A PRMT1 protein degradation agent modified with adamantane as a hydrophobic group and application thereof

Abstract

The application discloses a PRMT1 protein degrading agent modified with adamantane as a hydrophobic group and an application thereof, and belongs to the field of medicinal chemistry.The PRMT1 protein degrading agent has a structure as shown in formula I.The compound of formula I can effectively degrade PRMT1 protein in tumor cells, has a wide application prospect in the preparation of drugs for treating or preventing cancer, and is particularly suitable for the preparation of drugs for targeted treatment of breast cancer.

Description

Technical Field

[0001] This invention belongs to the field of medicinal chemistry, specifically relating to a PRMT1 protein degrader modified with adamantane as a hydrophobic group, its preparation method, and its application. Background Technology

[0002] Histone post-translational modifications are an important component of epigenetics, mainly including methylation, acetylation, phosphorylation, ubiquitination, and glycosylation. Among these, methylation plays a crucial role in various cellular life activities such as cell signal transduction, DNA damage and repair, and transcriptional regulation.

[0003] Protein methyltransferases are key enzymes in histone methylation modification and can be divided into two categories: protein lysine methyltransferases and protein arginine methyltransferases. There are nine protein arginine methyltransferases (PRMTs) (PRMT1-9), which can be classified into three types based on the products they catalyze: Type I includes PRMT1 / 2 / 3 / 4 / 6 / 8, catalyzing asymmetric dimethylation and monomethylation of arginine; Type II includes PRMT5 / 9, catalyzing symmetric dimethylation and monomethylation of arginine; and Type III contains only PRMT7, catalyzing only monomethylation of arginine. Among these, PRMT1 is the most important type I PRMT in mammalian cells. Its involvement in arginine methylation affects various cellular processes, including cell signal transduction, DNA damage and repair, transcriptional regulation, and RNA metabolism, playing a crucial role in regulating the development and progression of various human tumors. Numerous studies have shown that PRMT1 is abnormally expressed in various cancers, including breast cancer, colorectal cancer, lung cancer, and pancreatic cancer. PRMT1 is overexpressed in most tumors, suggesting that PRMT1 may be a potential biomarker or therapeutic target in cancer treatment.

[0004] Currently reported PRMT1 inhibitors are all pan-inhibitors with poor selectivity for PRMT1. Among them, GSK3368715, the only one to enter clinical trials, had its clinical trials terminated due to thrombotic toxicity and low clinical efficacy.

[0005] Targeted protein degradation (TPD) technology has facilitated a paradigm shift in therapeutic strategies, providing innovative avenues for drug design. Compared to traditional small molecule drugs, TPD drugs have a broader scope of action, can target "undruggable" targets, better overcome drug resistance issues, and improve selectivity for target proteins. Hydrophobic tags (HyTs) are bifunctional TPD molecules composed of a lipophilic small molecule tag group and a small molecule ligand of the target protein linked by a specific chemical chain. Hydrophobic tag technology increases the hydrophobicity of the target protein surface, thereby inducing instability and misfolding of the target protein, leading to its degradation by the proteasome. Hydrophobic tag strategies have been widely used in the design of degraders for multiple targets, such as EZH2, estrogen receptor (ER), and androgen receptor (AR). Hydrophobic tag-based degraders for ER have been approved for the treatment of breast cancer. Protein degraders for PRMT1 hold promise for improving selectivity and efficacy for PRMT1, reducing adverse clinical reactions, and enhancing clinical treatment outcomes. Summary of the Invention

[0006] The purpose of this invention is to provide a hydrophobic tag bifunctional molecule with PRMT1 protein degradation activity, its preparation method and uses.

[0007] In a first aspect, the present invention provides a compound of formula I or a pharmaceutically acceptable salt thereof.

[0008]

[0009] Linker is selected from:

[0010] Z1 is connected to the benzene ring.

[0011] Z1 and Z2 are each independently NHC(O) or C(O)NH;

[0012] n is a positive integer from 1 to 7.

[0013] In another preferred embodiment, Z1 is -NH-CO-, where NH is connected to a benzene ring.

[0014] In another preferred embodiment, Z2 is -NH-CO-, wherein CO is connected to a -CH2-adamantane ring.

[0015] In another preferred embodiment, n is 1, 2, 3, 4, 5, or 6.

[0016] In another preferred embodiment, the Linker is n can be 2, 3, 4, 5, or 6.

[0017] In another preferred embodiment, the Linker is selected from:

[0018]

[0019] In another preferred embodiment, the compound is selected from the group consisting of:

[0020]

[0021]

[0022] Preferably, the molecular structural formula of the PRMT1 degrading agent is:

[0023]

[0024] This compound can effectively induce the degradation of PRMT1 protein in a dose- and time-dependent manner.

[0025] A second aspect of the present invention provides a pharmaceutical composition comprising:

[0026] The compounds described in the first aspect or their pharmaceutically acceptable salts; and

[0027] Pharmaceutically acceptable carrier.

[0028] A third aspect of the invention provides the use of the compound described in the first aspect or a pharmaceutically acceptable salt thereof or a pharmaceutical composition described in the second aspect for the preparation of a PRMT1 degrading agent; or

[0029] Used to prepare drugs for treating diseases associated with abnormal PRMT1 protein expression.

[0030] In another preferred embodiment, the compound described in the first aspect or a pharmaceutically acceptable salt thereof, or the pharmaceutical composition described in the second aspect, is used to prepare a medicament for treating diseases associated with overexpression of the PRMT1 protein.

[0031] In another preferred embodiment, the compound described in the first aspect or a pharmaceutically acceptable salt thereof, or the pharmaceutical composition described in the second aspect, is used to prepare a medicament for treating cancers associated with abnormal PRMT1 protein expression.

[0032] In another preferred embodiment, the compound described in the first aspect or a pharmaceutically acceptable salt thereof, or the pharmaceutical composition described in the second aspect, is used to prepare a medicament for treating cancers associated with overexpression of the PRMT1 protein.

[0033] In another preferred embodiment, the disease is cancer.

[0034] In another preferred embodiment, the cancer is selected from: breast cancer, gastric cancer, lung cancer, melanoma, and acute myeloid leukemia.

[0035] In another preferred embodiment, the cancer is breast cancer.

[0036] A third aspect of the present invention provides an intermediate for preparing the compound described in the first aspect or a pharmaceutically acceptable salt thereof, having the following structure:

[0037]

[0038] This invention provides a protein degrading agent modified with adamantane as a hydrophobic group. This degrading agent can be used to prepare PRMT1 degrading agents, and is particularly suitable for the development of drugs for the treatment of breast cancer.

[0039] It should be understood that, within the scope of this invention, the above-described technical features of this invention and the technical features specifically described below (such as in the embodiments) can be combined with each other to form new or preferred technical solutions. Each feature disclosed in the specification can be replaced by any alternative feature that provides the same, equivalent, or similar purpose. Due to space limitations, they will not be described in detail here. Attached Figure Description

[0040] Figure 1 The results of detecting the degradation activity of compounds CM1-101 to CM1-112 on PRMT1 protein in MCF-7 cells by Western blotting are shown. In the figure, 101-112 are the abbreviations of compound numbers CM1-101 to CM1-112.

[0041] Figure 2 To detect the degradation activity of compound CM1-112 on PRMT1 protein in MCF-7 cells by Western blotting, the results showed that as the concentration of compound CM1-112 increased, the immunoblot in the imaging became shallower, and the protein degradation rate was higher.

[0042] Figure 3 To detect the degradation activity of compound CM1-112 on PRMT1 protein in MDA-MB-231 cells by Western blotting, the results showed that as the concentration of compound CM1-112 increased, the immunoblot in the imaging became shallower, and the protein degradation rate was higher.

[0043] Figure 4 The results show the detection results of the inhibitory effect of compound CM1-112 on the proliferation of MCF-7 tumor cells.

[0044] Figure 5 To detect the inhibitory effects of the compound on the asymmetric dimethylation of H4R3, the direct substrate of PRMT1, and the asymmetric dimethylation of arginine in the total intracellular protein by immunoblotting.

[0045] Figure 6 The results of staining cell clonal colonies with crystal violet reagent are shown. Figure 7To investigate the inhibitory effect of compound CM1-112 on the proliferation of MDA-MB-231 tumor cells by measuring cell confluence, the results showed that compound CM1-112 effectively inhibited the proliferation of MDA-MB-231 cells. Detailed Implementation

[0046] The inventors of this application, through extensive and in-depth research, have developed a protein degrading agent modified with adamantane as a hydrophobic group. This degrading agent can be used to prepare PRMT1 degrading agents, and is particularly suitable for the development of drugs for treating breast cancer. Based on this, the present invention was completed.

[0047] the term

[0048] In this invention, the halogen is F, Cl, Br or I.

[0049] In this invention, unless otherwise specified, the terms used have the general meanings known to those skilled in the art.

[0050] In this invention, the terms "C1-C6" refer to having 1, 2, 3, 4, 5, or 6 carbon atoms, "C1-C8" refers to having 1, 2, 3, 4, 5, 6, 7, or 8 carbon atoms, and so on. "3-8" refers to having 3, 4, 5, 6, 7, or 8 ring atoms, and so on.

[0051] In this invention, the term "alkyl" refers to a saturated linear or branched hydrocarbon moiety. For example, the term "C1-C6 alkyl" refers to a straight-chain or branched alkyl group having 1 to 6 carbon atoms, and includes, without limitation, methyl, ethyl, n-propyl, isopropyl, n-butyl, isobutyl, sec-butyl, tert-butyl, pentyl, and hexyl; preferably ethyl, n-propyl, isopropyl, n-butyl, isobutyl, sec-butyl, and tert-butyl.

[0052] "Alkylene" refers to a straight-chain or branched saturated aliphatic group, i.e., a divalent hydrocarbon group, having a specified number of carbon atoms and being attached to at least two other groups. The two groups attached to the alkylene can be the same or different atoms on the alkylene. For example, a straight-chain alkylene can be -(CH2). n - A divalent group, where n is 1, 2, 3, 4, 5, or 6. Representative alkylene groups include, but are not limited to, methylene, ethylene, propylene, isopropylene, butylene, isobutylene, sec-butylene, pentylene, and hexylene. Alkylenes can be substituted or unsubstituted.

[0053] The pharmaceutically acceptable salts described in this invention can be salts formed by anion and a positively charged group on a compound of Formula I. Suitable anions include chloride, bromide, iodide, sulfate, nitrate, phosphate, citrate, methanesulfonate, trifluoroacetate, acetate, malate, toluenesulfonate, tartrate, fumarate, glutamate, glucuronate, lactate, glutarate, or maleate. Similarly, salts can be formed by cations and negatively charged groups on a compound of Formula I. Suitable cations include sodium, potassium, magnesium, calcium, and ammonium ions, such as tetramethylammonium ions.

[0054] In another preferred embodiment, "pharmaceutically acceptable salt" refers to a salt formed by a compound of Formula I with an acid selected from the group consisting of: hydrofluoric acid, hydrochloric acid, hydrobromic acid, phosphoric acid, acetic acid, oxalic acid, sulfuric acid, nitric acid, methanesulfonic acid, aminosulfonic acid, salicylic acid, trifluoromethanesulfonic acid, naphthalenesulfonic acid, maleic acid, citric acid, acetic acid, lactic acid, tartaric acid, succinic acid, oxalic acid, pyruvic acid, malic acid, glutamic acid, p-toluenesulfonic acid, naphthalenesulfonic acid, ethanesulfonic acid, naphthalenedisulfonic acid, malonic acid, fumaric acid, propionic acid, oxalic acid, trifluoroacetic acid, stearic acid, pyric acid, hydroxymaleic acid, phenylacetic acid, benzoic acid, glutamic acid, ascorbic acid, p-aminobenzenesulfonic acid, 2-acetoxybenzoic acid, and hydroxyethanesulfonic acid; or a sodium, potassium, calcium, aluminum, or ammonium salt formed by a compound of Formula I with an inorganic base; or a methylamine, ethylamine, or ethanolamine salt formed by a compound of general Formula I with an organic base.

[0055] Pharmaceutical Composition

[0056] The present invention also provides a pharmaceutical composition comprising an active ingredient within a safe and effective range, and a pharmaceutically acceptable carrier.

[0057] The "active ingredient" mentioned in this invention refers to the compound of formula I described in this invention.

[0058] The "active ingredient" and pharmaceutical composition described in this invention can be used as a PRMT1 degrading agent.

[0059] The "active ingredient" and pharmaceutical composition described in this invention are used to prepare a medicament for treating diseases associated with abnormal PRMT1 protein expression. The diseases associated with abnormal PRMT1 protein expression are cancers related to abnormal PRMT1 protein expression.

[0060] The "active ingredient" and pharmaceutical composition described in this invention are used to prepare a medicament for treating diseases associated with PRMT1 protein overexpression. The diseases associated with abnormal PRMT1 protein expression are cancers related to PRMT1 protein overexpression.

[0061] The cancers mentioned are selected from: breast cancer, gastric cancer, lung cancer, melanoma, colon cancer, lymphoma (such as diffuse large B-cell lymphoma, follicular lymphoma), leukemia (such as acute myeloid leukemia), multiple myeloma, mesothelioma, malignant rhabdoid tumor, hepatocellular carcinoma, prostate cancer, bile duct and gallbladder cancer, bladder cancer; brain tumors, including neuroblastoma, schwannoma, glioma, glioblastoma and astrocytoma; cervical cancer, endometrial cancer, esophageal cancer, head and neck cancer, nasopharyngeal carcinoma, ovarian cancer, pancreatic cancer, renal cell carcinoma, rectal cancer, thyroid cancer, parathyroid tumors, uterine tumors and soft tissue sarcomas, cardiovascular diseases, neurodegenerative diseases, malaria, AIDS, gout, diabetes, renal failure, and chronic lung diseases.

[0062] "Safe and effective dose" refers to an amount of active ingredient sufficient to significantly improve the condition without causing serious side effects. Typically, the pharmaceutical composition contains 1-2000 mg of active ingredient per dose, more preferably 10-200 mg of active ingredient per dose. Preferably, "one dose" refers to one tablet.

[0063] "Pharmaceutically acceptable carriers" refer to one or more compatible solid or liquid fillers or gel substances that are suitable for human use and must have sufficient purity and sufficiently low toxicity. "Compatibility" here refers to the ability of the components in the composition to interact with and incorporate the active ingredient of the invention without significantly reducing the efficacy of the active ingredient. Examples of pharmaceutically acceptable carriers include cellulose and its derivatives (such as sodium carboxymethyl cellulose, sodium ethyl cellulose, cellulose acetate, etc.), gelatin, talc, solid lubricants (such as stearic acid, magnesium stearate), calcium sulfate, vegetable oils (such as soybean oil, sesame oil, peanut oil, olive oil, etc.), polyols (such as propylene glycol, glycerin, mannitol, sorbitol, etc.), emulsifiers (such as... Wetting agents (such as sodium dodecyl sulfate), colorants, flavoring agents, stabilizers, antioxidants, preservatives, pyrogen-free water, etc.

[0064] There are no particular limitations on the administration of the active ingredients or pharmaceutical compositions of the present invention. Representative administration methods include (but are not limited to): oral, intratumoral, rectal, parenteral (intravenous, intramuscular or subcutaneous), etc.

[0065] Solid dosage forms for oral administration include capsules, tablets, pills, powders, and granules.

[0066] Liquid dosage forms for oral administration include pharmaceutically acceptable emulsions, solutions, suspensions, syrups, or tinctures. In addition to the active ingredient, liquid dosage forms may contain inert diluents conventionally used in the art, such as water or other solvents, solubilizers and emulsifiers, e.g., ethanol, isopropanol, ethyl carbonate, ethyl acetate, propylene glycol, 1,3-butanediol, dimethylformamide, and oils, particularly cottonseed oil, peanut oil, corn germ oil, olive oil, castor oil, and sesame oil, or mixtures thereof. Besides these inert diluents, the composition may also contain adjuvants such as wetting agents, emulsifiers and suspending agents, sweeteners, flavoring agents, and fragrances.

[0067] In addition to the active ingredient, the suspension may contain suspending agents, such as ethoxylated isooctadecyl alcohol, polyoxyethylene sorbitol and dehydrated sorbitol esters, microcrystalline cellulose, aluminum methoxide and agar, or mixtures of these substances.

[0068] Compositions for parenteral injection may comprise physiologically acceptable sterile aqueous or anhydrous solutions, dispersions, suspensions, or emulsions, and sterile powders for reconstitution into sterile injectable solutions or dispersions. Suitable aqueous and non-aqueous carriers, diluents, solvents, or excipients include water, ethanol, polyols, and suitable mixtures thereof.

[0069] The compounds of this invention can be administered alone or in combination with other therapeutic agents (such as antitumor drugs).

[0070] When using the pharmaceutical composition, a safe and effective amount of the compound of the present invention is applied to the mammal (such as a human) requiring treatment. The dosage administered is the pharmaceutically considered effective dose. For a person weighing 60 kg, the daily dose is typically 1–2000 mg, preferably 20–500 mg. Of course, the specific dosage should also take into account factors such as the route of administration and the patient's health condition, which are all within the scope of the skills of a skilled physician.

[0071] The present invention is further illustrated below with reference to specific embodiments. It should be understood that these embodiments are for illustrative purposes only and are not intended to limit the scope of the invention. Experimental methods in the following embodiments, unless otherwise specified, are generally performed under conventional conditions (such as those described in Sambrook et al., Molecular Cloning: A Laboratory Manual (New York: Cold Spring Harbor Laboratory Press, 1989)) or as recommended by the manufacturer. Unless otherwise specified, the experimental materials used in the following embodiments were purchased from a regular biochemical reagent store. Nuclear magnetic resonance spectroscopy was performed using a Bruker 400 NMR spectrometer, and mass spectrometry was performed using an Agilent liquid chromatography-time-of-flight mass spectrometer. Unless otherwise stated, percentages and parts are weight percentages and parts by weight.

[0072] Unless otherwise defined, all technical and scientific terms used herein have the same meaning as are familiar to those skilled in the art. Furthermore, any methods and materials similar to or equivalent to those described herein may be applied to the methods of this invention. The preferred embodiments and materials described herein are for illustrative purposes only.

[0073] Example 1

[0074] Preparation of compound CM1-112

[0075]

[0076]

[0077] Preparation of compound Ma: Under nitrogen protection, p-nitrobenzaldehyde (3.00 g, 19.85 mmol) was dissolved in tetrahydrofuran (50 mL). Sodium hydride (0.52 g, 21.84 mmol) and diethylphosphonoacetate (4.59 g, 21.84 mmol) were added separately in an ice bath. The reaction was carried out at room temperature for 24 hours. After the reaction was completed by TLC monitoring, the solvent was evaporated, and the mixture was extracted three times with ethyl acetate (20 mL). The organic layer was collected and washed with saturated brine. The mixture was dried with anhydrous sodium sulfate, and the solvent was removed by concentration under reduced pressure to obtain the crude product, which was the desired compound Ma (4.985 g, yield 121.3%), a yellow solid. It could be used directly in the next reaction without further purification.

[0078] The results of the detection of compound Ma are as follows: 1 H NMR (400MHz, DMSO-d6) δ8.25(d,J=8.8Hz,2H),8.02(d,J=8.8Hz,2H),7.78(d,J=16.1Hz,1H),6.87(d,J=16.0Hz,1H),3.76(s,3H).MS(ESI):m / z[M+H] + 208.1.

[0079]

[0080] Preparation of compound Mb: Sodium hydride (1.39 g, 57.92 mmol), compound Ma (3.99 g, 19.31 mmol), and p-toluenesulfonylmethylisocyanate (4.55 g, 23.17 mmol) were dissolved in tetrahydrofuran / N,N-dimethylformamide DMF (100 mL, v:v = 1:1) under ice bath conditions. The reaction was carried out at room temperature for 2 hours. After the reaction was completed by TLC monitoring, the mixture was extracted three times with ethyl acetate (20 mL). The organic layer was collected and washed with saturated brine. The mixture was dried over anhydrous sodium sulfate and concentrated under reduced pressure to remove the solvent, yielding the crude product, compound Mb (4.00 g, yield 85.2%), a reddish-brown solid, which could be used directly in the next reaction without further purification.

[0081] The results of the detection of compound Mb are as follows: 1 H NMR (400MHz, CDCl3) δ8.19–8.11(m,2H),7.62–7.53(m,2H),7.48(dd,J=3.1,2.2Hz,1H),6.85(t,J=2.4Hz,1H),3.69(d,J=7.9Hz,3H).MS(ESI):m / z[M+H] + 247.1.

[0082]

[0083] Preparation of compound Mc: Compound Mb (4.00 g, 16.25 mmol), Boc anhydride (di-tert-butyl dicarbonate, 3.90 g, 17.87 mmol), triethylamine (2.30 g, 22.74 mmol), and 4-dimethylaminopyridine (DMAP) (198 mg, 0.16 mmol) were dissolved in dichloromethane (DCM) (80 mL) at room temperature and reacted for 5 hours. After the reaction was completed by TLC monitoring, the solvent was evaporated, and the mixture was extracted three times with dichloromethane (20 mL). The organic layer was collected and washed with saturated brine, dried over anhydrous sodium sulfate, and concentrated under reduced pressure to remove the solvent, yielding the crude product. The crude product was then obtained by silica gel column chromatography as a pale yellow solid (4.43 g, yield 64.6%).

[0084] The results of the detection of compound Mc are as follows: 1 H NMR (400MHz, CDCl3) δ8.26–8.19(m,2H),7.94(d,J=2.4Hz,1H),7.66–7.59(m,2H),7.33(d,J=2.4Hz,1H),3.78(s,3H),1.64(s,9H).MS(ESI):m / z[M+H] + 347.1.

[0085]

[0086] Preparation of compound Md: Under nitrogen protection, compound Mc (4.43 g, 28.92 mmol) was dissolved in anhydrous tetrahydrofuran (60 mL). Diisobutylaluminum hydride (DIBALH) (8.26 g, 72.29 mmol) was slowly added dropwise over 30 minutes at -78 °C, and the reaction was carried out for 6 hours at -78 °C. After the reaction was completed by TLC monitoring, it was quenched with methanol (5 mL), and saturated potassium sodium tartrate solution (100 mL) was added to the reaction solution. The mixture was stirred for 2 hours. The mixture was extracted three times with ethyl acetate (20 mL), and the organic layer was collected and washed with saturated brine. The solution was dried over anhydrous sodium sulfate, and the solvent was removed by concentration under reduced pressure to obtain the crude product. The crude product was then obtained by silica gel column chromatography as a yellow solid (1.83 g, yield 45.15%).

[0087] The results of the detection of compound Md are as follows: 1 H NMR (400MHz, CDCl3) δ8.18(d,J=9.0Hz,2H),7.69(d,J=9.0Hz,2H),7.42(s,1H),7.30(s,1H),4.57(s,2H),1.56(s,9H).MS(ESI):m / z[M+H] + 319.1.

[0088]

[0089] Preparation of compound Me: Compound Md (1.83 g, 5.81 mmol) was dissolved in dichloromethane (60 mL) at room temperature, and Dys-Martin oxidant (2.71 g, 6.39 mmol) was added. The mixture was stirred at room temperature for 2 hours. After the reaction was completed by TLC monitoring, the reaction solution was filtered through a Buchner funnel lined with diatomaceous earth. The filter cake was washed three times with dichloromethane, and anhydrous sodium sulfate was added to the filtrate for drying. The solvent was removed by concentration under reduced pressure to obtain the crude product. The desired compound Me (1.68 g, yield 92.39%) was obtained by silica gel column chromatography as a yellow solid.

[0090] The detection results for compound Me are as follows: 1 H NMR (400MHz, CDCl3) δ9.93 (s, 1H), 8.25 (d, J = 9.0Hz, 2H), 7.98 (d, J = 2.4Hz, 1H) ,7.72(d,J=9.0Hz,2H),7.45(d,J=2.4Hz,1H),1.66(s,9H).MS(ESI):m / z[M+H] + 317.1.

[0091]

[0092] Preparation of compound Mf: Compound Me (1.68 g, 5.31 mmol), sodium triacetoxyborohydride (1.69 g, 7.97 mmol), and tert-butyl 2-(methylamino)ethylcarbamate (1.11 g, 6.37 mmol) were dissolved in dichloromethane (50 mL) at room temperature and reacted for 24 hours. After the reaction was completed by TLC monitoring, the reaction solution was filtered through a Buchner funnel lined with diatomaceous earth. The filter cake was washed three times with dichloromethane, and anhydrous sodium sulfate was added to the filtrate for drying. The solvent was removed by concentration under reduced pressure to obtain the crude product. The crude product was obtained by silica gel column chromatography as a pale yellow solid, yielding compound Mf (1.88 g, 74.59% yield).

[0093] The results of the detection of compound Mf are as follows: 1 H NMR (400MHz, CDCl3) δ8.27–8.21(m,2H),7.83(d,J=8.8Hz,2H),7.46(d,J=2.4Hz,1H),7.21(d,J=2.5Hz,1H),3.4 0(s,2H),3.21(d,J=5.1Hz,2H),2.49(t,J=5.9Hz,2H),2.19(s,3H),1.63(s,9H),1.43(s,9H).MS(ESI):m / z[M+H] + 475.2.

[0094]

[0095] Preparation of compound Mg: Compound Mf (1.88 g, 3.96 mmol) was dissolved in methanol (50 mL) at room temperature, and 10% palladium on carbon (50 mg) was added. Hydrogen gas was introduced, and the reaction was carried out at room temperature for 10 hours. After the reaction was completed by TLC monitoring, the reaction solution was filtered through a Buchner funnel lined with diatomaceous earth. The filter cake was washed three times with methanol, and anhydrous sodium sulfate was added to the filtrate for drying. The solvent was removed by concentration under reduced pressure to obtain the crude product. The desired compound Mg (1.30 g, yield 76.86%) was obtained by silica gel column chromatography as a yellow solid.

[0096] The results of the detection of compound Mg are as follows: 1H NMR (400MHz, DMSO-d6) δ7.26(d,J=8.4Hz,2H),7.18–7.12(m,2H),6.56(d,J=8.5Hz,3H),5.06(s,2H),3.27(s ,2H),3.03(q,J=5.9Hz,2H),2.37(t,J=7.1Hz,2H),2.14(s,3H),1.56(s,9H),1.36(s,9H).MS(ESI):m / z[M+H] + 445.2.

[0097]

[0098] Preparation of compound M-12: 1-Adamantaneacetic acid (0.26 g, 1.37 mmol) was dissolved in DMF (5 mL) under ice bath conditions. N,N-diisopropylethylamine (DIPEA) (0.26 g, 2.05 mmol), 2-(7-azabenzotriazole)-N,N,N',N'-tetramethylurea hexafluorophosphate (HATU) (0.78 g, 2.05 mmol), and tert-butyl 15-amino-4,7,10,13-tetraoxapentadecanoate (0.5 g, 1.37 mmol) were added, and the mixture was reacted at room temperature for 1 hour. After the reaction was completed by TLC monitoring, the mixture was extracted three times with ethyl acetate (5 mL). The organic phase was collected, washed with saturated brine, dried over anhydrous sodium sulfate, concentrated under reduced pressure to remove the solvent, and then dissolved in dichloromethane (6 mL). Trifluoroacetic acid (3 mL) was added, and the mixture was reacted at room temperature for 5 hours. After the reaction was completed, the solvent was removed by concentration under reduced pressure, and the desired compound M-12 (0.7 g, yield 94.5%) was obtained by reversed-phase chromatography as a colorless and transparent oil.

[0099] The results of the detection of compound M-12 are as follows: 1 H NMR (400MHz, DMSO-d6) δ12.18(s,1H),7.74(t,J=5.7Hz,1H),3.59(t,J=6.4Hz,2H),3.50(d,J=4.6Hz,14H) ,3.37(s,3H),3.17(q,J=5.7Hz,2H),2.44(t,J=6.3Hz,2H),1.90(s,3H),1.82(s,2H),1.69–1.49(m,12H).

[0100] Preparation of compound CM1-112: Compound M12 (250 mg, 514.81 μmol), HATU (0.294 g, 772.21 μmol), DIPEA (0.100 g, 772.21 μmol), and compound Mg (0.228 g, 514.81 μmol) were dissolved in DMF (3 mL) under ice bath conditions and reacted at room temperature for 1 hour. After the reaction was monitored by TLC, the mixture was extracted three times with ethyl acetate (5 mL), the organic layer was collected and washed with saturated brine, dried over anhydrous sodium sulfate, concentrated under reduced pressure to remove the solvent, and the collected compound was dissolved in dichloromethane (4 mL). Trifluoroacetic acid (2 mL) was added, and the mixture was reacted at room temperature for 30 minutes. After the reaction was completed, the reaction solution was concentrated under reduced pressure to remove the solvent, and the desired compound CM1-112 (45 mg, yield 12.29%) was obtained by separation by reversed-phase chromatography as a brown oily liquid.

[0101] The results of the detection of compound CM1-112 are as follows: 1 H NMR (400MHz, Methanol-d4) δ7.63(d,J=8.5Hz,2H),7.33(d,J=8.6Hz,2H),7.14(s,1H),6.91(s,1H),4.47(s,2H),3.83(t,J=6.0Hz,2H),3.67–3. 56(m,16H),3.51(t,J=5.5Hz,2H),3.19(s,4H),2.65(s,3H),2.63(d,J=6 .0Hz,2H),1.94(d,J=8.1Hz,5H),1.79–1.61(m,12H).MS(ESI):m / z[M+H] + 712.4.

[0102] Following the above preparation method, the following compounds were prepared:

[0103]

[0104]

[0105]

[0106]

[0107] Example 2

[0108] In this embodiment, the degradation ability of the compounds CM1-101 to CM1-112 synthesized in Example 1 for PRMT1 protein was evaluated in the MCF-7 cell line by Western blotting.

[0109] 1. Experimental Methods

[0110] Cell treatment: Human breast cancer cells (MCF-7) were processed at 3 × 10⁻⁶ cells / year. 4 Human breast cancer cells were seeded at a density of cells / well in 12-well plates and incubated overnight at 37°C in a 5% CO2 cell culture incubator. The cells were treated with DMEM complete medium containing 5 μM of the compound for 72 hours, and a DMSO group was set up as a negative control.

[0111] Extraction of total cellular protein: The culture medium was removed from the culture plate, and the cells were washed with cold PBS. RIPA lysis buffer containing a cocktail protease inhibitor (1:100) was then added, and the cells were lysed at 4°C. The cells were centrifuged at 12000 rpm for 10 min at 4°C, and the supernatant was transferred to a new 1.5 mL centrifuge tube. The protein concentration was determined using a BCA protein assay kit. The remaining sample was added to an equal volume of 2×SDS-PAGE loading buffer, boiled at 96°C for 7 min, cooled to room temperature, and then loaded onto an SDS-PAGE gel.

[0112] Western blot: Samples were separated by electrophoresis using an 8-16% SDS-PAGE precast gel at 120V for 60 min. Wet transfer was performed at 4℃, 260mA for 90 min. After the transfer, the membrane was transferred to blocking buffer (TBST containing 5% nonfat milk and Tris buffer containing Tween 20) and blocked at room temperature for 1 h. The target molecular weight band was cut off, and the primary antibody used for the target protein was diluted 1:5000 using TBST containing 5% BSA. The mixture was then incubated overnight at 4℃. After labeling the primary antibody, the protein bands were collected and frozen. The protein bands were washed three times with 1×TBST for 10 min each time. The corresponding horseradish peroxidase-labeled (HRP) secondary antibody was added at a dilution of 1:5000 and incubated at room temperature for 1 h. After labeling the secondary antibody dilution buffer, collect and freeze it. Wash the protein bands three times with TBST (Tris-buffered saline containing Tween 20), 10 min each time. Add ECL chemiluminescent substrate (prepared fresh before use), expose and record the results.

[0113] 2. Experimental Results

[0114] Experimental results are as follows Figure 1 As shown, compounds, especially CM1-112, can effectively degrade PRMT1 protein in human breast cancer cells.

[0115] Example 3

[0116] Compound CM1-112 was selected to further investigate its ability to degrade PRMT1 protein in human breast cancer cells (MCF-7, MDA-MB-231).

[0117] 1. Experimental Methods

[0118] Cell treatment: Human breast cancer cells (MCF-7, MDA-MB-231) were processed at 3 × 10⁻⁶ cells / year. 4 Human breast cancer cells were seeded at a density of cells / well in 12-well plates and incubated overnight at 37°C in a 5% CO2 cell culture incubator. Human breast cancer cells were treated for 72 hours with DMEM complete medium containing compounds of varying concentrations, and a DMSO group was set up as a negative control.

[0119] The method described in Example 1 was used for detection by Western blotting.

[0120] 2. Experimental Results

[0121] Experimental results are as follows Figure 2 , Figure 3 As shown, Figure 2 The expression levels of PRMT1 protein in MCF-7 cells after treatment with different concentrations of compound CM1-112 for 72 hours were measured. Figure 3 The expression levels of PRMT1 protein in MDA-MB-231 cells were measured after 72 hours of treatment with different concentrations of compound CM1-112. The results indicate that the compound of this invention can effectively degrade PRMT1 protein in human breast cancer cells.

[0122] like Figure 4 As shown, the DCs measured in MCF-7 cells by compound CM1-112 50 =0.83μM, indicating that compound CM1-112 can effectively inhibit the proliferation of MCF-7 cells.

[0123] Example 4

[0124] Compound CM1-112 was selected to further investigate its inhibitory effect on asymmetric dimethylation mediated by endogenous PRMT1 protein in human breast cancer cells (MCF-7, MDA-MB-231).

[0125] 1. Experimental Methods

[0126] Cell treatment: Human breast cancer cells (MCF-7, MDA-MB-231) were processed at 5 × 10⁻⁶ cells / year. 4Human breast cancer cells were seeded at a density of cells / well in 12-well plates and incubated overnight at 37°C in a 5% CO2 cell culture incubator. Human breast cancer cells were treated with DMEM complete medium containing a concentration gradient of the compound for 48 hours, with a DMSO group serving as a negative control. Western blot analysis was used to detect the inhibitory effects of the compound on asymmetric dimethylation of Histone H4 protein (the direct substrate of PRMT1) and on asymmetric dimethylation of intracellular proteins.

[0127] 2. Experimental Results

[0128] Experimental results are as follows Figure 5 As shown, Figure 5 The levels of intracellular protein asymmetric dimethylation and the level of PRMT1 direct substrate H4R3 asymmetric dimethylation were measured after treating two types of breast cancer cells with different concentrations of compound CM1-112 for 48 hours. The results showed that compound CM1-112 could effectively inhibit the catalytic function of PRMT1 substrate asymmetric dimethylation in a concentration gradient-dependent manner.

[0129] Example 5

[0130] Compound CM1-112 was selected to investigate its inhibitory effect on the clonal colonies of human breast cancer cells (MDA-MB-231).

[0131] 1. Experimental Methods

[0132] Cell treatment: Human breast cancer cells (MDA-MB-231) were seeded at a density of 800 cells / well in 6-well plates and incubated overnight at 37°C in a 5% CO2 cell culture incubator. Cells were treated with DMEM complete medium containing 10 μM of compound for 10, 8, 6, 4, and 2 days, with a DMSO group as a negative control. Cell colonies were stained with crystal violet and the entire colony was photographed for recording.

[0133] 2. Experimental Results

[0134] Experimental results are as follows Figure 6 As shown, the number of cell colonies formed in human breast cancer cells (MDA-MB-231 cell line) after treatment with 10 μM compound CM1-112 for different number of days is illustrated. The experimental results indicate that 10 μM compound CM1-112 can effectively inhibit the formation of cell colonies in human breast cancer cells and effectively inhibit their proliferation in a time-dependent manner.

[0135] Example 6

[0136] Compound CM1-112 was selected to investigate its ability to inhibit the proliferation of human breast cancer cells (MDA-MB-231).

[0137] Cell treatment: Human breast cancer cells (MDA-MB-231) were processed at 5 × 10⁻⁶ cells / year. 3 Cells were seeded at a density of 1 cell per well in 6-well plates. After attachment, the cells were treated with DMEM complete medium containing 10 μM compound for 192 hours, and a DMSO group was set up as a negative control. The cells were incubated in an Incucyte live cell imaging system (the imaging system was placed in a 37°C, 5% CO2 cell culture incubator). The medium was changed every 48 hours. The live cell imaging system was set up to take pictures of four fixed fields of view of the 6-well plate every 12 hours.

[0138] Experimental results are as follows Figure 7 The figure shows the changes in cell confluence of human breast cancer cells (MDA-MB-231 cell line) treated with 10 μM compound CM1-112 over 192 hours. The confluence data were analyzed using the Incucyte live-cell imaging system's basic analysis program. The experimental results indicate that 10 μM compound CM1-112 can effectively slow the proliferation rate of human breast cancer cells in a time-dependent manner.

[0139] All documents mentioned in this invention are incorporated herein by reference as if each document were individually incorporated by reference. Furthermore, it should be understood that after reading the foregoing teachings of this invention, those skilled in the art can make various alterations or modifications to this invention, and these equivalent forms also fall within the scope defined by the appended claims.

Claims

1. A compound of formula I or a pharmaceutically acceptable salt thereof, Linker is selected from: Z1 is connected to the benzene ring. Z1 and Z2 are each independently NHC(O) or C(O)NH; n is a positive integer from 1 to 7.

2. The compound according to claim 1, characterized in that, Z1 is -NH-CO-, where NH is connected to a benzene ring.

3. The compound according to claim 1, characterized in that, Z2 is -NH-CO-, where CO is connected to a -CH2-adamantane ring.

4. The compound according to claim 1, characterized in that, n can be 1, 2, 3, 4, 5, or 6.

5. The compound according to claim 1, characterized in that, Linker is n can be 2, 3, 4, 5, or 6.

6. The compound according to claim 1, characterized in that, Linker is selected from:

7. The compound according to claim 1, characterized in that, The compounds are selected from the group consisting of: 。 8. A pharmaceutical composition, characterized in that, The pharmaceutical composition comprises: The compound of claim 1 or a pharmaceutically acceptable salt thereof; and Pharmaceutically acceptable carrier.

9. Use of the compound of claim 1 or a pharmaceutically acceptable salt thereof or the pharmaceutical composition of claim 8, characterized in that, Used to prepare PRMT1 degrading agents; or This is for the preparation of drugs for treating diseases associated with abnormal PRMT1 protein expression, wherein the abnormal PRMT1 protein expression is PRMT1 protein overexpression, and the disease is cancer, wherein the cancer is selected from: breast cancer, gastric cancer, lung cancer, melanoma, and acute myeloid leukemia.

10. An intermediate of the compound as described in claim 1 or a pharmaceutically acceptable salt thereof, having the following structure:

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