An isobenzofuranone compound, a preparation method and application thereof

Abstract

The application discloses an isobenzofuranone compound, a preparation method and application thereof. The structural formula of the isobenzofuranone compound is shown as formula (I), wherein R 1 ~R 4 are respectively selected from C1-C6 alkyl, hydrogen, halogen or methoxy; R 5 is selected from C1-C6 alkyl, aryl or halogenated aryl. The application discloses a new compound with anti-pancreatic cancer activity, which has great drug research and development potential and can be used as a lead compound for the development of anti-pancreatic cancer drugs. The isobenzofuranone compound is synthesized by using a non-metal oxidant high-valence iodine catalyst, without using expensive metal catalysts, and meets the requirements of green chemistry. The isobenzofuranone compound has high biological activity, small toxicity and high selectivity, is a new compound which is not easy to be drug-resistant, and can effectively inhibit the proliferation of pancreatic cancer cells.

Description

Technical Field

[0001] This invention belongs to the field of medicinal chemistry technology, specifically an isobenzofuranone compound, its preparation method, and its application. Background Technology

[0002] Pancreatic cancer (PC) is a common malignant tumor of the digestive tract, and its incidence and mortality rates are increasing year by year. Due to its poor prognosis, the mortality rate of PC is almost as high as its incidence rate, making it the seventh leading cause of cancer death in humans. According to a study of 28 European countries, it is projected that by 2025, PC will surpass breast cancer to become the third leading cause of cancer death.

[0003] Currently, drug treatments for pancreatic cancer mainly include chemotherapy, targeted therapy, and immunotherapy. Targeted therapy drugs can interfere with the growth and spread of cancer cells, such as inhibiting signaling pathways like EGFR and VEGF. Immunotherapy drugs can enhance the body's immune system's ability to attack cancer cells, such as anti-PD-1 antibodies. However, these treatments have drawbacks, including poor efficacy, significant side effects, high cost, and the inability to cure the cancer, leading to a high recurrence rate.

[0004] The exact causes of pancreatic cancer are not fully understood, but several factors are associated with its development. Smoking is one of the most significant risk factors; smokers are more than twice as likely to develop pancreatic cancer as non-smokers. Other possible risk factors include obesity, high blood sugar, pancreatitis, family history, and certain gene mutations. However, most pancreatic cancer patients do not have obvious risk factors, making prevention and early diagnosis more challenging. Summary of the Invention

[0005] Objective of the Invention: The first objective of this invention is to provide an isobenzofuranone compound with anti-pancreatic cancer activity. The second objective of this invention is to provide a method for preparing the above-mentioned compound and its applications.

[0006] Technical solution: The structural formula of the isobenzofuranone compound of the present invention is shown in formula (Ⅰ):

[0007]

[0008] Among them, R 1 ~R 4 They are selected from C1-C6 alkyl, hydrogen, halogen, or methoxy groups, respectively;

[0009] R 5 Selected from C1-C6 alkyl, aromatic or halogenated aromatic groups.

[0010] Among them, R 1 Selected from F or H;

[0011] R 2 Selected from methyl, Cl, or H;

[0012] R 3 Selected from methyl, F, H or methoxy;

[0013] R 4 Selected from methyl or H;

[0014] R 5 It is selected from methyl, ethyl, cyclopropyl, phenyl, naphthyl, methoxy-p-fluorophenyl, o-methylphenyl, or p-methylphenyl.

[0015] The structural formula of the compound is selected from the following compounds:

[0016]

[0017] The method for preparing the isobenzofuranone compound of the present invention includes the following steps: dissolving isocoumarin in dichloromethane, adding tetrabutylammonium bromide, PIFA and water, mixing evenly, and after the reaction is completed, separating the solvent, extracting and purifying to obtain the isobenzofuranone compound.

[0018] The molar ratio of isocoumarin, tetrabutylammonium bromide, PIFA, and water is 1:2-2.5:2-2.5:2-2.5. The reaction conditions after homogenization are room temperature and the reaction time is 2-12 hours.

[0019] In the preparation process of this invention, bromide anions are oxidized to bromide ions by PIFA, oxygen atoms in water attack the carbocation on the isocoumarin core, undergo proton migration and ring opening, lose one molecule of HBr and rearrange to form a five-membered ring, and the bromide ion attacks the hydrogen at the 3-position to obtain the bromoisobenzofuranone compound. The prepared bromoisobenzofuranone compound has high biological activity.

[0020] The application of the isobenzofuranone compound described in this invention in the preparation of drugs for treating pancreatic cancer.

[0021] The isobenzofuranone compound can inhibit the growth of pancreatic cancer cells.

[0022] The present invention provides a pharmaceutical composition in which the active ingredient comprises the above-mentioned isobenzofuranone compound or its stereoisomer, its prodrug, its pharmaceutically acceptable salt or its pharmaceutically acceptable solvate.

[0023] The pharmaceutical composition contains a pharmaceutically acceptable carrier.

[0024] Beneficial Effects: Compared with existing technologies, this invention has the following significant advantages: This invention discloses a novel class of compounds with anti-pancreatic cancer activity, possessing significant potential for drug development and serving as lead compounds for the development of anti-pancreatic cancer drugs. The isobenzofuranone compounds of this invention are synthesized using high-valent iodine catalysis, a non-metallic oxidant, eliminating the need for expensive metal catalysts and meeting the requirements of green chemistry. The isobenzofuranone compounds of this invention exhibit high biological activity, low toxicity, and high selectivity, making them novel compounds less prone to drug resistance and effectively inhibiting the proliferation of pancreatic cancer cells. Attached Figure Description

[0025] Figure 1 The 1H NMR spectrum of compound 12 of this invention;

[0026] Figure 2 The carbon NMR spectrum of compound 12 of this invention;

[0027] Figure 3 The 1H NMR spectrum of compound 13 of this invention;

[0028] Figure 4 This is the carbon NMR spectrum of compound 13 of the present invention. Detailed Implementation

[0029] The technical solution of the present invention will be further described below with reference to the accompanying drawings.

[0030] Example 1: Preparation of isobenzofuranone compounds

[0031]

[0032] Isocoumarin (1 eq.) was dissolved in 3 mL of dichloromethane (dcm), and then tetrabutylammonium bromide (TBAB, 2.2 eq.), bis(trifluoroacetyliodobenzene) (PIFA, 2.2 eq.), and water (2.2 eq.) were added and mixed thoroughly to obtain a mixture. The mixture was reacted at room temperature for 2.0–12.0 h, and then the solvent dichloromethane was removed by rotary evaporation. The mixture was extracted three times with ethyl acetate, and the purified product was obtained by column chromatography. In this example, a total of 18 compounds were prepared. The substituents of the compounds are shown in Table 1 (the substituents of the substrate isocoumarin correspond to the prepared compounds). The carbon and hydrogen spectral data are as follows:

[0033] Compound 1 (denoted as C1): white solid, yield 82%. 1H NMR (500MHz, CDCl3) δ = 8.37 (dd, J = 8.3, 1.4Hz, 2H), 8.06 (d, J = 7.8Hz, 1H), 7.92 (d, J =7.7Hz,1H),7.86(td,J=7.7,1.2Hz,1H),7.71–7.62(m,2H),7.53(t,J=7.9Hz,2H). 13 C NMR (125MHz, CDCl3)δ=185.66,166.70,146.34,138.25,137.49,134.32,131.92,131.49,131.37,128.57,123.62,88.80,19.50.ESI-MS, m / Z: [M+Na] + Theoretical value: 338.9633; Test value: 338.9627.

[0034] Compound 2 (denoted as C2): white solid, yield 72%. 1 H NMR (500MHz, CDCl3) δ8.38–8.33(m,2H),7.99(d,J=8.3Hz,1H),7.88(d,J=1.9 Hz,1H),7.81(dd,J=8.3,1.9Hz,1H),7.69–7.64(m,1H),7.54(t,J=7.8Hz,2H). 13 C NMR (125MHz, CDCl3) δ = 165.04, 146.90, 137.99, 135.79, 134.69, 131.56, 131.46, 128.75, 128.56, 125.55, 124.93. ESI-MS, m / Z: [M+Na] + Theoretical value: 372.9243; Test value: 372.9232.

[0035] Compound 3 (denoted as C3): white solid, yield 70%. 1 H NMR (500MHz, CDCl3) δ8.36(d,J=7.8Hz,2H),7.92(dt,J=8.4,4.6Hz,1H),7.75–7.63(m,2H),7.54(t,J=7.7Hz,2H),7.36(td,J=8.4,2.4Hz,1H). 13C NMR(125MHz, CDCl3)δ=186.31,167.90,165.84,165.02,151.48,151.39,134.56,131.44,131.33 ,128.65,128.62,127.97,127.88,119.72,119.53,119.10,114.78,114.58.ESI-MS,m / Z: [M+Na] + Theoretical value: 356.9539; Test value: 356.9526.

[0036] Compound 4 (denoted as C4): white solid, yield 62%. 1 H NMR (500MHz, CDCl3) δ=8.39–8.31(m,2H),7.89–7.81(m,2H),7.70–7.64(m,1H),7.57–7.51(m,2H),7.34–7.27(m,1H). 13 C NMR(126MHz, CDCl3)δ=186.61,158.46,150.85,138.21,138.15,134.93,131.8 8,131.71,129.00,123.78,123.74,118.50,118.36,85.88.ESI-MS,m / Z: [M+Na] + Theoretical value: 355.9539; Test value: 356.9528.

[0037] Compound 5 (denoted as C5): white solid, yield 75%. 1 H NMR (500MHz, CDCl3), δ = 9.48-9.39 (m, 1H, Ar-H), 7.64 (dt, J = 4.4, 2.7Hz, 1H, Ar-H), 7.52-7.4 5(m,1H,Ar-H),6.62-6.56(m,1H,Ar-H),2.96(d,J=6.6Hz,2H,-CH2),1.39-1.31(m,3H,-CH3); 13 C NMR (125MHz, CDCl3), δ = 171.39, 144.20, 138.82, 137.82, 131.81, 129.36, 115.20, 107.21, 28.83, 8.42.ESI-MS, m / Z: [M+Na] + Theoretical value: 352.9789; Test value: 352.9785.

[0038] Compound 6 (denoted as C6): white solid, yield 78%. 1H NMR (500MHz, CDCl3) δ = 8.36 (dd, J = 8.5, 1.3Hz, 2H), 7.93 (d, J = 7.9Hz, 1H), 7.70 (p, J = 0.8Hz, 1H), 7.68–7.61 (m, 2H), 7.55–7.49 (m, 2H), 2.51 (s, 3H). 13 C NMR (125MHz, CDCl3) δ = 195.26, 166.09, 146.85, 135.53, 131.37, 125.94, 125.86, 123.57, 87.30, 24.39.ESI-MS, m / Z: [M+Na] + Theoretical value: 352.9789; Test value: 352.9783.

[0039] Compound 7 (denoted as C7): white solid, yield 91%. 1 H NMR (500MHz, CDCl3) δ = 8.40–8.34 (m, 2H), 7.80 (d, J = 7.5Hz, 1H), 7.64 (dt, J = 7.4, 3.7Hz, 2H), 7.57 (t, J = 7.5Hz, 1H), 7.52 (t, J = 7.7Hz, 2H), 2.58 (s, 3H). 13 C NMR (126MHz, CDCl3) δ = 187.24, 166.59, 149.33, 147.41, 134.67, 132.77, 132.2 0,131.66,128.89,127.75,125.77,120.89,86.81,22.70.ESI-MS,m / Z:[M+Na] + Theoretical value: 352.9789; Test value: 352.9787.

[0040] Compound 8 (denoted as C8): white solid, yield 79%. 1 H NMR (500MHz, CDCl3) δ = 8.36 (d, J = 7.5Hz, 2H), 7.80 (d, J = 8.5Hz, 1H), 7.69–7.61 (m, 1H), 7.53(t,J=7.7Hz,2H),7.45(d,J=2.2Hz,1H),7.15(dt,J=8.5,1.6Hz,1H),4.01(s,3H). 13 C NMR (125MHz, CDCl3) δ = 187.10, 165.66, 151.60, 134.45, 131.98, 131.42, 128.67, 127.15, 119.62, 110.53, 56.38 .ESI-MS, m / Z: [M+Na] + Theoretical value: 368.9738; Test value: 368.9738.

[0041] Compound 9 (denoted as C9): white solid, yield 40%. 1 H NMR (500MHz, CDCl3) δ = 8.55–8.26 (m, 2H), 7.80 (d, J = 7.5Hz, 1H), 7.64 (dt, J = 7.4, 3.5Hz, 2H), 7.57 (t, J = 7.5Hz, 1H), 7.52 (t, J = 7.7Hz, 2H), 2.58 (s, 3H). 13 C NMR (126MHz, CDCl3) δ = 195.41, 166.24, 147.00, 135.67, 131.50, 126.09, 126.01, 123.72, 87.43, 24.54.ESI-MS, m / Z: [M+Na] + Theoretical value: 276.9476; Test value: 276.9471.

[0042] Compound 10 (denoted as C10): white solid, yield 60%. 1 H NMR (500MHz, CDCl3) δ = 7.91 (dd, J = 13.1, 7.8Hz, 2H), 7.80 (t, J = 7.6Hz, 1H), 7.6 4(t,J=7.5Hz,1H),3.10-3.38(m,1H),2.60-2.90(m,1H),1.16(t,J=7.2Hz,3H). 13 C NMR (125MHz, CDCl3) δ = 198.96, 166.37, 147.33, 135.68, 131.45, 125.99, 123.61, 87.16, 77.36, 30.36, 7.74.ESI-MS, m / Z: [M+Na] + Theoretical value: 290.9633; Test value: 290.9628.

[0043] Compound 11 (denoted as C11): white solid, yield 68%. 1 H NMR (500MHz, CDCl3) δ = 8.22 (dt, J = 8.0, 1.0 Hz, 1H), 7.85–7.72 (m, 2H), 7.48 (m, J = 8.2 ,5.7,2.7Hz,1H),2.44(tt,J=8.3,5.0Hz,1H),1.30–1.14(m,2H),1.14–0.96(m,2H). 13C NMR (125MHz, CDCl3) δ = 161.16, 154.95, 136.85, 135.56, 129.91, 128.07, 125.35, 120.12, 100.32, 13.90, 8.21.ESI-MS, m / Z: [M+H] + Theoretical value: 280.9813; Test value: 280.9809.

[0044] Compound 12 (denoted as C12): white solid, yield 85%. 1 H NMR (500MHz, CDCl3) δ = 7.91 (dd, J = 8.4, 4.7Hz, 1H), 7.57 (dd, J = 7.5, 2.2Hz, 1H), 7.35 (td, J = 8.5, 2.2Hz, 1H), 2.60 (s, 3H). 13 CNMR(125MHz, CDCl3)δ=195.00,168.13,166.06,164.95,149.91,149.82,1 28.38,128.29,119.98,119.79,113.77,113.56,24.47.ESI-MS,m / Z: [M+Na] + Theoretical value: 294.9382; Test value: 294.9386.

[0045] Compound 13 (denoted as C13): white solid, yield 79%. 1 H NMR(500MHz, CDCl3)δ=8.47–8.39(m,2H),8.06–8.01(m,1H),7.93(dt,J=7.6,0.9Hz, 1H), 7.86 (td, J=7.6, 1.1Hz, 1H), 7.67 (td, J=7.6, 0.9Hz, 1H), 7.21 (t, J=8.6Hz, 2H). 13 C NMR(125MHz, CDCl3)δ=185.24,166.24,148.49,135.51,134.41,134.34,131.39,12 8.13,128.10,127.35,125.82,123.11,116.10,115.93,86.42.ESI-MS,m / Z: [M+Na] + Theoretical value: 356.9539; Test value: 356.9546.

[0046] Compound 14 (denoted as C14): white solid, yield 79%. 1H NMR (500MHz, CDCl3) δ = 8.14 (dd, J = 7.9, 1.4Hz, 1H), 8.10 (dt, J = 7.9, 0.9Hz, 1H), 7.94 (dt, J = 7.7, 0.9Hz, 1H), 7.88 (td,J=7.7,1.2Hz,1H),7.70(td,J=7.6,0.9Hz,1H),7.47(td,J=7.6,1.4Hz,1H),7.36–7.29(m,2H),2.40(s,3H). 13 C NMR(125MHz, CDCl3)δ=191.36,166.44,148.52,139.73,135.66,132.89,132.43,13 1.88,131.39,130.47,126.67,125.88,125.51,123.46,21.27.ESI-MS,m / Z:[M+Na] + Theoretical value: 352.9789; Test value: 352.9781.

[0047] Compound 15 (denoted as C15): White solid, 70% yield. ¹H NMR (500 MHz, CDCl₃) δ = 8.31–8.24 (m, 2H), 8.05 (d, J = 7.8 Hz, 1H), 7.94–7.89 (m, 1H), 7.85 (td, J = 7.6, 1.1 Hz, 1H), 7.66 (td, J = 7.5, 0.9 Hz, 1H), 7.33 (d, J = 8.1 Hz, 2H), 2.46 (s, 3H). ¹³C NMR(125MHz, CDCl3)δ=186.39,166.47,148.83,145.73,135.38,131.57,131.22,1 29.41,129.23,127.38,125.70,123.18,86.66,77.36,22.01.ESI-MS,m / Z: [M+Na] + Theoretical value: 352.9789; Test value: 352.9788.

[0048] Compound 16 (denoted as C16): white solid, yield 74%. 1 H NMR (500MHz, CDCl3) δ = 9.07 (s, 1H), 8.28 (dd, J = 8.7, 1.6Hz, 1H), 8.12–8.04 (m, 2H), 7 .94(d,J=8.4Hz,2H),7.89(q,J=7.8Hz,2H),7.72–7.63(m,2H),7.60(t,J=7.5Hz,1H). 13C NMR(125MHz, CDCl3)δ=186.48,166.21,148.57,135.91,135.21,134.00,132.08,131.05,130.20 ,129.31,128.77,128.22,127.64,127.17,126.85,125.66,125.52,122.95.ESI-MS,m / Z: [M+Na] + Theoretical value: 388.9789; Test value: 388.9778.

[0049] Compound 17 (denoted as C17): white solid, yield 78%. 1 H NMR (500MHz, CDCl3) δ = 8.30–8.21 (m, 2H), 7.83 (d, J = 1.3Hz, 1H), 7.78 (d, J = 7. 8Hz,1H),7.49–7.39(m,1H),7.32(d,J=8.1Hz,2H),2.59(s,3H),2.45(s,3H). 13 C NMR (125MHz, CDCl3) δ = 186.60, 166.44, 149.23, 147.08, 145.67, 132.45, 131.56, 129. 38,129.31,127.55,125.47,120.63,86.68,77.36,22.46,22.00.ESI-MS,m / Z:[M+Na] + Theoretical value: 366.9946; Test value: 366.9942.

[0050] Compound 18 (denoted as C18): white solid, yield 75%. 1 H NMR(500MHz, CDCl3)δ=8.38–8.32(m,2H),7.98(d,J=8.3Hz,1H),7.86(d,J=1.8Hz,1H),7.80(dd,J=8.3,1.9Hz,1H),7.03–6.97(m,2H),3.92(s,3H).13C NMR(125MHz, CDCl3)δ=184.94,165.18,164.78,147.21,137.83,135.67,134.0 3,128.68,125.44,124.87,124.15,114.08,86.16,55.79.ESI-MS,m / Z:[M+Na] + Theoretical value: 402.9349; Test value: 402.9354.

[0051] Table 1 Substituent Information

[0052]

[0053] Example 2: Anti-pancreatic cancer activity test of isobenzofuranone compounds

[0054] Using 5-FU (5-fluorouracil), a commonly used anticancer drug in clinical practice, as a positive control, the in vitro antitumor activity of compounds C1-C18 against pancreatic cancer cells was tested using the CCK-8 assay. Human pancreatic cancer PNAC-1 cells were cultured in DMEM / 10% fetal bovine serum medium at 37°C with 5% CO2. Cells in the logarithmic growth phase were collected and seeded in 96-well plates at a density of 5 × 10⁶ cells / well. 3 Cells were incubated at 100 μL / well for 24 hours, then the solvent (DMSO, 1:2000 dilution, as a blank group) was replaced with different compounds and 5-FU (stock solution concentration 100 mM / L, working solution concentration 50 μM / L) and incubated for another 48 hours. Then, 10 μL of CCK-8 solution was added to each well and incubated for 2 hours. Each group was repeated in triplicate. Finally, the absorbance was measured at 450 nm using a microplate reader. Cell inhibition rate was calculated based on absorbance, and survival rate was calculated as: (OD of drug-treated group - OD of blank group) / (OD of positive control group (i.e., 5-FU-treated group) - OD of blank group) * 100%. Specific results are shown in Table 2.

[0055] Table 2. Cell viability test at 100 μm concentration

[0056] Compound serial number Compound Name Cell viability (% control group) 1 Blank group 100±2.896 2 Positive control group 41.35±1.821 3 C1 136.30±3.246 4 C2 82.39±6.127 5 C3 153.83±1.257 6 C4 73.60±2.290 7 C5 89.10±3.070 8 C6 91.56±2.512 9 C7 183.20±1.121 10 C8 113.50±0.147 11 C9 180.01±2.510 12 C10 190.20±0.469 13 C11 194.50±1.144 14 C12 7.22±1.163 15 C13 17.70±2.670 16 C14 163.60±4.623 17 C15 18.66±0.547 18 C16 81.02±0.940 19 C17 38.02±1.078 20 C18 101.64±2.126

[0057] Cell viability assay at 100 μm concentration: At a concentration of 100 μm, compared to the positive control, compounds C12 / C13 / C15 / C17 exhibited significantly stronger inhibitory activity against pancreatic cancer cells than 5-FU. Compound C12 showed the most prominent activity, reducing the survival rate of pancreatic cancer cells to below 10%. Compounds C13 / C15 / C17 also showed high inhibitory activity against cancer cell proliferation, with significant differences compared to the positive control group. This fully demonstrates that the compounds of this invention have a good ability to inhibit cancer cell proliferation and can be developed as a potential anti-pancreatic cancer drug.

Claims

1. An isobenzofuranone compound, characterized in that, The structural formula of the compound is shown in formula (Ⅰ): ； The substituents are as follows: ； Specifically, the structural formulas of the compounds are selected from the following compounds: 。 2. A method for preparing the isobenzofuranone compound according to claim 1, comprising the following steps: dissolving isocoumarin in dichloromethane (DCM), adding tetrabutylammonium bromide (TBAB), PIFA and water, mixing evenly, and after the reaction is completed, separating the solvent, extracting and purifying to obtain the isobenzofuranone compound; The reaction route is as follows: ; The substituents of isocoumarin are as follows: 。 3. The method for preparing the isobenzofuranone compound according to claim 2, characterized in that, The molar ratio of isocoumarin, tetrabutylammonium bromide, PIFA and water is 1:2-2.5:2-2.5:2-2.

5.

4. The method for preparing the isobenzofuranone compound according to claim 2, characterized in that, The reaction conditions after uniform mixing are room temperature and the reaction time is 2-12 hours.

5. The use of the isobenzofuranone compound of claim 1 in the preparation of a drug for treating pancreatic cancer.

6. The application according to claim 5, characterized in that, The isobenzofuranone compound can inhibit the growth of pancreatic cancer cells.

7. A pharmaceutical composition, characterized in that, The active ingredient comprises the isobenzofuranone compound of claim 1 or a pharmaceutically acceptable salt thereof.

8. The pharmaceutical composition according to claim 7, characterized in that, The pharmaceutical composition contains a pharmaceutically acceptable carrier.

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