A PROTAC drug molecule targeting Fam20C and its preparation method and application

Abstract

The present application relates to the technical field of anti-tumor drugs, and in particular to a PROTAC drug molecule targeting Fam20C, and a preparation method and application thereof; the PROTAC drug has a molecular structure as shown in Formula 1, in which the R group includes at least one of the following groups: The PROTAC drug molecule can increase its specificity for targeting Fam20C, thereby enabling the PROTAC drug molecule to treat brain gliomas, so as to fill the gap in the prior art of PROTAC drugs targeting Fam20C in the treatment of brain gliomas.

Description

Technical Field

[0001] The present application relates to the technical field of anti-tumor drugs, and in particular to a PROTAC drug molecule targeting Fam20C and a preparation method and application thereof. Background Art

[0002] Gliomas are tumors that originate from glial cells in the brain, accounting for 40% to 60% of all primary central nervous system tumors. Common types of gliomas include diffuse astrocytomas, oligodendrogliomas, etc. The causes of gliomas are not very clear, but are usually related to environmental changes and gene mutations, such as exposure to high-dose ionizing radiation and gene mutations associated with rare syndromes. Gliomas can occur at any age, but the incidence rate in men is significantly higher than that in women, almost reaching 1.5:1. Current treatments for gliomas include surgical resection, radiotherapy, and chemotherapy. Surgical treatment can relieve symptoms of intracranial hypertension and solve related symptoms caused by gliomas, but for some patients with recurrent or metastatic gliomas, personalized treatments such as targeted therapy or immunotherapy are also needed. Fam20C is a Golgi protein. Fam20C expressed in various tissues has been shown through functional annotation of substrates to be not only involved in biomineralization, but also plays an important role in apoptosis and migration, wound healing, endopeptidase inhibitor activity and lipid metabolism disorders. At the same time, current studies have shown that Fam20C has several links with many types of human diseases, including Rayne syndrome (RS), cancer (triple-negative breast cancer, lung adenocarcinoma, bladder urothelial carcinoma, low-grade glioma of the brain, gastric adenocarcinoma) and diabetes. In addition, Fam20C has been found to enhance the metastasis of several types of human malignancies, such as breast cancer and colorectal cancer, which indicates that Fam20C may be a therapeutic target for the current development of brain glioma-related drugs.

[0003] At present, targeted protein degradation (TPD) is an emerging treatment method that may provide new solutions to the problems encountered by traditional small molecule targeted therapy (such as drug resistance and "undruggability"). Protein degradation targeting chimeras (PROTACs) are a new treatment method with a unique mechanism of action (MOA), which mainly induces the degradation of target proteins by hijacking the endogenous ubiquitin-proteasome system (UPS). Therefore, if PROTAC drugs targeting Fam20C can be developed, it may become an effective strategy for treating cancer metastasis. Summary of the invention

[0004] The present application provides a PROTAC drug molecule targeting Fam20C and a preparation method and application thereof to solve the following technical problem: how to fill the gap in the prior art of PROTAC drugs targeting Fam20C in the treatment of brain gliomas.

[0005] In a first aspect, the present application provides a PROTAC drug molecule targeting Fam20C, wherein the PROTAC drug has a molecular structure as shown in Formula 1,

[0006]

[0007] In Formula 1, the R group includes at least one of the following groups:

[0008]

[0009] Optionally, the R group is The PROTAC drug molecule has a molecular structure as shown in Formula 2,

[0010]

[0011] In a second aspect, the present application provides a method for preparing the PROTAC drug molecule of the first aspect, the method comprising:

[0012] 3,4,5-trimethoxyaniline and N,N-diisopropylethylamine are mixed, and then 2,4-dichloropyrimidine is added to carry out a first reaction to obtain a first intermediate;

[0013] The p-phenylenediamine and the first intermediate are mixed, and then 1,4-dioxane and acetic acid are added to carry out a second reaction to obtain a second intermediate;

[0014] Adding 4-(chlorosulfonyl)benzoic acid to the second intermediate to carry out a third reaction, followed by purification to obtain a third intermediate;

[0015] Conducting a fourth reaction of 4-fluoroisobenzofuran-1,3-dione and 3-amino-2,6-piperidindione, followed by purification, to obtain a fourth intermediate;

[0016] Mixing R-group tert-butyl formate and the fourth intermediate, and then adding N,N-diisopropylethylamine to carry out a fifth reaction to obtain a fifth intermediate having an R group;

[0017] Performing a sixth reaction on a hydrochloric acid solution of 1,4-dioxane and the fifth intermediate to obtain a sixth intermediate;

[0018] 2-(7-azabenzotriazole)-N,N,N',N'-tetramethyluronium hexafluorophosphate, the sixth intermediate and the third intermediate are mixed, and tetrahydrofuran and N,N-diisopropylethylamine are added to carry out the seventh reaction, followed by purification to obtain a PROTAC drug molecule.

[0019] Optionally, the first intermediate is 2-chloro-N-(3,4,5-trimethoxyphenyl)pyrimidine-4-amine; the second intermediate is N2-(4-aminophenyl)-N4-(3,4,5-trimethoxyphenyl)pyrimidine-2,4-diamine; and the third intermediate is 4-(N-(4-((4-((3,4,5-trimethoxyphenyl)amino)pyrimidine-2-yl)amino)phenyl)aminosulfonyl)benzoic acid.

[0020] Optionally, the fourth intermediate is 2-(2,6-dioxopiperidin-3-yl)-4-fluoroisoindoline-1,3-dione; and the fifth intermediate is tert-butyl 4-((2-(2,6-dioxopiperidin-3-yl)-1,3-dioxoisoindoline-4-yl)amino)R-formate.

[0021] Optionally, the temperature of the first reaction is 80° C., and the time of the first reaction is 8 h to 12 h;

[0022] The temperature of the second reaction is 110° C., and the time of the second reaction is 2 h;

[0023] The temperature of the third reaction is 20° C. to 30° C., and the time of the third reaction is 0.5 h.

[0024] Optionally, the temperature of the fourth reaction is 120° C., and the time of the fourth reaction is 12 h;

[0025] The temperature of the fifth reaction is 90° C., and the time of the fifth reaction is 2 h;

[0026] The temperature of the sixth reaction is 60° C., and the time of the sixth reaction is 8 h to 12 h;

[0027] The temperature of the seventh reaction is 30° C., and the time of the seventh reaction is 8 h to 12 h.

[0028] Optionally, the yield of the PROTAC drug molecule is 61% to 78%.

[0029] In a third aspect, the present application provides an application of a PROTAC drug molecule targeting Fam20C, the application comprising: using the PROTAC drug molecule described in the first aspect for preparing a therapeutic drug for glioblastoma cells.

[0030] In a fourth aspect, the present application provides a pharmaceutical composition for treating glioblastoma cells, wherein the pharmaceutical composition comprises the PROTAC drug molecule described in the first aspect or a pharmaceutically acceptable salt thereof.

[0031] The above technical solution provided by the embodiment of the present application has the following advantages compared with the prior art:

[0032] The embodiment of the present application provides a PROTAC drug molecule targeting Fam20C, wherein the PROTAC drug has a molecular structure as shown in Formula 1, Formula 1, wherein the R group includes at least one of the following groups:

[0033] Based on the existing PROTAC drug structure, the most commonly used E3 ubiquitin ligase CRBN ligand was selected for matching. Due to its small molecular weight advantage, it is not only easy to synthesize and prepare, but also in a more suitable chemical space for oral absorption. By optimizing the Linker structure of the PROTAC drug molecule and introducing an R group with an amino group, its specificity for targeting Fam20C can be increased to promote the specificity of PROTAC drug molecules targeting brain glioma cells. In addition, subsequent experiments have shown that the difference in R groups has a significant effect on activity. When the R group is an alkyl group, different chain lengths will also affect the inhibitory activity. As the length of the carbon chain increases, the inhibition rate of the compound on tumor cell growth tends to decrease; in addition, when the carbon chain length is 3 carbons, the inhibitory activity against SW-1783 cells is the best. BRIEF DESCRIPTION OF THE DRAWINGS

[0034] The accompanying drawings, which are incorporated in and constitute a part of this specification, illustrate embodiments consistent with the present application and, together with the description, serve to explain the principles of the present application.

[0035] In order to more clearly illustrate the embodiments of the present application or the technical solutions in the prior art, the drawings required for use in the embodiments or the description of the prior art will be briefly introduced below. Obviously, for ordinary technicians in this field, other drawings can be obtained based on these drawings without paying any creative labor.

[0036] Figure 1 A schematic diagram of the molecular structure of a PROTAC drug molecule provided in an embodiment of the present application;

[0037] Figure 2 for Figure 1 continuation of

[0038] Figure 3A schematic flow chart of a method for preparing a PROTAC drug molecule provided in an embodiment of the present application;

[0039] Figure 4 A schematic diagram of the actual process of preparing a PROTAC drug molecule provided in an embodiment of the present application; wherein: Figure 4 Here, a is the first reaction, b is the second reaction, c is the third reaction, d is the fourth reaction, e is the combined reaction of the fifth and sixth reactions, and f is the seventh reaction. DETAILED DESCRIPTION

[0040] In order to make the purpose, technical solution and advantages of the embodiments of the present application clearer, the technical solution in the embodiments of the present application will be clearly and completely described below in conjunction with the drawings in the embodiments of the present application. Obviously, the described embodiments are part of the embodiments of the present application, not all of the embodiments. Based on the embodiments in the present application, all other embodiments obtained by ordinary technicians in this field without making creative work are within the scope of protection of this application.

[0041] Various embodiments of the present application may be presented in the form of a range; it should be understood that the description in the form of a range is only for convenience and brevity and should not be understood as a rigid limitation on the scope of the present application; therefore, it should be considered that the range description has specifically disclosed all possible sub-ranges and single numerical values ​​within the range; for example, it should be considered that the range description from 1 to 6 has specifically disclosed sub-ranges, such as from 1 to 3, from 1 to 4, from 1 to 5, from 2 to 4, from 2 to 6, from 3 to 6, etc., as well as single numbers within the range, such as 1, 2, 3, 4, 5 and 6, which applies regardless of the range; in addition, whenever a numerical range is indicated in this document, it is meant to include any cited numbers (fractions or integers) within the indicated range.

[0042] In this article, the terms including "including" and the like mean "including but not limited to". Relational terms such as "first" and "second" are used only to distinguish one entity or operation from another entity or operation, and do not necessarily require or imply any such actual relationship or order between these entities or operations. "At least one" means one or more, and "plurality" means two or more; "at least one", "at least one of the following" or similar expressions refer to any combination of these items, including any combination of single items or plural items; for example, "at least one of a, b, or c", or "at least one of a, b, and c" can all represent: a, b, c, ab (i.e., a and b), ac, bc, or abc, where a, b, c can be single or multiple. Unless otherwise specified, the various raw materials, reagents, instruments and equipment used in this application can be purchased on the market or prepared by existing methods.

[0043] Figure 1 The molecular structure diagram of a PROTAC drug molecule provided in the examples of the present application is exemplarily shown;

[0044] like Figure 1 and Figure 2 As shown, the PROTAC drug provided in the embodiment of the present application has a molecular structure as shown in Formula 1,

[0045]

[0046] In Formula 1, the R group includes at least one of the following groups:

[0047]

[0048]

[0049] In some optional embodiments, the R group is The PROTAC drug molecule has a molecular structure as shown in Formula 2,

[0050]

[0051] In these embodiments, based on the formula 1 structure of the PROTAC drug molecule, the specific R group is refined as follows: The optimal choice of this group can be further clarified to further improve its specificity in targeting Fam20C and the anti-proliferative ability of ROTAC drug molecules against glioblastoma cells.

[0052] Figure 3A schematic flow chart of a method for preparing a PROTAC drug molecule provided in an embodiment of the present application is shown exemplarily;

[0053] Figure 4 The actual process diagram of a method for preparing a PROTAC drug molecule provided in an embodiment of the present application is exemplarily shown;

[0054] Based on a general inventive concept, such as Figure 3 and Figure 4 As shown, the present application provides a method for preparing the PROTAC drug molecule, the method comprising:

[0055] S1. 3,4,5-trimethoxyaniline and N,N-diisopropylethylamine are mixed, and then 2,4-dichloropyrimidine is added to carry out a first reaction to obtain a first intermediate;

[0056] S2. mixing p-phenylenediamine and the first intermediate, and then adding 1,4-dioxane and acetic acid to carry out a second reaction to obtain a second intermediate;

[0057] S3. adding 4-(chlorosulfonyl)benzoic acid to the second intermediate to carry out a third reaction, followed by purification to obtain a third intermediate;

[0058] S4. performing a fourth reaction on 4-fluoroisobenzofuran-1,3-dione and 3-amino-2,6-piperidindione, followed by purification to obtain a fourth intermediate;

[0059] S5. The R-group tert-butyl formate and the fourth intermediate are mixed, and then N,N-diisopropylethylamine is added to carry out a fifth reaction to obtain a fifth intermediate having an R group;

[0060] S6. performing a sixth reaction on a hydrochloric acid solution of 1,4-dioxane and the fifth intermediate to obtain a sixth intermediate;

[0061] S7. 2-(7-azabenzotriazole)-N,N,N',N'-tetramethyluronium hexafluorophosphate, the sixth intermediate and the third intermediate are mixed, and tetrahydrofuran and N,N-diisopropylethylamine are added to carry out the seventh reaction, followed by purification to obtain a PROTAC drug molecule.

[0062] This method is a method for preparing the above-mentioned PROTAC drug molecule. The specific structure of the PROTAC drug molecule can refer to the above-mentioned embodiment. Since this method adopts part or all of the technical solutions of the above-mentioned embodiment, it at least has all the beneficial effects brought by the technical solutions of the above-mentioned embodiment, which will not be described one by one here.

[0063] In some optional embodiments, the first intermediate is 2-chloro-N-(3,4,5-trimethoxyphenyl)pyrimidine-4-amine; the second intermediate is N2-(4-aminophenyl)-N4-(3,4,5-trimethoxyphenyl)pyrimidine-2,4-diamine; and the third intermediate is 4-(N-(4-((4-((3,4,5-trimethoxyphenyl)amino)pyrimidin-2-yl)amino)phenyl)aminosulfonyl)benzoic acid.

[0064] In some optional embodiments, the fourth intermediate is 2-(2,6-dioxopiperidin-3-yl)-4-fluoroisoindoline-1,3-dione; the fifth intermediate is tert-butyl 4-((2-(2,6-dioxopiperidin-3-yl)-1,3-dioxoisoindoline-4-yl)amino)R-formate.

[0065] In these embodiments, the first intermediate can be 2-chloro-N-(3,4,5-trimethoxyphenyl)pyrimidine-4-amine, the second intermediate can be N2-(4-aminophenyl)-N4-(3,4,5-trimethoxyphenyl)pyrimidine-2,4-diamine, the third intermediate can be 4-(N-(4-((4-((3,4,5-trimethoxyphenyl)amino)pyrimidine-2-yl)amino)phenyl)aminosulfonyl)benzoic acid, the fourth intermediate can be 2-(2,6-dioxopiperidin-3-yl)-4-fluoroisoindoline-1,3-dione and the fifth intermediate can be 4-((2-(2,6-dioxopiperidin-3-yl)-1,3-dioxoisoindoline-4-yl)amino)R-tert-butyl carboxylate, and the intermediate structures of each stage of the preparation method can be clearly defined.

[0066] In some optional embodiments, the temperature of the first reaction is 80° C., and the time of the first reaction is 8 h to 12 h;

[0067] The temperature of the second reaction is 110° C., and the time of the second reaction is 2 h;

[0068] The temperature of the third reaction is 20° C. to 30° C., and the time of the third reaction is 0.5 h.

[0069] In some optional embodiments, the temperature of the fourth reaction is 120° C., and the time of the fourth reaction is 12 h;

[0070] The temperature of the fifth reaction is 90° C., and the time of the fifth reaction is 2 h;

[0071] The temperature of the sixth reaction is 60° C., and the time of the sixth reaction is 8 h to 12 h;

[0072] The temperature of the seventh reaction is 30° C., and the time of the seventh reaction is 8 h to 12 h.

[0073] In these embodiments, the temperature of the first reaction can be 80°C, the time of the first reaction can be 8h~12h, and the temperature of the second reaction can be 110°C, the time of the second reaction can be 2h, and the temperature of the third reaction can be 20°C~30°C, the time of the third reaction can be 0.5h, and the temperature of the fourth reaction can be 120°C, the time of the fourth reaction can be 12h, and the temperature of the fifth reaction can be 90°C, the time of the fifth reaction can be 2h, and the temperature of the sixth reaction can be 60°C, the time of the sixth reaction can be 8h~12h, and the temperature of the seventh reaction can be 30°C, and the time of the seventh reaction can be 8h~12h. These reaction temperatures and reaction conditions can be used to prepare a sufficient amount of PROTAC drug molecules.

[0074] The time of the first reaction can be 8 h, 9 h, 10 h, 11 h or 12 h.

[0075] The temperature of the third reaction can be 20°C, 21°C, 22°C, 23°C, 24°C, 25°C, 26°C, 27°C, 28°C, 29°C or 30°C.

[0076] The sixth reaction time can be 8 h, 9 h, 10 h, 11 h or 12 h.

[0077] The seventh reaction time can be 8 h, 9 h, 10 h, 11 h or 12 h.

[0078] Based on a general inventive concept, an embodiment of the present application provides an application of a PROTAC drug molecule targeting Fam20C, the application comprising: using the PROTAC drug molecule in the preparation of a therapeutic drug for brain glioma cells.

[0079] This application is realized based on the above-mentioned PROTAC drug molecule. The specific structure of the PROTAC drug molecule can refer to the above-mentioned embodiment. Since this application adopts part or all of the technical solutions of the above-mentioned embodiment, it has at least all the beneficial effects brought by the technical solutions of the above-mentioned embodiment, which will not be described one by one here.

[0080] Based on a general inventive concept, an embodiment of the present application provides a pharmaceutical composition for treating glioblastoma cells, wherein the pharmaceutical composition comprises the PROTAC drug molecule or a pharmaceutically acceptable salt thereof.

[0081] The pharmaceutical composition is realized based on the above-mentioned PROTAC drug molecule. The specific structure of the PROTAC drug molecule can refer to the above-mentioned embodiment. Since the pharmaceutical composition adopts part or all of the technical solutions of the above-mentioned embodiment, it at least has all the beneficial effects brought by the technical solutions of the above-mentioned embodiment, which will not be described one by one here.

[0082] It should be noted that the PROTAC drug molecule and its pharmaceutically acceptable salt in the pharmaceutical composition can be prepared into the following pharmaceutical forms:

[0083] Tablets, capsules, emulsions, creams, ointments, gels, suppositories, finely dispersed powders or aerosols or sprays for inhalation, sterile aqueous or oily solutions or suspensions or sterile emulsions for extra-gastrointestinal (including intravenous, intramuscular or infusion). In addition, liquid preparations containing PROTAC drug molecules and pharmaceutically acceptable salts thereof can be prepared using sterile water or water-propylene glycol solutions as solvents; the active components of PROTAC drug molecules and pharmaceutically acceptable salts thereof can also be dissolved in aqueous polyethylene glycol solutions; aqueous solutions for oral administration can also be prepared by dissolving the active components of PROTAC drug molecules and pharmaceutically acceptable salts thereof in water and adding appropriate colorants, flavoring agents, stabilizers and thickeners as needed; aqueous suspensions for oral use can be prepared by dispersing the finely dispersed active components of PROTAC drug molecules and pharmaceutically acceptable salts thereof in water together with viscous substances, which can be natural synthetic gums, resins, methylcellulose, carboxymethylcellulose and other suspending agents known in the pharmaceutical field.

[0084] It should be noted that the pharmaceutical composition may be in unit dosage form, for example, the pharmaceutical composition is divided into unit doses containing appropriate amounts of active ingredients. The unit dosage form may be a packaged preparation, the package including separate quantities of the preparation, such as boxed tablets, capsules, and powders in vials or ampoules. The unit dosage form may also be a capsule, cachet or tablet or any of these packaged forms may be in appropriate quantities.

[0085] It should be noted that the pharmaceutical composition can also be used in combination with other anti-glioma drugs, for example, such combination therapy can be achieved by administering various therapeutic components simultaneously, sequentially or separately. The application of such combination forms includes the pharmaceutical composition within the effective dosage range and other pharmaceutically active agents within the permitted dosage range.

[0086] The present application is further described below in conjunction with specific examples. The experimental methods in the following examples that do not specify specific conditions are usually measured according to industry standards; if there are no corresponding industry standards, they are carried out according to common international standards, conventional conditions, or conditions recommended by the manufacturer.

[0087] Example 1

[0088] like Figure 3 and Figure 4 As shown, a method for preparing the PROTAC drug molecule comprises:

[0089] 3,4,5-Trimethoxyaniline (Intermediate 2, 4.000 g, 21.83 mmol) was placed in a three-necked round-bottom flask, and anhydrous ethanol was added until it was completely dissolved. N,N-diisopropylethylamine (DIPEA, 7.6 mL, 43.66 mmol) was added while the solution was stirred to maintain the reaction temperature at room temperature. After continuous stirring for 0.5 h, the reaction apparatus was moved to an oil bath, and 2,4-dichloropyrimidine (Intermediate 1, 3.252 g, 21.83 mmol) was added while the solution was stirred. , at this time, the solution changes from white to yellow-brown, the reaction temperature is raised to 80°C, and refluxed overnight for the first reaction; after the reaction is completed, the temperature is cooled to room temperature, ethyl acetate is added to the mixture for suction filtration, and the filtrate is spin-dried at 40°C to obtain a crude product, which is separated and purified by silica gel column chromatography (volume ratio of petroleum ether: ethyl acetate = 5:1 to 3:1) to obtain the first intermediate 2-chloro-N-(3,4,5-trimethoxyphenyl)pyrimidine-4-amine (intermediate 3, 2.582 g, 8.73 mmol, 40%) as a light yellow solid.

[0090] The first intermediate (2.582 g, 8.73 mmol) and p-phenylenediamine (intermediate 4, 1.416 g, 13.10 mmol) were placed in a round-bottom flask, and 15.0 mL of 1,4-dioxane solution was added. The temperature was raised to 60°C under stirring to carry out the second reaction. The reaction system turned yellow. Acetic acid (0.55 mL, 9.60 mmol) was then added, and the temperature was raised to 110°C. The reaction was continued for 2 h, and the reaction system turned from yellow to dark brown. After the reaction was completed, the reaction system solution was lifted, and the temperature was lowered to room temperature. Then, the second intermediate N2-(4-aminophenyl)-N4-(3,4,5-trimethoxyphenyl)pyrimidine-2,4-diamine (intermediate 5, 2.502 g, 6.81 mmol, 78%) was separated and purified by silica gel column chromatography (volume ratio of petroleum ether: ethyl acetate = 1:1) to obtain a gray-brown solid.

[0091] In a 250.0 mL round-bottom flask under argon protection, the second intermediate (2.502 g, 6.81 mmol) was dissolved in 10.0 mL of ultra-dry pyridine, the round-bottom flask was placed on an ice bath at 0°C, and 4-(chlorosulfonyl)benzoic acid (intermediate 6, 1.502 g, 6.81 mmol) was slowly added under stirring to turn the reaction system red, the ice bath was removed, and the temperature was raised to room temperature to carry out the third reaction. After 0.5 h of reaction, a small amount of water was added to the reaction to quench the reaction, and then the reaction solution was placed in a 250 mL separatory funnel, washed with saturated copper sulfate solution, and ethyl acetate was added and allowed to stand for a while to produce stratification. At this time, the color of the aqueous phase of the reaction system was blue-black. When the aqueous phase of the reaction system showed light blue, the organic phase was separated and dried with Na2SO4 and filtered to remove the volatile solvent to obtain a crude product. The product was purified by silica gel column chromatography (volume ratio of dichloromethane:methanol=10:1) to obtain the third intermediate 4-(N-(4-((4-((3,4,5-trimethoxyphenyl)amino)pyrimidin-2-yl)amino)phenyl)aminosulfonyl)benzoic acid (Intermediate 7, 2.930 g, 5.31 mmol, 78%) as a pink solid.

[0092] 4-Fluoroisobenzofuran-1,3-dione (intermediate 8, 5.000 g, 30.12 mmol) was dissolved in acetic acid (20.0 mL), and 3-amino-2,6-piperidinedione (intermediate 9, 4.960 g, 30.12 mmol) was added to a three-necked round-bottom flask to carry out the fourth reaction, the reaction temperature was raised to 120°C, and the reaction time was controlled to 12 h. After the reaction was completed, ice water was added to the mixture obtained by the reaction, and the crude product was filtered to obtain a crude product, which was separated and purified by silica gel column chromatography (volume ratio of petroleum ether: ethyl acetate = 2:1) to obtain the fourth intermediate 2-(2,6-dioxopiperidin-3-yl)-4-fluoroisoindoline-1,3-dione (intermediate 10, 7.490 g, 27.12 mmol, 90%) as a gray solid.

[0093] The fourth intermediate (intermediate 10, 0.300 g, 1.09 mmol) was dissolved in 10.0 mL of N-methylpyrrolidone (NMP), and then R-butyl formate (0.42 mL, 2.17 mmol) was added to a 100.0 mL round-bottom flask, and N, N-diisopropylethylamine (0.38 mL, 2.17 mmol) was added under stirring to raise the reaction temperature to 90 ° C for the fifth reaction. The reaction system showed yellow-green color. After the reaction was completed, the solution was cooled to room temperature and then diluted with 100.0 mL of ethyl acetate. The obtained solution was washed with aqueous solution and protective salt water, the organic phase was dried with Na2SO4, filtered, the filtrate was collected, and the volatile substances were removed under reduced pressure. The product was separated and purified by silica gel column chromatography (volume ratio of petroleum ether:ethyl acetate=1:1) to obtain the fifth intermediate tert-butyl 4-((2-(2,6-dioxopiperidin-3-yl)-1,3-dioxoisoindolin-4-yl)amino)R-formate (0.368 g, 0.83 mmol) as a yellow fluorescent solid.

[0094] The fifth intermediate (0.368 g, 0.83 mmol) was placed in a 100.0 mL round-bottom flask, and 8.0 mL of HCl in 1,4-dioxane was added, followed by stirring at 60 ° C for 3 h and the sixth reaction was carried out. After the reaction was completed, the temperature was lowered to room temperature, and then rotary evaporation was performed at 45 ° C to remove volatile substances to obtain the sixth intermediate (intermediate 11a-11p, 0.286 g, 0.83 mmol) as a yellow fluorescent solid.

[0095] The third intermediate (0.458 g, 0.83 mmol, 1.0 eq) was dissolved in 2.0 mL of DMF, HATU (0.946 g, 2.49 mmol, 3.0 eq) and the sixth intermediate (0.3 mmol, 1.0 eq) were added to a round-bottom flask for the seventh reaction, and the mixture obtained by the reaction was dissolved in 5.0 mL of tetrahydrofuran, and N, N-diisopropylethylamine (0.22 mL, 1.25 mmol, 1.5 eq) was added and stirred at 30 ° C overnight. After the reaction was completed, the volatile substances were removed by decompression, the mixture was added to a 250.0 mL separatory funnel, 100.0 mL of water was added, and then 3×100.0 mL of ethyl acetate was used for extraction, and Na2SO4 was used for drying. After filtration, the volatile substances such as the filtrate were removed by decompression to obtain a crude product. Finally, the PROTAC drug molecule shown in Formula 1 was obtained by purification and separation by thin layer chromatography (PTLC).

[0096] Among them, the R group of the PROTAC drug molecule is ( ), denoted as PROTAC drug molecule 15a.

[0097] Example 2

[0098] Based on the contents disclosed in Example 1, the following modifications are further made:

[0099] The R group of the PROTAC drug molecule is ( ), and finally obtained PROTAC drug molecule 15b.

[0100] Example 3

[0101] Based on the contents disclosed in Example 1, the following modifications are further made:

[0102] The R group of the PROTAC drug molecule is ( ), and finally obtained PROTAC drug molecule 15c.

[0103] Example 4

[0104] Based on the contents disclosed in Example 1, the following modifications are further made:

[0105] The R group of the PROTAC drug molecule is ( ), and finally obtained PROTAC drug molecule 15d.

[0106] Example 5

[0107] Based on the contents disclosed in Example 1, the following modifications are further made:

[0108] The R group of the PROTAC drug molecule is ( ), and finally obtained PROTAC drug molecule 15e.

[0109] Example 6

[0110] Based on the contents disclosed in Example 1, the following modifications are further made:

[0111] The R group of the PROTAC drug molecule is ( ), and finally obtained the PROTAC drug molecule 15f.

[0112] Example 7

[0113] Based on the contents disclosed in Example 1, the following modifications are further made:

[0114] The R group of the PROTAC drug molecule is ( ), and finally obtained PROTAC drug molecule 15g.

[0115] Example 8

[0116] Based on the contents disclosed in Example 1, the following modifications are further made:

[0117] The R group of the PROTAC drug molecule is ( ), and finally the PROTAC drug molecule was obtained for 15h.

[0118] Example 9

[0119] Based on the contents disclosed in Example 1, the following modifications are further made:

[0120] The R group of the PROTAC drug molecule is ( ), and finally obtained the PROTAC drug molecule 15i.

[0121] Example 10

[0122] Based on the contents disclosed in Example 1, the following modifications are further made:

[0123] The R group of the PROTAC drug molecule is ( ), and finally obtained PROTAC drug molecule 15j.

[0124] Embodiment 11

[0125] Based on the contents disclosed in Example 1, the following modifications are further made:

[0126] The R group of the PROTAC drug molecule is ( ), and finally obtained PROTAC drug molecule 15k.

[0127] Example 12

[0128] Based on the contents disclosed in Example 1, the following modifications are further made:

[0129] The R group of the PROTAC drug molecule is ( ), and finally obtained PROTAC drug molecule 15l.

[0130] Example 13

[0131] Based on the contents disclosed in Example 1, the following modifications are further made:

[0132] The R group of the PROTAC drug molecule is ( ), and finally obtained PROTAC drug molecule 15m.

[0133] Embodiment 14

[0134] Based on the contents disclosed in Example 1, the following modifications are further made:

[0135] The R group of the PROTAC drug molecule is ( ), and finally obtained the PROTAC drug molecule 15n.

[0136] Embodiment 15

[0137] Based on the contents disclosed in Example 1, the following modifications are further made:

[0138] The R group of the PROTAC drug molecule is ( ), and finally obtained the PROTAC drug molecule 15o.

[0139] Example 16

[0140] Based on the contents disclosed in Example 1, the following modifications are further made:

[0141] The R group of the PROTAC drug molecule is ( ), and finally obtained the PROTAC drug molecule 15p.

[0142] Related experiments and effect data:

[0143] 1. The PROTAC drug molecules obtained in each example were collected and analyzed using nuclear magnetic resonance hydrogen spectrum. The results are as follows:

[0144] PROTAC drug molecule 15a, compound 15a, yellow solid, 55%. 1 H NMR (600MHz, DMSO-d6) δ11.09(s,1H),10.14(s,1H),9.31(s,1H),8.96(s,1H),8.85(t,J=5.4Hz,1H),7.98(d,J=5.8Hz,1H),7.94-7.91 (d,2H),7.80-7.77(d,2H),7.63(d,J=8.5Hz,2H),7.54(dd,J=8.6,7.1Hz,1H),7.20(d,J=8.6Hz,1H),7.06(s,2H),7.00(m,J=8.0,6.1Hz ,3H),6.82(t,J=6.1Hz,1H),6.15(d,J=5.8Hz,1H),5.04(dd,J=12.8,5.5Hz,1H),3.66(s,6H),3.61(s,3H),3.49(m,J=5.8,4.2Hz,2H),3 .45(m,J=5.5Hz,2H),2.87(m,J=17.1,13.9,5.5Hz,1H),2.58(m,J=17.1,3.0Hz,1H),2.52(m,J=13.2Hz,1H),2.01(m,J=5.3,2.4Hz,1H). 13CNMR(100MHz,DMSO)δ172.76,170.04,168.80,167.27,165.23,160.12,159.46,155.81, 152.56,152.56,146.26,146.26,141.65,138.31,137.29,136.90,136.20,132.24,132. 24,132.17,131.02,127.98,127.98,126.69,126.69,122.03,122.03,120.08,120.08,117.14,110.39,109.19,97.39,60.11,55.66,55.66,48.54,36.76,32.05,30.96,22.15.

[0145] PROTAC drug molecule 15b, yellow solid, 60%. 1 H NMR (600MHz, DMSO-d6) δ11.09(s,1H),10.14(s,1H),9.28(s,1H),8.95(s,1H),8.70(t,J=5.7Hz,1H),7.98(d,J=5.7Hz,1H),7.95 (d,J=8.6Hz,2H),7.79(d,J=8.5Hz,2H),7.63(d,J=8.5Hz,2H),7.54(dd,J=8.6,7.1Hz,1H),7.12–7.04(m,3H),7.00(dd,J=8.0,3. 0Hz,3H),6.72(t,J=6.2Hz,1H),6.14(d,J=5.8Hz,1H),5.04(dd,J=12.9,5.5Hz,1H),3.66(s,6H),3.61(s,3H),3.37(m,J=11.9,5 .4Hz,2H),3.32(m,J=6.3Hz,2H),2.92–2.83(m,1H),2.60(m,J=3.3Hz,1H),2.08(s,2H),2.03–1.99(m,1H),1.78(p,J=6.7Hz,2H). 13C NMR (100MHz, DMSO) δ172.79,170.06,168.70,167.28,165.66,160.16,159.41,155.68,15 2.59,152.59,146.33,146.33,141.76,138.06,137.32,136.85,136.17,132.24,132.24, 131.04,128.00,128.00,126.74,126.74,122.13,122.13,120.12,120.12,117.15,110.59,109.38,98.89,97.48,60.14,59.75,55.68,55.68,48.54,41.27,30.97,22.16,14.07.

[0146] PROTAC drug molecule 15c, yellow solid, 62%. 1 H NMR (600MHz, DMSO-d6) δ11.10(s,1H),10.15(s,1H),9.30(s,1H),8.96(s,1H),8.65(t,J=5.7Hz,1H),7.98(d,J=5.7 Hz,1H),7.93(d,J=8.3Hz,2H),7.78(d,J=8.2Hz,2H),7.63(d,J=8.4Hz,2H),7.53(t,J=7.9Hz,1H),7.13–7.04(m,3H) ,7.02–6.97(m,3H),6.56(s,1H),6.14(d,J=6.0Hz,1H),5.04(dd,J=12.9,5.4Hz,1H),3.66(s,6H),3.61(s,3H),3.33 –3.27(m,4H),2.90–2.84(m,1H),2.60(m,J=3.5Hz,1H),2.01(m,J=13.2,5.6,2.8Hz,1H),1.59(m,J=9.5,7.2Hz,4H). 13C NMR(100MHz,DMSO-d6)δ172.82,170.10,168.93,167.30,165.06,160.15,159.47,152.59,1 52.59,146.41,146.41,141.62,138.39,137.31,136.92,136.25,132.20,132.20,131.05,1 27.98,127.98,126.71,126.71,122.06,122.06,120.11,120.11,117.22,110.40,109.04,98.90,97.40,60.15,55.83,55.68,55.68,48.56,41.54,30.98,29.60,26.35,26.23,22.17.

[0147] PROTAC drug molecule 15d, yellow solid, 45%. 1 H NMR (600MHz, DMSO-d6) δ11.09(s,1H),10.14(s,1H),9.30(s,1H),8.96(s,1H),8.62(t,J=5.6Hz,1H),7.97(d,J=5.7Hz,1H),7.93(d,J=8.2Hz,2 H),7.77(d,J=8.1Hz,2H),7.63(d,J=8.5Hz,2H),7.54(dd,J=8.5,7.1Hz,1H),7.11–7.04(m,3H),7.00(dd,J=7.9,3.5Hz,3H),6.53(t,J=5.9Hz, 1H),6.14(d,J=5.8Hz,1H),5.04(dd,J=12.9,5.5Hz,1H),3.66(s,6H),3.61(s,3H),3.25(m,J=18.9,6.7Hz,4H),2.87(m,J=17.1,13.9,5.5Hz,1 H),2.58(m,J=17.9Hz,1H),2.53(m,J=10.5Hz,1H),2.04–1.99(m,1H),1.59(q,J=7.4Hz,2H),1.54(q,J=7.4Hz,2H),1.37(q,J=8.9,5.0Hz,2H). 13C NMR (100MHz, DMSO) δ172.74,170.02,168.91,167.26,165.00,160.11,159.53,152.55,152.5 5,146.39,146.39,141.32,138.40,137.26,136.87,136.21,132.19,132.19,132.15,131.02 ,130.00,127.92,127.92,126.65,126.65,122.01,122.01,120.07,120.07,117.10,110.34,109.01,97.39,60.09,55.65,55.65,48.52,42.13,30.95,30.74,28.62,28.38,23.72,22.05.

[0148] PROTAC drug molecule 15e, yellow solid, 66%. 1 H NMR (600MHz, DMSO-d6) δ11.09(s,1H),10.11(s,1H),9.28(s,1H),8.95(s,1H),8.59(t,J=5.6Hz,1H),7.97(d,J=5.7Hz,1H),7.92(d,J=8.5Hz ,2H),7.77(d,J=8.6Hz,2H),7.62(d,J=8.4Hz,2H),7.54(dd,J=8.6,7.1Hz,1H),7.06(d,J=8.8Hz,3H),7.00(dd,J=8.0,6.2Hz,3H),6.52(t,J= 6.0Hz,1H),6.14(d,J=5.7Hz,1H),5.04(dd,J=12.9,5.5Hz,1H),3.66(s,6H),3.61(s,3H),3.27(q,J=6.7Hz,2H),3.24–3.21(q,2H),2.91–2. 85(m,1H),2.60–2.56(m,1H),2.53(m,J=11.6,3.2Hz,1H),2.03–1.99( m,1H),1.58–1.54(q,2H),1.50(q,J=7.0Hz,2H),1.34(m,J=5.5Hz,4H). 13C NMR (100MHz, DMSO) δ172.84,170.12,168.99,167.33,165.01,160.17,159.51,155.83,152.62 ,152.62,146.44,146.44,141.61,138.47,137.34,136.95,136.26,132.20,132.20,131.08,12 7.99,127.99,126.73,126.73,122.09,122.09,120.13,120.13,117.14,110.40,109.05,98.92,97.41,60.16,55.69,55.69,54.89,48.59,41.82,31.01,28.91,28.65,26.20,26.07,22.19.

[0149] PROTAC drug molecule 15f, yellow solid, 63%. 1 H NMR (600MHz, DMSO-d6) δ11.09(s,1H),10.13(s,1H),9.30(s,1H),8.96(s,1H),8.59(t,J=5.6Hz,1H),7.97(d,J=5.7Hz,1H),7.92(d,J=8.4H z,2H),7.77(d,J=8.3Hz,2H),7.63(d,J=8.4Hz,2H),7.55(dd,J=8.6,7.0Hz,1H),7.06(d,J=8.6Hz,3H),7.00(t,J=7.4Hz,3H),6.51(t,J=6.0 Hz,1H),6.14(d,J=5.8Hz,1H),5.04(dd,J=12.9,5.4Hz,1H),3.66(s,6H),3.61(s,3H),3.28–3.20(m,4H),2.87(m,J=17.0,13.9,5.5Hz,1H) ,2.60–2.56(m,1H),2.53(m,J=8.9Hz,1H),2.01(m,J=12.2,4.9,4.4Hz,1H),1.55(q,J=7.1Hz,2H),1.49(q,J=7.0Hz,2H),1.34–1.30(m,6H). 13CNMR(100MHz,DMSO)δ172.75,170.12,168.98,167.27,164.90,160.28,159.19,152.55,152.55, 152.06,146.58,146.58,146.42,143.66,141.56,135.89,132.43,132.43,131.16,127.92,127.9 2,126.65,126.65,125.53,124.08,122.03,122.03,120.08,120.08,117.55,110.34,109.00,97.36,60.10,55.65,55.65,48.53,41.80,30.75,28.93,28.86,28.62,28.43,26.39,26.24,22.15.

[0150] PROTAC drug molecule 15g, yellow solid, 68%. 1 H NMR (600MHz, DMSO-d6) δ11.10(s,1H),10.13(s,1H),9.28(s,1H),8.95(s,1H),8.59(t,J=5.7Hz,1H),7.98(d,J=6.0Hz,1H),7.93(d,J=8.8Hz, 2H),7.78(d,J=8.8Hz,2H),7.63(d,J=8.5Hz,2H),7.55(t,J=7.8Hz,1H),7.09–7.04(m,3H),7.02–6.98(m,3H),6.50(t,J=5.9Hz,1H),6.14(d, J=5.7Hz,1H),5.05(dd,J=12.9,5.4Hz,1H),3.66(s,6H),3.61(s,3H),3.28–3.24(q,2H),3.22(q,J=7.1,6.7Hz,2H),2.88(m,J=13.2,7.0Hz,2 H),2.61–2.57(m,1H),2.53(m,J=4.5Hz,1H),2.02(m,J=11.8,5.9Hz,1H),1.55(q,J=7.5Hz,2H),1.48(q,J=7.1Hz,2H),1.28(m,J=9.3Hz,8H). 13C NMR (100MHz, DMSO) δ172.77,170.05,168.95,167.28,164.91,160.13,159.47,155.79,152.57,15 2.57,146.43,146.43,141.57,138.44,137.30,136.91,136.24,132.17,132.17,131.03,127.93,1 27.93,126.67,126.67,122.05,122.05,120.08,120.08,117.13,110.36,109.01,98.87,97.39,60.11,55.66,55.66,54.86,48.55,41.82,39.40,30.97,28.88,28.67,28.65,26.38,26.26,22.15.

[0151] PROTAC drug molecule 15h, yellow solid, 60%. 1 H NMR (600MHz, DMSO-d6) δ11.10(s,1H),10.13(s,1H),9.28(s,1H),8.95(s,1H),8.58(t,J=5.6Hz,1H),7.97(d,J=5.8Hz,1H),7.93(d,J=8.5Hz,2 H),7.77(d,J=8.5Hz,2H),7.63(d,J=8.4Hz,2H),7.56(dd,J=8.6,7.0Hz,1H),7.09–7.05(m,3H),7.00(dd,J=8.0,6.6Hz,3H),6.50(t,J=5.9Hz, 1H),6.14(d,J=5.8Hz,1H),5.05(dd,J=12.8,5.5Hz,1H),3.66(s,6H),3 .61(s,3H),3.27–3.24(q,2H),3.21(q,J=6.7Hz,2H),2.91–2.85(m,1H) ,2.59(m,J=17.0,3.0Hz,1H),2.56–2.51(m,1H),2.02(m,J=12.8,5.2,2 .3Hz,1H),1.54(q,J=7.2Hz,2H),1.50–1.46(q,2H),1.31–1.24(m,10H). 13C NMR (100MHz, DMSO) δ172.76,170.05,168.94,167.28,164.89,160.12,159.46,155.79,152.56,152. 56,146.42,146.42,141.56,138.43,137.30,136.91,136.23,132.16,132.16,131.02,127.92,127. 92,126.66,126.66,122.06,122.06,120.07,120.07,117.13,110.35,109.01,98.86,97.36,60.10,55.65,55.65,54.86,48.54,41.82,39.94,39.31,30.96,28.89,28.66,28.66,26.41,26.26,22.14.

[0152] PROTAC drug molecule 15i, yellow solid, 67%. 1 H NMR (600MHz, DMSO-d6) δ11.10(s,1H),10.16(s,1H),9.36(s,1H),8.97(s,1H),8.61(t,J=5.6Hz,1H),7.97(d,J=5.7Hz,1H),7.94(d,J=8.2Hz ,2H),7.79(d,J=8.2Hz,2H),7.65(d,J=8.4Hz,2H),7.55(dd,J=8.6,7.1Hz,1H),7.10–7.04(m,3H),7.03–6.99(m,3H),6.49(t,J=5.8Hz,1H), 6.17(d,J=5.8Hz,1H),5.05(dd,J=12.8,5.5Hz,1H),3.66(s,6H),3.62(s,3H),3.27–3.23(q,2H),3.21(q,J=6.5Hz,2H),2.91–2.87(m,1H),2 .59(m,J=17.0,3.1Hz,1H),2.53(m,J=13.3,4.4Hz,1H),2.05–2.00(m,1H),1.54(q,J=7.2Hz,2H),1.47(q,J=7.0Hz,2H),1.29–1.21(m,12H). 13C NMR (100MHz, DMSO) δ172.77,170.05,168.96,167.29,164.90,160.15,159.41,155.63,152.58,152.5 8,146.43,146.43,141.59,138.43,137.31,136.89,136.23,132.21,132.21,132.17,131.06,127.93, 127.93,126.67,126.67,122.07,122.07,120.09,120.09,117.11,110.36,109.03,97.43,60.11,55.66,55.66,54.86,48.56,41.84,30.98,28.92,28.90,28.71,28.65,28.65,26.43,26.28,22.16,15.13.

[0153] PROTAC drug molecule 15j, yellow solid, 55%. 1 H NMR (600MHz, DMSO-d6) δ11.10(s,1H),10.14(s,1H),9.29(s,1H),8.96(s,1H),8.69(t,J=5.6Hz,1H),7.98(d,J=5.8Hz,1H),7.94 (d,J=8.3Hz,2H),7.78(d,J=8.3Hz,2H),7.63(d,J=8.2Hz,2H),7.52(dd,J=8.6,7.1Hz,1H),7.12–7.07(m,3H),7.00(dd,J=7.8,4. 4Hz,3H),6.60(t,J=5.9Hz,1H),6.14(d,J=5.8Hz,1H),5.04(dd,J=12.9,5.5Hz,1H),3.67(s,6H),3.63(t,2H),3.61(s,3H),3.56 (t,J=6.0Hz,2H),3.44(m,J=17.6,5.8Hz,4H),2.91–2.85(m,1H),2.59(m,J=17.3,3.4Hz,1H),2.53(m,J=13.4,4.6Hz,1H),2.01(m J=10.6,5.5,3.2,2.8Hz,1H). 13C NMR (100MHz, DMSO) δ172.75,170.03,168.92,167.25,165.18,160.13,159.47,155.79,152 .57,152.57,146.39,146.39,141.69,138.10,137.32,136.92,136.17,132.17,132.17,132 .06,131.01,128.01,128.01,126.68,126.68,122.10,122.10,120.09,120.09,117.38,110.66,109.25,97.37,68.71,68.60,60.12,55.66,55.66,54.86,48.57,41.66,30.96,22.13.

[0154] PROTAC drug molecule 15k, yellow solid, 62%. 1 H NMR (600MHz, DMSO-d6) δ11.10(s,1H),10.14(s,1H),9.31(s,1H),8.97(s,1H),8.67(t,J=5.6Hz,1H),7.97(d,J=5.7Hz,1H),7.94(d,J=8 .2Hz,2H),7.77(d,J=8.2Hz,2H),7.63(d,J=8.4Hz,2H),7.56–7.52(m,1H),7.10–7.05(m,3H),7.01(dd,J=15.2,7.8Hz,3H),6.58(t,J=5 .9Hz,1H),6.15(d,J=5.8Hz,1H),5.05(dd,J=12.8,5.5Hz,1H),3.67(s,6H),3.61(s,3H),3.58(t,J=5.4Hz,2H),3.54(m,J=2.6Hz,4H),3 .51(t,J=5.9Hz,2H),3.42–3.38(m,4H),2.87(m,J=16.9,13.7,5.5Hz,1H),2.60–2.56(m,1H),2.53(m,J=4.6Hz,1H),2.05–1.99(m,1H). 13C NMR (100MHz, DMSO) δ172.75,170.04,168.92,167.26,165.10,160.12,159.37,155.61,152.57 ,152.57,146.36,146.36,141.67,138.10,137.29,136.85,136.16,132.21,132.21,132.07,13 1.03,127.97,127.97,126.68,126.68,122.09,122.09,120.09,120.09,117.34,110.65,109.26,97.43,69.65,69.57,68.84,68.73,60.11,55.66,55.66,48.56,41.67,40.15,30.96,22.12.

[0155] PROTAC drug molecule 15l, yellow solid, 45%. 1 H NMR (600MHz, DMSO-d6) δ11.08(s,1H),10.11(s,1H),9.26(s,1H),8.93(s,1H),8.64(t,J=5.6Hz,1H),7.97(d,J=5.7Hz,1H),7.94(d, J=8.3Hz,2H),7.78(d,J=8.2Hz,2H),7.63(d,J=8.5Hz,2H),7.55(t,J=7.9Hz,1H),7.14–7.05(m,3H),7.03(d,J=7.0Hz,1H),7.00(d,J =8.7Hz,2H),6.57(t,J=5.9Hz,1H),6.14(d,J=5.8Hz,1H),5.05(dd,J=12.8,5.5Hz,1H),3.67(s,6H),3.62(s,3H),3.57(t,J=5.5Hz, 2H),3.50(m,J=5.0,4.3Hz,10H),3.40(m,J=17.7,5.7Hz,4H),2.92–2.84(m,1H),2.62–2.51(m,2H),2.02(m,J=13.4,6.5,3.4Hz,1H). 13CNMR(100MHz,DMSO)δ172.78,170.05,168.93,167.28,165.12,160.13,159.47,155.79,152.58,15 2.58,146.38,146.38,141.68,138.11,137.35,136.94,136.18,132.15,132.15,132.08,131.00,1 27.99,127.99,126.70,126.70,122.13,122.13,120.07,120.07,117.37,110.67,109.26,98.89,97.33,69.75,69.58,68.84,68.68,60.12,55.66,55.66,54.87,54.87,48.57,41.68,30.98,22.15.

[0156] PROTAC drug molecule 15m, yellow solid, 56%. 1 H NMR (600MHz, DMSO-d6) δ11.11(s,1H),10.18(s,1H),9.35(s,1H),9.00(s,1H),7.98(d,J=5.7Hz,1H),7.79(d,J=8.2Hz,2 H),7.71–7.65(m,3H),7.63(d,J=8.3Hz,2H),7.38(d,J=7.1Hz,1H),7.28(d,J=8.4Hz,1H),7.07(s,2H),7.03(d,J=8.8Hz ,2H),6.16(d,J=5.8Hz,1H),5.10(dd,J=12.8,5.5Hz,1H),3.87–3.77(m,2H),3.69(m,J=11.7Hz,2H),3.66(s,6H),3.61( s,3H),3.44–3.41(m,2H),2.90–2.84(m,1H),2.58(m,J=17.1,3.4Hz,1H),2.53(m,J=13.2,4.7Hz,1H),2.04–1.99(m,1H). 13C NMR (100MHz, DMSO) δ172.75,169.92,167.66,166.97,166.30,160.16,159.36,155.55,152 .58,152.58,149.25,149.25,140.45,139.71,137.25,136.82,135.90,133.58,132.23,132 .23,131.17,127.79,127.79,126.91,126.91,123.83,122.04,122.04,120.19,120.19,117.04,115.34,97.46,60.13,55.67,55.67,54.87,48.84,31.52,30.94,29.43,28.99,22.02.

[0157] PROTAC drug molecule 15n, yellow solid, 45%. 1 H NMR (600MHz, DMSO-d6) δ11.08(s,1H),10.13(s,1H),9.29(s,1H),8.95(s,1H),8.54(d,J=7.7Hz,1H),7.97(dd,J=7.0,4.9Hz ,3H),7.78(d,J=8.2Hz,2H),7.71–7.66(m,1H),7.63(d,J=8.4Hz,2H),7.34(t,J=8.5Hz,2H),7.07(s,2H),7.00(d,J=8.6Hz,2 H),6.14(d,J=5.8Hz,1H),5.09(dd,J=12.8,5.5Hz,1H),4.03–3.97(m,1H),3.71(d,J=11.6Hz,2H),3.67(s,6H),3.61(s,3H) ,3.00(t,J=12.2Hz,2H),2.90–2.83(m,1H),2.62–2.52(m,2H),2.06–2.00(m,1H),1.91(d,J=11.9Hz,2H),1.82–1.73(m,2H). 13C NMR (100MHz, DMSO) δ172.79,170.02,167.08,166.32,164.50,160.13,155.98,152.57,152.5 7,149.84,141.63,141.63,138.37,137.32,136.94,135.75,133.65,132.32,132.32,131.00, 128.14,128.14,126.66,126.66,125.02,124.05,122.09,122.09,120.07,120.07,116.50,114.62,113.50,98.41,97.37,60.13,55.67,55.67,50.03,49.12,46.93,31.27,28.99,22.08.

[0158] PROTAC drug molecule 15o, yellow solid, 59%. 1 H NMR (600MHz, DMSO-d6) δ11.11(s,1H),10.15(s,1H),9.30(s,1H),8.96(s,1H),8.51(d,J=7.6Hz,1H),8.01–7.92(m,3H),7.77(d,J=8.5Hz, 2H),7.66–7.60(d,2H),7.61–7.58(m,1H),7.14(d,J=8.7Hz,1H),7.12–7.02(m,3H),7.00(d,J=8.9Hz,2H),6.45(d,J=7.9Hz,1H),6.14(d,J =5.8Hz,1H),5.05(dd,J=12.9,5.5Hz,1H),3.95(m,J=8.2,5.0,4.1Hz,1H),3.83–3.78(m,1H),3.66(s,6H),3.60(s,3H),2.88(m,J=17.1,13 .7,5.5Hz,1H),2.59(m,J=16.9,3.6Hz,1H),2.53(m,J=13.3Hz,1H),2.02(m,J=10.8,5.6,2.9Hz,1H),1.84–1.70(m,6H),1.66–1.59(m,2H). 13C NMR (100MHz, DMSO) δ172.77,170.02,169.36,167.19,164.63,160.13,159.47,155.41,152 .56,152.56,145.46,145.46,141.56,138.52,137.29,136.90,136.46,132.17,132.17,132 .13,131.03,128.23,128.23,126.55,126.55,122.09,122.09,120.10,120.10,117.48,110.67,109.17,97.40,60.10,55.65,55.65,48.57,46.63,31.51,29.41,27.93,27.17,22.14.

[0159] PROTAC drug molecule 15p, yellow solid, 64%. 1 H NMR (600MHz, DMSO-d6) δ11.10(s,1H),10.20(s,1H),9.31(s,1H),8.97(s,1H),7.97(d,J=5.7Hz,1H),7.82(d,J=8.4Hz,2H),7.79( d,J=8.5Hz,2H),7.69–7.61(m,3H),7.32(d,J=7.0Hz,1H),7.28(d,J=8.5Hz,1H),7.07(s,2H),7.01(d,J=8.8Hz,2H),6.14(d,J=5. 7Hz,1H),5.09(dd,J=12.8,5.6Hz,1H),4.06(m,J=6.2Hz,2H),3.82(m,2H),3.65(s,6H),3.61(s,3H),3.24(m,J=5.3Hz,3H),3.20– 3.09(m,3H),2.88(m,J=13.2Hz,2H),2.85(m,J=12.8,4.3Hz,1H),2.61–2.57(m,1H),2.56–2.51(m,1H),2.02(m,J=7.2,4.0Hz,1H). 13C NMR (100MHz, DMSO) δ172.75,169.98,167.36,167.04,166.28,160.13,159.47,155.80,152.57 ,152.57,149.87,149.87,141.54,137.28,137.01,136.91,135.73,133.63,132.19,132.19,13 1.05,128.56,128.56,126.74,126.74,123.94,121.95,121.95,120.14,120.14,116.58,114.62,97.41,60.12,55.67,55.67,54.87,48.79,47.90,34.72,33.32,31.52,30.94,29.42,22.06.

[0160] 2. For the PROTAC drug molecules obtained above, the anti-proliferative effect on human glioblastoma cells (SW-1783) was investigated in vitro. The results are shown in Table 1. Among them, the detection method of the anti-proliferative effect on human glioblastoma cells (SW-1783): MTT method was used to detect the anti-proliferative activity of cells; for specific methods, refer to Eur J Med Chem. 2021 Jan 15: 210: 113088. doi: 10.1016 / j.ejmech.2020.113088.

[0161] Table 1 Antiproliferative effects of various PROTAC drug molecules on human glioma SW-1783 cells

[0162]

[0163] In Table 1, a Each compound was tested 3 times, and the values ​​are mean ± error. b The cell action time is 24h and 48h.

[0164] As can be seen from Table 1, PROTAC drug molecule 15b with molecular structure 2 has the strongest anti-proliferative activity against human astrocytoma cells (SW-1783), which shows that a PROTAC drug molecule provided in the examples of the present application can be used as a new targeted therapeutic drug for brain glioma.

[0165] The above is only a specific implementation of the present application, so that those skilled in the art can understand or implement the present application. It will be apparent to those skilled in the art that various modifications to these embodiments are possible, and the general principles defined in the present application can be implemented in other embodiments without departing from the spirit or scope of the present application. Therefore, the present application will not be limited to these embodiments shown in the present application, but will conform to the widest scope consistent with the principles and novel features applied for by the present application.

Claims

1. A PROTAC drug molecule targeting Fam20C, characterized in that: The PROTAC drug has a molecular structure as shown in Formula 1, In Formula 1, the R group is:

2. The PROTAC drug molecule according to claim 1, characterized in that The R group is The PROTAC drug molecule has a molecular structure as shown in Formula 2, 3. A method for preparing a PROTAC drug molecule according to claim 1 or 2, characterized in that: The method comprises: Mixing 3,4,5-trimethoxyaniline and N,N-diisopropylethylamine, and then adding 2,4-dichloropyrimidine to carry out a first reaction to obtain a first intermediate; The p-phenylenediamine and the first intermediate are mixed, and then 1,4-dioxane and acetic acid are added to carry out a second reaction to obtain a second intermediate; Adding 4-(chlorosulfonyl)benzoic acid to the second intermediate to carry out a third reaction, followed by purification to obtain a third intermediate; Conducting a fourth reaction of 4-fluoroisobenzofuran-1,3-dione and 3-amino-2,6-piperidindione, followed by purification, to obtain a fourth intermediate; Mixing R-group tert-butyl formate and the fourth intermediate, and then adding N,N-diisopropylethylamine to carry out a fifth reaction to obtain a fifth intermediate having an R group; Performing a sixth reaction on a hydrochloric acid solution of 1,4-dioxane and the fifth intermediate to obtain a sixth intermediate; Mixing 2-(7-azabenzotriazole)-N,N,N',N'-tetramethyluronium hexafluorophosphate, the sixth intermediate and the third intermediate, adding tetrahydrofuran and N,N-diisopropylethylamine to carry out a seventh reaction, and then purifying to obtain a PROTAC drug molecule; The first intermediate is 2-chloro-N-(3,4,5-trimethoxyphenyl)pyrimidin-4-amine; the second intermediate is N2-(4-aminophenyl)-N4-(3,4,5-trimethoxyphenyl)pyrimidine-2,4-diamine; the third intermediate is 4-(N-(4-((4-((3,4,5-trimethoxyphenyl)amino)pyrimidin-2-yl)amino)phenyl)aminosulfonyl)benzoic acid; The fourth intermediate is 2-(2,6-dioxopiperidin-3-yl)-4-fluoroisoindoline-1,3-dione; the fifth intermediate is tert-butyl 4-((2-(2,6-dioxopiperidin-3-yl)-1,3-dioxoisoindoline-4-yl)amino)R-formate.

4. The method according to claim 3, characterized in that The temperature of the first reaction is 80° C., and the time of the first reaction is 8 h to 12 h; The temperature of the second reaction is 110° C., and the time of the second reaction is 2 h; The temperature of the third reaction is 20° C. to 30° C., and the time of the third reaction is 0.5 h.

5. The method according to claim 3, characterized in that: The temperature of the fourth reaction is 120° C., and the time of the fourth reaction is 12 h; The temperature of the fifth reaction is 90° C., and the time of the fifth reaction is 2 h; The temperature of the sixth reaction is 60° C., and the time of the sixth reaction is 8 h to 12 h; The temperature of the seventh reaction is 30° C., and the time of the seventh reaction is 8 h to 12 h.

6. The method according to claim 3, characterized in that The yield of the PROTAC drug molecule is 61% to 78%.

7. An application of a PROTAC drug molecule targeting Fam20C, characterized in that: Use of the PROTAC drug molecule according to claim 1 or 2 in the preparation of a therapeutic drug for brain glioma cells.

8. A pharmaceutical composition, characterized in that The pharmaceutical composition comprises the PROTAC drug molecule according to claim 1 or 2 or a pharmaceutically acceptable salt thereof.

Images