Compounds targeting ubiquitination degradation of Pin1 protein and pharmaceutical compositions and applications thereof

Abstract

The present invention discloses a compound for targeted ubiquitination degradation of Pin1 protein, a pharmaceutical composition and application thereof. Its structure is shown in Formula I, and it has the activity of inhibiting Pin1 protein activity and degrading Pin1 protein, and can effectively inhibit the malignant proliferation of leukemia MV-4-11 cells. Therefore, the present invention can be used for diseases related to abnormal expression of Pin1 protein, such as various cancers.

Description

Technical Field

[0001] The present invention relates to the field of pharmaceutical chemistry, and in particular to a class of compounds for targeted ubiquitination degradation of Pin1 protein, and a pharmaceutical composition and application thereof. Background Art

[0002] Leukemia is a type of malignant clonal disease of hematopoietic stem cells and is a blood tumor. In addition to the characteristics of ordinary tumors that they grow quickly, are not easy to die, and can metastasize to other organs, unlike solid tumors such as liver cancer and gastric cancer, leukemia cannot be treated by surgical resection, and the therapeutic effects of chemotherapy and radiotherapy cannot be guaranteed, so it is more difficult to treat. Because the pancreas is a gland located between the stomach and the spine, deep in the abdomen, patients have atypical early symptoms and hidden onset; most patients are already in the late stage of the disease when diagnosed and lose the opportunity for surgery, resulting in a poor prognosis for pancreatic cancer. Most patients survive only about 1 year after diagnosis. Difficult diagnosis and poor treatment are the main reasons why pancreatic cancer is called the "king of cancer."

[0003] Abnormal activation or overexpression of Pin1 protein function usually (1) Proline isomerase Pin1 regulates multiple cancer-related pathways. In various tumors, abnormal activation of Pin1 can directly or indirectly cause the activation of 60 oncoproteins and the inhibition of more than 30 inhibitory factors, thereby promoting tumor formation, including hematological tumors and pancreatic cancer. Pin1 gene knockout mice develop normally and tumor formation is significantly inhibited. Therefore, Pin1 is a highly potential anti-tumor target.

[0004] Ubiquitin-mediated protein degradation is the most important negative regulation of intracellular proteins. The ubiquitin-proteasome system (UPS) is responsible for cleaning up misused or harmful proteins in cells. It is the "cleaner" in cells and maintains the balance of intracellular proteins. Proteolysis targeting chimeras (PROTACs) are a class of heterobifunctional molecules that can simultaneously bind to proteins of interest and E3 ubiquitin ligases, ubiquitinate and degrade proteins of interest. It has the characteristics of catalytic degradation and low dosage.

[0005] Therefore, by designing and synthesizing an inverse agonist with stronger activity against Pin1 protein, and applying the proteolysis targeting chimera (PROTACs) technology to Pin1 protein, it can provide important references for Pin1 as a drug development target, the treatment of leukemia, and the feasibility of the development and application of PROTAC technology. Summary of the invention

[0006] In view of the above problems, one of the objects of the present invention is to provide a novel compound or a pharmaceutically acceptable salt, hydrate or solvate thereof for targeting ubiquitination degradation of Pin1 protein.

[0007] Based on the above purpose, the present invention provides the following technical solutions:

[0008] In one aspect, the present invention provides a compound having a structure as shown in Formula I or a pharmaceutically acceptable salt, hydrate or solvate thereof:

[0009]

[0010] in:

[0011] R 1 Selected from

[0012] R 2 (-CH2-) n , n is selected from an integer of 1-4; that is, R 2 It can be (-CH2-)1, (-CH2-)2, (-CH2-)3, (-CH2-)4;

[0013] L is selected from C 4-18 Alkyl, alkoxy or -R 3 -C(=O)NH-R 4 - a group consisting of; L is selected from C 4-18 When alkyl, it includes but is not limited to C4 alkyl, C5 alkyl, C6 alkyl, C7 alkyl, C8 alkyl, C9 alkyl, C 10 Alkyl, C 11 Alkyl, C 12 Alkyl, C 13 Alkyl, C 14 Alkyl, C 15 Alkyl, C 16 Alkyl, C 17 Alkyl, C 18 alkyl.

[0014] R 3 Selected from C 1-6 Alkyl, specifically, C1 alkyl, C2 alkyl, C3 alkyl, C4 alkyl, C5 alkyl, C6 alkyl;

[0015] R 4 Selected from C 3-14 Alkyl or -CH2-(-CH2-O-CH2-) m -CH2-, m is an integer selected from 1 to 4; R 4 Specifically, it can be C3 alkyl, C4 alkyl, C5 alkyl, C6 alkyl, C7 alkyl, C8 alkyl, C9 alkyl, C 10 Alkyl, C 11 Alkyl, C 12 Alkyl, C 13 Alkyl, C 14Alkyl, -CH2-(-CH2-O-CH2-)1-CH2-, -CH2-(-CH2-O-CH2-)2-CH2-, -CH2-(-CH2-O-CH2-)3-CH2-, -CH2-(-CH2-O-CH2-)4-CH2-; R 4 Preferred C 12 Alkyl or -CH2-(-CH2-O-CH2-)3-CH2-;

[0016] X is -CH2-, -NH- or -O-;

[0017] A Choose any of the following groups:

[0018] A-1 is particularly preferred.

[0019] In addition, generally speaking, L has no directionality and can be connected to amide on the left side and to A on the right side, or it can be connected to A on the left side and to amide on the right side. However, in some preferred cases, when L is -R 3 -C(=O)NH-R 4 -, preferably R 3 The side is connected to the amide, R 4 The side is connected to X.

[0020] Preferably, the compound or its pharmaceutically acceptable salt, hydrate or solvate has the structure shown in Formula II:

[0021]

[0022] L is selected from C 4-18 Alkyl, alkoxy or -R 3 -C(=O)NH-R 4 - the groups formed;

[0023] R 3 Selected from C 1-6 alkyl;

[0024] R 4 Selected from C 3-14 Alkyl or -CH2-(-CH2-O-CH2-) m -CH2-, m is an integer selected from 1 to 4, m is preferably 3;

[0025] X is -CH2-, -NH- or -O-;

[0026] A Choose any of the following groups:

[0027]

[0028] Preferably, X is -NH-.

[0029] Preferably, the compound or its pharmaceutically acceptable salt, hydrate or solvate has a structure shown in Formula III:

[0030]

[0031] X is -CH2-, -NH- or -O-;

[0032] n is an integer selected from 1 to 4;

[0033] A Choose any of the following groups:

[0034]

[0035] Preferably, the compound or its pharmaceutically acceptable salt, hydrate or solvate has a structure shown in Formula IV:

[0036]

[0037] n is an integer selected from 1 to 4;

[0038] L is selected from C 4-18 Alkyl, alkoxy or -R 3 -C(=O)NH-R 4 - the groups formed;

[0039] R 3 Selected from C 1-6 alkyl;

[0040] R 4 Selected from C 3-14 Alkyl or -CH2-(-CH2-O-CH2-) m -CH2-, m is an integer selected from 1 to 4;

[0041] X is -CH2-, -NH- or -O-.

[0042] Preferably, the compound or its pharmaceutically acceptable salt, hydrate or solvate has the structure shown in Formula V:

[0043]

[0044] L is selected from C 4-18 Alkyl, alkoxy or -R 3 -C(=O)NH-R 4 - the groups formed;

[0045] R 3 Selected from C 1-6 alkyl;

[0046] R 4 Selected from C 3-14Alkyl or -CH2-(-CH2-O-CH2-) m -CH2-, m is an integer selected from 1 to 4;

[0047] X is -CH2-, -NH- or -O-.

[0048] More preferably, X is -NH-.

[0049] More preferably, L is -CH2CH2CONH(-CH2-) 12 -.

[0050] Preferably, the compound or its pharmaceutically acceptable salt, hydrate or solvate is selected from:

[0051]

[0052]

[0053]

[0054] Another aspect of the present invention provides a use of the above compound or a pharmaceutically acceptable salt, hydrate or solvate thereof in the preparation of a drug for preventing or treating a disease associated with abnormal expression of Pin1 protein activity.

[0055] Preferably, the diseases associated with abnormal expression of Pin1 protein activity include: tumors, hyperglycemia, diabetes, obesity, hyperlipidemia, hypercholesterolemia, hyperlipoproteinemia, hypertriglyceridemia, hypertension, hyperinsulinemia, hyperuricemia, Parkinson's disease, and Alzheimer's disease.

[0056] Another aspect of the present invention provides a use of the above compound or its pharmaceutically acceptable salt, hydrate or solvate in the preparation of a drug for preventing or curing tumors or preventing postoperative recurrence of tumors.

[0057] Preferably, the tumor is one or more of: non-small cell lung cancer, malignant melanoma, prostate cancer, kidney cancer, bladder cancer, ovarian cancer, colon cancer, rectal cancer, breast cancer, cervical cancer, lung cancer, laryngeal cancer, nasopharyngeal cancer, pancreatic cancer or multiple myeloma, B lymphoma, leukemia.

[0058] Another aspect of the present invention provides a composition comprising any one of the above compounds or a pharmaceutically acceptable salt, hydrate or solvate thereof.

[0059] Based on the above scheme, the present invention has the following beneficial effects:

[0060] The compound provided by the present invention has the ability to inhibit the activity of Pin1 protein and degrade Pin1 protein, and can effectively inhibit the malignant proliferation of leukemia MV-4-11 cells. Therefore, the present invention can be used for diseases related to abnormal expression of Pin1 protein, such as various cancers.

[0061] This compound can effectively inhibit Pin1 protein and has the function of degrading Pin1 target protein. Its protein degradation mechanism is that the molecule simultaneously binds to Pin1 protein and E3 ubiquitin ligase, and then ubiquitinates and degrades the target protein. This compound can significantly inhibit the proliferation of tumor cells and can provide an important reference for the research and development of Pin1 as a drug development target. The treatment of leukemia and the feasibility of the development and application of PROTAC technology. BRIEF DESCRIPTION OF THE DRAWINGS

[0062] Figure 1 The chemical structures of fragments S1-S6.

[0063] Figure 2 The inhibitory effect of Sulfopin and compound 1 on the proliferation of various acute myeloid leukemia cells. DETAILED DESCRIPTION

[0064] The following will explain the implementation methods of the present application in detail through examples, so that the implementation process of how the present application uses technical means to solve technical problems and achieve technical effects can be fully understood and implemented accordingly.

[0065] The raw materials and equipment used in this application, unless otherwise specified, are all commonly used raw materials and equipment in this field, and are all commercially available products. The methods used in this application, unless otherwise specified, are all conventional methods in this field.

[0066] There are many other feasible technical solutions in the present application, which are not listed one by one here. The technical solutions claimed in the claims of the present application are all feasible.

[0067] "Comprising" or "including" is intended to indicate that compositions (e.g., media) and methods include the listed elements, but do not exclude other elements. When used to define compositions and methods, "consisting essentially of" means excluding other elements that have any significance for the combination for the stated purpose. Thus, a composition consisting essentially of the elements defined herein does not exclude other materials or steps that do not materially affect the basic and novel features of the claimed application. "Consisting of" means excluding trace elements and substantial method steps of other components. Embodiments defined by each of these transition terms are within the scope of the present application.

[0068] The present invention will be further described below in conjunction with the accompanying drawings.

[0069] Example 1: Synthesis of M6

[0070]

[0071] Step 1: Synthesize M2

[0072] Compound M1 (1.03 g, 10 mmol) and mono-tert-butyl succinate (1.74 g, 10 mmol) were dissolved in N, N-dimethylformamide (10 ml), and 2-(7-azabenzotriazole)-N, N, N', N'-tetramethyluronium hexafluorophosphate (3.8 g, 10 mmol) and N, N-diisopropylethylamine (2.6 ml, 15 mmol) were added, and stirred at room temperature for 2 hours. After the reaction was completed, water and ethyl acetate were added for extraction, and the organic phases were combined, washed with water and saturated brine, dried over anhydrous sodium sulfate, the solvent was spin-dried, and purified by column chromatography to obtain a white solid M2 (2.07 g, 80% yield). 1 HNMR(500MHz,Chloroform-d)δ6.40–6.28(m,1H),4.02–3.90(m,1H),3.11(d,J=5.9Hz,2H),3 .06(d,J=6.8Hz,2H),2.57(t,J=6.6Hz,2H),2.44(t,J=6.6Hz,2H),1.42(s,9H),0.85(s,6H). 13 C NMR(126MHz,Chloroform-d)δ173.69,172.32,81.00,67.95,46.43,36.83,31.21,30.96,28.04,22.61.UPLC-MS(ESI)for[M+H] + 260.18.

[0073] Step 2: Synthesize M3

[0074] The intermediate M2 (1.55 g, 6 mmol) was dissolved in dichloromethane (10 mL), and then the Dess-Martin oxidation reagent (2.5 g, 6 mmol, dissolved in 20 mL dichloromethane) was slowly added. The mixed solution was stirred at room temperature for 5 hours. After the reaction was complete, sodium thiosulfate solution (20 mL, 0.5 M) was added until the system was clear, and then washed with water (20 mL), saturated brine (20 mL), concentrated and purified by column chromatography to obtain compound M3 as a white solid (1.33 g, 86% yield). 1H NMR (500MHz, Chloroform-d) δ9.42(s,1H),3.34(d,J=6.5Hz,2H),2.53(t,J=6.7Hz,2H),2.38(t,J=6.8Hz,2H),1.42(s,9H),1.07(s,6H). 13 CNMR(126MHz,Chloroform-d)δ205.30,172.24,171.96,80.80,47.50,44.15,31.25,30.82,28.04,19.61.UPLC-MS(ESI)for[M+H] + 258.18.

[0075] Step 3: Synthesize M4

[0076] Compound M3 (1.29 g, 5 mmol) was dissolved in 1,2-dichloroethane (8 mL), followed by the addition of 3-aminocyclopentane hydrochloride (CAS: 51642-03-6, 858 mg, 5 mmol) and sodium triacetoxyborohydride (3.18 g, 15 mmol), and the mixed solution was stirred at room temperature for 1 day. After the reaction was complete, saturated sodium bicarbonate solution was added to quench, extracted with ethyl acetate (3×10 mL), and the organic phases were combined and washed with water and saturated brine, respectively. The solvent was dried and purified by column chromatography to obtain compound M4 as a white solid (2.8 g, 67% yield). 1 H NMR(500MHz,Chloroform-d)δ6.53(s,1H),3.55–3.47(m,1H),3.36–3.30(m,1H),3.23–3.14(m,2H),3.09–2.98 (m,3H),2.60–2.53(m,2H),2.44–2.37(m,3H),2.36–2.31(m,2H),2.20–2.08(m,1H),1.43(s,9H),0.90(s,6H). 13 C NMR(126MHz,Chloroform-d)δ172.50,172.28,80.73,56.84,56.14,55.80,50.39 ,47.58,35.26,31.36,30.96,29.79,28.11,24.33,24.20.UPLC-MS(ESI)for[M+H] + 377.21.

[0077] Step 4: Synthesize M5

[0078] Intermediate M4 (1.88 g, 5 mmol) was dissolved in dichloromethane (10 mL), and acetyl chloride (355 μL, 5 mmol) and triethylamine (837 μL, 6 mmol) were slowly added under ice bath conditions. The mixed solution was stirred at room temperature for 1 hour. After the reaction was complete, water was added to quench, and dichloromethane (3×10 mL) was used for extraction. The organic phases were combined and washed with water (10 mL) and saturated brine (10 mL). The organic phase was dried and purified by column chromatography to obtain compound M5 as a white solid (2.08 g, 92% yield). 1 HNMR(500MHz,Chloroform-d)δ6.26–6.13(m,1H),4.17–4.06(m,2H),3.97–3.86(m,1H),3.74–3.62(m,2H),3.34(d,J=15.8Hz,1H),3.22(d . 13 C NMR(126MHz,Chloroform-d)δ172.77,172.59,167.95,81.10,58.48,57.66,50.40,49.10 ,48.20,42.29,38.04,31.30,30.63,28.07,26.58,23.87,23.80.UPLC-MS(ESI)for[M+H] + 453.18.

[0079] Step 5: Synthesize M6

[0080] Intermediate M5 (452 ​​mg, 1 mmol) was dissolved in dichloromethane (10 ml), trifluoroacetic acid (3 ml) was added, and stirred at room temperature for 2 hours. After monitoring the reaction was complete, the reaction was stopped, and the solvent was dried to obtain viscous intermediate M6, which was directly used in the next step without purification. UPLC-MS (ESI) for [M+H] + 397.22.

[0081] Example 2: Synthesis of Compound 1

[0082]

[0083] Intermediate M6 (1 mmol) and fragment S1 (chemical structure of S1-S6 see Figure 1) (1mmol) was dissolved in N,N-dimethylformamide (4ml), and 2-(7-azabenzotriazole)-N,N,N',N'-tetramethyluronium hexafluorophosphate (1mmol) and N,N-diisopropylethylamine (1.5mmol) were added, and the mixture was reacted at room temperature for 2 hours. After the reaction was completed, water (10ml) was added, and the mixture was extracted with ethyl acetate (3×10ml). The organic phases were combined, washed with water and saturated brine, dried over anhydrous sodium sulfate, and the solvent was removed by rotation. Compound 1 was obtained as a yellow solid with a yield of 84%. 1 H NMR(500MHz,Methanol-d4)δ7.55(d,J=8.4Hz,1H),6.97(d,J=2.2Hz,1H),6.82(dd,J=8.4,2.2Hz,1H) ,5.03(dd,J=12.7,5.5Hz,1H),4.32–4.21(m,2H),4.15–3.98(m,1H),3.68–3.49(m,2H),3.40–3.32(m, 2H),3.23–3.17(m,2H),3.16–3.04(m,5H),2.90–2.80(m,1H),2.78–2.65(m,2H),2.57–2.51(m,2H),2 .49(s,4H),2.13–2.06(m,1H),1.71–1.59(m,2H),1.52–1.40(m,4H),1.37–1.26(m,16H),0.98(s,6H). 13 C NMR(126MHz,Methanol-d4)δ173.90,173.30,172.99,170.37,168.97,168.29, 167.93,154.79,134.55,124.83,116.54,115.19,105.34,57.90,57.82,50.52, 49.17,48.90,42.78,42.08,39.10,37.50,30.81,30.59,29.21,29.02,28.99, 28.42,26.68,26.57,26.12,22.66,22.46.HPLC>95%; HRMS(ESI-TOF)m / z:[M+H] + Calculate for C 40 H 60 ClN6O9S + ,835.3826;found,835.3832.

[0084] Example 3: Synthesis of Compound 2

[0085]

[0086] The intermediate M6 (1 mmol) and the fragment S2 (1 mmol) were dissolved in N, N-dimethylformamide (4 ml), and 2-(7-azabenzotriazole)-N, N, N', N'-tetramethyluronium hexafluorophosphate (1 mmol) and N, N-diisopropylethylamine (1.5 mmol) were added, and the mixture was reacted at room temperature for 2 hours. After the reaction was completed, water (10 ml) was added, and the mixture was extracted with ethyl acetate (3×10 ml). The organic phases were combined, washed with water and saturated brine, dried over anhydrous sodium sulfate, and the solvent was removed by rotation. Compound 2 was purified and separated by column chromatography to obtain a yellow solid with a yield of 87%. 1 H NMR (500MHz, Methanol-d4) δ7.54(t,J=7.8Hz,1H),7.03(d,J=7.3Hz,2H),5.05(dd,J=12.4,5 .5Hz,1H),4.32–4.22(m,2H),4.12–4.01(m,1H),3.67–3.50(m,2H),3.37(d,J=15.9Hz,1H),3. 33(d,J=4.7Hz,3H),3.17–3.02(m,5H),2.91–2.80(m,1H),2.79–2.66(m,2H),2.58–2.47(m,6 H),2.15–2.06(m,1H),1.69–1.61(m,2H),1.51–1.40(m,4H),1.39–1.26(m,16H),0.98(s,6H). 13 C NMR(126MHz,Methanol-d4)δ173.90,173.26,172.99,170.27,169.40,168.96,167 .94,146.91,135.83,132.51,116.60,110.33,109.56,57.91,57.82,50.53,49.18, 48.79,42.09,42.02,39.11,37.51,30.83,30.60,29.22,29.18,29.04,29.00,28. 98,28.88,26.58,26.52,26.12,22.66,22.41.HPLC>95%; HRMS(ESI-TOF)m / z:[M+H] + Calculate for C 40 H 60 ClN6O9S + ,835.3826;found,835.3826.

[0087] Example 4: Synthesis of Compound 3

[0088]

[0089] The intermediate M6 (1 mmol) and the fragment S3 (1 mmol) were dissolved in N, N-dimethylformamide (4 ml), and 2-(7-azabenzotriazole)-N, N, N', N'-tetramethyluronium hexafluorophosphate (1 mmol) and N, N-diisopropylethylamine (1.5 mmol) were added, and the mixture was reacted at room temperature for 2 hours. After the reaction was completed, water (10 ml) was added, and the mixture was extracted with ethyl acetate (3×10 ml). The organic phases were combined, washed with water and saturated brine, dried over anhydrous sodium sulfate, and the solvent was removed by rotation. Compound 3 was purified and separated by column chromatography to obtain a yellow solid with a yield of 69%. 1 H NMR(500MHz,Chloroform-d)δ9.24(s,1H),7.57(d,J=8.3Hz,1H),7.07–6.98(m,2H),6.91(t,J=5.2Hz,1 H),6.76(dd,J=8.3,2.2Hz,1H),4.99–4.89(m,1H),4.18–4.06(m,2H),3.96–3.88(m,1H),3.86–3.78(m, 2H),3.75–3.71(m,2H),3.68–3.60(m,9H),3.56–3.51(m,2H),3.44–3.36(m,4H),3.34–3.19(m,2H),3.1 8–2.96(m,4H),2.89–2.80(m,1H),2.78–2.69(m,2H),2.59–2.42(m,6H),2.19–2.06(m,1H),0.95(s,6H). 13 C NMR(126MHz,Chloroform-d)δ173.53,172.87,171.80,169.35,168.19,168.05 ,167.46,153.99,134.39,125.62,117.94,115.78,107.08,70.38,70.18,70.06 ,69.55,68.89,58.14,57.57,50.50,49.09,48.08,43.02,42.41,39.45,37.73, 31.66,31.46,31.23,26.55,23.83,22.80.HPLC>95%; HRMS(ESI-TOF)m / z:[M+H] + Calculate for C 36 H 52 ClN6O 12 S +,827.3047;found,827.3047.

[0090] Example 5: Synthesis of Compound 4:

[0091]

[0092] The intermediate M6 (1 mmol) and the fragment S4 (1 mmol) were dissolved in N, N-dimethylformamide (4 ml), and 2-(7-azabenzotriazole)-N, N, N', N'-tetramethyluronium hexafluorophosphate (1 mmol) and N, N-diisopropylethylamine (1.5 mmol) were added, and the mixture was reacted at room temperature for 2 hours. After the reaction was completed, water (10 ml) was added, and the mixture was extracted with ethyl acetate (3×10 ml). The organic phases were combined, washed with water and saturated brine, dried over anhydrous sodium sulfate, and the solvent was removed by rotation. Compound 4 was purified and separated by column chromatography to obtain a yellow solid with a yield of 73%. 1 H NMR(500MHz,Methanol-d4)δ7.55(d,J=8.4Hz,1H),6.96(d,J=2.1Hz,1H),6.82(dd,J=8.4,2.1Hz, 1H),5.04(dd,J=12.6,5.5Hz,1H),4.32–4.22(m,2H),4.12–4.03(m,1H),3.68–3.49(m,2H),3.41–3 .33(m,2H),3.22–3.17(m,2H),3.16–3.03(m,5H),2.90–2.81(m,1H),2.78–2.65(m,2H),2.58–2.47 (m,6H),2.13–2.05(m,1H),1.69–1.61(m,2H),1.50–1.41(m,4H),1.39–1.23(m,14H),0.98(s,6H). 13 C NMR(126MHz,Methanol-d4)δ173.45,172.86,172.53,169.94,168.51,167.83,16 7.46,154.32,134.10,124.38,116.07,114.75,104.89,57.43,57.36,50.07,48. 71,48.44,42.32,41.65,38.64,37.06,30.36,30.12,28.75,28.70,28.58,28.53 ,27.96,26.23,26.11,25.67,22.20,22.00.HPLC>95%; HRMS(ESI-TOF)m / z:[M+H] + Calculate for C39 H 58 ClN6O9S + ,821.3669;found,821.3667.

[0093] Example 6: Synthesis of Compound 5

[0094]

[0095] The intermediate M6 (1 mmol) and the fragment S5 (1 mmol) were dissolved in N, N-dimethylformamide (4 ml), and 2-(7-azabenzotriazole)-N, N, N', N'-tetramethyluronium hexafluorophosphate (1 mmol) and N, N-diisopropylethylamine (1.5 mmol) were added, and the mixture was reacted at room temperature for 2 hours. After the reaction was completed, water (10 ml) was added, and the mixture was extracted with ethyl acetate (3×10 ml). The organic phases were combined, washed with water and saturated brine, dried over anhydrous sodium sulfate, and the solvent was removed by rotation. Compound 5 was purified and separated by column chromatography to obtain a yellow solid with a yield of 85%. 1 H NMR(500MHz,Methanol-d4)δ7.55(d,J=8.4Hz,1H),6.97(d,J=2.2Hz,1H),6.82(dd,J=8.4,2.2Hz,1H),5.0 3(dd,J=12.6,5.5Hz,1H),4.34–4.22(m,2H),4.12–4.04(m,1H),3.62–3.58(m,1H),3.58–3.49(m,1H),3.40 –3.32(m,2H),3.22–3.17(m,2H),3.15–3.04(m,5H),2.88–2.79(m,1H),2.76–2.66(m,2H),2.56–2.50(m,2 H),2.49(s,4H),2.12–2.05(m,1H),1.69–1.61(m,2H),1.50–1.40(m,4H),1.37–1.29(m,12H),0.99(s,6H). 13C NMR(126MHz,Methanol-d4)δ173.89,173.28,172.97,170.37,168.96,168.28,167 .92,154.83,134.57,124.81,116.51,115.16,105.29,57.89,57.82,50.51,49.17, 48.90,42.74,42.07,39.07,37.50,30.81,30.58,29.13,29.11,29.02,28.99,28. 94,28.42,26.66,26.54,26.12,22.65,22.45.HPLC>95%; HRMS(ESI-TOF)m / z:[M+H] + Calculate for C 38 H 56 ClN6O9S + ,807.3513;found,807.3519.

[0096] Example 7: Synthesis of Compound 6

[0097]

[0098] The intermediate M6 (1 mmol) and the fragment S6 (1 mmol) were dissolved in N, N-dimethylformamide (4 ml), and 2-(7-azabenzotriazole)-N, N, N', N'-tetramethyluronium hexafluorophosphate (1 mmol) and N, N-diisopropylethylamine (1.5 mmol) were added, and the mixture was reacted at room temperature for 2 hours. After the reaction was completed, water (10 ml) was added, and the mixture was extracted with ethyl acetate (3×10 ml). The organic phases were combined, washed with water and saturated brine, dried over anhydrous sodium sulfate, and the solvent was removed by rotation. Compound 6 was purified and separated by column chromatography to obtain a yellow solid with a yield of 89%. 1H NMR (500MHz, Methanol-d4) δ7.55(d,J=8.4Hz,1H),6.97(d,J=2.2Hz,1H),6.82(dd,J=8.4,2.2Hz,1 H),5.03(dd,J=12.6,5.4Hz,1H),4.32–4.22(m,2H),4.11–4.02(m,1H),3.69–3.49(m,2H),3.40–3.3 2(m,2H),3.20(t,J=7.1Hz,2H),3.16–3.04(m,5H),2.90–2.80(m,1H),2.77–2.64(m,2H),2.58–2.4 6(m,6H),2.13–2.05(m,1H),1.69–1.61(m,2H),1.51–1.41(m,4H),1.40–1.26(m,10H),0.98(s,6H). 13 CNMR(126MHz,Methanol-d4)δ173.90,173.30,172.99,170.38,168.97,168.29,1 67.93,154.82,134.56,124.82,116.52,115.19,105.32,57.90,57.82,50.52,49 .17,48.91,42.75,42.07,39.06,37.51,30.81,30.59,29.12,29.02,28.95,28.8 8,28.42,26.65,26.52,26.12,22.66,22.45.HPLC>95%; HRMS(ESI-TOF)m / z:[M+H] + Calculate for C 37 H 54 ClN6O9S + ,793.3356;found,793.3358.

[0099] Test Example 1: IC50 Test Method

[0100] According to the growth rate of different cells, cells in the logarithmic growth phase are inoculated into non-edge wells in a 96-well plate in a certain number. The volume of the cell suspension is 80 μL. 100 μL of sterilized 1×PBS is added to the edge wells and placed in a cell culture incubator for equilibrium for at least 30 minutes. The compound is diluted into 8 concentration gradients, and 3 replicate wells are set for each concentration. After dilution with the corresponding culture medium, it is added to the 96-well plate inoculated with cells in a volume of 20 μL. 0.2% DMSO is used as the control group, and pure culture medium is used as the blank control. The final volume in the plate is 100 μL. After the cell plate is cultured for 3-7 days, 20 μL of MTS solution is added to each well, incubated in a cell culture incubator for a certain period of time, and the absorbance values ​​at 490nM and 690nM (reference wavelength) are detected using a multifunctional microplate reader. The final collected absorbance value OD=OD 490 -OD 690 Cell survival percentage (%) = [(OD value of experimental group - OD value of blank group) / (OD value of control group - OD value of blank group)] × 100 (%). Using GraphPad Prism 9.0 software, nonlinear regression fitting of the IC values ​​of the compounds was performed. 50 value.

[0101] The above method was used to measure the inhibitory activity of compounds 1-6 against leukemia MV-4-11 cells, and the results are shown in Table 1. The above method was also used to measure the inhibitory activity of compound 1 and Sulfopin against a variety of different AML cells, and the cell lines used included MV-4-11, MOLM-13, HL-60, THP-1, Kasumi-1, BDCM, and OCI-AML3. The results are shown in Figure 2 The above results indicate that the compound of the present application has better tumor cell inhibition activity than Sulfopin.

[0102] Test Example 2: Degradation ratio test method

[0103] The cultured MV-4-11 cells were inoculated into 12-well plates at 500,000 cells / well, and a certain concentration of compound was added for 24 hours, with 0.2% DMSO as a control, and the plate volume was 1 mL. After collecting the cells, the expression of the target protein was detected by WB, the grayscale of protein expression was analyzed by Image Lab software, and the internal reference protein Actin was used for normalization analysis to calculate the protein degradation efficiency. The results are shown in Table 1.

[0104] Table 1

[0105]

[0106] The contents not described in detail in this application specification belong to common knowledge of those skilled in the art.

[0107] As mentioned in the entire specification and claims, "including" is an open term and should be interpreted as "including but not limited to". "Substantially" means that within an acceptable error range, those skilled in the art can solve the technical problem within a certain error range and basically achieve the technical effect.

[0108] It should also be noted that the terms "include", "comprises" or any other variations thereof are intended to cover non-exclusive inclusion, so that a product or system including a series of elements includes not only those elements, but also other elements not explicitly listed, or also includes elements inherent to such a product or system. In the absence of more restrictions, the elements defined by the sentence "comprises a ..." do not exclude the existence of other identical elements in the product or system including the elements.

[0109] The above description shows and describes several preferred embodiments of the present application, but as mentioned above, it should be understood that the present application is not limited to the form disclosed herein, and should not be regarded as excluding other embodiments, but can be used in various other combinations, modifications and environments, and can be modified within the scope of the invention concept described herein through the above teachings or the technology or knowledge of the relevant field. The changes and modifications made by those skilled in the art do not depart from the spirit and scope of the present application, and should be protected by the claims attached to the present application.

Claims

1. A compound having a structure shown in Formula I or a pharmaceutically acceptable salt thereof: in: R 1 Selected from R 2 (-CH2-) n , n is an integer selected from 1 to 4; L is -R 3 -C(=O)NH-R 4 -; R 3 Selected from C 2-4 alkyl; R 4 Selected from C 8-14 Alkyl or -CH2-(-CH2-O-CH2-) m -CH2-, m is an integer selected from 3 to 4; X is -CH2-, -NH- or -O-; A Choose any of the following groups:

2. The compound according to claim 1 or a pharmaceutically acceptable salt thereof, characterized in that: It has the structure shown in formula II: L is -R 3 -C(=O)NH-R 4 -; R 3 Selected from C 2-4 alkyl; R 4 Selected from C 8-14 Alkyl or -CH2-(-CH2-O-CH2-) m -CH2-, wherein m is 3; X is -CH2-, -NH- or -O-; A Choose any of the following groups:

3. The compound according to claim 1 or 2 or a pharmaceutically acceptable salt thereof, characterized in that: X is -NH-.

4. The compound according to claim 1 or a pharmaceutically acceptable salt thereof, characterized in that: It has the structure shown in formula III: X is -CH2-, -NH- or -O-; n is an integer selected from 1 to 4; A Choose any of the following groups:

5. The compound according to claim 1 or a pharmaceutically acceptable salt thereof, characterized in that: It has the structure shown in Formula IV: n, L, X are as defined in claim 1.

6. The compound according to claim 1 or a pharmaceutically acceptable salt thereof, characterized in that: It has the structure shown in formula V: L, X are as defined in claim 1.

7. The compound according to claim 6 or a pharmaceutically acceptable salt thereof, characterized in that: X is -NH-.

8. The compound according to claim 6 or a pharmaceutically acceptable salt thereof, characterized in that: L is -CH2CH2CONH(-CH2-) 12 -.

9. The compound according to claim 1 or 2 or a pharmaceutically acceptable salt thereof, characterized in that: The compound is selected from:

10. Use of the compound according to any one of claims 1 to 9 or a pharmaceutically acceptable salt thereof in the preparation of a medicament for preventing or treating a disease associated with abnormal expression of Pin1 protein activity.

11. The use according to claim 10, characterized in that The disease associated with abnormal expression of Pin1 protein activity is selected from the group consisting of: tumor, hyperglycemia, diabetes, obesity, hyperlipidemia, hypercholesterolemia, hyperlipoproteinemia, hypertriglyceridemia, hypertension, hyperinsulinemia, hyperuricemia, Parkinson's disease, and Alzheimer's disease.

12. Use of the compound according to any one of claims 1 to 9 or a pharmaceutically acceptable salt thereof in the preparation of a drug for treating or preventing tumors, or a drug for preventing postoperative tumor recurrence; the tumor is leukemia.

13. A composition, characterized in that Containing the compound according to any one of claims 1 to 9 or a pharmaceutically acceptable salt thereof.

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