Targeted protein degradation agent taking artemisinin analogue as hydrophobic label as well as preparation method and application of targeted protein degradation agent

Abstract

The invention discloses a targeted protein degradation agent taking an artemisinin analogue as a hydrophobic label as well as a preparation method and application of the targeted protein degradation agent. The structural formula of the targeted protein degradation agent is shown as a formula (II), a formula (III) or a formula (IV). The targeted protein degradation agent disclosed by the invention has the activity of inhibiting human breast cancer cells and also has good degradation activity. The targeted protein degradation agent can also be used for preparing drugs for targeted degradation of estrogen receptors or anti-breast cancer drugs.

Description

Technical Field

[0001] The present invention relates to the field of medical technology, and in particular to a targeted protein degradation agent using an artemisinin analog as a hydrophobic tag, and a preparation method and application thereof. Background Art

[0002] In recent years, breast cancer has become one of the main factors threatening human life. At present, the treatment of breast cancer is mainly traditional chemotherapy, but there are still shortcomings, such as low bioavailability, large drug toxicity and side effects, and certain damage to the immune system. Therefore, choosing a more effective treatment method is crucial for the treatment of breast cancer patients. Targeted protein degradation strategy, as a revolutionary drug development strategy, has great development potential in overcoming drug resistance. Taking estrogen receptor α (ERα) as an example, it plays an important role in the occurrence and development of breast cancer and is an important molecular target for breast cancer treatment. In the clinical treatment of breast cancer, how to overcome drug resistance has also become a key criterion for the therapeutic effect of this target drug. For the strategy of targeted degradation of ERα targets, the ubiquitin-proteasome system is the main degradation pathway. The most representative technology is the protein degradation targeting chimera (PROTAC), which directly binds to the E3 ubiquitin ligase through the ternary complex E3 ligand to degrade the protein by ubiquitin tag. Representative ERα degraders such as ARV-471 have entered Phase III clinical studies.

[0003] For PROTAC technology, its structural characteristics lead to its large molecular weight and lack of tissue specificity, resulting in poor oral absorption and membrane permeability, which still faces huge challenges in terms of oral drugability and toxic side effects. Therefore, its poor pharmacokinetic properties are the main obstacle to its drugability. Therefore, exploring new degradation strategies, comparing the advantages and disadvantages of various degradation strategies, and studying the mechanism of action and biological activity of different types of degraders are of great significance to the therapeutic effect of targeted drugs. Summary of the invention

[0004] In view of the defects in the prior art, the present invention proposes a targeted protein degradation agent with artemisinin analogs as hydrophobic agent tags, and its preparation method and application. In the bifunctional molecule, the present invention uses artemisinin analogs as hydrophobic groups, and selects benzothiophene compounds (targeting ERα), methylpyridopyrimidine compounds (targeting CDK4 / 6), and suberoylanilide hydroxamic acid compounds (targeting HDAC6) as the basic skeleton in the target protein ligand structure, and then adds hydrophobic tags of artemisinin analogs to the compounds through the side chain introduction method, and designs and synthesizes ERα degradation molecules, CDK4 / 6 degradation molecules, and HDAC6 degradation molecules. By selecting linkers of different lengths, the purpose of not affecting the binding of two proteins in space and maintaining their binding is achieved. By selecting different target protein ligands, the influence of artemisinin analogs as hydrophobic tags in different proteins is verified.

[0005] The present invention provides a hydrophobic tag based on artemisinin compounds, the structural formula of which is shown in formula (I):

[0006]

[0007] The present invention provides a targeted protein degradation agent, comprising the hydrophobic tag, a target protein ligand and a linker; the hydrophobic tag and the target protein ligand are connected via the linker; the target protein ligand is any one of a benzothiophene compound, a methylpyridopyrimidine compound, and a suberoylanilide hydroxamic acid compound.

[0008] Furthermore, the linker is any one of an alkyl chain and an alkoxy group.

[0009] Furthermore, the targeted protein degradation agent has a structure as shown in the following formula (II), formula (III) or formula (IV):

[0010] Where n is any integer from 1 to 10;

[0011] Where n is any integer from 1 to 10;

[0012] Wherein n is any integer from 1 to 10.

[0013] Furthermore, the compound of formula (II) includes but is not limited to the following compounds:

[0014] (3R,6R,9S,11R,11aR)-N-(2-(4-(6-hydroxy-2-(4-hydroxyphenyl)benzo[b]thiophene-3-carbonyl)phenoxy)ethyl)-3,6,9-trimethyloctahydro-3H,11H-3,11-epoxy[1,2]dioxazo[3,4-d]isobenzofuran-9-carboxamide (RA2), (3R,6R,9S,11R,11aR)-N-(3-(4-(6-hydroxy-2-(4-hydroxyphenyl)benzo[b]thiophene-3-carbonyl)phenoxy)propyl)-3,6,9-trimethyloctahydro-3H,11H-3,11-epoxy[1,2]dioxazo[3,4-d]isobenzofuran-9-carboxamide (RA3), (3R,6R,9S,11R,11aR)-N-(4-(4-(6-hydroxy-2-(4-hydroxyphenyl)benzo[b]thiophene-3-carbonyl)phenoxy)butyl)-3,6,9-trimethyloctahydro-3H,11H-3,11-epoxy[1,2]dioxazo[3,4-d]isobenzofuran-9-carboxamide (RA4), (3R,6R,9S,11R,11aR)-N-(6-(4-(6-hydroxy-2-(4-hydroxyphenyl)benzo[b]thiophene-3-carbonyl)phenoxy)hexyl)-3,6,9-trimethyloctahydro-3H,11H-3,11-epoxy[1,2]dioxazo[3,4-d]isobenzofuran-9-carboxamide (RA6).

[0015] Furthermore, the compound of formula (III) includes but is not limited to the following compounds:

[0016] (3R,6R,9S,11R,11aR)-N-(3-((6-acetyl-8-cyclopentyl-5-methyl-7-oxo-7,8-dihydropyrido[2,3-d]pyrimidin-2-yl)amino)propyl)-3,6,9-trimethyloctahydro-3H,11H-3,11-epoxy[1,2]dioxepane[3,4-d]isobenzofuran-9-carboxamide (PA).

[0017] Furthermore, the compound of formula (IV) includes but is not limited to the following compounds:

[0018] N 1 -Hydroxy-N 8 -(4-((3R,6R,9S,11R,11aR)-3,6,9-trimethyloctahydro-3H,11H-3,11-epoxy[1,2]dioxepano[3,4-d]isobenzofuran-9-carboxamido)butanamido)phenyl)octanediamide (SA).

[0019] The present invention also provides a method for synthesizing the targeted protein degradation agent, comprising the following steps:

[0020] 1) Synthesis of artemisinin analogs;

[0021] 2) Synthesis of a target protein ligand with a linker, and then subjecting the artemisinin analogue described in step 1) to an amidation reaction with the target protein ligand to obtain a targeted protein degrader.

[0022] Furthermore, the step 1) of synthesizing the artemisinin analogue specifically comprises the following steps:

[0023] S1. Compound 1 was dissolved in tetrahydrofuran, cooled to 0°C, and then boron trifluoride ether was added dropwise to react for 24 hours. After the reaction was completed, the reaction was quenched with saturated sodium bicarbonate, extracted, dried, distilled, and purified to obtain compound 2;

[0024] S2. Compound 2 was dissolved in carbon tetrachloride, bromine was slowly added dropwise at 0°C, and the mixture was stirred. After the reaction was completed, ether was added to dilute the mixture, sodium thiosulfate was added to quench the mixture, the mixture was dried, and the mixture was distilled under reduced pressure to obtain compound 3;

[0025] S3. Compound 3 was dissolved in dichloromethane, triethylamine was added, and the reaction was stirred until the reaction was completed, and the excess triethylamine was neutralized, washed, dried, filtered, concentrated, and purified to obtain compound 4;

[0026] S4. Dissolve compound 4 in 1,4-dioxane solution, add an aqueous solution containing aminosulfonic acid, stir overnight at 0°C, add NaC1O2 and water, stir, add Na2SO3, dilute, extract, wash, dry and evaporate to obtain compound 5, which is an artemisinin analogue.

[0027] Furthermore, the step 2) of synthesizing the target protein ligand with a linker specifically comprises the following steps:

[0028] 1) When the target protein ligand is a benzothiophene compound, the method for synthesizing the targeted protein degradation agent comprises the following steps:

[0029] S1.4-acetylbenzoic acid is stirred with oxalyl chloride at room temperature to obtain an acyl chloride intermediate, the intermediate is reacted with compound 7 and anhydrous aluminum chloride to obtain compound 8, compound 8 is dissolved in an oxyethanol / water solution, sodium acetate is added and stirred at 100°C to obtain compound 9, compound 9 is reacted with (n-bromo-n-alkyl)carbamic acid tert-butyl ester to obtain compound 10, boron tribromide is used to demethylate and demethylate the Boc group to obtain compound 11; wherein n is 1 to 10; and compound 11 is a benzothiophene compound with a linker;

[0030] S2. Using HATU as a catalyst and DIPEA as a base, the compound 11 is subjected to an amidation reaction with the compound 5 in a DMF solvent;

[0031] 2) When the target protein ligand is a methylpyrido-pyrimidine compound, the method for synthesizing the targeted protein degradation agent comprises the following steps:

[0032] S1. Compound 13 and compound 14 were mixed and reacted under argon, DIPEA and DMSO, extracted, dried, filtered and concentrated to obtain compound 15;

[0033] S2. dissolving the compound 15 in dichloromethane, adding TFA dropwise at 0°C, stirring, extracting, washing, drying, filtering and concentrating to obtain compound 16;

[0034] S3. The above compound 5 is dissolved in DMF, EDCI, HOBT and triethylamine are added, and the mixture is stirred under an inert gas. The compound 16 is added and stirred at room temperature. After the reaction is complete, the mixture is extracted, dried and purified.

[0035] 3) When the target protein ligand is a suberoylanilide hydroxamic acid compound, the method for synthesizing the targeted protein degradation agent comprises the following steps:

[0036] S1. Compound 18 is reacted with compound 19 at room temperature to obtain compound 20;

[0037] S2. removing the Boc group of the compound 20 to obtain compound 21;

[0038] S3. subjecting the compound 21 to an amide condensation reaction with the compound 22 to obtain a compound 23;

[0039] S4. removing the Boc group of the compound 23 to obtain compound 24;

[0040] S5. subjecting the compound 24 to an amide condensation reaction with the compound 5 to obtain a compound 25;

[0041] S6. React hydroxylamine hydrochloride with sodium hydroxide to generate hydroxylamine, and react the hydroxylamine with the compound 25 under alkaline conditions.

[0042] The present invention also provides a pharmaceutical composition comprising the hydrophobic tag or the targeted protein degrader.

[0043] Furthermore, it also includes pharmaceutically acceptable carriers and pharmaceutically acceptable dosage forms.

[0044] The present invention also provides the use of the hydrophobic tag or the targeted protein degrading agent, including the preparation of drugs for targeted degradation of hormone receptors or the preparation of drugs against breast cancer.

[0045] In summary, compared with the prior art, the present invention achieves the following technical effects:

[0046] The present invention finds that artemisinin compounds can be used as a hydrophobic tag, and are coupled with a variety of specific POIs (such as ERα, CDK6 and HDAC6) to synthesize a series of targeted protein degraders. The obtained target compounds all show a certain ability to bind to POIs, and by using artemisinin as a hydrophobic tag, not only can the compounds obtain MCF-7 cell inhibitory activity that is superior to the parent compound, but also the compounds can have good degradation activity.

[0047] The present invention introduces a simple synthesis of hydrophobic tags with a high yield. Compared with PROTAC, the molecular weight of the target molecule obtained is smaller, and it is better in terms of cell membrane permeability and drugability. By introducing artemisinin compounds as hydrophobic tags, a successful technical means for selective degradation of target proteins can be achieved. Its modular design strategy makes the rational design of binding pockets more widely applicable, and it is possible to obtain active compounds for non-drugable targets. At the same time, it was found that artemisinin compounds can be used as a class of hydrophobic tags as a modular strategy in the future, which also provides ideas and references for the development of new targeted protein degraders. BRIEF DESCRIPTION OF THE DRAWINGS

[0048] In order to more clearly illustrate the technical solutions of the embodiments of the present invention, the following briefly introduces the drawings required for use in the embodiments. It should be understood that the following drawings only show certain embodiments of the present invention and therefore should not be regarded as limiting the scope. For ordinary technicians in this field, other related drawings can be obtained based on these drawings without creative work.

[0049] Figure 1 This is a protein immunoblot analysis of ERα in MCF-7 cells after 10 μM of raloxifene, artemisinin, RA2, RA3, RA4, and RA6 were used for 24 hours in the examples of the present invention;

[0050] Figure 2 This is a protein immunoblot analysis of CDK6 in MCF-7 cells after the compound PA was treated with 0.1 μM, 1.0 μM, and 10 μM for 24 hours in an embodiment of the present invention;

[0051] Figure 3 This is a protein immunoblot analysis of HDAC6 in MCF-7 cells after being treated with 0.1 μM, 1.0 μM, and 10 μM compound SA for 24 hours in an example of the present invention. DETAILED DESCRIPTION

[0052] In order to enable those skilled in the art to better understand the scheme of the present invention, the technical scheme in the embodiments of the present invention will be clearly and completely described below in conjunction with the drawings in the embodiments of the present invention. Obviously, the described embodiments are only part of the embodiments of the present invention, not all of the embodiments. Based on the embodiments of the present invention, all other embodiments obtained by ordinary technicians in this field without creative work should fall within the scope of protection of the present invention.

[0053] Example 1

[0054] The preparation method of the benzothiophene compound containing a hydrophobic tag of the present invention comprises the following steps:

[0055] (1) Synthesis of artemisinin analogs

[0056] The synthetic route is shown in the following reaction formula, which specifically includes the following steps:

[0057]

[0058] 1) Synthesis of (3R,6R,12R,12aR)-3,6,9-trimethyl-3,4,5,5a,6,7,8,8a-octahydro-12H-3,12-epoxy[1,2]dioxacyclo[4,3-I]isochromene (Compound 2)

[0059] 1 (4.0 g, 14.17 mmol) was dissolved in 50 mL of tetrahydrofuran, cooled to 0°C, and then slowly added with boron trifluoride etherate (2.09 mL, 18.37 mmol) and reacted for 24 h. After the reaction, the reaction was quenched with saturated sodium bicarbonate. The mixture was extracted with ethyl acetate and dried over anhydrous sodium sulfate. The mixture was evaporated under reduced pressure and purified by column chromatography using petroleum ether:ethyl acetate = 50:1 as the eluent to obtain compound 2 as a white solid (2.8 g) with a yield of 74.2%.

[0060] 2) Synthesis of (3R, 6R, 12R, 12aR)-9-bromo-3,6,9-trimethyldecahydro-12H-3,12-epoxy[1,2]dioxirane[4,3-I]isochrome-10-ol (Compound 3)

[0061] Compound 2 (2.0 g, 7.5 mmol) was dissolved in 40 mL of carbon tetrachloride, bromine (11.3 mL, 11.3 mmol, 1.2 equiv) was slowly added dropwise at 0°C, and stirred for 30 minutes, 20 mL of distilled water was added, and stirred for another 2 hours, after the reaction was completed, ether was added to dilute, sodium thiosulfate was added to quench the excess bromine, dried over anhydrous sodium sulfate, and directly carried out the next step of reaction after reduced pressure distillation. Compound 3 (1.7 g, yield 81.8%) was obtained.

[0062] 3) Synthesis of (3R, 6R, 9S, 11R, 11aR)-3,6,9-trimethyloctahydro-3H, 11H-3,11-epoxy[1,2]dioxepane[3,4-d]isobenzofuran-9-carbaldehyde (Compound 4)

[0063] Compound 3 (2.5 g, 6.9 mmol) was dissolved in 150 mL of dichloromethane, and triethylamine (TEA) (17.2 mmol, 2.5 mL, 2.5 equiv) was then added. The reaction mixture was stirred at room temperature until the reaction was complete as monitored by TLC. The mixture was then treated with HCl (0.6 N) to neutralize the excess TEA. The organic layer was washed with NaHCO 3 , dried over anhydrous Na 2 SO 4 , filtered and concentrated to give a crude product, which was purified by column chromatography to give compound 4 (1.8 g, 91% yield) as fine milky white crystals.

[0064] 4) Synthesis of (3R, 6R, 9S, 11R, 11aR)-3,6,9-trimethyloctahydro-3H, 11H-3,11-epoxy[1,2]dioxacyclo[3,4-d]isobenzofuran-9-carboxylic acid (Compound 5)

[0065] Compound 4 (1.3 g, 4.6 mmol) was dissolved in 1,4-dioxane (50 mL) solution, and then an aqueous solution (17.5 mL) containing sulfamic acid (1.8 g, 18.77 mmol) was added. The mixture was stirred at 0 ° C for 20 h, and then NaClO2 (1.7 g, 18.77 mmol) and water (12.5 mL) were added. After stirring for 30 minutes, Na2SO3 (1.16 g) was added, and the mixture was diluted with water (50 mL). The mixture was extracted with ether (3×100 ml), and the organic layer was washed with brine, dried over anhydrous Na2SO4 and evaporated. The product (1.1 g, 80%) was obtained as a brown solid.

[0066] (2) Synthesis of benzothiophene derivatives with linkers of different lengths

[0067] The synthetic route is shown in the following reaction formula, which specifically includes the following steps:

[0068]

[0069] The specific synthesis is that 4-acetylbenzoic acid is stirred with oxalyl chloride at room temperature to obtain an acyl chloride intermediate, the intermediate is reacted with 6-methoxy-2-(4-methoxyphenyl)benzothiophene (compound 7) and anhydrous aluminum chloride at room temperature to obtain compound 8, compound 8 is dissolved in an anaerobic ethanol / water (2:1) solution, and then sodium acetate is added and stirred at 100°C to obtain compound 9, compound 9 is reacted with (n-bromo-n-alkyl) tert-butyl carbamates of different chain lengths to obtain compound 10, and then boron tribromide is used to demethylate and demethylate the Boc group to obtain benzothiophene derivatives 11 (n=2, 3, 4, 6, 8) with linkers of different lengths.

[0070] (3) Synthesis of benzothiophene compounds with hydrophobic tags of artemisinin analogs (for targeting ERα)

[0071] After obtaining benzothiophene derivatives 11a-d with side chains of different lengths, these four compounds were subjected to amidation reaction with artemisinin analogs (compound 5) respectively. The synthetic route is shown in the following reaction formula:

[0072]

[0073] Using HATU as a catalyst and DIPEA as a base, in the presence of DMF solvent, we synthesized final products 12a-d (n=2, 3, 4, 6, 8) with artemisinin tags and linkers of different lengths. Compared with PROTAC, the raw materials of this type of compound are cheap and easy to obtain, the synthesis is fast, and the operation is relatively simple.

[0074] The following examples are used to further illustrate the present invention, but should not be construed as limiting the present invention. Unless otherwise specified, the technical means used in the examples are conventional means well known to those skilled in the art.

[0075] (4) Synthesis of methylpyrido-pyrimidine compounds with hydrophobic tags of artemisinin analogs (targeting CDK4 / 6)

[0076] The synthetic route is shown in the following reaction formula, which specifically includes the following steps:

[0077]

[0078] 1) Synthesis of tert-butyl (3-((6-acetyl-8-cyclopentyl-5-methyl-7-oxo-7,8-dihydropyrido[2,3-d]pyrimidin-2-yl)amino)propyl)carbamate (Compound 15)

[0079] Under argon atmosphere, compound 13 (100 mg, 0.33 mmol) and 5 mL of anhydrous DMSO were added to a 50 ml round-bottom flask, and a mixed solution of compound 14 (57.0 mg, 0.33 mmol) and DIPEA (187.0 mg, 0.5 mmol) in DMSO (3 mL) was added dropwise over 3 min, and the mixture was reacted at room temperature for 2 h. After the reaction, the reaction solution was extracted with 25 mL of ethyl acetate and saturated sodium chloride solution (25 mL×3), and the organic layer was dried over anhydrous Na2SO4, filtered and concentrated to obtain a crude product, which was purified by column chromatography to obtain white solid compound 15 (120 mg, 81.9%).

[0080] 2) Synthesis of 6-acetyl-2-((3-aminopropyl)amino)-8-cyclopentyl-5-methylpyrido[2,3-d]pyrimidin-7(8H)-one (Compound 16)

[0081] Compound 15 (100 mg, 0.23 mmol) was dissolved in dichloromethane, TFA was added dropwise at 0°C, and then stirred at 0°C for 4 h. The resulting solution was poured into ice water and extracted with CH2Cl2 (3×25 mL). The organic layers were combined, washed with saturated NaHCO3 (3×40 mL), and dried over anhydrous Na2SO4. The organic layer was dried over anhydrous Na2SO4, filtered and concentrated to obtain a crude product, Compound 16, which was directly used in the next step.

[0082] 3) Synthesis of (3R, 6R, 9S, 11R, 11aR)-N-(3-((6-acetyl-8-cyclopentyl-5-methyl-7-oxo-7,8-dihydropyrido[2,3-d]pyrimidin-2-yl)amino)propyl)-3,6,9-trimethyloctahydro-3H, 11H-3,11-epoxy[1,2]dioxirane[3,4-d]isobenzofuran-9-carboxamide (Compound 17)

[0083] Compound 5 (78.2 mg, 0.26 mmol) was placed in a 50 mL single-mouth bottle, dissolved in 10 mL of DMF, EDCI (60.4 mg, 0.32 mmol), HOBT (42.6 mg, 0.32 mmol) and triethylamine (53.0 mg, 0.52 mmol) were added, and stirred for half an hour under an inert atmosphere, and then compound 16 (90.0 mg, 0.26 mmol) was added, and stirred at room temperature for 4 h. After TLC confirmed that the reaction was complete, it was extracted with dichloromethane and dried over anhydrous sodium sulfate. The organic phase was spin-dried and mixed, and separated and purified by silica gel column chromatography (the eluent ratio was v 二氯甲烷 / v 甲 Alcohol = 100:1-10:1), to obtain 81 mg of white solid compound 17 with a yield of 49.9%.

[0084] (6) Synthesis of suberoylanilide hydroxamic acid compounds (targeting HDAC6) with hydrophobic tags of artemisinin analogs

[0085] The synthetic route is shown in the following reaction formula, which specifically includes the following steps:

[0086]

[0087] Tert-butyl (4-aminophenyl)carbamate (Compound 18) was stirred with methyl 8-chloro-8-oxooctanoate (Compound 19) at room temperature to obtain Compound 20, and then the Boc group was removed with trifluoroacetic acid at 0°C with stirring to obtain Compound 21. Compound 21 and Boc-4-aminobutyric acid (Compound 22) were subjected to amide condensation using EDCI, HOBT and triethylamine to obtain Compound 23, and then the Boc group was removed with HCl at 45°C to obtain Compound 24. Compound 24 and Compound 5 were subjected to amide condensation to obtain Compound 25, and then hydroxylamine hydrochloride was reacted with sodium hydroxide to generate hydroxylamine, and hydroxylamine was reacted with Compound 25 under alkaline conditions to obtain the final product Compound 26.

[0088] Example 2

[0089] Preparation of (3R,6R,9S,11R,11aR)-N-(2-(4-(6-hydroxy-2-(4-hydroxyphenyl)benzo[b]thiophene-3-carbonyl)phenoxy)ethyl)-3,6,9-trimethyloctahydro-3H,11H-3,11-epoxy[1,2]dioxazo[3,4-d]isobenzofuran-9-carboxamide (RA2):

[0090]

[0091] Compound 5 (100 mg, 0.33 mmol) was placed in a 50 ml single-mouth bottle, dissolved in 10 mL of DMF, HATU (152.9 mg, 0.4 mmol) and DIPEA (86.6 mg, 0.66 mmol) were added, and stirred for half an hour under an inert atmosphere. Then compound 11a (n=2, 135.9 mg, 0.33 mmol) was added, and stirred at room temperature overnight. After TLC confirmed that the reaction was complete, it was extracted with dichloromethane and dried over anhydrous sodium sulfate. The organic phase was spin-dried and mixed, and separated and purified by silica gel column chromatography (the eluent ratio was v 二氯甲烷 / v 甲醇 =100:1-10:1) to give 84 mg of yellow solid compound 12a with a yield of 37.1%.

[0092] 11H NMR (400 MHz, Methanol-d4) δ 7.66 (d, J = 8.8 Hz, 2H), 7.36 (d, J = 8.8 Hz, 1H), 7.24 (d, J = 2.3 Hz, 1H), 7.17 (d, J = 8.6 Hz, 2H), 6.83 (dd, J = 8.8, 2.3 Hz, 1H), 6.79 (d, J = 8.9 Hz, 2H), 6.61 (d, J = 8.6 Hz, 2H), 5.67 (s, 1H), 4.04 (tq, J = 10.2, 6.0, 5.2 Hz, 2H), 3.63 (dt, J = 14.1, 5.1 Hz, 1H), 3.46 (ddd, J = 14.0, 6.6, 4.5 Hz, 1H), 2.22 (ddd, J = 14.5, 12.9, 3.9 Hz, 1H), 2.08 (dd, J = 13.0, 6.4 Hz, 1H), 2.01 (dt, J = 14.5, 3.8 Hz, 1H), 1.90 (dt, J = 13.7, 4.4 Hz, 1H), 1.71 (ddd, J = 12.8, 6.2, 3.1 Hz, 1H), 1.64 (s, 3H), 1.33 (s, 4H), 1.25 (dd, J = 12.8, 4.2 Hz, 1H), 1.18 (dd, J = 13.1, 3.2 Hz, 1H), 0.97 (td, J = 12.9, 12.5, 2.8 Hz, 2H), 0.83 (d, J = 6.3 Hz, 4H). 13C NMR (101 MHz, CD3OD) δ 192.86, 173.66, 161.94, 156.61, 154.15, 141.24, 138.80, 131.64, 130.87, 129.15, 128.74, 128.54, 123.36, 122.07, 113.89, 113.42, 112.69, 105.31, 102.36, 96.23, 86.38, 85.04, 64.76, 50.17, 36.82, 35.39, 35.14, 30.66, 24.09, 23.35, 22.78, 22.72, 17.44. ESI-HRMS m / z calcd for C38H39NO9SNa + 708.2243, found 722.2240 [M+Na] + . HPLC purity 98%

[0093] Example 3

[0094] Preparation of (3R,6R,9S,11R,11aR)-N-(3-(4-(6-hydroxy-2-(4-hydroxyphenyl)benzo[b]thiophene-3-carbonyl)phenoxy)propyl)-3,6,9-trimethyloctahydro-3H,11H-3,11-epoxy[1,2]dioxazo[3,4-d]isobenzofuran-9-carboxamide (RA3):

[0095]

[0096] Compound 5 (100 mg, 0.33 mmol) was placed in a 50 ml single-mouth bottle, dissolved in 10 mL of DMF, HATU (152.9 mg, 0.4 mmol) and DIPEA (86.6 mg, 0.66 mmol) were added, and stirred for half an hour under an inert atmosphere. Then compound 11b (n=3, 140.6 mg, 0.33 mmol) was added, and stirred at room temperature overnight. After TLC confirmed that the reaction was complete, it was extracted with dichloromethane and dried over anhydrous sodium sulfate. The organic phase was spin-dried and mixed, and separated and purified by silica gel column chromatography (the eluent ratio was v 二氯甲烷 / v 甲醇 =100:1-10:1) to obtain 80 mg of yellow solid compound 12b with a yield of 34.6%.

[0097] 1 H NMR (400MHz, Methanol-d4) δ7.67(d,J=8.9Hz,2H),7.36(d,J=8.7Hz,1H),7.25(d,J=2.3Hz,1H),7.18(dt,J=9.5,2.6Hz,2H),6.8 5(dt,J=4.3,2.1Hz,1H),6.82(d,J=9.1Hz,2H),6.62(d,J=8.6Hz,2H),5.69(s,1H),4.01(t,J=5.9Hz,2H),3.34(s,2H),2.23(ddd ,J=14.5,12.9,3.9Hz,1H),2.10(dd,J=12.8,6.5Hz,1H),2.06–1.88(m,5H),1.79(ddd,J=12.7,6.2,3.0Hz,1H),1.64(s,3H),1.3 4(s,3H),1.28(s,2H),1.19(s,1H),1.09–1.03(m,1H),1.00(dd,J=13.0,2.9Hz,1H),0.96(d,J=6.3Hz,1H),0.87(d,J=6.3Hz,3H). 13C NMR (101MHz, CD3OD) δ194.20,174.56,163.35,157.85,155.40,142.35,140. 03,132.92,132.11,130.23,129.97,129.93,129.86,124.66,123.33,115.1 3,114.66,113.99,106.54,103.61,97.39,87.64,86.37,66.05,51.30,36.6 5,36.39,36.00,31.97,28.38,25.48,24.84,24.07,24.00,18.72.ESI-HRMS m / z calcd for C 39 H 41 NO9S + 722.2400, found 722.2408[M+Na] + .HPLC purity 95%.

[0098] Example 4

[0099] Preparation of (3R,6R,9S,11R,11aR)-N-(4-(4-(6-hydroxy-2-(4-hydroxyphenyl)benzo[b]thiophene-3-carbonyl)phenoxy)butyl)-3,6,9-trimethyloctahydro-3H,11H-3,11-epoxy[1,2]dioxazo[3,4-d]isobenzofuran-9-carboxamide (RA4):

[0100]

[0101] Compound 5 (100 mg, 0.33 mmol) was placed in a 50 mL single-mouth bottle, dissolved in 10 mL of DMF, HATU (152.9 mg, 0.4 mmol) and DIPEA (86.6 mg, 0.66 mmol) were added, and stirred for half an hour under an inert atmosphere. Then compound 11d (n=4, 145.3 mg, 0.33 mmol) was added and stirred at room temperature overnight. After TLC confirmed that the reaction was complete, it was extracted with dichloromethane and dried over anhydrous sodium sulfate. The organic phase was spin-dried and mixed, and separated and purified by silica gel column chromatography (the eluent ratio was v 二氯甲烷 / v 甲醇 =100:1-10:1) to obtain 85 mg of yellow solid compound 12c with a yield of 36.1%.

[0102] 11H NMR (400 MHz, Methanol-d4) δ 7.67 (d, J = 8.9 Hz, 2H), 7.41 (d, J = 8.7 Hz, 1H), 7.28 (d, J = 2.4 Hz, 1H), 7.19 (d, J = 8.7 Hz, 2H), 6.87 (dd, J = 8.8, 2.2 Hz, 1H), 6.75 (d, J = 8.9 Hz, 2H), 6.64 (d, J = 8.7 Hz, 2H), 5.73 (s, 1H), 3.94 (t, J = 6.2 Hz, 2H), 3.24 (t, J = 6.9 Hz, 2H), 2.24 (ddd, J = 14.6, 13.0, 3.9 Hz, 1H), 2.14 (dd, J = 13.0, 6.4 Hz, 1H), 2.05–1.98 (m, 1H), 1.89 (ddp, J = 13.6, 10.0, 3.5 Hz, 2H), 1.75 (p, J = 6.5, 6.0 Hz, 2H), 1.68 (s, 3H), 1.64 (d, J = 4.8 Hz, 1H), 1.48–1.42 (m, 1H), 1.39 (dd, J = 11.1, 5.6 Hz, 1H), 1.35 (s, 2H), 1.31 (d, J = 9.5 Hz, 2H), 1.29–1.20 (m, 1H), 1.08 (qd, J = 13.2, 2.9 Hz, 2H), 0.95 (d, J = 6.3 Hz, 1H), 0.90 (d, J = 6.3 Hz, 3H). 13 13C NMR (101 MHz, CD3OD) δ 194.14, 174.52, 163.49, 157.81, 155.38, 142.46, 140.04, 132.97, 132.13, 130.03, 129.88, 124.68, 123.41, 115.14, 114.69, 113.99, 113.94, 106.59, 103.61, 97.41, 87.68, 86.39, 67.51, 51.32, 38.25, 36.67, 36.41, 32.01, 26.13, 25.69, 25.48, 24.93, 24.07, 24.03, 18.75. ESI-HRMS m / z calcd for C 40 H 43 NO9SNa + 736.2556, found 736.2558 [M+Na] + . HPLC purity 99%.

[0103] Example 5

[0104] Preparation of (3R,6R,9S,11R,11aR)-N-(6-(4-(6-hydroxy-2-(4-hydroxyphenyl)benzo[b]thiophene-3-carbonyl)phenoxy)hexyl)-3,6,9-trimethyloctahydro-3H,11H-3,11-epoxy[1,2]dioxazo[3,4-d]isobenzofuran-9-carboxamide (RA6):

[0105]

[0106] Compound 5 (100 mg, 0.33 mmol) was placed in a 50 mL single-mouth bottle, dissolved in 10 mL of DMF, HATU (152.9 mg, 0.4 mmol) and DIPEA (86.6 mg, 0.66 mmol) were added, and stirred for half an hour under an inert atmosphere. Then compound 11c (n=6, 154.7 mg, 0.33 mmol) was added, and stirred at room temperature overnight. After TLC confirmed that the reaction was complete, it was extracted with dichloromethane and dried over anhydrous sodium sulfate. The organic phase was spin-dried and mixed, and separated and purified by silica gel column chromatography (the eluent ratio was v 二氯甲烷 / v 甲醇 =100:1-10:1) to obtain 78 mg of yellow solid compound 12d with a yield of 31.9%.

[0107] 11H NMR (400 MHz, Acetone-d6) δ 7.67 (d, J = 8.8 Hz, 2H), 7.39 (d, J = 8.8 Hz, 1H), 7.26 (d, J = 2.3 Hz, 1H), 7.19 (d, J = 8.6 Hz, 2H), 6.86 (dd, J = 8.8, 2.3 Hz, 1H), 6.76 (d, J = 8.9 Hz, 2H), 6.63 (d, J = 8.6 Hz, 2H), 5.73 (s, 1H), 3.91 (t, J = 6.4 Hz, 2H), 3.18 (t, J = 7.0 Hz, 2H), 2.24 (ddd, J = 14.6, 13.0, 3.9 Hz, 1H), 2.13 (dd, J = 13.0, 6.3 Hz, 1H), 2.01 (dt, J = 14.5, 3.9 Hz, 1H), 1.88 (dddd, J = 16.3, 9.3, 7.3, 3.6 Hz, 2H), 1.71 (q, J = 6.9 Hz, 2H), 1.66 (s, 3H), 1.51 (q, J = 6.6 Hz, 2H), 1.44 (d, J = 8.6 Hz, 2H), 1.39 (dd, J = 11.7, 6.8 Hz, 3H), 1.34 (s, 3H), 1.32 (d, J = 8.6 Hz, 1H), 1.30–1.20 (m, 1H), 1.08 (qd, J = 13.0, 11.9, 3.6 Hz, 2H), 0.95 (td, J = 12.4, 11.8, 2.5 Hz, 1H), 0.89 (d, J = 6.3 Hz, 3H). 13 13C NMR (101 MHz, CD3OD) δ 192.94, 173.14, 162.40, 156.58, 154.14, 141.11, 138.77, 131.70, 130.88, 128.73, 128.70, 128.64, 123.41, 122.12, 113.88, 113.42, 112.67, 105.31, 102.35, 96.15, 86.42, 85.13, 66.58, 50.06, 37.26, 35.41, 35.20, 30.77, 27.76, 27.33, 25.02, 24.21, 24.06, 23.65, 22.81, 22.75, 17.48. ESI-HRMS m / z calcd for C 42 H 47 NO9SNa + 764.2869, found 764.2861 [M+Na] + ..HPLC purity 97%.

[0108] Example 6

[0109] Preparation of (3R,5R)-N-(3-(4-(6-hydroxy-2-(4-hydroxyphenyl)benzo[b]thiophene-3-carbonyl)phenoxy)propyl)adamantane-1-carboxamide (RAdam):

[0110]

[0111] 1-Adamantanecarboxylic acid (25.8 mg, 0.14 mmol) was placed in a 25 mL single-mouth bottle, dissolved in 5 mL of DMF, HATU (65.26 mg, 0.17 mmol) and DIPEA (37 mg, 0.28 mmol) were added, and stirred for half an hour under an inert atmosphere. Then, compound 11a (n=3, 60 mg, 0.14 mmol) was added, and stirred at room temperature overnight. After TLC confirmed that the reaction was complete, it was extracted with dichloromethane and dried over anhydrous sodium sulfate. The organic phase was spin-dried and mixed, and separated and purified by silica gel column chromatography (the eluent ratio was v 二氯甲烷 / v 甲醇 =100:1-10:1), to obtain 55 mg of a yellow solid compound with a yield of 67.5%.

[0112] 1 H NMR (400MHz, Methanol-d4) δ7.71(d,J=8.5Hz,2H),7.40(d,J=8.8Hz,1H),7.28(d,J=2.2Hz,1H),7.20(d,J=8.3Hz,2H),6.87(dd,J=8.8,2.2 Hz,1H),6.82(d,J=8.6Hz,2H),6.65(d,J=8.3Hz,2H),3.33(s,4H),2.00(s,3H),1.94(t,J=6.4Hz,2H),1.84(s,6H),1.75(q,J=12.3Hz,6H). 13 CNMR(101MHz,CD3OD)δ194.28,179.60,163.48,157.83,155.38,142.38,140.03,132.93,132.10,130.13,129.95,1 29.88,124.67,123.30,115.10,114.64,113.93,106.51,65.86,40.39,38.79,36.19,36.16,28.64,28.23.ESI-HRMS m / zcalcd for C 35 H 35 NO5S + 604.2134, found 604.2134[M+Na] +.HPLC purity99%.

[0113] Example 7

[0114] Preparation of (3R, 6R, 9S, 11R, 11aR)-N-(3-((6-acetyl-8-cyclopentyl-5-methyl-7-oxo-7,8-dihydropyrido[2,3-d]pyrimidin-2-yl)amino)propyl)-3,6,9-trimethyloctahydro-3H, 11H-3,11-epoxy[1,2]dioxirane[3,4-d]isobenzofuran-9-carboxamide (PA):

[0115]

[0116] Compound 4 (78.2 mg, 0.26 mmol) was placed in a 50 mL single-mouth bottle, dissolved in 10 mL of DMF, EDCI (60.4 mg, 0.32 mmol), HOBT (42.6 mg, 0.32 mmol) and triethylamine (53.0 mg, 0.52 mmol) were added, and stirred for half an hour under an inert atmosphere. Then compound 16 (90.0 mg, 0.26 mmol) was added and stirred at room temperature for 4 h. After TLC confirmed that the reaction was complete, it was extracted with dichloromethane and dried over anhydrous sodium sulfate. The organic phase was spin-dried and mixed, and separated and purified by silica gel column chromatography (the eluent ratio was v 二氯甲烷 / v 甲醇 =100:1-10:1) to give 81 mg of white solid compound 17 with a yield of 49.9%.

[0117] 1 H NMR(400MHz,Chloroform-d)δ8.71(d,J=74.1Hz,1H),7.95(s,0H),7.19(s,0H),5.94–5 .80(m,1H),5.75(s,1H),3.69–3.25(m,4H),2.53(s,3H),2.33(s,3H),2.28(dd,J=12.4, 6.5Hz,2H),2.14–1.96(m,5H),1.79(d,J=4.1Hz,9H),1.67(s,2H),1.53(h,J=5.7,4.1H z,2H),1.47(s,3H),1.42–1.28(m,1H),1.18(s,1H),1.01(s,2H),0.97(d,J=6.3Hz,3H). 13C NMR (101MHz, CDCl3) δ203.03,161.69,155.75,104.02,97.42,86.70,51.18,48.31,37 .03,36.99,32.32,31.59,28.05,26.06,26.00,25.29,24.45,19.79,13.94.ESI-HRMS m / z calcd for C 33 H 46 N5O7 + 624.3397, found 624.3402[M+H] + .HPLC purity 98%.

[0118] Example 8

[0119] Preparation of N1-hydroxy-N8-(4-(((3R, 6R, 9S, 11R, 11aR)-3,6,9-trimethyloctahydro-3H, 11H-3,11-epoxy[1,2]dioxacyclo[3,4-d]isobenzofuran-9-carboxamide)butyramido)phenyl)octanediamide (SA):

[0120]

[0121] Hydroxylamine hydrochloride (377.8 mg, 5.44 mmol) was dissolved in 10 mL of methanol, and NaOH (249.0 mg, 6.23 mmol) was added. The mixture was stirred at 40 ° C for 20 min, and then the reaction mixture was cooled to 0 ° C, and then the mixed solution was filtered. At 0 ° C, the filtrate was slowly added dropwise to a solution of compound 25 (100 mg, 0.16 mmol) in THF (10 mL), and the reaction was allowed to react at room temperature for 1 h. After the reaction was completed, the solvent was concentrated by vacuum distillation, and then the solid was dissolved in water, and the pH was adjusted to 7 with 2 mol / L HCl, and the solid was filtered to obtain a solid. It was separated and purified by silica gel column chromatography (the eluent ratio was v 二氯甲烷 / v 甲醇 =100:1-10:1) to give 50 mg of white solid compound 26 with a yield of 48.5%.

[0122] 1H NMR (400MHz, DMSO-d6) δ10.33(s,1H),9.84(d,J=8.2Hz,1H),9.78(s,1H),8.65(s,1H),8.01(s,1H),7.49(d ,J=3.3Hz,4H),5.76(d,J=22.6Hz,1H),3.10(q,J=6.3Hz,2H),2.27(dt,J=14.4,7.3Hz,4H),2.04(dd,J=11.7 ,5.6Hz,2H),1.93(t,J=7.4Hz,2H),1.75(q,J=6.9Hz,3H),1.56(s,4H),1.49(d,J=7.2Hz,2H),1.40(s,1H),1 .35(d,J=7.3Hz,4H),1.31–1.24(m,6H),1.23(d,J=5.4Hz,3H),0.92(d,J=6.0Hz,2H),0.86(d,J=6.3Hz,3H). 13 C NMR(101MHz,CD3OD_SPE)δ177.17,173.03,172.41,171.55,134.65,134.62,120.19,103.10,96.97,86.41,86.04,49.82,38.49,36.6 6,36.59,36.41,33.86,32.30,32.25,31.36,28.51,28.43,25.35,25.19,24.64,24.32,24.22,22.31,20.83,18.75,13.04.ESI-HRMS m / z calcd for C 33 H 48 N4O9Na + 667.3319, found 667.3323[M+Na] + .HPLC purity 98%.

[0123] Example 9 Cell Viability Test

[0124] MCF-7 cells were cultured in DMEM liquid medium containing phenol red and 10% fetal bovine serum. When the cell density reached 80% to 90%, the cells were digested and the cell suspension was plated in a 96-well cell culture plate using DMEM medium without phenol red and 10% FBS. After the cells were completely attached to the wall, the original culture medium was discarded and 100 μL of fresh compound solution prepared in DMEM medium containing 10% FBS was added to each well. The compound concentration gradient was 1×10 -7.5 M, 1×10 -7 M, 1×10 -6.5 M, 1×10-6 M, 1×10 - 5.5 M, 1×10 -5 M, 1×10 -4.5 M, 1×10 -4 M. After 4 days of drug treatment, take out the culture plate, aspirate the culture medium, add 100 μL CCK8 working solution to each well, and incubate in a 37°C, 5% CO2 incubator for 1.5-2 hours. Read the plate on an ELISA reader, select 450nm as the reference wavelength, analyze the experimental results, and calculate the IC 50 , the results are shown in Table 1.

[0125] Table 1 Effects of target compounds on the viability of breast cancer cells MCF-7

[0126]

[0127]

[0128] a IC 50 The data are the mean ± SD of at least three independent experiments.

[0129] This example evaluates the effects of the seven compounds in Table 1 on the viability of MCF-7 cells. The artemisinin-tagged compounds RA2, RA3, RA4, RA6, RAdam, PA, and SA all have a certain effect on the viability of MCF-7 cells, among which compound RA3 has the greatest effect, and the IC 50 The value was 2.28±0.80μM.

[0130] Example 10 ER degradation activity assay

[0131] In order to prove that the benzothiophene derivatives with hydrophobic tags synthesized by the present invention have the ability to degrade estrogen receptor (ERα), and to analyze the effect of chain length on protein degradation activity. The structural formula of the benzothiophene derivatives with hydrophobic tags is as follows:

[0132]

[0133] In this example, raloxifene, artemisinin, and RA2, RA3, RA4, and RA6 (10 μM) were used to treat MCF-7 for 24 h, and then all proteins were extracted. The effects of the compounds on ERα degradation were analyzed by Western blotting and compared with raloxifene. The results are shown in FIG. Figure 1 shown.

[0134] pass Figure 1It can be concluded that the series of compounds of the present invention have good ERα down-regulating activity, and all four compounds significantly reduce the ERα level, among which compound RA3 shows the highest efficacy, reaching a degradation efficiency of 73.1%. It is worth noting that neither raloxifene nor artemisinin with d-ring contraction alone significantly reduces the ERα level, which illustrates the necessity of conjugated structure in effective degradation. It can also be seen from the results that the degradation activity of the compound on ERα is also related to the length of its side chain. When the chain length increases to a certain length, the degradation activity of the target product decreases. Therefore, the appropriate side chain length is also important for the degradation activity of ERα. By comparing the two types of compounds containing adamantyl tags and artemisinin tags, it was found that the introduction of artemisinin tags can indeed enable the compounds to obtain better ERα degradation activity, verifying the feasibility of artemisinin tags as hydrophobic tags.

[0135] Example 11 Determination of CDK6 degradation activity

[0136] In order to prove that the synthesized methylpyrido-pyrimidine compound with a hydrophobic tag has the ability to degrade cyclin-dependent kinase 6 (CDK6), this example used compound PA at concentrations of 0.1 μM, 1.0 μM, and 10 μM to treat MCF-7 for 24 hours, then extracted all proteins, and analyzed the effect of the compound on the degradation of CDK6 by protein immunoblotting. The structural formula of the methylpyrido-pyrimidine compound with a hydrophobic tag is as follows:

[0137]

[0138] The results are as follows Figure 2 As shown, PA significantly reduced the level of CDK6 in MCF-7 cells, and 10 μM PA induced CDK6 degradation by 73%.

[0139] Example 12 HDAC 6 degradation activity assay

[0140] In order to prove that the synthesized suberoylanilide hydroxamic acid compound with a hydrophobic tag has the ability to degrade histone deacetylase 6 (HDAC 6), this example used compound SA at concentrations of 0.1 μM, 1.0 μM, and 10 μM to treat MCF-7 for 24 hours, then extracted all proteins, and analyzed the effect of the compound on the degradation of HDAC 6 by western blotting.

[0141] The results are as follows Figure 3 As shown, SA significantly reduced the level of HDAC 6 in MCF-7 cells, and 10 μM SA induced CDK6 degradation by 69%.

[0142] The above embodiments are preferred implementation modes of the present invention, but the implementation modes of the present invention are not limited to the above embodiments. Any other changes, modifications, substitutions, combinations, and simplifications that do not deviate from the spirit and principles of the present invention should be equivalent replacement methods and are included in the protection scope of the present invention.

Claims

1. A hydrophobic tag based on artemisinin compounds, It is characterized in that Its structural formula is shown in formula (I):

2. A targeted protein degrader, It is characterized in that It comprises the hydrophobic tag and the target protein ligand and a linker as described in claim 1; the hydrophobic tag and the target protein ligand are connected through the linker; the target protein ligand is any one of benzothiophene compounds, methylpyridopyrimidine compounds, and suberoylanilide hydroxamic acid compounds.

3. The targeted protein degradation agent according to claim 2, It is characterized in that The linker is any one of an alkyl chain and an alkoxy group.

4. The targeted protein degradation agent according to any one of claims 2 to 3, It is characterized in that The targeted protein degradation agent has a structure as shown in the following formula (II), formula (III) or formula (IV): Where n is any integer from 1 to 10; Where n is any integer from 1 to 10; Wherein n is any integer from 1 to 10.

5. A method for synthesizing the targeted protein degradation agent according to any one of claims 2 to 4, It is characterized in that The steps include: 1) Synthesis of artemisinin analogs; 2) Synthesis of a target protein ligand with a linker, and then subjecting the artemisinin analogue described in step 1) to an amidation reaction with the target protein ligand to obtain a targeted protein degrader.

6. The method according to claim 5, It is characterized in that The step 1) of synthesizing artemisinin analogs specifically comprises the following steps: S1. Compound 1 was dissolved in tetrahydrofuran, cooled to 0°C, and then boron trifluoride ether was added dropwise to react for 24 hours. After the reaction was completed, the reaction was quenched with saturated sodium bicarbonate, extracted, dried, distilled, and purified to obtain compound 2; S2. Compound 2 was dissolved in carbon tetrachloride, bromine was slowly added dropwise at 0°C, and the mixture was stirred. After the reaction was completed, ether was added to dilute the mixture, sodium thiosulfate was added to quench the mixture, the mixture was dried, and the mixture was distilled under reduced pressure to obtain compound 3; S3. Compound 3 was dissolved in dichloromethane, triethylamine was added, and the reaction was stirred until the reaction was completed, and the excess triethylamine was neutralized, washed, dried, filtered, concentrated, and purified to obtain compound 4; S4. Dissolve compound 4 in 1,4-dioxane solution, add aqueous solution containing aminosulfonic acid, stir overnight at 0°C, add NaClO 2 and water, stir, add Na 2 SO 3 , dilute, extract, wash, dry and evaporate to obtain compound 5, which is an artemisinin analogue.

7. The method according to claim 5, It is characterized in that The step 2) synthesis of the target protein ligand with a linker specifically comprises the following steps: 1) When the target protein ligand is a benzothiophene compound, the method for synthesizing the targeted protein degradation agent comprises the following steps: S1.4-acetylbenzoic acid is stirred with oxalyl chloride at room temperature to obtain an acyl chloride intermediate, the intermediate is reacted with compound 7 and anhydrous aluminum chloride to obtain compound 8, compound 8 is dissolved in an oxyethanol / water solution, sodium acetate is added and stirred at 100°C to obtain compound 9, compound 9 is reacted with (n-bromo-n-alkyl)carbamic acid tert-butyl ester to obtain compound 10, boron tribromide is used to demethylate and demethylate the Boc group to obtain compound 11; wherein n is 1 to 10; and compound 11 is a benzothiophene compound with a linker; S2. Using HATU as a catalyst and DIPEA as a base, the compound 11 is subjected to an amidation reaction with the compound 5 according to claim 6 in a DMF solvent; 2) When the target protein ligand is a methylpyrido-pyrimidine compound, the method for synthesizing the targeted protein degradation agent comprises the following steps: S1. Compound 13 and compound 14 were mixed and reacted under argon, DIPEA and DMSO, extracted, dried, filtered and concentrated to obtain compound 15; S2. dissolving the compound 15 in dichloromethane, adding TFA dropwise at 0°C, stirring, extracting, washing, drying, filtering and concentrating to obtain compound 16; S3. dissolving the compound 5 according to claim 6 in DMF, adding EDCI, HOBT and triethylamine, stirring under inert gas, adding the compound 16, stirring at room temperature, extracting, drying and purifying after the reaction is complete; 3) When the target protein ligand is a suberoylanilide hydroxamic acid compound, the method for synthesizing the targeted protein degradation agent comprises the following steps: S1. Compound 18 is reacted with compound 19 at room temperature to obtain compound 20; S2. removing the Boc group of the compound 20 to obtain compound 21; S3. subjecting the compound 21 to an amide condensation reaction with the compound 22 to obtain a compound 23; S4. removing the Boc group of the compound 23 to obtain compound 24; S5. subjecting the compound 24 to an amide condensation reaction with the compound 5 according to claim 6 to obtain a compound 25; S6. React hydroxylamine hydrochloride with sodium hydroxide to generate hydroxylamine, and react the hydroxylamine with the compound 25 under alkaline conditions.

8. A pharmaceutical composition, It is characterized in that Comprising the hydrophobic tag of claim 1 or the targeted protein degradation agent of any one of claims 2-4.

9. The pharmaceutical composition according to claim 8, It is characterized in that Also included are pharmaceutically acceptable carriers and pharmaceutically acceptable dosage forms.

10. Use of the hydrophobic tag according to claim 1 or the targeted protein degradation agent according to any one of claims 2 to 4, It is characterized in that Applications include the preparation of drugs that target the degradation of hormone receptors or the preparation of drugs against breast cancer.

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