butane,CuI (0.2eq), Pd(PPh3)2Cl2(0.1eq), DMF as a solvent, protected with argon, and reacted at 80°C overnight. After the completion of the reaction, the mixture was extracted three times with ethyl acetate and water, the organic phases were combined, dried over anhydrous sodium sulfate, the solvent was spin-dried, and a 200-300 mesh silica gel column was used for separation and purification.

Add hex-5-ynoic acid, TATU, DIAE to a 100 mL round-bottomed flask, pre-activate for 10 minutes, add intermediate A1, and react at room temperature for 3 hours. After the completion of the reaction, the mixture was extracted three times with dichloromethane, the organic phases were combined, dried over anhydrous sodium sulfate, the solvent was spin-dried, and a 200-300 mesh silica gel column was used for separation and purification.

Example 1 Synthesis of Formulas 1 to 27

Synthesis of formula 1:

In a 5 mL round bottom flask were added the Pomalidomide terminal derivative, intermediate A10, CuSO4, sodium ascorbate, water and tert-butanol. After stirring at 70 degrees Celsius for 6 hours under the protection of argon, 15 mL of saturated aqueous sodium bicarbonate solution was added to quench the reaction, the mixture was extracted three times with 15 mL × 3 dichloromethane, the organic phases were combined, dried over anhydrous sodium sulfate, and spun Use a 200-300 mesh silica gel chromatographic column to separate and purify the dry solvent, and the mobile phase is dichloromethane:methanol=20:1, to obtain the compound represented by formula 1 with a yield of 61%.

Formulas 2 to 7 are synthesized with reference to the method of Formula 1:

In a 10mL round-bottom flask, intermediate A8, TATU (1.2eq), DIAE (2eq) were added, and intermediate A1 (1.2eq) was added for pre-activation for 10 minutes, and the reaction was carried out at room temperature for 3 hours. After the completion of the reaction, the mixture was extracted three times with dichloromethane, the organic phases were combined, dried over anhydrous sodium sulfate, the solvent was spin-dried, and a 200-300 mesh silica gel column was used for separation and purification.

The synthetic method of formula 8-formula 27 refers to formula 7:

Example 2 Biological activity test of compounds represented by formula 1 to formula 27 at the level of Western Blot

Cell whole protein extraction:

Cell collection: scrape the treated cells in the culture medium, suspend well and centrifuge at 300g for 5 minutes to collect, wash once with PBS, and discard the PBS.

Lyse cells: add 150 μl of 2×Loading Buffer to each sample, shake well and mix well, denature at 95°C for 15 minutes, store at -20°C after mixing or use directly for Western Blot detection.

The formula of 5×Loading Buffer is: 250mM Tris-HCl (pH6.8), 10% (W/V) SDS, 0.5% (W/V) bromophenol blue, 50% (V/V) glycerol, 5% ( W/V) β-Mercaptoethanol (2-ME). 2×Loading Buffer is prepared by adding 1.5 times the volume of dd water to 5×Loading Buffer.

The specific steps of Western Blot detection are as follows:

1) Prepare SDS-PAGE gel of appropriate concentration. Refer to Table 18.3 on page 883 of "Molecular Cloning Experiment Guide" (Science Press, 2nd Edition) to prepare a separating gel with a suitable concentration, and refer to Table 18.4 on page 883 to prepare a stacking gel with a concentration of 4%.

2) Prepare the sample. The protein samples were prepared according to the experimental requirements, denatured at 95°C for 15 minutes, centrifuged, mixed and loaded into the sample wells of SDS-PAGE gel. Adjust the sample volume appropriately according to the protein quantification results, usually the sample volume per well is 4 μl.

3) Electrophoresis. When the power is turned on, the voltage of the protein sample in the stacking gel is 83 volts. When the protein sample enters the separating gel, we adjust the voltage to 110 volts to continue electrophoresis. The electrophoresis was terminated when the bromophenol blue had almost completely escaped the PAGE gel.

4) Transfer film. After electrophoresis, remove the gel and install the membrane transfer device in the following order: (negative electrode), filter paper, gel, activated PVDF membrane, filter paper, (positive electrode). Remember that there must be no air bubbles between the gel and the PVDF membrane. Then clamp the transfer device and place it in the transfer buffer, and finally put it in an ice box, and place it in a 4°C freezer with a constant voltage of 100V for 1.5 hours.

5) closed. After transfer, the PVDF membrane was taken out, and the membrane was immersed in TBST buffer containing 5% nonfat dry milk and shaken for 1 hour at room temperature.

6) Primary antibody incubation. After blocking, wash 3 times with TBST buffer, then add primary antibody in appropriate dilution ratio, overnight at 4°C. The primary antibody was recovered, and the PVDF membrane was washed three times with TBST buffer for 10 minutes each time.

7) Secondary antibody incubation. Discard TBST buffer, add secondary antibody (mouse antibody or rabbit antibody, determined by primary antibody) at a certain dilution ratio (usually 1:3000-1:5000), shake at room temperature for 1 hour. The secondary antibody was discarded, and the PVDF membrane was washed three times with TBST buffer for 10 minutes each time. Finally, wash with TBST buffer for 10 minutes.

8) Color development and tableting. The ECL chromogenic substrate was evenly covered on the PVDF membrane, and the color was developed at room temperature for 0.5 to 15 minutes.

It can be seen from Figures 3-5 that the compounds in this example can obviously induce the degradation of EZH2 in cells, and the EC50 of formula 10 on two EZH2 wild-type DLBCL cell lines are ₅₀ nM and 100 nM, respectively.

Embodiment 3MTT experimental process:

MTT experimental reagents:

Reagents: RPIM 1640medium; DMEM medium; 100× non-essential amino acids (NEAA); 100× penicillin-streptomycin mixture; 50 mM β-mercaptoethanol; calf serum (FBS, inactivated beforehand).

A medium (500ml): RPIM 1640medium (450ml)+100xNEAA (5ml)+100xpenicillin-streptomycin mixture (5ml)+calf serum (50ml)+50mM beta mercaptoethanol (0.5ml).

B medium (500ml): DMEM medium (450ml)+100×NEAA (5ml)+100×penicillin-streptomycin mixture (5ml)+calf serum (50ml)+50mM β-mercaptoethanol (0.5ml). .

CCK-8 kit (Cell Counting Kit-8)

1) Collect log-phase cells, and adjust the cell suspension concentration to 6.6×10 ⁴ /ml with A medium.

2) 2-fold serial dilution of small molecules in medium A to a concentration of 100 nM to 2 nM. Configured as a small molecule solution.

3) 45 μL of the cell suspension was added to a 96-well plate (the edge wells were filled with sterile PBS, 3000 cells/well). Negative controls (45 μL of cell suspension and 45 μL of A medium) were set in each plate, and 3 replicate wells were set in each group.

4) After incubating at 37°C and 5% CO ₂ for 1 hour, add 45 μL of the corresponding small molecule solution to each well of the 96-well plate. Then incubate for 72-96 hours at 37°C, 5% _CO2 . In the system, the concentration of small molecules was diluted by 2-fold gradient, and the concentration was from 50nM to 1nM.

10 μl of cck-8 solution was added to each well, and the culture was continued for 4 h. The absorbance value of each well was measured by direct enzyme-linked immunosorbent assay OD490nm.

The experimental results are shown in Table 1.

Table 1: Inhibition of large B diffuse lymphoma cell lines by series of compounds.

Wherein, N.D indicates that no inhibitory activity was detected.

In EZH2 wild-type DLBCL cell lines DOHH2 cells and LY7 cells, the compounds shown in formula 7, formula 8, formula 9, formula 10, formula 11, and formula 12 have more obvious cell activity than the control compound EPZ6438. Inhibition, indicating that degraders can be used in cell lines that are ineffective with traditional inhibitors.

And it can be seen from Fig. 6-Fig. 8 that formula 10 and formula 12 showed activity superior to or comparable to the inhibitor in various other cell lines.

Embodiment 4 animal experiment method

1) Cell culture

DOHH2 cells were cultured in RPMI1640 medium containing 10% fetal bovine serum. DOHH2 cells in exponential growth phase were collected and resuspended in PBS to an appropriate concentration for subcutaneous tumor inoculation in CB17SCID mice.

2) Animal modeling

The experimental mice were subcutaneously inoculated with 5×106DoHH2 cells on the right back, and the cells were resuspended in 1:1 PBS and Matrigel (0.1ml/mice). Size and weight of mice were randomized into groups. The day of tumor cell inoculation was defined as day 0.

3) Random grouping

Before the start of dosing, all animals were weighed and tumor volumes were measured with vernier calipers. Since the tumor volume will affect the effectiveness of the treatment, a randomization design was used to group the mice according to their tumor volume to ensure that the tumor volume between the different groups is similar. Grouping was performed using StudyDirector™ (version number 3.1.399.19, vendor Studylog Systems, Inc., S. San Francisco, CA, USA).

The number of animals in each group and the detailed administration routes, doses and schedules are shown in Table 2 below.

Table 2:

Note: The administration volume is 10ul/g; that is, 250ul/25g mice.

From Figure 9 and Figure 10, it can be seen that formula 10 is superior to the human B-cell lymphoma DoHH2 cell line subcutaneous xenografted CB17 SCID female mouse model and the human B-cell lymphoma OCI-LY7 cell line mouse model. inhibitory effect.

In addition, the terms "first" and "second" are only used for descriptive purposes, and should not be construed as indicating or implying relative importance or implying the number of indicated technical features. Thus, a feature delimited with "first", "second" may expressly or implicitly include at least one of that feature. In the description of the present invention, "plurality" means at least two, such as two, three, etc., unless otherwise expressly and specifically defined.

In the description of this specification, description with reference to the terms "one embodiment," "some embodiments," "example," "specific example," or "some examples", etc., mean specific features described in connection with the embodiment or example , structure, material or feature is included in at least one embodiment or example of the present invention. In this specification, schematic representations of the above terms are not necessarily directed to the same embodiment or example. Furthermore, the particular features, structures, materials or characteristics described may be combined in any suitable manner in any one or more embodiments or examples. Furthermore, those skilled in the art may combine and combine the different embodiments or examples described in this specification, as well as the features of the different embodiments or examples, without conflicting each other.

Although the embodiments of the present invention have been shown and described above, it should be understood that the above-mentioned embodiments are exemplary and should not be construed as limiting the present invention. Embodiments are subject to variations, modifications, substitutions and variations.

Claims (16)

1. A compound that is a compound of formula I or a stereoisomer, geometric isomer, tautomer, nitrogen oxide, hydrate, solvate, metabolite, pharmaceutically acceptable salt, or prodrug thereof:

X-Y-Z

formula I.

Wherein X represents a ligand of EZH2 protein, Z represents a ligand of E3 ligase, and Y represents a chain connecting X and Z.

2. The compound of claim 1, wherein X is an optionally substituted structure shown below:

3. the compound of claim 1, wherein Z is an optionally substituted structure as shown below:

4. the compound of claim 1, wherein Y is an optionally substituted structure shown below:

wherein n1 is an integer of 0-3, m is an integer of 1-5, and n2 is an integer of 1-12.

5. The method of any one of claims 2 to 4, wherein X, Y, Z is independently optionally substituted by F, Cl, Br, CN, NO₂、OR^b、-NR^cR^d、R^bO-C_1-4Alkylene radical, R^dR^cN-C_1-4Alkylene, -C (═ O) R^a、-C(＝O)OR^b、-C(＝O)NR^cR^d、C_1-12Alkyl radical, C_1-6Haloalkyl, C_3-6Cycloalkyl radical, C_3-6cycloalkyl-C_1-4Alkylene, heterocyclic group consisting of 3 to 8 atoms, (heterocyclic group consisting of 3 to 8 atoms) -C_1-4Alkylene radical, C_6-10Aryl radical, C_6-10aryl-C_1-4Alkylene, heteroaryl of 5 to 8 atoms or (heteroaryl of 5 to 8 atoms) -C_1-4The substitution of an alkylene group is carried out,

wherein R is^a、R^b、R^c、R^dEach independently is H, D, C_1-6Haloalkyl, C_1-6Alkyl radical, C_2-6Alkenyl radical, C_2-6Alkynyl, C_3-6Carbocyclyl, C_3-6carbocyclyl-C_1-4Alkylene, heterocyclic group consisting of 3 to 12 atoms, (heterocyclic group consisting of 3 to 12 atoms) -C_1-4Alkylene radical, C_6-10Aryl radical, C_6-10aryl-C_1-4Alkylene, heteroaryl of 5 to 10 atoms, (heteroaryl of 5 to 10 atoms) -C_1-4Alkylene, or R^c、R^dAnd together with the nitrogen atom to which they are attached, form a heterocyclic group of 3 to 8 atoms or a heteroaryl group of 5 to 8 atoms, wherein said C_1-6Alkyl radical, C_2-6Alkenyl radical, C_2-6Alkynyl, C_3-6Carbocyclyl, C_3-6carbocyclyl-C_1-4Alkylene, heterocyclic group consisting of 3 to 12 atoms, (heterocyclic group consisting of 3 to 12 atoms) -C_1-4Alkylene radical, C_6-10Aryl radical, C_6-10aryl-C_1-4Alkylene, heteroaryl of 5 to 10 atoms, (heteroaryl of 5 to 10 atoms) -C_1-4Alkylene, heterocyclyl of 3-8 atoms and heteroaryl of 5-8 atoms are each independently unsubstituted or substituted with 1,2,3 or 4 substituents independently selected from D, F, Cl, CN, OH, NH₂、C_1-6Alkyl radical, C_1-6Haloalkyl, C_1-6Alkoxy or C_1-6An alkylamino group.

6. A compound which is a compound shown as below or a stereoisomer, geometric isomer, tautomer, nitrogen oxide, hydrate, solvate, metabolite, pharmaceutically acceptable salt or prodrug thereof,

7. a pharmaceutical composition comprising a compound according to any one of claims 1 to 6.

8. The pharmaceutical composition of claim 7, further comprising a pharmaceutically acceptable carrier, excipient, diluent, adjuvant, vehicle, or combination thereof.

9. The pharmaceutical composition of claim 7, further comprising other drugs for treating or preventing tumors.

10. Use of a compound according to any one of claims 1 to 6 or a pharmaceutical composition according to any one of claims 7 to 9 in the manufacture of a medicament for degrading EZH2 or inhibiting EZH 2.

11. Use of a compound according to any one of claims 1 to 6 or a pharmaceutical composition according to any one of claims 7 to 9 for degrading EZH 2.

12. Use of a compound according to any one of claims 1 to 6 or a pharmaceutical composition according to any one of claims 7 to 9 in the manufacture of a medicament for the treatment or prophylaxis of a disease associated with EZH 2.

13. The use according to claim 12, wherein the EZH 2-associated disease is lymphoma or malignant rhabdomyoma, optionally the lymphoma comprises at least one selected from hodgkin's lymphoma, non-hodgkin's lymphoma (NHL), diffuse large B-cell lymphoma (DLBCL), breast cancer and prostate cancer.

14. Use of a compound according to any one of claims 1 to 6 or a pharmaceutical composition according to any one of claims 7 to 9 for combating tumours.

15. The use according to claim 14, wherein the tumor is a lymphoma or a malignant rhabdomyoma.

16. The use according to claim 15, wherein the lymphoma comprises at least one selected from hodgkin's lymphoma, non-hodgkin's lymphoma (NHL), diffuse large B-cell lymphoma (DLBCL), breast cancer and prostate cancer.

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