Supramolecular targeting chimera, methods of construction and use thereof

Abstract

The present application relates to a kind of supramolecular targeting chimera and its construction method and application.The supramolecular targeting chimera of the present application has high efficient tissue selectivity on the level of active animal, such as liver, lung selectivity, significantly improve the bioavailability of traditional small molecule drugs in liver and lung tissue, indicating that it has good safety and broad application prospect.

Description

Technical Field

[0001] This invention relates to the field of biomedical materials, and in particular to a supramolecular targeted chimera, its construction method, and its application in protein degradation and post-translational modification regulation. Background Technology

[0002] Chemically induced proximity (CIP) is a technique that uses bifunctional small chemical molecules or molecular gels to bring two proteins closer together, thereby regulating protein degradation, homeostasis, and post-translational modifications, and ultimately achieving precise intervention in cell fate. Protein-targeting chimeras (PROTACs) based on small molecules have become important tools in the fields of basic chemical biology and drug discovery. A PROTAC is a bifunctional molecule that binds to a target protein at one end and to an E3 ubiquitin ligase at the other. When a PROTAC binds to both proteins simultaneously, a ternary complex is formed. Once this complex is formed, the E3 ligase catalyzes the sequential attachment of ubiquitin residues to the target protein, achieving ubiquitination labeling and initiating its degradation. PROTAC technology has been widely used in the chemical and biomedical fields and has entered the clinical research stage due to its significant advantages, including catalytic degradation, high selectivity, and continued degradation activity even for "undruggable proteins." However, when using traditional small chemical molecules for targeted intervention with protein degraders, especially at the in vivo animal level and in clinical translational studies, challenges such as lack of tissue specificity and time controllability are often faced, which may lead to off-target regulation and toxic side effects. Therefore, developing novel chemically induced proximity technologies with higher specificity and controllability remains an important research direction. With a deeper understanding of the molecular design and mechanism of action of PROTACs, the development of novel protein-targeting chimeras is expected to play a greater role in precision medicine and drug development, providing new strategies and tools for treating a variety of diseases. Summary of the Invention

[0003] The purpose of this invention is to provide a method for designing and constructing supramolecular targeted chimeras, and a tissue-specific protein post-translational modification regulation method and application developed based on this technology. The tissue-specific protein post-translational modification regulation system of this invention includes metal coordination-guided supramolecular nanoparticles, exhibiting high liver and lung selectivity at the living organism level, enabling efficient retention in lung tissue or liver, and achieving high spatiotemporal resolution protein post-translational modification regulation.

[0004] The solution provided by this invention is as follows:

[0005] A supramolecular targeting chimera, the supramolecular targeting chimera comprising a target protein binding ligand.

[0006] According to an embodiment of the present invention, the supramolecular targeted chimera is obtained by self-assembly of a pre-supramolecular targeted chimera with polyethylene glycol and biomacromolecules to form nanoparticles; or, nanoparticles are directly formed by self-assembly of a metal-organic cage, polyethyleneimine, target protein binding ligand, ethylene glycol, and biomacromolecules.

[0007] The pre-supramolecular targeting chimera is constructed by using supramolecular nanoparticles assembled from metal-organic cages and polyethyleneimine through host-guest interactions as the main framework, and then binding target protein binding ligands to the surface of the supramolecular nanoparticles through host-guest interactions with the supramolecular surface.

[0008] The metal-organic cage is selected from compounds with the structure shown in Formula A:

[0009]

[0010] In formula A, Ra is selected from one of the following segments, or a chain formed by two or more segments: C containing one, two or more heteroatoms 2-12 Alkylene, wherein the heteroatom is selected from S or N, C 6-20 aryl, 5-20 methyl aryl, C 2-12 Alkylene, 3-20 membered heterocyclic or C 3-20 Cycloalkylene;

[0011] This is the connection point, where m is an integer selected from 1 to 100;

[0012] The structure of the polyethylene glycol is shown in Formula B:

[0013]

[0014] Wherein, Rc is selected from -O-adamantane, -OCO-adamantane, -COO-adamantane, -NHCO-adamantane, -CONH-adamantyl, azide, -OC 2-12 Alkyne group; n is an integer selected from 1 to 300;

[0015] Rb is selected from one of the following segments, or a chain formed by two or more segments: C containing one, two or more heteroatoms 2-12 Alkylene, wherein the heteroatom is selected from S or N, C 2-12 Alkylene, C 6-20 arylene, 5-20 quinone heteroarylene, 3-20 quinone heterocyclic or C 3-20 Cycloalkylene;

[0016] The polyethyleneimine contains repeating units as shown in Formula C;

[0017]

[0018] Rd is selected from cyclodextrin groups, azide groups, and -OC groups. 2-12 Alkyne, -OCO-adamantyl, -COO-adamantane, -NHCO-adamantyl, -CONH-adamantyl, -O-adamantyl.

[0019] According to an embodiment of the present invention, the metal-organic cage can be prepared by referring to, for example, the methods described in Angew. Chem. Int. Ed. 2021, 60, 5429-5435 or Liu J., Sheng JH, Shao LH, et al. Tetraphenylethylene-featured fluorescent supramolecular nanoparticles for intracellular trafficking of protein delivery and neuroprotection.

[0020] According to an embodiment of the present invention, in formula A, Ra is selected from one of the following segments, or a chain formed by two or more segments: C containing one, two or more heteroatoms 2-6 Alkylene, wherein the heteroatom is selected from S or N, C 6-14 aryl, 5-14 methyl aryl, C 2-6 Alkylene, 3-14 membered heterocyclic or C 3-14 Cycloalkylene;

[0021] For example, R is Where m is selected from integers from 1 to 80, for example, 1, 3, 5, 10, 20, 30, 40.

[0022] According to an embodiment of the present invention, the polyethylene glycol may be prepared by the method described in the literature Dong RJ, Chen HY, Wang D.L., et al. Supramolecular fluorescent nanoparticles for targeted cancer imaging[J]. ACS Macro Lett. 2012, 1, 1208-1211.

[0023] According to an embodiment of the present invention, the polyethylene glycol has the following structural units:

[0024]

[0025] Wherein, Rc is selected from -O-adamantyl, -OCO-adamantyl, -COO-adamantyl, -NHCO-adamantyl, -CONH-adamantyl, azide, -OC 2-6 Alkyne group; n is an integer selected from 5 to 100, for example, 10, 20, 30, 40.

[0026] According to an embodiment of the present invention, the molecular weight of the polyethylene glycol is 100-10000, for example 200-8000, such as 1900.

[0027] According to an embodiment of the present invention, the polyethylene glycol is selected from compounds having at least one structure of the following formulas B-1, B-2 and B-3:

[0028]

[0029] In equations B-1, B-2, and B-3, n may be the same or different, and is independently selected from integers from 1 to 200, for example, 10, 20, 30, 40, 44, and 50.

[0030] According to an exemplary embodiment of the present invention, the polyethylene glycol is selected from compounds having the structure of formula B-1, that is, polyethylene glycol containing adamantyl alkyl group; for example, the polyethylene glycol is the polyethylene glycol containing adamantyl alkyl group as in B-1 above, where n is 44, the molecular weight is 1900, and it is named Ada-PEG1900.

[0031] According to an embodiment of the present invention, the molecular weight of the polyethyleneimine is 100-50000, for example 600-50000, preferably 600, 1800, 5000, 10000, or 25000.

[0032] According to a preferred embodiment of the present invention, the polyethyleneimine may be prepared by means of, for example, the method described in Angew. Chem. Int. Ed. 2021, 60, 5429-5435.

[0033] According to a preferred embodiment of the present invention, the polyethyleneimine is selected from polyethyleneimine PEI-βCD containing cyclodextrin groups. Preferably, the polyethyleneimine PEI-βCD has a structure as shown in Formula C-1, where βCD is a cyclodextrin group.

[0034]

[0035] According to an embodiment of the present invention, the structure of βCD in formula C-1 is as shown in formula D:

[0036]

[0037] According to an embodiment of the present invention, the target protein binding ligand is selected from protein inhibitors or activators known in the art, such as inhibitors or activators of target proteins BRD4, EGFR, BTK, and ACSL4.

[0038] According to an embodiment of the present invention, the target protein is selected from at least one of the proteins including, but not limited to, BRD4, EGFR, BTK, and ACSL4.

[0039] According to an embodiment of the present invention, the biomacromolecule is selected from nucleic acids, proteins, and biomacromolecule drugs.

[0040] As an example, the biomolecules are selected from BSA and catalase (CAT).

[0041] According to an embodiment of the present invention, in the supramolecular targeting chimera, a metal-organic cage serves as the main framework and, under the action of polyethylene glycol and polyethyleneimine, assembles with the target protein binding ligand and the target protein to form a supramolecular targeting chimera.

[0042] According to an embodiment of the present invention, the self-assembly refers to the assembly of the components in the supramolecular targeted chimera through, for example, host-guest interactions, click chemical modification, or cross-linking.

[0043] According to an embodiment of the present invention, the mass ratio of the polyethyleneimine, the metal-organic cage and the polyethylene glycol is 100:(1-50):(50-200), for example 100:(5-30):(80-150), such as 100:(5-20):(100-120).

[0044] According to an embodiment of the present invention, the mass ratio of the polyethyleneimine to the biomacromolecule is 100:(1-500), for example 100:(5-230), such as 100:(8-50).

[0045] According to an embodiment of the present invention, the mass ratio of the polyethyleneimine to the target protein binding ligand is 0.5:(1-500), for example 1:(2-300), or for example 1:(5-100).

[0046] According to an exemplary embodiment of the present invention, the target protein binding ligand is selected, for example, from compounds represented by formulas I-1 to I-6:

[0047]

[0048] Where X is oxygen or nitrogen; R1, R2, R3, R4, R5, and R6 are the first modifying groups, which may be the same or different, and are independently selected from -O-adamantyl, -OCO-adamantyl, -COO-adamantyl, -NHCO-adamantyl, -CONH-adamantyl, -azido, and -OC. 2-6 alkynyl group; when R7 represents a single or double bond. When it is a single bond, R7 is selected from H; when it is a double bond, R7 is selected from carbonyl; R8 is selected from -NH- and -O-.

[0049] n can be the same or different, and can be selected independently from 0 to 300, for example, from 1 to 100, such as 1, 2, 3, 4, 5, 6, 7, 8, 9 or 10, 20, 30, 40.

[0050] In some embodiments of the present invention, the target protein binding ligand is selected from the following compounds:

[0051]

[0052]

[0053] In E-1, E-2, E-3, E-4, E-5, E-6, E-7 and E-8, n is the same or different and is independently selected from integers from 0 to 40, for example, 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 20, 30 and 40.

[0054] According to an embodiment of the present invention, the target protein binding ligand may be prepared by referring to the literature ACSMed.Chem.Lett.2021,12,720-725, or Angew.Chem.Int.Ed.2021,60,5429-5435, or by synthetic methods known in the art.

[0055] According to an exemplary embodiment of the present invention, the target protein binding ligand is selected from compounds having the structure of formula E-1, that is, target protein binding ligands containing adamantyl groups; for example, when n is 3, the target protein binding ligand containing adamantyl groups is named JQ1-Ada.

[0056] As an exemplary implementation, the target protein binding ligand is selected from the following compounds:

[0057]

[0058]

[0059] According to an embodiment of the present invention, the supramolecular targeted chimera is administered intravenously.

[0060] According to an embodiment of the present invention, the supramolecular targeting chimera is named SupTAC.

[0061] According to an embodiment of the present invention, the supramolecular targeting chimeric protein-binding ligands are of two or more types. When multiple target protein-binding ligands are used, the simultaneous degradation of multiple target proteins can be achieved within the cell.

[0062] The present invention also provides a method for constructing the above-mentioned supramolecular targeted chimera, the method comprising: self-assembling the metal-organic cage, target protein binding ligand, polyethylene glycol and polyethyleneimine and biomacromolecules as described above to obtain nanoparticles, which are the supramolecular targeted chimeras;

[0063] Alternatively, the metal-organic cage, target protein binding ligand, and polyethyleneimine can be self-assembled to obtain a pre-supramolecular targeting chimera; then, the biomacromolecules and polyethylene glycol described above can be added to the pre-supramolecular targeting chimera for assembly.

[0064] According to an embodiment of the present invention, the construction method specifically includes the following steps:

[0065] (1) Prepare storage solutions for biomacromolecules, target protein binding ligands, metal-organic cages, polyethyleneimine, and polyethylene glycol respectively.

[0066] (2) Mix the storage solution of polyethyleneimine, the storage solution of biomacromolecule, the storage solution of target protein binding ligand, the storage solution of polyethylene glycol and the storage solution of metal-organic cage in step (1) together to obtain a mixture. The supramolecular targeted chimera is obtained by self-assembly of metal-organic cage, polyethylene glycol and polyethyleneimine and biomacromolecule drug.

[0067] Alternatively, the storage solutions of polyethyleneimine, target protein binding ligands, and metal-organic cages are first mixed and stirred to obtain a pre-supramolecular targeting chimera. Then, the storage solutions of biomacromolecules and polyethylene glycol are added to the pre-supramolecular targeting chimera, and a supramolecular targeting chimera is obtained through self-assembly.

[0068] According to an embodiment of the present invention, the method further includes a step of centrifugation purification after self-assembly to disperse the supramolecular targeting chimera in a buffer solution.

[0069] According to an embodiment of the present invention, in step (1), the storage solution of the metal-organic cage refers to dissolving the metal-organic cage in an organic solvent, where the metal-organic cage has the meaning as described above. Preferably, the organic solvent can be an organic solvent known in the art, such as dimethyl sulfoxide (DMSO).

[0070] According to an embodiment of the present invention, in step (1), the storage solution of polyethyleneimine refers to the polyethyleneimine dissolved in a solvent, wherein the polyethyleneimine has the meaning as described above. Preferably, the solvent may be a solvent known in the art, such as pure water.

[0071] According to an embodiment of the present invention, in step (1), the polyethylene glycol storage solution refers to the polyethylene glycol dissolved in a solvent, where the polyethylene glycol has the meaning as described above. Preferably, the solvent can be a solvent known in the art, such as pure water.

[0072] According to an embodiment of the present invention, in step (1), the storage solution of the adamantyl target protein binding ligand refers to dissolving the adamantyl target protein binding ligand in an organic solvent, wherein the adamantyl target protein binding ligand has the meaning as described above. Preferably, the organic solvent may be an organic solvent known in the art, such as one selected from dimethyl sulfoxide.

[0073] According to an embodiment of the present invention, in step (2), the mass ratio of polyethyleneimine, metal-organic cage and polyethylene glycol is 100:(1-50):(50-200), such as 100:(5-30):(80-150), for example 100:(5-20):(100-120).

[0074] According to an embodiment of the present invention, in step (2), the mass ratio of the polyethyleneimine to the biomacromolecule is 100:(1-500), for example 100:(5-230), such as 100:(8-50).

[0075] According to an embodiment of the present invention, in step (2), the mass ratio of the polyethyleneimine to the target protein binding ligand is 0.5:(1-500), for example 1:(2-300), or for example 1:(5-100).

[0076] According to an embodiment of the present invention, in step (2), the concentration of the biomacromolecule in the mixture is 0.1-1 mg / mL, such as 0.125-0.5 mg / mL, for example 0.125 mg / mL.

[0077] According to an embodiment of the present invention, in step (2), the concentration of the target protein binding ligand in the mixture is 1-200 μM, for example 2-150 μM or 5-100 μM, such as 2.5 μM, 10 μM, 25 μM, or 100 μM.

[0078] According to an embodiment of the present invention, in step (2), the concentration of polyethyleneimine in the mixture is 0.005-4 mg / mL, for example 0.01-2 mg / mL, for example 0.03 mg / mL or 1 mg / mL.

[0079] According to an embodiment of the present invention, in step (2), the concentration of the metal-organic cage in the mixture is 0.025-0.8 mg / mL, for example 0.25-0.5 mg / mL, such as 0.01 mg / mL or 0.1 mg / mL.

[0080] According to an embodiment of the present invention, in step (2), the concentration of polyethylene glycol in the mixture is 0.4-5 mg / mL, for example 1-3 mg / mL, such as 1.02 mg / mL.

[0081] According to an embodiment of the present invention, in step (2), the mixing conditions are: stirring at room temperature for 0.25-2 hours.

[0082] According to an embodiment of the present invention, room temperature refers to 0-30°C, such as 20-26°C.

[0083] According to an embodiment of the present invention, in step (2), during centrifugal purification, the centrifugation speed is 10000-15000 r / min.

[0084] According to an embodiment of the present invention, in step (2), the buffer solution may be a buffer solution known in the art, such as PBS buffer.

[0085] According to an exemplary embodiment of the present invention, the construction method specifically includes the following steps: self-assembling a metal-organic cage Ada-MOC containing adamantyl groups, polyethylene glycol Ada-PEG1900 containing adamantyl groups, a target protein binding ligand Ada-POI containing adamantyl groups, polyethyleneimine PEI-βCD containing cyclodextrin groups, and biomacromolecules to obtain the supramolecular targeted chimera.

[0086] The present invention also provides the application of the above supramolecular targeted chimera in selective protein degradation in liver and / or lung tissue.

[0087] According to an exemplary embodiment of the present invention, the protein includes, but is not limited to, BRD4, EGFR, BTK, and ACSL4 proteins.

[0088] According to an exemplary embodiment of the present invention, the supramolecular targeting chimera selectively degrades proteins in liver and / or lung tissue for non-disease diagnostic and therapeutic purposes.

[0089] The present invention also provides the use of the above-mentioned supramolecular targeted chimera in the preparation of medicaments for treating diseases.

[0090] According to an exemplary embodiment of the present invention, the drug is used to treat diseases of the liver and / or lungs, such as pneumonia, hepatitis, lung cancer, and liver cancer.

[0091] Beneficial effects

[0092] This invention utilizes supramolecular nanoparticles assembled from metal-organic cages and cyclodextrin-based polyethyleneimine via host-guest interactions as the main framework, and incorporates ligands for adamantyl alkyl target proteins. The target protein ligands are bound to the surface of the nanoparticles through host-guest interactions with cyclodextrin, constructing a pre-supramolecular targeted chimera. Then, polyethylene glycol and biomolecules (proteins) are added to increase the circulation time and tissue targeting of the supramolecular proximity system in the blood, resulting in a supramolecular targeted chimera. The supramolecular proximity system constructed in this invention exhibits tissue selectivity, is simple to manufacture, and uses inexpensive and readily available raw materials. Transmission electron microscopy reveals that the formed supramolecular targeted chimeras have uniform particle size. Experimental results show that the supramolecular targeted chimera of this invention can efficiently degrade target proteins within cells, and the simultaneous degradation of multiple proteins can be achieved by cross-linking different target protein binding ligands. Furthermore, phosphorylation of target proteins can be regulated through proximity phosphorylation kinases. In vivo experiments showed that this supramolecular degradation system can selectively accumulate in the liver and lungs, achieving specific degradation of target proteins in the liver and lungs. This provides a new method for developing tissue-specific protein degradation systems and a new strategy for the treatment of liver and lung diseases.

[0093] Terminology Definitions and Explanations

[0094] Term "C" 2-12"Alynyl" should be understood to refer to a straight-chain or branched monovalent hydrocarbon group containing one or more triple bonds and having 2 to 12 carbon atoms, optionally "C2-C6 alkynyl". The term "C2-C6 alkynyl" should be understood to optionally refer to a straight-chain or branched monovalent hydrocarbon group containing one or more triple bonds and having 2, 3, 4, 5, or 6 carbon atoms, particularly 2 or 3 carbon atoms ("C2-C3-alkynyl"). The alkynyl group is, for example, ethynyl, propynyl, propynyl-2-alkynyl, butynyl-1-alkynyl, butynyl-2-alkynyl, butynyl-3-alkynyl, pentynyl-1-alkynyl, pentynyl-2-alkynyl, pentynyl-3-alkynyl, pentynyl-4-alkynyl, hexynyl-1-alkynyl, hexynyl-2-alkynyl, hexynyl-3-alkynyl, hexynyl-4-alkynyl, hexynyl-5-alkynyl, 1-methylpropynyl-2-alkynyl, 2-methylbutynyl-3-alkynyl, 1-methylbutynyl-3-alkynyl, etc. -yneyl, 1-methylbut-2-yneyl, 3-methylbut-1-yneyl, 1-ethylprop-2-yneyl, 3-methylpentan-4-yneyl, 2-methylpentan-4-yneyl, 1-methylpentan-4-yneyl, 2-methylpentan-3-yneyl, 1-methylpentan-3-yneyl, 4-methylpentan-2-yneyl, 1-methylpentan-2-yneyl, 4-methylpentan-1-yneyl, 3-methylpentan-1-yneyl, 2 -Ethylbut-3-ynyl, 1-Ethylbut-3-ynyl, 1-Ethylbut-2-ynyl, 1-Prop-2-ynyl, 1-Isopropylp-2-ynyl, 2,2-Dimethylbut-3-ynyl, 1,1-Dimethylbut-3-ynyl, 1,1-Dimethylbut-2-ynyl, or 3,3-Dimethylbut-1-ynyl. In particular, the ynyl group is ethynyl, prop-1-ynyl, or prop-2-ynyl.

[0095] Term "C" 3-20 "Cycloalkylene" should be understood as referring to a saturated divalent monocyclic, bicyclic, or polycyclic hydrocarbon ring (also called a fused ring hydrocarbon ring) with 3-20 carbon atoms. Bicyclic or polycyclic alkylene rings include fused cycloalkylene rings, bridged cycloalkylene rings, and spirocycloalkylene rings; fused rings refer to fused ring structures formed by two or more cyclic structures sharing two adjacent ring atoms (i.e., sharing a bond). Bridged rings refer to fused ring structures formed by two or more cyclic structures sharing two non-adjacent ring atoms. Spirocyclic rings refer to fused ring structures formed by two or more cyclic structures sharing a single ring atom. For example, the C... 3-20 Cycloalkylene groups can be C 3-8 Monocyclic cycloalkyl groups, such as cyclopropyl, cyclobutyl, cyclopentyl, cyclohexyl, cycloheptyl, cyclooctyl, or C... 7-12 Heterocyclic cycloalkyl groups, such as decahydronaphthalene rings.

[0096] The term "3-20 membered heterocyclic group" refers to a saturated divalent monocyclic or bicyclic hydrocarbon ring containing 1-5 heteroatoms independently selected from N, O, and S, preferably a "3-12 membered heterocyclic group". The term "3-12 membered heterocyclic group" also refers to a saturated monovalent monocyclic or bicyclic hydrocarbon ring containing 1-5, preferably 1-3, heteroatoms selected from N, O, and S. The heterocyclic group can be attached to the remainder of the molecule via any one of the carbon atoms or a nitrogen atom (if present). In particular, the heterocyclic group may include, but is not limited to: 4-membered rings, such as azapyrocyclobutane or oxopyrocyclobutane; 5-membered rings, such as tetrahydrofuranyl, dioxacyclopentenyl, pyrrolylyl, imidazoalkyl, pyrrololinyl; or 6-membered rings, such as tetrahydropyranyl, piperidinyl, morpholinyl, dithiaalkyl, thiomorpholinyl, piperazineyl, or trithiaalkyl; or 7-membered rings, such as diazapyrocycloheptane.

[0097] Term "C" 6-20 "Aromatic" should be understood to refer to a monocyclic, bicyclic, tricyclic, or more cyclic hydrocarbon ring with 6 to 20 carbon atoms, exhibiting divalent aromaticity or partial aromaticity. Preferably, "C" is used. 6-14 "Aspartic", the term "C" 6-14 "Arylidene" should be understood to preferably represent a monocyclic, bicyclic, or tricyclic hydrocarbon ring ("C") having 6, 7, 8, 9, 10, 11, 12, 13, or 14 carbon atoms, exhibiting divalent aromaticity or partial aromaticity. 6-14 arylene), particularly a ring with 6 carbon atoms (“C6 arylene”), such as phenylene; or biphenyl, or a ring with 9 carbon atoms (“C9 arylene”), such as indenylene or indenylene, or a ring with 10 carbon atoms (“C9 arylene”). 10 Arylenes, such as tetrahydronaphthylene, dihydronaphthylene, or naphthylene, or a ring with 13 carbon atoms (“C”). 13 Aryl groups, such as fluorene groups, or rings with 14 carbon atoms (“C”). 14 (e.g., aryl), such as anthracene.

[0098] The term "5-20-membered heteroaryl" should be understood to include divalent monocyclic, bicyclic, or tricyclic aromatic ring systems or more aromatic ring systems having 5 to 20 ring atoms and containing 1 to 5 heteroatoms independently selected from N, O, and S, for example, "5-14-membered heteroaryl". The term "5-14-membered heteroaryl" should also be understood to include divalent monocyclic, bicyclic, or tricyclic aromatic ring systems having 5, 6, 7, 8, 9, 10, 11, 12, 13, or 14 ring atoms, particularly 5, 6, 9, or 10 carbon atoms, and containing 1 to 5, preferably 1 to 3, heteroatoms independently selected from N, O, and S. Furthermore, in each case, it may be benzo-fused. Specifically, the heteroaryl group is selected from thiopheneyl, furanyl, pyrrolyl, oxazolyl, thiazolyl, imidazolyl, pyrazolyl, isoxazolyl, isothiazolyl, oxadiazolyl, triazolyl, thiazolyl-4H-pyrazolyl, etc. Attached Figure Description

[0099] To more clearly illustrate the embodiments of the present invention, the accompanying drawings of the experiments required in the embodiments will be briefly introduced below. The drawings described below are only some embodiments of the present invention and are not intended to limit the present invention.

[0100] Figure 1 The NMR spectrum of JQ1-Ada;

[0101] Figure 2 The NMR spectrum of POM-Ada;

[0102] Figure 3 The NMR spectrum of IBR-Ada;

[0103] Figure 4 The NMR spectrum of EX229-Ada;

[0104] Figure 5 The NMR spectrum of GEF-Ada;

[0105] Figure 6 The NMR spectrum of TGZ-Ada;

[0106] Figure 7 TEM image of SupTAC;

[0107] Figure 8 After incubating HeLa cells with JQ1-Ada, POM-Ada, and JQ1-Ada+POM-Ada, Western blot experiments were performed to investigate the degradation of intracellular BRD4 by different components.

[0108] Figure 9We incubated HeLa cells with SupTAC while simultaneously incubating them with the proteasome inhibitor MG132 and the E3 ubiquitin ligase competitor Pomalidomide to investigate the mechanism by which BRD4-SupTAC degrades intracellular BRD4.

[0109] Figure 10 We incubated HeLa cells with different concentrations of EGFR-SupTAC and then performed Western blot (WB) experiments to investigate the degradation of intracellular EGFR by different concentrations.

[0110] Figure 11 After incubating HeLa cells with different concentrations of BRD4-EGFR-SupTAC, Western blot experiments were performed to investigate the simultaneous degradation of intracellular BRD4 and EGFR by different concentrations of SupTAC.

[0111] Figure 12 After incubating HeLa cells with EX229-Ada, IBR-Ada, and EX229-Ada+IBR-Ada, Western blot experiments were performed to investigate the phosphorylation of intracellular BTK by different components.

[0112] Figure 13 Mice were injected via tail vein with SupTAC containing BSA-Cy5 ((a) Ada-MOC:PEI-βCD = 6:1; (c) Ada-MOC:PEI-βCD = 10:1, mass ratio) and fluorescence images of major organs were obtained. (b) and (d) Quantification of BRD4 levels in different tissues of mice were obtained. SupTAC selectively degraded BRD4 in the liver (b) and lung (d), respectively.

[0113] Figure 14 This is a schematic diagram of the construction process of the supramolecular targeted chimera of this application and a schematic diagram of its specific degradation of target proteins in the liver and lungs. Detailed Implementation

[0114] The technical solution of the present invention will be further described in detail below with reference to specific embodiments. It should be understood that the following embodiments are merely illustrative and explanatory of the present invention, and should not be construed as limiting the scope of protection of the present invention. All technologies implemented based on the above content of the present invention are covered within the scope of protection intended by the present invention.

[0115] Unless otherwise stated, the raw materials and reagents used in the following examples are commercially available products or can be prepared by known methods.

[0116] Preparation Example 1

[0117] The synthesis of JQ1-Ada is as follows: Compound 2 was synthesized according to the method described in the reference (ACS Med. Chem. Lett. 2021, 12, 720-725). Compound 1 (100 mg, 0.25 mmol), compound 2 (98 mg, 0.3 mmol), and DMAP (3 mg, 0.025 mmol) were dissolved in 10 mL of dichloromethane and placed in an ice-water bath for 15 minutes. DCC (77.25 mg, 0.375 mmol) was dissolved in 5 mL of dichloromethane and slowly added dropwise to the above mixture. After 15 minutes, the mixture was allowed to return to room temperature and reacted overnight. The solution was evaporated to dryness and purified by column chromatography (DCM / MeOH = 50:1) to obtain the target compound JQ1-Ada (yield 60%).

[0118]

[0119] Preparation Example 2

[0120] The synthesis of POM-Ada was as follows: Compound 4 was synthesized according to the method described in the reference (Angew. Chem. Int. Ed. 2021, 60, 5429-5435). Compound 3 (100 mg, 0.37 mmol), compound 4 (226 mg, 0.45 mmol), and K2CO3 (102 mg, 0.74 mmol) were dissolved in 15 mL of acetonitrile and heated to 85°C overnight. The solution was evaporated to dryness and purified by column chromatography (DCM / MeOH = 50:1) to obtain the target compound POM-Ada (yield 58%).

[0121]

[0122] Preparation Example 3

[0123] The synthesis of EX229-Ada was as follows: Compound 5 (10.5 mg, 0.024 mmol), compound 6 (13 mg, 0.036 mmol), HOAT (4.9 mg, 0.036 mmol), EDCI (6.9 mg, 0.036 mmol), and N-methylmorpholine (12.2 mg, 0.12 mmol) were dissolved in 10 mL of dichloromethane. After placing the mixture in an ice-water bath for 15 minutes, DCC (77.25 mg, 0.375 mmol) was dissolved in 0.5 mL of DMF and slowly added dropwise to the above mixture. The reaction was allowed to proceed for 24 hours. The mixture was extracted three times with dichloromethane, evaporated to dryness, and purified by column chromatography (DCM / MeOH = 25:1) to obtain the target compound EX229-Ada (yield 48%).

[0124]

[0125] Preparation Example 4

[0126] The synthesis of IBR-Ada was as follows: Compound 7 (10 mg, 0.0226 mmol), compound 4 (13.8 mg, 0.0271 mmol), and K2CO3 (6.86 mg, 0.05 mmol) were dissolved in 1 mL of acetonitrile and reacted at 85 °C for 48 hours. The solution was evaporated to dryness and purified by column chromatography (DCM / MeOH = 25:1) to obtain the target compound IBR-Ada (yield 36%).

[0127]

[0128] Preparation Example 5

[0129] The synthesis of GEF-Ada was as follows: Compound 8 (118.3 mg, 0.37 mmol), compound 4 (226 mg, 0.45 mmol), and K2CO3 (102 mg, 0.74 mmol) were dissolved in 15 mL of acetonitrile, heated to 85°C, and reacted overnight. The solution was evaporated to dryness and purified by column chromatography (DCM / MeOH = 50:1) to obtain the target compound GEF-Ada (yield 68%).

[0130]

[0131] Preparation Example 6

[0132] The synthesis of TGZ-Ada was as follows: Compound 9 (163.17 mg, 0.37 mmol), compound 4 (226 mg, 0.45 mmol), and K2CO3 (102 mg, 0.74 mmol) were dissolved in 15 mL of acetonitrile, heated to 85°C, and reacted overnight. The solution was evaporated to dryness and purified by column chromatography (DCM / MeOH = 50:1) to obtain the target compound TGZ-Ada (yield 68%).

[0133]

[0134] Example 1

[0135] The construction method of the pre-supramolecular proximity system PEI-βCD / Ada-MOC / POI-Ada is as follows:

[0136] (1) Synthesis of PEI-βCD and Ada-MOC: The synthesis of PEI-βCD and Ada-MOC can be carried out by referring to the methods in the literature (specifically Angew. Chem. Int. Ed. 2021, 60, 5429-5435 or Liu J., Sheng JH, Shao LH, et al. Tetraphenylethylene-featured fluorescent supramolecular nanoparticles for intracellular trafficking of protein delivery and neuroprotection[J]. Angew. Chem. Int. Ed. 2021, 60, 26740-26746), and the structures shown below are obtained respectively.

[0137]

[0138] (2) Synthesis of Ada-PEG1900: The synthesis of adamantane-modified PEG was carried out by referring to the method in the literature (specifically Dong RJ, Chen HY, Wang D.L., et al. Supramolecular fluorescent nanoparticles for targeted cancer imaging[J].ACS Macro Lett.2012,1,1208-1211), and the molecular weight of the PEG was 1900, which was denoted as Ada-PEG1900. The structure is shown below.

[0139]

[0140] (3) Preparation of BSA-Cy5, PEI-βCD, Ada-MOC and Ada-PEG1900 storage solutions: BSA-Cy5 protein storage solution was prepared in PBS buffer (0.01M, pH7.2) at a concentration of 10 mg / mL; PEI-βCD and Ada-PEG1900 storage solutions were prepared in pure water at concentrations of 10 mg / mL and 10 mg / mL, respectively; Ada-MOC storage solution was prepared in DMSO at a concentration of 10 mg / mL; JQ1-Ada, POM-Ada, IBR-Ada and EX229-Ada storage solutions were prepared in DMSO at a concentration of 1 mM.

[0141] (4) In a clean 2 mL centrifuge tube, PEI-βCD, JQ1-Ada, POM-Ada, and Ada-MOC stock solutions were added dropwise to achieve a final PEI-βCD concentration of 0.03 mg / mL, an Ada-MOC concentration of 0.01 mg / mL, a JQ1-Ada concentration of 10 μM, and a POM-Ada concentration of 2.5 μM. The resulting mixture was stirred at room temperature for 1 hour to obtain the pre-supramolecular targeting chimera PEI-βCD / Ada-MOC / JQ1-Ada / POM-Ada (i.e., BRD4-SupTAC).

[0142] Example 2

[0143] In a clean 2mL centrifuge tube, PEI-βCD, GEF-Ada, POM-Ada, and Ada-MOC stock solutions were added dropwise to achieve a final PEI-βCD concentration of 0.03 mg / mL and an Ada-MOC concentration of 0.01 mg / mL. Different concentrations of GEF-Ada (1×10⁻⁶) were then added. 6 nM) and POM-Ada(1×10 6 The concentration ratio of the two (nM) was 4:1. The resulting mixture was stirred at room temperature for 1 hour to obtain the pre-supramolecular targeted chimera EGFR-SupTAC.

[0144] Example 3

[0145] In a clean 2 mL centrifuge tube, PEI-βCD, JQ1-Ada, GEF-Ada, POM-Ada, and Ada-MOC stock solutions were added dropwise to achieve a final PEI-βCD concentration of 0.03 mg / mL and an Ada-MOC concentration of 0.01 mg / mL. JQ1-Ada, GEF-Ada, and POM-Ada were then added, each at a concentration of 1 mM. The resulting mixture was stirred at room temperature for 1 hour to obtain the pre-supramolecular targeted chimeric BRD4-EGFR-SupTAC nanoparticles.

[0146] Example 4

[0147] In a clean 2 mL centrifuge tube, PEI-βCD, IBR-Ada, EX229-Ada, and Ada-MOC stock solutions were added dropwise to achieve final concentrations of PEI-βCD (0.03 mg / mL), Ada-MOC (0.01 mg / mL), IBR-Ada (10 μM), and EX229-Ada (2.5 μM). The resulting mixture was stirred at room temperature for 1 hour to obtain the pre-supramolecular targeted chimera.

[0148] Example 5

[0149] In a clean 2 mL centrifuge tube, PEI-βCD, BSA-Cy5, Ada-PEG1900, JQ1-Ada, POM-Ada, and Ada-MOC stock solutions were added dropwise to achieve final concentrations of PEI-βCD (1 mg / mL), BSA-Cy5 (0.125 mg / mL), Ada-PEG1900 (0.8 mg / mL), Ada-MOC (0.17 mg / mL), JQ1-Ada (100 μM), and POM-Ada (25 μM). The resulting mixture was stirred at room temperature for 1 hour, then centrifuged at 15000 rpm for 15 minutes. The nanoparticles were resuspended in PBS buffer to achieve a final PEI-βCD concentration of 1 mg / mL, thus obtaining the supramolecular targeted chimera.

[0150] Example 6

[0151] In a clean 2 mL centrifuge tube, PEI-βCD, BSA-Cy5, Ada-PEG1900, JQ1-Ada, POM-Ada, and Ada-MOC stock solutions were added dropwise to achieve final concentrations of PEI-βCD (1 mg / mL), BSA-Cy5 (0.125 mg / mL), Ada-PEG1900 (1.02 mg / mL), Ada-MOC (0.1 mg / mL), JQ1-Ada (100 μM), and POM-Ada (25 μM). The resulting mixture was stirred at room temperature for 1 hour, then centrifuged at 15000 rpm for 15 minutes. The nanoparticles were resuspended in PBS buffer to achieve a final PEI-βCD concentration of 1 mg / mL, thus obtaining the supramolecular targeted chimera.

[0152] Comparative Example 1

[0153] The construction method of the PEI-βCD / Ada-MOC / JQ1-Ada system in this comparative example is basically the same as that in Example 1, except that:

[0154] POM-Ada is not added in steps (3) and (4).

[0155] Comparative Example 2

[0156] The construction method of the PEI-βCD / Ada-MOC / POM-Ada system in this comparative example is basically the same as that in Example 1, except that JQ1-Ada is not added.

[0157] Comparative Example 3

[0158] The construction method of the PEI-βCD / Ada-MOC / EX229-Ada system in this comparative example is basically the same as that in Example 4, except that IBR-Ada is not added.

[0159] Comparative Example 4

[0160] The construction method of the PEI-βCD / Ada-MOC / IBR-Ada system in this comparative example is basically the same as that in Example 4, except that EX229-Ada is not added.

[0161] Test Example 1

[0162] The TEM test results for the pre-supramolecular targeted chimera are as follows:

[0163] The pre-supramolecular targeted chimera constructed in Example 1 was diluted 10 times with pure water, and 5 μL was dropped onto a copper grid. After standing at room temperature overnight to dry, TEM testing was performed to obtain the following results. Figure 7 The TEM image shown. From Figure 7 The test results show that the obtained pre-supramolecular targeted chimera has a spherical morphology, uniform particle size and nanoscale, with particle size basically between 100nm and 150nm.

[0164] Test Example 2

[0165] (A) Take 100 μL of the pre-supramolecular targeting chimera prepared in Example 1 and incubate it with HeLa cells. After 18 hours, lyse the cells and perform Western blotting to analyze the expression level of BRD4 in the cells. The test results are as follows: Figure 7 The fourth group from the far right is shown in the image. Three additional control groups were selected: Group 1 on the left was incubated with 100 μL of PBS buffer (PBS group); Group 2 was incubated with 100 μL of the system prepared in Comparative Example 1 (PEI-βCD / Ada-MOC / JQ1-Ada group); and Group 3 was incubated with 100 μL of the system prepared in Comparative Example 2 (PEI-βCD / Ada-MOC / POM-Ada group). Figure 8 It can be seen that the systems of Comparative Examples 1 and 2 can hardly degrade the BRD4 protein, while the pre-supramolecular targeting chimera PEI-βCD / Ada-MOC / JQ1-Ada / POM-Ada (i.e., BRD4-SupTAC) constructed in this invention can effectively degrade the BRD4 protein.

[0166] (B) Take 100 μL of the pre-supramolecular targeting chimera prepared in Example 1 and incubate it with HeLa cells. After 18 hours, lyse the cells and perform Western blotting to analyze the expression level of BRD4 in the cells. The test results are as follows: Figure 9The fourth group from the far right is shown in the diagram. Three additional control groups were selected: Group 1 cells were incubated with 100 μL of PBS buffer (PBS group); Group 2 cells were incubated with 5 μM MG132 and 100 μL of the pre-supramolecular targeting chimera prepared in Example 1 (BRD4-SupTAC+MG132 group); Group 3 cells were incubated with 50 μM pomalidomide and 100 μL of the pre-supramolecular targeting chimera prepared in Example 1 (BRD4-SupTAC+Pomalidomide). Figure 9 It can be seen that after adding MG132 and Pomalidomide, BRD4-SAPCE can hardly degrade BRD4 protein, while the pre-supramolecular targeting chimera BRD4-SupTAC constructed in this invention can effectively degrade BRD4 protein.

[0167] (C) Different volumes of the pre-supramolecular targeting chimera prepared in Example 2 were incubated with HeLa cells. After 18 hours, the cells were lysed for Western blotting analysis to determine the intracellular EGFR expression level. The test results are as follows: Figure 10 As shown in the figure. The results indicate that the degradation efficiency of the pre-supramolecular targeted chimera EGFR-SupTAC constructed in this invention for EGFR increases with increasing Ada-MOC concentration (systems with different Ada-MOC concentrations can be prepared by referring to the preparation method in Example 2).

[0168] (D) Different volumes of the pre-supramolecular targeting chimera prepared in Example 3 were incubated with HeLa cells. After 18 hours, the cells were lysed for Western blotting analysis to determine the expression levels of BRD4 and EGFR within the cells. The test results are as follows: Figure 11 As shown in the figure. The results show that the simultaneous degradation efficiency of the pre-supramolecular targeted chimera BRD4-EGFR-SupTAC constructed in this invention for BRD4 and EGFR increases with the increase of Ada-MOC concentration (systems with different Ada-MOC concentrations can be prepared by referring to the preparation method in Example 3).

[0169] (E) Following the reagent manual, BTK plasmid was transfected into HeLa cells using Lipofectamine 8000 reagent. After incubation for 6 hours, the medium was replaced with fresh medium. Then, 100 μL of the pre-supramolecular targeting chimera prepared in Example 4 was incubated. After 12 hours, cells were lysed for Western blotting analysis of intracellular p-BTK expression levels. The test results are as follows: Figure 12The fifth group from the far right is shown in the diagram. Four additional control groups were selected: Group 1 cells were incubated with 100 μL of PBS buffer (PBS group); Group 2 cells were incubated with 100 μL of the system prepared in Comparative Example 3 (PEI-βCD / Ada-MOC / EX229-Ada group); Group 3 cells were incubated with 100 μL of the system prepared in Comparative Example 4 (PEI-βCD / Ada-MOC / IBR-Ada group); and Group 4 was the BTK plasmid group (BTK group). Figure 12 It can be seen that the supramolecular components in Comparative Examples 3 and 4 can hardly induce BTK phosphorylation, while the supramolecular targeted chimera PEI-βCD / Ada-MOC / EX229-Ada / IBR-Ada constructed in this invention can effectively promote BTK phosphorylation.

[0170] (F) BALB / c mice (4-6 weeks old) were injected intravenously with 200 μL of the supramolecular system encapsulating BSA-Cy5 protein prepared in Example 5 and the supramolecular system encapsulating BSA-Cy5 protein prepared in Example 6, respectively. BALB / c mice injected only with PBS served as a blank control group. Six hours later, the mice were anesthetized and sacrificed. Imaging was performed on the major organs (heart, liver, spleen, lung, and kidney) using an excitation wavelength of 640 nm and a 680 nm filter. The test results are as follows: Figure 13 As shown in a and c.

[0171] (G) BALB / c mice (4-6 weeks old) were injected intravenously with 200 μL of the supramolecular system encapsulating BSA-Cy5 protein prepared in Example 5 and the supramolecular system encapsulating BSA-Cy5 protein prepared in Example 6, respectively. BALB / c mice injected with only PBS served as a blank control group. After 24 hours, the mice were sacrificed, and the BRD4 content in the major organs of the mice was determined as follows:

[0172] Heart, liver, spleen, lung, and kidney samples were collected into 2 mL centrifuge tubes. After mincing the tissue, 500 μL of lysis buffer was added, and the mixture was ground for 2 minutes. The tubes were then incubated on ice for 30 minutes. The samples were then centrifuged at 15000 rpm for 15 minutes, the supernatant was discarded, and protein concentration was determined using the BCA method. 40 μg of total protein was then used for gel electrophoresis and Western blotting. The results are shown below. Figure 13 As shown. From Figure 13 As can be seen, compared with the control group, the level of BRD4 in liver and lung tissues was significantly reduced, while no significant changes were observed in other tissues. This indicates that the supramolecular proximity degradation system of the present invention can selectively accumulate in the liver and lungs, thereby achieving specific degradation of target proteins in the liver and lungs.

[0173] In this invention, when polyethyleneimine, polyethylene glycol, or the metal-organic cage contains other groups with host-guest interactions or click chemistry modifications and cross-linking effects, it exhibits similar cross-linking functionality and yields the same results. For example, when polyethyleneimine, polyethylene glycol, or the metal-organic cage contains azides and terminal alkynes, a click reaction occurs. Nucleic acids, proteins, and other biomolecules can also be loaded onto the metal-organic cage; these will not be elaborated upon here. The selected groups include, but are not limited to, the azides and terminal alkynes listed above; therefore, the aforementioned cross-linking effects can be achieved through click chemistry and other mechanisms.

[0174] The exemplary embodiments of the present invention have been described above. However, the scope of protection of this application is not limited to the above embodiments. Any modifications, equivalent substitutions, improvements, etc., made by those skilled in the art within the spirit and principles of the present invention should be included within the scope of protection of the present invention.

Claims

1. A supramolecular targeted chimera, characterized in that, The supramolecular targeting chimera includes a target protein binding ligand.

2. The supramolecular targeted chimera according to claim 1, characterized in that, The supramolecular targeted chimera is a nanoparticle obtained by self-assembly of a pre-supramolecular targeted chimera with polyethylene glycol and biomacromolecules; or, nanoparticles formed directly by self-assembly of a metal-organic cage, polyethyleneimine, target protein binding ligand, ethylene glycol, and biomacromolecules. The pre-supramolecular targeting chimera is constructed by using supramolecular nanoparticles assembled from metal-organic cages and polyethyleneimine through host-guest interactions as the main framework, and then binding target protein binding ligands to the surface of the supramolecular nanoparticles through host-guest interactions with the supramolecular surface. The metal-organic cage is selected from compounds with the structure shown in Formula A: In formula A, Ra is selected from one of the following segments, or a chain formed by two or more segments: C containing one, two or more heteroatoms 2-12 Alkylene, wherein the heteroatom is selected from S or N, C 6-20 aryl, 5-20 methyl aryl, C 2-12 Alkylene, 3-20 membered heterocyclic or C 3-20 Cycloalkylene; This is the connection point, where m is an integer selected from 1 to 100; The structure of the polyethylene glycol is shown in Formula B: Wherein, Rc is selected from -O-adamantane, -OCO-adamantane, -COO-adamantyl, -NCO-adamantane, azide, and -OC. 2-12 Alkyne group; n is an integer selected from 1 to 300; Rb is selected from one of the following segments, or a chain formed by two or more segments: C containing one, two or more heteroatoms 2-12 Alkylene, wherein the heteroatom is selected from S or N, C 2-12 Alkylene, C 6-20 arylene, 5-20 quinone heteroarylene, 3-20 quinone heterocyclic or C 3-20 Cycloalkylene; The polyethyleneimine contains repeating units as shown in Formula C; Rd is selected from cyclodextrin groups, azide groups, and -OC groups. 2-12 Alkyne group, -OCO-adamantyl group, -COO-adamantyl group, -NCO-adamantyl group, -O-adamantyl group.

3. The supramolecular targeted chimera according to claim 1 or 2, characterized in that, In formula A, Ra is selected from one of the following segments, or a chain formed by two or more segments: C containing one, two or more heteroatoms 2-6 Alkylene, wherein the heteroatom is selected from S or N, C 6-14 aryl, 5-14 methyl aryl, C 2-6 Alkylene, 3-14 membered heterocyclic or C 3-14 Cycloalkylene; For example, R is Where m is selected from integers from 1 to 80; Preferably, the polyethylene glycol has the following structural units: Wherein, Rc is selected from -O-adamantyl, -OCO-adamantyl, -COO-adamantyl, -NHCO-adamantyl, -CONH-adamantyl, azide, -OC 2-6 Alkyne group; n is an integer selected from 5 to 100; Preferably, the molecular weight of the polyethylene glycol is 100-10000; Preferably, the polyethylene glycol is selected from compounds having at least one structure of the following formulas: B-1, B-2, and B-3. In equations B-1, B-2, and B-3, n may be the same or different, and is independently selected from integers from 1 to 200. Preferably, the molecular weight of the polyethyleneimine is 100-50000. Preferably, the polyethyleneimine is selected from polyethyleneimine PEI-βCD containing cyclodextrin groups. Preferably, the polyethyleneimine PEI-βCD has a structure as shown in Formula C-1, where βCD is a cyclodextrin group: Preferably, the structure of βCD in formula C-1 is as shown in formula D: Preferably, the target protein binding ligand is selected from protein inhibitors or activators known in the art, such as inhibitors or activators of target proteins BRD4, EGFR, BTK, and ACSL4. Preferably, the target protein is selected from at least one of BRD4, EGFR, BTK, and ACSL4 proteins; Preferably, the biomacromolecule is selected from nucleic acids, proteins, and biomacromolecule drugs; Preferably, the biomolecule is selected from BSA and catalase (CAT).

4. The supramolecular targeted chimera according to any one of claims 1-3, characterized in that, In the supramolecular targeting chimera, a metal-organic cage serves as the main framework and, under the action of polyethylene glycol and polyethyleneimine, assembles with the target protein binding ligand and the target protein to form a supramolecular targeting chimera. Preferably, the assembly refers to the assembly of the components in the supramolecular targeted chimera through, for example, host-guest interactions, click chemical modification, or cross-linking.

5. The supramolecular targeted chimera according to any one of claims 1-4, characterized in that, The mass ratio of the polyethyleneimine, metal-organic cage and polyethylene glycol is 100:(1-50):(50-200); Preferably, the mass ratio of the polyethyleneimine to the biomacromolecule is 100:(1-500), for example 100:(5-230); Preferably, the mass ratio of the polyethyleneimine to the target protein binding ligand is 0.5:(1-500).

6. The supramolecular targeted chimera according to any one of claims 1-5, characterized in that, The target protein binding ligand is selected from compounds represented by formulas I-1 to I-6 as follows: Where X is oxygen or nitrogen, and R1, R2, R3, R4, R5, and R6 are the first modifying groups, which may be the same or different, and are independently selected from -O-adamantyl, -OCO-adamantyl, -COO-adamantyl, -NHCO-adamantyl, -CONH-adamantyl, -azido, and -OC. 2-6 acetylinyl group; when Represents a single or double bond. When it is a single bond, R7 is selected from H; when it is a double bond, R7 is selected from carbonyl; R8 is selected from -NH- and -O-. n are integers that are either the same or different and are independently selected from 0 to 300; Preferably, the target protein binding ligand is selected from the following compounds: In E-1, E-2, E-3, E-4, E-5, E-6, E-7 and E-8, n is the same or different and is independently selected from integers from 0 to 40.

7. The supramolecular targeted chimera according to claim 6, characterized in that, The target protein binding ligand is selected from the following compounds:

8. The method for constructing the supramolecular targeted chimera according to any one of claims 1-7, characterized in that, The construction method includes: self-assembling a metal-organic cage, a target protein binding ligand, polyethylene glycol, polyethyleneimine, and a biomacromolecule to obtain nanoparticles, which are the supramolecular targeted chimeras. Alternatively, the metal-organic cage, target protein binding ligand, and polyethyleneimine are first self-assembled to obtain a pre-supramolecular targeting chimera; then, biomacromolecules and polyethylene glycol are added to the pre-supramolecular targeting chimera for assembly.

9. The use of the supramolecular targeting chimera according to any one of claims 1-7 in the selective protein degradation in liver and / or lung tissue; preferably, the protein is selected from BRD4, EGFR, BTK, and ACSL4 proteins.

10. Use of the supramolecular targeting chimera according to any one of claims 1-7 in the preparation of a medicament for treating diseases; Preferably, the drug is used to treat diseases of the liver and / or lungs, such as pneumonia, hepatitis, lung cancer, and liver cancer.

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