NAPG proteolysis targeting chimera capable of resisting multiple myeloma as well as preparation method and application of NAPG proteolysis targeting chimera

Abstract

The invention discloses an NAPG proteolysis targeting chimera capable of resisting multiple myeloma as well as a preparation method and application of the NAPG proteolysis targeting chimera. The NAPG protein hydrolysis targeting chimera is prepared by connecting (5-phenyl-1H-benzo [d] imidazole-2-yl) methylamine or 4-(2-(aminomethyl)-1H-benzo [d] imidazole-5-yl) benzoic acid and a derivative thereof or connecting the (5-phenyl-1H-benzo [d] imidazole-2-yl) methylamine or 4-(2-(aminomethyl)-1H-benzo [d] imidazole-5-yl) benzoic acid and a derivative thereof with lenalidomide, thalidomide, hydroxyl thalidomide and a pomalidomide E3 ligase ligand through a linker. The compound provided by the invention can effectively degrade multiple myeloma (MM) cells and NAPG protein in exosomes secreted by the MM cells. The degradation effect affects related signal channels to further inhibit proliferation and osteoclast differentiation of MM cells, so that the activity of resisting multiple myeloma is exerted. The compound has a good degradation effect on NAPG protein and has the potential of being developed into a novel NAPG degradation agent medicine.

Description

Technical Field

[0001] This invention belongs to the field of biomedical technology, specifically relating to a class of PROTAC molecules and their application in the preparation of NAPG-targeting inhibitors, NAPG-targeting degraders, or drugs for the prevention and treatment of cancer. Background Technology

[0002] Multiple myeloma (MM) is a malignant plasma cell disease, accounting for approximately 10% of all hematologic malignancies. Myeloma originates from the asymptomatic precancerous proliferation of monoclonal plasma cells derived from postgerminal central B cells. Multistep genetic and microenvironmental changes lead these cells to transform into malignant tumors. Myeloma most commonly progresses from clinically undetermined monoclonal gammopathy (often called MGUS) to smoldering myeloma, and finally to symptomatic myeloma. Currently, novel proteasome inhibitors, emerging immunomodulatory drugs, monoclonal antibodies (mAbs), antibody-drug conjugates (ADCs), bispecific T-cell conjugates (BiTEs), chimeric antigen-T-cell therapy (CAR-T), peptide-drug conjugates, selective nuclear export inhibitors, and small molecule targeted therapies are novel and developing treatments for MM. Over the past decade, these newly introduced treatment regimens have significantly improved treatment outcomes and survival rates for MM. However, the majority of these patients eventually relapse, highlighting the need for new treatment approaches.

[0003] NAPG protein is a core member of the soluble NSF attachment protein (SNAPs) family and plays a crucial role in membrane fusion. Given the importance of SNAPs in vesicle transport and exosome secretion, NAPG is likely involved in regulating exosome biogenesis or release. Literature reports indicate that NAPG protein expression is closely related to the development and progression of neurological diseases, vascular diseases, and lung cancer. Based on this background, this invention, through in-depth research of clinical databases in previous work, found that in multiple myeloma (MM) patients, the overall survival of patients in the high NAPG expression group was significantly shorter than that of the low expression group. Figure 1 -A) suggests that high NAPG expression is associated with poor prognosis. To verify the pro-cancer effect of NAPG in MM, we overexpressed NAPG in MM cell lines (such as CAG and KMS28PE) and observed a significant increase in cell proliferation. Figure 1 -B). Furthermore, in the MM xenograft model, the tumor weight and volume of mice with high NAPG expression were significantly larger than those of control mice ( Figure 1-C). Notably, we detected high levels of NAPG protein in both MM cell lines and exosomes derived from their conditioned media, suggesting that NAPG not only exists intracellularly but may also mediate pro-cancer signaling via exosomes. These in vitro and in vivo experimental results (including elevated NAPG levels in both cells and exosomes) strongly indicate a high positive correlation between NAPG expression levels and the development and progression of MM. Given the key pathogenic role of NAPG in MM (possibly through intracellular mechanisms and exosome pathways) and the lack of a functional pocket that traditional small molecule drugs can target due to its protein structure, developing effective inhibitors is difficult. Therefore, this study successfully validated the feasibility of NAPG as a targetable degradation agent using chemical probe technology. Based on this finding, developing a targeted protein degrader (TPD) capable of simultaneously and effectively degrading NAPG protein in both intracellular and secreted exosomes of MM cells, thereby comprehensively inhibiting NAPG-driven pro-cancer signaling and blocking its potential propagation via exosomes, thus inhibiting MM cell proliferation, is of great significance for the clinical treatment of MM.

[0004] PROTAC technology aims to artificially induce the degradation of a given target protein (POI) by hijacking the UPS (Proteosome-on-Proteosome). PROTACs are heterobifunctional molecules comprising an E3 ligase ligand fused to a POI ligand via an intermediate chemical linker. Following the formation of the E3-PROTAC-POI ternary complex, the E3 ligase induces ubiquitination of the target protein and subsequent proteasome degradation. Compared to traditional small molecules that primarily function by blocking the catalytic activity of druggable proteins through occupation-driven pharmacology, PROTACs eliminate proteins through an event-driven mode of action, thereby removing all possible functions associated with them (i.e., enzymes, scaffolds, regulators, etc.). Currently, the number of PROTACs targeting MM (Multiple Myeloma) reported in the literature is limited, mainly targeting the degradation of proteins such as BRD4, BTK, MCL1, HPK1, Bcl-xL, CDK6, IKZF1, and HDAC6 to inhibit MM cell proliferation and achieve anti-cancer effects. The NAPG PROTAC disclosed in this paper provides a novel mechanism for treating NAPG-mediated diseases. Summary of the Invention

[0005] The purpose of this invention is to find novel NAPG protein degrading agents, and a series of NAPG protein hydrolysis-targeting chimeric compounds have been designed accordingly. Therefore, this invention provides NAPG protein hydrolysis-targeting chimeric compounds having general formula (I or II) that can be used as NAPG protein degrading agents.

[0006] According to a first embodiment of the present invention, a compound having general formula (I or II) or its stereoisomers, tautomers, or pharmaceutically acceptable salts, hydrates, solvates, polymorphs, or prodrugs, or its NAPG protein hydrolysis-targeting chimeric derivatives are provided:

[0007]

[0008]

[0009] In formula (I), R1 represents a mono- or poly-substituted group on the benzene ring, and R1 is selected from any one or more of the following: alkyl groups having 1 to 6 carbon atoms, alkoxy groups having 1 to 6 carbon atoms, trifluoromethyl groups, halogen groups, nitro groups, cyano groups, hydroxy groups, carboxyl groups, ester groups, amino groups, tetrazolium groups, sulfonamide groups, and aromatic or heterocyclic groups obtained by fusion with the benzene ring;

[0010] In formula (II), R2 is selected from C1-C4 alkyl, ester, hydroxyl, amino, -(CH2)n-OH, -(CH2)n-NH2, n = 1 to 2;

[0011] In formula (I / II), the E3 ligase ligand is lenalidomide, thalidomide, 4-hydroxy-thalidomide, 5-hydroxy-thalidomide, pomalidomide, or their derivatives and isomers.

[0012] Compounds of general formula (I / II) are NAPG protein hydrolysis-targeting chimeras. They can act as degradative agents for NAPG proteins.

[0013] Preferably, the E3 ligase ligand in formula (I / II) is lenalidomide (III):

[0014]

[0015] In general formula (I / II), the linker is a (divalent) linking group or bridging group. Generally, the linker is one or a combination of two or more flexible linking groups or bridging groups.

[0016] Preferably, the flexible linker is selected from one of the following formulas:

[0017]

[0018] In this application, n = 4-15 means n = 4, 5, 6, 7, 8, 9, 10, 11, 12, 13, 14, 15.

[0019] According to a first embodiment of the present invention, a method for preparing the NAPG protein hydrolysis targeting chimera (I) is provided, the method comprising:

[0020] First, using lanadomine as a starting material, it undergoes an acid-amine condensation reaction with a commercially available N-Boc-protected amino fatty acid using N-methylmorpholine as an acid-binding agent and DMF as a solvent to obtain compound 3. Under acidic conditions, the tert-butyloxycarbonyl protecting group is removed to obtain compound 4 in hydrochloride form. This hydrochloride is then condensed with a commercially available dianhydride under conditions using DIPEA as an acid-binding agent and DMF as a solvent to generate compound 5. Finally, compound 5 is reacted with compound 11 under the same acid-amine condensation conditions to obtain the target compound I. Compound I has the following structural formula:

[0021]

[0022] Wherein: R1 represents a mono- or poly-substituted group on the benzene ring, and R1 is selected from any one or more of the following: alkyl groups having 1 to 6 carbon atoms, alkoxy groups having 1 to 6 carbon atoms, trifluoromethyl groups, halogen groups, nitro groups, cyano groups, hydroxyl groups, carboxyl groups, ester groups, amino groups, tetrazolium groups, sulfonamide groups, and aromatic or heterocyclic groups obtained by fusion with the benzene ring.

[0023] More specifically, the preparation method of the NAPG protein hydrolysis targeting chimera (I), i.e., compound (I), includes the following steps:

[0024]

[0025] a. Using linaliamide, i.e., compound 1, as the starting material, and compound 2, with N-methylmorpholine as the acid-binding agent and DMF as the solvent, the acid-amine condensation reaction is carried out to obtain compound 3, which is used for later use;

[0026] b. Take compound 3 obtained in step a and remove the tert-butyloxycarbonyl protecting group under acidic conditions to obtain compound 4 in hydrochloride form, for later use;

[0027] c. Take compound 4 obtained in step b and dianhydride and condense them with DIPEA as an acid-binding agent and DMF as a solvent to generate compound 5 for later use;

[0028] d. Using 4-bromo-o-phenylenediamine, i.e., compound 6, as the starting material, it undergoes a cyclization reaction with glycolic acid under acidic conditions to generate compound 8, which is then set aside.

[0029] e. Compound 8 obtained in step d is reacted with substituted boric acid under alkaline conditions with 1,4-dioxane as solvent via a metal-catalyzed Suzuki coupling reaction to obtain compound 10, which is then set aside for later use.

[0030] f. Take compound 10 obtained in step e and remove the protecting group under acidic conditions to obtain compound 11, for later use;

[0031] g. Compound 5 from step c and compound 11 are subjected to the same acid-amine condensation conditions as described above to obtain target compound I.

[0032] A more preferred embodiment is the method for preparing the NAPG protein hydrolysis targeting chimera 1, i.e., compound (I), which includes:

[0033] a. Add nalidomide, compound 2, polypeptide condensation reagent COMU, and N-methylmorpholine to DMF as solvent and react at room temperature for 6-15 hours. Monitor the reaction progress by TLC. After the reaction is complete, dilute with ethyl acetate and extract with saturated Na2HCO3 solution, water, and saturated NaCl solution in sequence. Combine the organic phases, dry with anhydrous Na2SO4, filter, collect the filtrate, and let it stand at 0℃ for 12 hours. A large amount of white solid precipitates. Filter and collect the residue to obtain compound 3.

[0034] b. Dissolve compound 3 prepared above in dichloromethane, add trifluoroacetic acid, stir at room temperature for 2 hours, remove the solvent under reduced pressure, dilute the mixture with a mixed solvent of dichloromethane and methanol, filter, collect the filter residue, and obtain compound 4 without purification.

[0035] c. Add dianhydride compound 5, compound 4 prepared above, acid-binding agent DIPEA, and DMF as solvent, and react at room temperature for 6-20 hours. After the reaction is complete, dilute with water, extract with organic solvent (ethyl acetate and water), separate the organic solvent (ethyl acetate) layer with a separatory funnel, evaporate to dryness under vacuum, and purify by column chromatography to obtain compound 6.

[0036] d. 4-Bromo-o-phenylenediamine (i.e., compound 6) was dissolved in dichloromethane, and BOC-glycine, EDCI, and HOBt were added. Triethylamine was added as an acid-binding agent. The mixture was stirred at room temperature for 12 hours, the solvent was removed under reduced pressure, and the mixture was extracted three times with water and ethyl acetate. The organic phases were combined, glacial acetic acid was added, and the mixture was heated to reflux at 65°C for 10 hours. After the reaction was completed, saturated Na2CO3 was added to adjust the pH to 8, and the mixture was extracted three times with ethyl acetate. The organic phases were combined, dried over anhydrous Na2SO4, filtered, and concentrated under reduced pressure to obtain the crude product. The crude product was purified by silica gel column chromatography using a petroleum ether-ethyl acetate mixed solvent with a volume ratio of 4:1 as the eluent to obtain compound 8.

[0037] e. The above-prepared compound 8 was added sequentially to commercially available compound 9, Pd(PPh3)4, and potassium acetate. The mixture was heated under nitrogen protection at 105°C and refluxed for 10 hours. After the reaction was completed, the mixture was cooled to room temperature, and the precipitate was filtered off with diatomaceous earth. The filtrate was concentrated under reduced pressure to obtain the crude product, which was then purified by silica gel column chromatography using a 30:1 volume ratio of dichloromethane ether-methanol mixed solvent as the eluent to obtain compound 10.

[0038] f. Dissolve the above-prepared compound 10 in a 1,4-dioxane solution of 4N hydrochloric acid, stir at room temperature for 2 hours, remove the solvent under reduced pressure, dilute the mixture with a mixed solvent of dichloromethane and methanol, filter, collect the filter residue, and obtain compound 11.

[0039] g. Take a round-bottom flask, add compound 5, COMU and N-methylmorpholine prepared above in sequence, add DMF as solvent, stir at room temperature for 2 minutes, then add compound 11 and DMAP, stir at room temperature for 10 hours. After the reaction is completed, separate and purify by silica gel column chromatography, using a 10:1 volume ratio of dichloromethane-methanol mixed solvent as eluent to obtain target product I.

[0040] According to a second embodiment of the present invention, a method for preparing the NAPG protein hydrolysis targeting chimera (II) is provided, the method comprising:

[0041] (1) Using linadouamine as a raw material, and commercially available bromoacyl chloride as a solvent, an acid-amine condensation reaction was carried out to obtain compound 13.

[0042] (2) Compound 13 and compound 18 were subjected to an SN2 nucleophilic substitution reaction under alkaline conditions using DMF as a solvent to obtain target compound II. Compound II has the following structural formula:

[0043]

[0044] Wherein: R2 is selected from any one of C1-C4 alkyl, ester, hydroxyl, amino, -(CH2)n-OH, -(CH2)n-NH2, n = 1 to 2;

[0045] More specifically, the preparation method of the NAPG protein hydrolysis targeting chimera (II) includes the following steps:

[0046] a. Using commercially available lenadoamine as a raw material, and commercially available bromoacyl chloride, in THF as a solvent, an acid-amine condensation reaction was carried out to obtain compound 13.

[0047] b. Using commercially available 4-bromo-o-phenylenediamine as a base, a cyclization reaction occurs with commercially available fatty acids under acidic conditions to produce...

[0048] Compound 15;

[0049] c. The 15 prepared above is reacted with commercially available 4-methoxycarbonylphenylboronic acid under alkaline conditions to form 1,4-dioxane.

[0050] Using a hexacyclic ring as a solvent, compound 17 was obtained by a metal-catalyzed Suzuki coupling reaction.

[0051] d. The protecting group of the above-obtained compound 17 was removed under alkaline conditions to obtain compound 18;

[0052] e. Compound 13 and compound 18 obtained above were subjected to an SN2 nucleophilic substitution reaction under alkaline conditions with DMF as solvent to obtain target compound II.

[0053] As a more preferred embodiment, the method for preparing the estrogen receptor protein hydrolysis targeting chimera (II), i.e., compound (II), includes:

[0054] a. Take a round-bottom flask, add commercially available lenalidomide and bromoacyl chloride in sequence, add THF as solvent, heat under reflux at 70°C for 6 hours, after the reaction is completed, filter, wash with ethyl acetate, collect the filter residue, and obtain 13;

[0055] b. Take a 20mL microwave reaction flask, add 4-bromo-o-phenylenediamine and fatty acid, add 4N HCl, use BiotageInitiator+ microwave synthesizer, turn on the instrument and click Organic Synthesis, set the temperature to 110℃, wavelength to High, gas pressure to 1 bar, pre-stirring time to 5 minutes, reaction time to 1 hour, the reaction is carried out in a closed reaction flask, after the reaction is completed, cool to room temperature, add saturated NaHCO3 to adjust the pH until a large amount of yellow solid precipitates, filter to obtain crude product, and separate and purify by silica gel column chromatography to obtain compound 15;

[0056] c. The compound 15 prepared above was dissolved in a mixed solvent of 1,4-dioxane and water, and commercially available 4-methoxycarbonylphenylboronic acid, i.e., compound 16, Pd(PPh3)4, and K2CO3 were added sequentially. The mixture was heated under nitrogen protection at 105°C for 10 hours. After the reaction was completed, the mixture was cooled to room temperature, and the precipitate was filtered off with diatomaceous earth. The filtrate was concentrated under reduced pressure to obtain the crude product, which was then purified by silica gel column chromatography to obtain 17.

[0057] d. Dissolve the above-prepared compound 17 in a mixed solvent of tetrahydrofuran and water, add NaOH, heat at 50°C for 12 hours, after the reaction is complete, add 1N HCl to adjust to acidity (pH=2), filter, collect the filter residue, and obtain compound 18;

[0058] e. Take a 25 mL round-bottom flask, add compound 13, compound 18 and K2CO3, use DMF as solvent, heat at 90 °C under reflux for 12 hours. After the reaction is completed, the target compound II is obtained by silica gel column chromatography.

[0059] According to a preferred embodiment of the present invention, compounds P1 and P2 are provided as compounds of general formulas (I and II):

[0060]

[0061] This invention has conducted in-depth chemical and pharmacological activity studies on a series of protein hydrolysis-targeting chimeric compounds, and found that the two types of compounds P1 and P2 have good inhibitory effects on NAPG protein, which are promising for development into new drugs for the treatment of multiple myeloma, and provide a new approach for developing new drugs for the treatment of multiple myeloma and enriching the variety of clinical drugs.

[0062] The NAPG protein hydrolysis targeting chimera provided by this invention has good NAPG protein degradation activity and can be used to prepare drugs against multiple myeloma.

[0063] This application also provides a pharmaceutical composition comprising a therapeutically effective amount of the compounds (I and II) of this application, their isomers or pharmaceutically acceptable salts thereof, and a pharmaceutically acceptable carrier.

[0064] This application also provides the use of the compounds (I and II), their isomers, or pharmaceutically acceptable salts thereof in the preparation of medicaments for treating diseases associated with NAPG protein degradation-targeting chimeras.

[0065] This application also provides a method for treating diseases associated with NAPG protein degradation-targeting chimeras, including administering a therapeutically effective amount of the compound of this application, its isomers, or pharmaceutically acceptable salts thereof to a mammal (preferably a human) in need of such treatment.

[0066] This application also provides the use of the compound of this application, its isomers or pharmaceutically acceptable salts thereof in the treatment of diseases associated with NAPG protein degradation-targeting chimeras.

[0067] This application also provides isomers of the compounds shown in this application or pharmaceutically acceptable salts thereof for the treatment of diseases associated with NAPG protein degradation-targeting chimeras.

[0068] In some of the schemes of this application, the NAPG protein degradation targeting chimeric-related diseases is selected from tumors or cancer.

[0069] In some embodiments of this application, the estrogen receptor protein degradation targeting chimeric-related diseases are selected from multiple myeloma.

[0070] Related definitions

[0071] Unless otherwise stated, the following terms and phrases as used herein are intended to have the following meanings. A particular term or phrase should not be considered uncertain or unclear unless specifically defined, but should be understood in its ordinary sense. When a trade name appears herein, it is intended to refer to the corresponding product or its active ingredient.

[0072] The term "pharmaceutically acceptable" as used herein refers to compounds, materials, compositions, and / or dosage forms that, within the bounds of reliable medical judgment, are suitable for use in contact with human and animal tissues without undue toxicity, irritation, allergic reactions, or other problems or complications, in proportion to a reasonable benefit / risk ratio.

[0073] The term "pharmaceutically acceptable salt" refers to the salt of the compounds in this application, prepared by reacting a compound with a relatively non-toxic acid or base, as discovered in this application, with a specific substituent. When the compounds in this application contain relatively acidic functional groups, a base addition salt can be obtained by contacting such compounds with a sufficient amount of base in a pure solution or a suitable inert solvent. When the compounds in this application contain relatively basic functional groups, an acid addition salt can be obtained by contacting such compounds with a sufficient amount of acid in a pure solution or a suitable inert solvent. Certain specific compounds in this application contain both basic and acidic functional groups, and thus can be converted into either a base or an acid addition salt.

[0074] The pharmaceutically acceptable salts of this application can be synthesized from parent compounds containing an acid radical or a base using conventional chemical methods. Generally, such salts are prepared by reacting these compounds, in their free acid or base form, with a stoichiometric amount of a suitable base or acid in water or an organic solvent or a mixture of both.

[0075] The compounds of this application may exist in specific geometric or stereoisomeric forms. This application envisions all such compounds, including cis and trans isomers, (-)- and (+)- enantiomers, (R)- and (S)- enantiomers, diastereomers, (D)- isomers, (2)- isomers, and racemic mixtures thereof, as well as other mixtures, such as mixtures enriched with enantiomers or diastereomers, all of which are within the scope of this application. Additional asymmetric carbon atoms may be present in substituents such as alkyl groups. All such isomers and mixtures thereof are included within the scope of this application.

[0076] The compounds and intermediates of this application may also exist in different tautomer forms, and all such forms are included within the scope of this application. The terms "tautomer" or "tautomer form" refer to structural isomers with different energies that can interconvert via low energy barriers. For example, proton tautomers (also known as proton transfer tautomers) include interconversions via proton migration, such as keto-enol and imine-enamine isomerization. A specific example of a proton tautomer is the imidazole moiety, in which a proton can migrate between two ring nitrogens. Valence tautomers include interconversions via the recombination of some bonding electrons. The compounds of this application may contain atomic isotopes in non-natural proportions on one or more atoms constituting the compound. For example, the compound may be labeled with radioactive isotopes such as tritium (2H), iodine-125 (125I), or C-14 (14C). For example, deuterium can be used to replace hydrogen to form deuterated drugs. The bond between deuterium and carbon is stronger than that between ordinary hydrogen and carbon. Compared with undeuterated drugs, deuterated drugs have advantages such as reduced toxicity, increased drug stability, enhanced efficacy, and prolonged drug biological half-life. All isotopic transformations of the compounds in this application, regardless of radioactivity, are included within the scope of this application.

[0077] The terms “optional” or “optionally” refer to events or conditions that may occur but are not required to occur as described below, and the description includes both cases where said events or conditions occur and cases where said events or conditions do not occur.

[0078] The term "targeting chimera" refers to a bifunctional molecule containing two small molecule ligands: one with high affinity for the target protein and a second for recruiting an E3 ligase that ubiquitinates the protein and targets it for proteolysis via the proteasome.

[0079] The compounds of this application can be prepared by a variety of synthetic methods known to those skilled in the art, including the specific embodiments listed below, embodiments formed by combining them with other chemical synthetic methods, and equivalent substitutions known to those skilled in the art. Preferred embodiments include, but are not limited to, the embodiments of this application.

[0080] The solvents used in this application are commercially available. Compounds are named according to conventional naming principles in the art or using ChemDraw@ software; commercially available compounds are named using supplier directory names.

[0081] Beneficial technical effects of the present invention:

[0082] The bifunctional NAPG protein hydrolysis targeting chimera synthesized in this invention has good pharmacokinetic, bioavailability and in vivo efficacy properties, and can be developed into a novel NAPG protein degrading agent for the treatment of multiple myeloma.

[0083] The compounds I and II provided in this invention achieve effective treatment of multiple myeloma by targeting and degrading the NAPG protein. Their mechanism of action is based on PROTAC technology, inducing the ubiquitination and degradation of the NAPG protein through the formation of an E3-PROTAC-NAPG ternary complex, thereby inhibiting tumor cell proliferation and blocking exosome-mediated oncogenic signaling. The development of these compounds provides a new strategy for the treatment of multiple myeloma and also lays the foundation for the treatment of other diseases related to the NAPG protein. Attached Figure Description

[0084] Figure 1 This is the result of an experiment on NAPG-regulated malignant proliferation of MM.

[0085] Figure 2 These are the results of the NAPG protein hydrolysis activity test.

[0086] Figure 3 The figure shows the experimental results of the effects of compounds P1 and P2 of the present invention on the proliferation of MM cells.

[0087] Figure 4 This is the H-NMR spectrum of compound P1 of the present invention.

[0088] Figure 5 This is the H-NMR spectrum of compound P2 of the present invention. Detailed Implementation

[0089] The technical solution of the present invention will be illustrated below with examples. The scope of protection sought by the present invention includes, but is not limited to, the following embodiments.

[0090] In order to find NAPG protein degrading agents with better efficacy, based on the current understanding of this type of drug and relevant literature, two types of NAPG protein degrading agents were synthesized. The NAPG protein degradation activity of the obtained target compounds was tested, and it was found that both compounds, P1 and P2, have the effect of degrading NAPG protein.

[0091] The structure of the target compound has been confirmed by 1H-NMR.

[0092] Example 1: Preparation of compound P1, the steps are as follows:

[0093]

[0094] Step a: Take a 25 mL round-bottom flask and add commercially available lenalidomide (170 mg, 0.66 mmol), N-Boc-6-aminocaproic acid (231 mg, 1 mmol), COMU (402 mg, 0.95 mmol), and N-methylmorpholine (202 mg, 2 mmol) in sequence. Add DMF (2 mL) as a solvent and stir at room temperature. After the reaction is complete, dilute with ethyl acetate (10 mL) and add saturated Na2HCO3 in sequence. Extracted with 3 solution (20 mL), water (20 mL), and saturated NaCl solution (20 mL), the organic phases were combined, dried over anhydrous Na2SO4, filtered, and the filtrate was collected. The mixture was allowed to stand at 0 °C for 12 hours, and a large amount of white solid precipitated out. The mixture was then filtered, and the residue was collected to obtain (7-((2-(2,6-dioxopiperidin-3-yl)-1-oxoisoindoline-4-yl)amino)-7-oxoheptyl)tert-butyl carbamate (3) (289 mg, 90%), which was a white solid. 1 H NMR (500MHz, DMSO) δ11.04(s,1H),9.84(s,1H),7.83(d,J=7.2Hz,1H),7.50(q,J=7.4Hz,2H),5.28-5.05(m,1H),4.48-4.24(m,2H),3.00-2. 85(m,1H),2.63(t,J=14.5Hz,1H),2.42(t,J=7.3Hz,2H),2.33-2.24(m,2H),2.04(dd,J=17.8,12.1Hz,1H),1.91(s,1H),1.90-1.77(m,2H).

[0095] Step b: The compound 3 (243.2 mg, 0.5 mmol) prepared above was dissolved in dichloromethane (15 mL), and trifluoroacetic acid (5 mL) was added. The mixture was stirred at room temperature for 2 hours, and the solvent was removed under reduced pressure. The mixture was diluted with a mixed solvent of dichloromethane and methanol (10 mL, volume ratio 20:1), filtered, and the residue was collected. No purification was required to obtain 7-amino-N-(2-(2,6-dioxopiperidin-3-yl)-1-oxoisoindoline-4-yl)heptamide (4) (164.2 mg, 85%), which was a white solid.

[0096] Step c: Take a 25 mL round-bottom flask and add commercially available glutaric anhydride (115 mg, 1.1 mmol), compound 4 (372 mg, 1 mmol), and DIPEA (390 mg, 3 mmol) in sequence. Add DMF (1 mL) as solvent and stir at room temperature for 8-12 hours. After the reaction is complete, 5-((7-((2-(2,6-dioxopiperidin-3-yl)-1-oxoisoindoline-4-yl)amino)-7-oxoheptyl)amino)-5-oxovalerate (5) (467.7 mg, 85%) is obtained by silica gel column chromatography. The product is a white solid. 1 HNMR(500MHz,DMSO)δ11.03(s,1H),9.85(s,1H),7.83(d,J=6.3Hz,2H),7.49(q, J=7.4Hz,2H),5.14(dd,J=13.3,5.1Hz,1H),4.37(q,J=17.5Hz,2H),3.04(dt,J=1 2.9,6.9Hz,4H),2.97-2.86(m,1H),2.36(t,J=7.5Hz,2H),2.18(t,J=7.3Hz,2H) ,2.07(t,J=7.4Hz,2H),1.78-1.55(m,4H),1.50-1.36(m,2H),1.33-1.28(m,2H).

[0097] Step d: Dissolve 4-bromo-o-phenylenediamine (9.4 g, 50 mmol) in dichloromethane (30 mL), add BOC-glycine (8.8 g, 50 mmol), EDCI (19.1 g, 100 mmol), and HOBt (10.2 g, 75 mmol), and add triethylamine (20 mL, 150 mmol) as an acid-binding agent. Stir the mixture at room temperature for 8-12 hours, remove the solvent under reduced pressure, extract three times with water and ethyl acetate, combine the organic phases, add glacial acetic acid (20 mL), heat to reflux at 65 °C for 10 hours, after the reaction is complete, add saturated Na2CO3 to adjust the pH to 8, extract three times with ethyl acetate, combine the organic phases, dry with anhydrous Na2SO4, filter, concentrate under reduced pressure to obtain crude product, and purify by silica gel column chromatography to obtain ((5-bromo-1H-benzo[d]imidazol-2-yl)methyl)tert-butyl carbamate (8) (6.5 g, 40%), the product is a white solid. 1 H NMR(500MHz,DMSO)δ9.51-9.31(m,1H),7.81(d,J=24.1Hz,1H),7.61-7.29(m,1 H),7.10(d,J=6.5Hz,1H),3.76(d,J=23.2,6.1Hz,2H),1.39(d,J=19.5Hz,9H).

[0098] Step e: Dissolve the prepared 8 (163 mg, 0.5 mmol) in a mixed solvent of 1,4-dioxane and water (20 mL, volume ratio 20:1), and add commercially available 4-methoxycarbonylphenylboronic acid (180 mg, 1 mmol), Pd(PPh3)4 (28 mg, 5%), and potassium acetate (142 mg, 1.5 mmol) in sequence. Heat under nitrogen protection at 105 °C under reflux for 10 hours. After the reaction is complete, cool to room temperature, filter off the precipitate with diatomaceous earth, concentrate the filtrate under reduced pressure to obtain the crude product, and purify by silica gel column chromatography to obtain methyl 4-(2-(((tert-butyloxycarbonyl)amino)methyl)-1H-benzo[d]imidazol-5-yl)benzoate (10) (102.9 mg, 54%), which is a white solid. 1 H NMR(500MHz,DMSO)δ12.37(d,J=11.6Hz,1H),7.77(s,1H),7.60(dd,J=16.5,10.5Hz,3H),7.5 2-7.45(m,3H),7.44-7.40(m,1H),5.72(t,J=5.4Hz,1H),4.70(d,J=5.7Hz,2H),1.33(s,9H).

[0099] Step f: Dissolve the 10 (381 mg, 1.0 mmol) prepared above in 20 mL of 1,4-dioxane solution in 4N hydrochloric acid, stir at room temperature for 2 hours, remove the solvent under reduced pressure, dilute the mixture with a mixed solvent of dichloromethane and methanol, filter, collect the filter residue, and obtain methyl 4-(2-(aminomethyl)-1H-benzo[d]imidazol-5-yl)benzoate (11) (109.7 mg, 78%), which is a white solid. 1 H NMR (500MHz, DMSO) δ8.81 (s, 1H), 8.13-7.58 (m, 7H), 3.95 (d, J = 3.2Hz, 2H), 3.88 (d, J = 2.3Hz, 3H).

[0100] Step g: Take a 25 mL round-bottom flask and add compound 5 (730 mg, 1.5 mmol), COMU (620 mg, 1.45 mmol), and N-methylmorpholine (303 mg, 3 mmol) in sequence. Add DMF (0.5 mL) as solvent and stir at room temperature for 2 minutes. Then add compound 11 (281 mg, 1 mmol) and DM AP (6 mg, 5%) and stir at room temperature for 10 hours. After the reaction is complete, the product is purified by silica gel column chromatography to obtain methyl 4-(2-((5-((6-((2-(2,6-dioxopiperidin-3-yl)-3-oxoisoindoline-4-yl)amino)-6-oxohexyl)amino)-5-oxopentanyl)methyl)-1H-benzo[d]imidazol-5-yl)benzoate (P1) (651.9 mg, 58%), which is a white solid.

[0101] The structure of P1 was confirmed by 1H-NMR: 1 H NMR (500MHz, DMSO) δ12.45(s,1H),11.03(s,1H),9.85(s,1H),8.54(s,1H),8.22(d,J=6.7Hz,1H),8.03(d,J=7.9Hz,2 H),7.89(s,1H),7.83(dd,J=14.7,7.8Hz,3H),7.52-7.45(m,2H),6.97(d,J=6.7Hz,1H),5.14(dd,J=13.2,4.8Hz,1H), 4.50(d,J=5.6Hz,2H),3.88(s,3H),3.05(d,J=6.3Hz,2H),2.92(t,J=13.0Hz,2H),2.40-2.30(m,4H),2.20(t,J=7.4H z,2H),2.09(t,J=7.4Hz,2H),1.75(dd,J=26.2,18.8Hz,2H),1.65-1.56(m,2H),1.48-1.38(m,2H),1.35-1.27(m,2H). 13C NMR (126MHz, DMSO) δ174.62,173.33,172.78,172.09,171.83,171.52,168.30,166.60,1 54.26,146.12,134.30,134.17,133.30,133.12,130.26,129.03,128.17,127.47,125.7 2,121.71,119.39,63.73,52.79,52.57,52.01,47.05,42.78,39.98,38.83,38.77,37.43,36.20,35.28,35.03,34.95,33.52,31.68,29.44,26.60,25.31,23.10,21.84,21.21.

[0102] Example 2:

[0103] The preparation of compound P2 follows these steps:

[0104]

[0105] Step a: Add commercially available lenalidomide (170 mg, 0.66 mmol) and 6-bromohexanoyl chloride (214 mg, 1 mmol) sequentially, add THF (10 mL) as solvent, heat under reflux at 60-80 °C for 6 hours. After the reaction is complete, filter, wash with ethyl acetate, collect the filter residue, and obtain 6-bromo-N-(2-(2,6-dioxopiperidin-3-yl)-1-oxoisoindoline-4-yl)hexanoamide (13) (392 mg, 90%), which is a white solid. 1 H NMR (500MHz, DMSO) δ11.04(s,1H),9.80(s,1H),7.81(d,J=7.2Hz,1H),7.50(q, J=7.6Hz,2H),5.16(dd,J=13.3,4.9Hz,1H),4.37(q,J=17.4Hz,2H),3.56(t,J=6 .6Hz,2H),2.97-2.88(m,1H),2.61(d,J=17.2Hz,1H),2.39-2.30(m,3H),2.07-2 .01(m,1H),1.88-1.81(m,2H),1.65(dd,J=14.8,7.4Hz,2H),1.48-1.41(m,2H).

[0106] Step b: Dissolve 4-bromo-o-phenylenediamine (9.4 g, 50 mmol) in dichloromethane (30 mL), add BOC-glycine (8.8 g, 50 mmol), EDCI (19.1 g, 100 mmol), and HOBt (10.2 g, 75 mmol), and add triethylamine (20 mL, 150 mmol) as an acid-binding agent. Stir the mixture at room temperature for 10 hours, remove the solvent under reduced pressure, extract three times with water and ethyl acetate, combine the organic phases, add glacial acetic acid (20 mL), heat to reflux at 65 °C for 10 hours, after the reaction is complete, add saturated Na2CO3 to adjust the pH to 8, extract three times with ethyl acetate, combine the organic phases, dry with anhydrous Na2SO4, filter, concentrate under reduced pressure to obtain crude product, and purify by silica gel column chromatography to obtain ((5-bromo-1H-benzo[d]imidazol-2-yl)methyl)tert-butyl carbamate (8) (6.5 g, 40%), the product is a white solid. 1 H NMR(500MHz,DMSO)δ9.51-9.31(m,1H),7.81(d,J=24.1Hz,1H),7.61-7.29(m,1 H),7.10(d,J=6.5Hz,1H),3.76(d,J=23.2,6.1Hz,2H),1.39(d,J=19.5Hz,9H).

[0107] Step c: The compound 8 (163 mg, 0.5 mmol) prepared above was dissolved in a mixed solvent of 1,4-dioxane and water (20 mL, volume ratio 20:1). Commercially available 4-methoxycarbonylphenylboronic acid (180 mg, 1 mmol), Pd(PPh3)4 (28 mg, 5%), and potassium acetate (142 mg, 1.5 mmol) were added sequentially. The mixture was heated under nitrogen protection at 105 °C and refluxed for 10 hours. After the reaction was completed, the mixture was cooled to room temperature, and the precipitate was filtered off with diatomaceous earth. The filtrate was concentrated under reduced pressure to obtain a crude product. The crude product was purified by silica gel column chromatography to obtain methyl 4-(2-(((tert-butyloxycarbonyl)amino)methyl)-1H-benzo[d]imidazol-5-yl)benzoate (10) (102.9 mg, 54%), which was a white solid. 1 H NMR(500MHz,DMSO)δ12.37(d,J=11.6Hz,1H),7.77(s,1H),7.60(dd,J=16.5,10.5Hz,3H),7.5 2-7.45(m,3H),7.44-7.40(m,1H),5.72(t,J=5.4Hz,1H),4.70(d,J=5.7Hz,2H),1.33(s,9H).

[0108] Step d: Add 10 (381 mg, 1 mmol) sequentially, add NaOH (400 mg, 10 mmol), and add a mixed solvent of methanol and water (volume ratio 4:1, 20 mL) as solvent. Heat under reflux at 50 °C for 12 hours. After the reaction is complete, remove methanol under reduced pressure, add 1 N HCl until pH is 5, and extract three times with ethyl acetate (10 mL) and water (10 mL). Combine the organic phases, remove the solvent under reduced pressure, and obtain 4-(2-(((tert-Butoxycarbonyl)amino)methyl)-1H-benzo[d]imidazol-5-yl)benzoic acid (14) (304 mg, 83%), which is a white solid. 1 H NMR (500MHz, DMSO) δ8.07(d,J=4.8Hz,1H),8.03(dd,J=15.3,5.5Hz,3H),7.89(s,4H),7.86(s,1H),4.70(d,J=4.7Hz,2H),1.41(s,9H).

[0109] Step e: Compound 14 (367 mg, 1 mmol), compound 13 (436 mg, 1.1 mmol), K2CO3 (414 mg, 3 mmol), and DMF (10 mL) were added sequentially. The mixture was heated under reflux at 90 °C for 12 hours. After the reaction was completed, the product was purified by silica gel column chromatography to obtain 6-((2-(2,6-dioxopiperidin-3-yl)-1-oxoisoindoline-4-yl)amino)-6-oxohexyl4-(2-(((tert-butoxycarbonyl)amino)methyl)-1H-benzo[d]imidazol-5-yl)benzoate (15) (361 mg, 51%), which was a white solid. 1 H NMR (500MHz, DMSO) δ12.39(s,1H),11.03(s,1H),9.80(s,1H),8.02(d,J=7.9Hz,2H),7.82(d,J=7.1H z,3H),7.61(s,1H),7.51(dd,J=15.3,8.1Hz,4H),5.15(dd,J=13.2,4.9Hz,1H),4.37(d,J=8.3Hz,3H ),4.32(d,J=6.2Hz,2H),2.91(t,J=13.1Hz,1H),2.65-2.58(m,2H),2.43-2.40(m,3H),2.36(d,J=13 .3Hz,2H),2.01(d,J=9.3Hz,2H),1.79(s,2H),1.73-1.68(m,2H),1.50(d,J=6.9Hz,2H),1.43(s,9H).

[0110] Step f: The compound 15 (361 mg, 0.5 mmol) prepared above was dissolved in 20 mL of 1,4-dioxane solution in 4 M hydrochloric acid. The mixture was stirred at room temperature for 2 hours, the solvent was removed under reduced pressure, the mixture was diluted with a mixed solvent of dichloromethane and methanol, filtered, and the residue was collected to obtain 6-((2-(2,6-dioxopiperidin-3-yl)-1-oxoisoindoline-4-yl)amino)-6-oxohexyl4-(2-(aminomethyl)-1H-benzo[d]imidazol-5-yl)benzoate (P2) (560 mg, 90%), which was a white solid.

[0111] The structure of P2 was confirmed by 1H-NMR: 1 H NMR (500MHz, DMSO) δ10.14(d,J=18.3Hz,1H),8.02(s,2H),7.83(d,J=6.6Hz,3H),7.49(s,4H),5.12(d,J=8.7Hz,1H),4.59(s,2H),4.46( d,J=17.4Hz,2H),4.35(dd,J=22.8,12.0Hz,3H),2.38(d,J=7.1Hz,2H),2.02(s,2H),1.78(s,2H),1.69(s,2H),1.60(s,2H),1.46(s,4H).

[0112] Example 3

[0113] NAPG protein hydrolysis activity assay:

[0114] The NAPG protein hydrolysis activity test was conducted, and the experimental procedure is as follows:

[0115] 1. Take ARP1 cells in the logarithmic growth phase and administer at a dose of 1.5 × 10⁻⁶. 6 Spread the seeded material into 6-well plates and incubate overnight;

[0116] 2. Add compounds P1 and P2 (DMSO ratio of 0.1%) prepared at different gradients of 0, 1, 10, 20, 50, and 100 μM and incubate in an incubator for 24 h;

[0117] 3. Collect cells from each group and wash once with PBS; add cell lysis buffer containing 1×Protease Inhibitor coc ktail, lyse thoroughly on ice, centrifuge at 14000 rpm for 10 min at 4℃ and collect the supernatant.

[0118] 4. After determining the protein concentration using the BCA kit, the NAPG protein level was detected by Western blot.

[0119] Test results are as follows Figure 2 As shown, in ARP1 cells, compounds P1 and P2 were added at concentration gradients of 0, 1, 10, 20, 50, and 100 μM, respectively. After treatment for 24 hours, the expression level of the target NAPG protein was quantitatively detected by Western blotting. Both compounds P1 and P2 showed strong, concentration-dependent degradation ability of the target protein. The degradation level of compound P1 increased with increasing concentration from 0 to 20 μM, while compound P2 did not show significant degradation ability from 0 to 10 μM. When the concentration reached 20 μM, it showed significant degradation ability and had higher activity than compound P1. Both compounds could achieve near-complete degradation at ≥20 μM.

[0120] Example 4: Effects of compounds P1 and P2 of the present invention on MM cell proliferation

[0121] Multiple myeloma (MM) cell line XG1 wild-type cells were collected. Succinate dehydrogenase in the mitochondria of living cells can reduce exogenous MTT to water-insoluble blue-purple formazan crystals, which are then deposited in the cells. Dead cells lack this function. Dimethyl sulfoxide (DMSO) can dissolve the formazan in the cells. The absorbance was measured at 570 nm using a microplate reader. Within a certain cell number range, the amount of MTT crystals formed is directly proportional to the cell number. The number of live cells was determined based on the measured absorbance value (OD); the higher the OD, the stronger the cell viability.

[0122] Specific steps: 1) XG1 wild-type cell lines in the logarithmic growth phase were seeded in sterile 96-well plates with 3 replicates per group. Compounds P1 and P2 with 6 concentration gradients were added and incubated in a 37°C, 5% CO2 incubator for 48 hours.

[0123] 2) Add freshly prepared MTT reagent to each well and incubate at 37°C in a 5% CO2 incubator for 4 hours;

[0124] 3) Centrifuge in a centrifuge, adding 150 μL of DMSO to each well;

[0125] 4) Measure the OD of each well at 570 nm using an ELISA reader. Calculate the survival rate of myeloma cells using the drug according to the following formula: Survival rate = (Experimental group OD - Blank group OD) / (Control group OD - Blank group OD) × 100%; plot the curve and solve for the IC50. 50 .

[0126] The results are as follows (see details) Figure 3The effects of the two compounds on the proliferation of the XG1 wild-type cell line were tested. In the human multiple myeloma cell line (XG1), compounds P1 and P2 were added at concentration gradients of 0, 1, 10, 20, 50, and 100 μM, respectively. After culturing for 48 hours, cell viability was detected by the MTT assay. Compounds P1 and P2 showed dose-dependent inhibition of cell proliferation, with compound P1 exhibiting the highest IC50 value. 50 =14.8 μM, IC50 of compound P2 50 =18.8μM.

[0127] The synthesized compounds P1 and P2 exhibit good anti-proliferative activity against MM cells and can target and degrade NAPG protein to inhibit MM cell proliferation, showing promising potential for the treatment of multiple myeloma.

[0128] The above description is merely a representative embodiment of the present invention and is not intended to limit the present invention in any way. Any modifications, equivalent changes, or alterations made to the above embodiments based on the technical essence of the present invention shall still fall within the scope of the present invention.

Claims

1. A NAPG proteolytic targeting chimera with anti-multiple myeloma properties, characterized in that, It has compounds with the structure shown in general formula I or general formula II: R1 is selected from any one of the mono- or poly-substituted groups on the benzene ring; R2 is selected from C1-C4 alkyl, ester, hydroxyl, amino, -(CH2)n-OH, -(CH2)n-NH2, n = 1 to 2; Linker is selected from alkyl long chains with a length of generally 4 to 15 carbon atoms or containing heteroatoms; The E3 ligase ligand is selected from lenalidomide, thalidomide, hydroxythalidomide, or pomalidomide.

2. The NAPG protein hydrolysis targeting chimera according to claim 1, characterized in that, The E3 ligase ligand is pomalidomide.

3. The NAPG protein hydrolysis targeting chimera according to claim 1, characterized in that, The flexible linker is selected from one of the following formulas:

4. The method for preparing the NAPG protein hydrolysis targeting chimera according to claim 1, characterized in that, The preparation method of compound I includes the following steps: First, using lanadoamine as a raw material, it undergoes an acid-amine condensation reaction with commercially available N-Boc-protected amino fatty acids using N-methylmorpholine as an acid binder and DMF as a solvent to obtain compound (3). Under acidic conditions, the tert-butyloxycarbonyl protecting group is removed to obtain compound (4) in hydrochloride form. Then, it is condensed with commercially available dianhydrides under conditions using DIPEA as an acid binder and DMF as a solvent to generate compound (5). Finally, it is obtained by reacting compound 11 under the same acid-amine condensation conditions as described above. Compound I has the following structural formula: Wherein: R1 represents a mono- or poly-substituted group on the benzene ring, and R1 is selected from any one or more of the following: alkyl groups having 1 to 6 carbon atoms, alkoxy groups having 1 to 6 carbon atoms, trifluoromethyl groups, halogen groups, nitro groups, cyano groups, hydroxy groups, carboxyl groups, ester groups, amino groups, tetrazolium groups, sulfonamide groups, and aromatic or heterocyclic groups obtained by fusion with the benzene ring.

5. The method for preparing the NAPG protein hydrolysis targeting chimera according to claim 4, characterized in that, The preparation method of compound I includes the following steps: a. Using linaliamide, i.e., compound 1, as the starting material, and compound 2, with N-methylmorpholine as the acid-binding agent and DMF as the solvent, the acid-amine condensation reaction is carried out to obtain compound 3, which is used for later use; b. Take compound 3 obtained in step a and remove the tert-butyloxycarbonyl protecting group under acidic conditions to obtain compound 4 in hydrochloride form, for later use; c. Take compound 4 obtained in step b and dianhydride and condense them with DIPEA as an acid-binding agent and DMF as a solvent to generate compound 5 for later use; d. Using 4-bromo-o-phenylenediamine, i.e., compound 6, as the starting material, it undergoes a cyclization reaction with glycolic acid under acidic conditions to generate compound 8, which is then set aside. e. Compound 8 obtained in step d is reacted with substituted boric acid under alkaline conditions with 1,4-dioxane as solvent via a metal-catalyzed Suzuki coupling reaction to obtain compound 10, which is then set aside. f. Take compound 10 obtained in step e and remove the protecting group under acidic conditions to obtain compound 11, for later use; g. Compound 5 from step c and compound 11 are subjected to the same acid-amine condensation conditions as described above to obtain target compound I.

6. The method for preparing the NAPG protein hydrolysis targeting chimera according to claim 1, characterized in that, The preparation method of compound II includes the following steps: Using linadolamine as a raw material, and bromoacyl chloride as a solvent, an acid-amine condensation reaction was carried out to obtain compound 13; Compound 13 and compound 18 were subjected to an SN2 nucleophilic substitution reaction under basic conditions using DMF as a solvent to obtain target compound II, which has the following structural formula: Wherein: R2 is selected from any one of C1-C4 alkyl, ester, hydroxyl, amino, -(CH2)n-OH, -(CH2)n-NH2, n = 1 to 2.

7. The method for preparing the NAPG protein hydrolysis targeting chimera according to claim 6, characterized in that, The preparation method of compound II includes the following steps: a. Using linadoumine, i.e., compound 1, as a raw material, and bromoacyl chloride, i.e. compound 12, in THF as a solvent, an acid-amine condensation reaction is carried out to obtain compound 13. b. Compound 6 is formed by the cyclization reaction of 4-bromo-o-phenylenediamine with fatty acid, compound 14, under acidic conditions to produce compound 15. c. Compound 15 prepared above is reacted with 4-methoxycarbonylphenylboronic acid, i.e., compound 16, under alkaline conditions with 1,4-dioxane as solvent via a metal-catalyzed Suzuki coupling reaction to obtain compound 17. d. The protecting group of the above-obtained compound 17 was removed under alkaline conditions to obtain compound 18; e. Compound 13 and compound 18 obtained above were subjected to an SN2 nucleophilic substitution reaction under alkaline conditions with DMF as solvent to obtain target compound II.

8. The NAPG proteolytic targeting chimera with anti-multiple myeloma properties according to claim 1, characterized in that, It is a compound with the following structure:

9. A pharmaceutically acceptable salt, isomer, metabolite, prodrug, solvate, or hydrate of the NAPG protein hydrolysis targeting chimera of claim 1.

10. The use of a pharmaceutically acceptable salt, isomer, metabolite, prodrug, solvate, or hydrate of the NAPG protein hydrolysis targeting chimera of claim 1 in the preparation of a medicament for the treatment of multiple myeloma.

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