A quinazoline-fused benzothiocyanate tetracyclic compound that is a structural analog of berberine, its preparation method, and its use in the preparation of drugs for treating breast cancer.

Abstract

This invention discloses a quinazoline-fused benzothiocyanate tetracyclic compound, a structural analog of berberine, its preparation method, and its use in the preparation of drugs for treating breast cancer. The general structural formula of the compound is shown in Formula 3, which can be considered a simulation and optimization of the dominant structure of the natural product berberine. The preparation method involves using benzodithiocyclopentanone compounds and quinazoline-derived dipolar compounds as raw materials, undergoing a highly efficient [4+3] cyclization reaction in an organic solvent under the action of a phosphine reagent. This method is mild, simple, and has broad substrate applicability, enabling the rapid construction of a compound library containing more than 25 target compounds with excellent yields. In vitro antitumor evaluation shows that the compounds of this invention exhibit significant antiproliferative and antimigration activity against triple-negative breast cancer cells, and possess a high selectivity index.

Description

Technical Field

[0001] This invention belongs to the field of pharmaceutical technology. Specifically, it relates to a quinazoline fused benzothiocyanate tetracyclic compound that is a structural analog of berberine, its preparation method, and its use in the preparation of drugs for treating breast cancer. Background Technology

[0002] Natural products and their structural analogs have always been an important source for anticancer drug development. Berberine is an isoquinoline alkaloid isolated from medicinal plants such as Coptis chinensis, possessing a characteristic fused tetracyclic structure. Due to its broad pharmacological activities, including antitumor effects against various human cancer cell lines, berberine has attracted the research interest of scientists. Its nitrogen-containing tetracyclic skeleton is considered a superior structure for drug discovery, providing a valuable template for designing novel antitumor compounds.

[0003] However, directly developing berberine and its derivatives into chemotherapy drugs targeting specific cancers such as triple-negative breast cancer (TNBC) faces several limitations, including moderate pharmacological activity, potential side effects due to insufficient selectivity for cancerous and normal cells, and lack of efficacy against the highly invasive and metastatic TNBC phenotype. TNBC is a clinically refractory subtype of breast cancer characterized by negativity of estrogen receptor (ER), progesterone receptor (PR), and human epidermal growth factor receptor 2 (HER2). This "triple-negative" status makes it insensitive to endocrine therapy and HER2-targeted therapy, resulting in limited treatment options and poor prognosis for patients. Currently, there is an urgent clinical need for novel chemical entities that can effectively and selectively combat TNBC.

[0004] Modifying the core structure of natural products is an effective strategy to overcome the limitations of the parent compound and discover new drugs with superior pharmacological properties. Against this backdrop, although the tetracyclic structure of berberine is recognized for its biological activity, the exploration of novel tetracyclic systems that retain its core structural features and are modified to regulate target binding properties and physicochemical properties, ultimately enhancing therapeutic potential, remains a promising but still insufficiently researched area.

[0005] Therefore, in this invention, we aim to provide a novel class of tetracyclic compounds based on the tetracyclic structure of berberine. These compounds are designed as structural analogs of berberine and can be prepared through a simple synthetic strategy, thus yielding a new type of chemical structure that shows great potential in the treatment of TNBC. Summary of the Invention

[0006] To address the shortcomings of existing technologies, particularly the limitations of berberine-based natural products in their anti-TNBC activity and selectivity, and the cumbersome and inefficient steps involved in constructing complex tetracyclic heterocyclic molecules using current synthetic methods, this invention aims to provide a novel class of berberine structural analogues—quinazoline-fused benzothiazole tetracyclic compounds. Furthermore, this invention provides an efficient and universal synthetic method for these compounds, as well as their application in the preparation of anti-TNBC drugs. These compounds exhibit significant anti-proliferative and anti-migration activity against TNBC cells, along with a high selectivity index, providing valuable lead compounds for the development of novel TNBC therapeutics.

[0007] To achieve the above objectives, the present invention adopts the following technical solution: A method for synthesizing a quinazoline-fused benzothiocyanate tetracyclic compound, a berberine structural analog, includes reacting a benzodithiocyclopentanone compound of general formula 1 with a quinazoline-derived dipolar compound of general formula 2 in an organic solvent at a certain temperature and under the action of a phosphine reagent for a period of time, thereby obtaining the quinazoline-fused benzothiocyanate tetracyclic compound through a [4 + 3] cyclization reaction.

[0008] In the formula, R 1 The following are possible meanings: hydrogen, 4-methyl, 5-methyl, 5-methoxy, 6-methoxy, 5-fluorine, 5-chloro, 5-bromine, 6-bromine, 7-bromine, 5-iodine, 5-nitro, 5-methoxycarboxyl; R 2 For hydrogen, 6-methoxy, 6-fluoro, 7-methoxy, 7-fluoro, 7-chloro, 8-chloro; R 3 It is phenyl, 4-methylphenyl, 4-methoxyphenyl, 4-chlorophenyl, 4-tert-butylphenyl, thiophene, or benzyl.

[0009] Furthermore, the phosphine reagent is a trialkylphosphine or a triarylphosphine; preferably a methyldiphenylphosphine.

[0010] Furthermore, the organic solvent is a polar aprotic solvent or a halogenated hydrocarbon solvent; preferably dichloromethane.

[0011] Furthermore, the reaction temperature is from 25 degrees to the reflux temperature of the solvent; preferably 35 degrees.

[0012] Furthermore, the molar ratio of the benzodithiocyclopentanone 1 to the quinazoline-derived dipolar compound 2 is 1:1 to 1:1.5; preferably 1:1.

[0013] Furthermore, the reaction time is 1 to 5 hours; preferably 3 hours.

[0014] The method has the advantages of mild conditions, wide substrate applicability, and high reaction efficiency (most yields >85%), and can rapidly construct a compound library containing more than 25 products.

[0015] The present invention also provides a quinazoline-fused benzothiocyanate tetracyclic compound prepared by the above-described synthetic method, the general structural formula of which is shown in Formula 3 below:

[0016] In the formula, R 1 The radicals are hydrogen, methyl, methoxy, fluorine, chlorine, bromine, iodine, nitro, and methoxyformyl; R 2 For hydrogen, methoxy, fluorine, chlorine; R 3 It can be phenyl, 4-methylphenyl, 4-methoxyphenyl, 4-chlorophenyl, 4-tert-butylphenyl, thiophene, or benzyl. Specific structures are as follows: Figure 1 As shown.

[0017] The in vitro antitumor activity assay was performed using the MTT assay to evaluate the inhibitory activity and selectivity of the compounds of this invention on the proliferation of TNBC cell lines (MDA-MB-231, Hs578T), estrogen receptor-positive (ER+) breast cancer cell line (MCF-7), and normal human embryonic kidney cells (HEK-293).

[0018] Experimental results show that the compounds of this invention exhibit selective inhibitory effects on TNBC cell lines. Specifically, their toxicity to MCF-7 and HEK-293 cells is lower than that to TNBC cells, demonstrating good selectivity. Among the compounds, compounds 3aa, 3ba, and 3ma exhibit the best activity. Compound 3ma shows the most prominent activity, with a half-maximal inhibitory concentration (IC50) against MDA-MB-231 cells. 50 As low as 3.4 m M, and the selectivity index (SI) for MDA-MB-231 cells was 7.3, indicating that its cytotoxicity to cancer cells was 7.3 times that to normal cells, a value superior to the control drug 5-fluorouracil (5-FU, SI = 0.7). Compounds 3aa and 3ba also showed good activity and selectivity, with IC50 values ​​for MDA-MB-231 cells. 50 The values ​​are 5.2. m M and 6.5 m M. The experimental results are shown in Table 1.

[0019] Table 1. IC50 values ​​of quinazoline-fused benzothiocyanate tetracyclic compounds against various breast cancer cell lines and normal cell lines. 50 Value and selectivity index

[0020] Metastasis in breast cancer is a major cause of poor prognosis in patients. To verify the anti-metastatic potential of the compounds of this invention, we selected the compound 3ma with the best activity and evaluated its effect on the migration ability of MDA-MB-231 cells through wound healing assays and Transwell assays.

[0021] In wound healing experiments, the 3ma-treated group showed dose-dependent inhibition of cell migration after 48 hours, compared with the untreated control group. In 10 m At concentration M, the inhibitory effect of this compound on cell migration was statistically significant. P <0.05). Transwell assay results also confirmed the anti-migration effect of 3-ma. Compared with the control group, after 2.5... m M, 5 m M and 10 m Treatment with M concentrations of 3ma reduced the transmembrane migration rate of MDA-MB-231 cells by approximately 10%, 19%, and 30%, respectively, exhibiting a dose-response relationship. These two experiments demonstrate that the compound 3ma provided in this invention can inhibit the migration of TNBC cells, suggesting its significant potential for intervention in breast cancer metastasis.

[0022] Compared with the prior art, the beneficial effects of the present invention are as follows: (1) This invention creatively designs and synthesizes a novel class of quinazoline fused benzothiazide compounds. By referencing the tetracyclic skeleton of berberine, the two pharmacophores, quinazoline (found in anti-breast cancer drugs such as lapatinib) and benzothiazide (found in drugs such as diltiazem), are combined to form a fused tetracyclic structure, resulting in berberine structural analogs with novel structures and good activity.

[0023] (2) A novel organophosphorus-promoted [4+3] cyclization strategy was developed, enabling rapid and efficient construction of complex tetracyclic molecules from readily available raw materials. This method has good atom economy and is easy to operate, laying the foundation for subsequent research on this type of molecule.

[0024] (3) In vitro antitumor activity experiments showed that several compounds of the present invention exhibited significant inhibitory activity against TNBC cell lines (such as MDA-MB-231), and their efficacy was superior to that of the control drug 5-FU.

[0025] (4) The compounds of the present invention exhibit good selectivity for TNBC cells, with a selectivity index higher than that of the control drug 5-FU. For example, the selectivity index of compound 3ma for MDA-MB-231 cells is 7.3, indicating that it is expected to have a longer safety window in clinical applications.

[0026] (5) Wound healing scratch assay and Transwell assay confirmed that the preferred compounds of the present invention (such as 3ma) can inhibit the migration of highly invasive MDA-MB-231 cells in a dose-dependent manner, indicating that such compounds can not only inhibit tumor growth, but also have potential anti-tumor metastasis ability, which is crucial for improving the prognosis of TNBC patients.

[0027] In summary, this invention not only provides a class of novel and efficiently prepared compounds, but more importantly, these compounds exhibit advantages such as proliferation inhibition, high selectivity, and anti-migration potential in targeting refractory TNBC, and have high prospects for development into novel anti-TNBC drugs. Attached Figure Description

[0028] The accompanying drawings, which are included to provide a further understanding of the invention and form part of this application, illustrate exemplary embodiments of the invention and do not constitute an undue limitation thereof. The drawings are described below: Figure 1 The structure of quinazoline-fused benzothiocyanate tetracyclic compounds; Figure 2 (A) Wound healing assay of MDA-MB-231 cells treated with compound 3ma; (B) Percentage migration of compound 3ma; (C) Transwell assay of MDA-MB-231 cells treated with compound 3ma (24 hours); (D) Number of MDA-MB-231 cells that migrated after 24 hours of treatment with compound 3ma. (Note: Compared with the control group, ** P <0.01, * P <0.05). Detailed Implementation

[0029] Example 1

[0030] A solution of benzodithiocyclopentanone 1 (0.1 mmol) and quinazoline-derived dipolar compound 2 (0.1 mmol) in dichloromethane (2 mL) was mixed with methyldiphenylphosphine (0.1 mmol), and the mixture was stirred at 35°C for 3 hours until the reaction was stopped. The mixture was evaporated under reduced pressure and separated by column chromatography to obtain compound 3, which is the quinazoline-fused benzothiocyanate heterocyclic compound.

[0031] Compound 3aa, yield 97%, mp 229-231 °C. 1 H NMR (400 MHz, CDCl3): d 8.11 (d, J = 6.8 Hz, 2H), 7.68 (d,J = 7.6 Hz, 1H), 7.44-7.29 (m, 6H), 7.25-7.22 (m,2H), 6.98-6.96 (m, 1H), 6.91 (s, 1H), 6.55 (s, 1H), 2.49 (s, 3H). 13 C NMR (100MHz, CDCl3): d 169.7, 146.7, 146.6, 137.6, 137.5, 135.8, 134.1, 133.3, 131.4,130.2, 129.8, 129.4, 129.0, 127.9, 127.3, 125.8, 123.9, 72.0, 21.9. HRMS(ESI) m / z: [M + H] + calcd. for C 22 H 18 N3O3S2, 436.0784; found, 436.0784.

[0032] Compound 3ba, yield 95%, mp 202-204 °C. 1 H NMR (400 MHz, CDCl3): d 8.10 (d, J = 7.6 Hz, 2H), 7.48 (s, 1H), 7.42 (d, J = 8.0 Hz, 2H), 7.30-7.26 (m, 1H), 7.24-7.15 (m, 4H), 6.98-6.95 (m, 1H), 6.87 (s, 1H), 6.51 (s, 1H), 2.48 (s,3H), 2.31 (s, 3H). 13 C NMR (100 MHz, CDCl3): d 169.8, 147.1, 146.6, 140.9,137.1, 137.0, 135.7, 134.2, 134.1, 132.2, 130.2, 129.8, 129.4, 128.0, 127.5,125.4, 125.3, 123.8, 71.9, 21.9, 21.2. HRMS (ESI) m / z: [M + H] + calcd. forC 23 H 20N3O3S2, 450.0941; found, 450.0949.

[0033] Compound 3ca, yield 97%, mp 255-257 °C. 1 H NMR (400 MHz, CDCl3): d 8.10 (d, J = 7.6 Hz, 2H), 7.43 (d, J = 8.0 Hz, 2H), 7.31-7.18 (m, 5H), 7.00-6.97 (m,1H), 6.87 (d, J = 8.4Hz, 1H), 6.83 (s, 1H), 6.58 (s, 1H), 3.76 (s, 3H), 2.49(s, 3H). 13 C NMR (100 MHz, CDCl3): d 169.6, 161.1, 146.8, 146.6, 138.7, 137.6,137.2, 134.1, 130.2, 129.7, 129.4, 127.9, 127.3, 125.7, 123.9, 119.2, 119.1,116.6, 71.8, 55.7, 22.0. HRMS (ESI) m / z: [M + H] + calcd. for C 23 H 20 N3O4S2,466.0890; found, 466.0897.

[0034] Compound 3da, yield 93%, mp 228-230 °C. 1 H NMR (400 MHz, CDCl3): d 8.10 (d, J = 8.0 Hz, 2H), 7.65 (s, 1H), 7.44 (d, J = 8.0 Hz, 2H), 7.34 (d, J = 8.4 Hz,1H), 7.29-7.23 (m, 4H), 7.01-6.99 (m, 1H), 6.92 (s, 1H), 6.52 (s, 1H), 2.49(s, 3H). 13 C NMR (100 MHz, CDCl3): d168.4, 163.4 ( J = 252.0 Hz), 146.8,146.5, 139.7 ( J = 7.6 Hz), 137.9 ( J = 8.0 Hz), 137.5, 133.9, 130.3, 130.0,129.4, 127.9, 127.5, 125.9, 124.3 ( J = 3.8 Hz), 123.6, 120.4 ( J = 21.4 Hz), 118.8 ( J = 24.5 Hz), 72.1, 22.0. HRMS (ESI) m / z: [M + H] + calcd. forC 22 H 17 FN3O3S2, 454.0690; found, 454.0689.

[0035] Compound 3ea, yield 89%, mp 234-236 °C. 1 H NMR (400 MHz, CDCl3): d 8.10 (d, J = 8.4 Hz, 2H), 7.44 (d, J = 7.6 Hz, 2H), 7.40 (d, J = 8.0 Hz, 1H), 7.32-7.23(m, 4H), 7.08 (t, J = 8.4 Hz, 1H), 7.01-6.98 (m, 1H), 6.90 (s, 1H), 6.56 (s, 1H), 2.50 (s, 3H). 13 C NMR (100 MHz, CDCl3): d 168.4, 146.8, 146.4, 138.9,137.4, 136.9, 136.6, 133.8, 133.2, 131.4, 130.2, 130.0, 129.3, 127.8, 127.5,127.4, 125.9, 123.6, 72.1, 21.9. HRMS (ESI) m / z: [M + H] + calcd. forC 22 H 17ClN3O3S2, 470.0394; found, 470.0399.

[0036] Compound 3fa, yield 86%, mp 239-241 °C. 1 H NMR (400 MHz, DMSO- d 6 ): d 8.11 (d, J = 8.0 Hz, 2H), 7.80 (s, 1H), 7.68 (d, J = 8.4 Hz, 1H), 7.57 (d, J = 8.0 Hz,2H), 7.50-7.47 (m, 1H), 7.29-7.26 (m, 2H), 7.23 (d, J = 6.8 Hz, 3H), 6.92-6.89 (m, 1H) 2.47 (s, 3H). 13 C NMR (100 MHz, DMSO- d 6 ): d 168.0, 148.6, 147.0,139.5, 137.2, 137.1, 136.3, 134.7, 133.6, 130.6, 130.3, 129.8, 128.7, 128.6,127.6, 125.2, 124.2, 123.4, 71.7, 21.8. HRMS (ESI) m / z: [M + H] + calcd. forC 22 H 17 BrN3O3S2, 513.9889; found, 513.9888.

[0037] Compound 3ga, yield 83%, mp 257-259 °C. 1 H NMR (400 MHz, DMSO- d 6 ): d 8.10 (d, J = 8.0 Hz, 2H), 7.93 (s, 1H), 7.83 (d, J = 7.6 Hz, 1H), 7.57 (d, J= 8.0 Hz,2H), 7.48-7.46 (m, 1H), 7.27-7.25 (m, 2H), 7.20 (s, 2H), 7.06 (d, J = 8.0 Hz,1H), 6.91-6.89 (m, 1H), 2.47 (s, 3H). 13 C NMR (100 MHz, DMSO- d 6 ): d 168.0,148.3, 147.0, 142.1, 140.3, 139.2, 137.6, 137.0, 133.7, 130.6, 130.3, 129.8,128.9, 128.7, 127.5, 125.5, 124.4, 96.5, 71.7, 21.7. HRMS (ESI) m / z: [M + H] + calcd. for C 22 H 17 IN3O3S2, 561.9751; found, 561.9756.

[0038] Compound 3ha, yield 79%, mp 222-224 °C. 1 H NMR (400 MHz, CDCl3): d 8.50 (s, 1H), 8.20 (d, J = 8.4 Hz, 1H), 8.11 (d, J = 7.6 Hz, 2H), 7.54 (d, J = 6.8 Hz, 1H), 7.47 (d, J = 8.0 Hz, 2H), 7.29 (s, 3H), 7.07 (s, 1H), 7.03-7.00 (m, 1H), 6.52 (s, 1H), 2.51 (s, 3H). 13 C NMR (100 MHz, CDCl3): d 167.5, 148.4, 147.2,146.0, 138.8, 137.8, 137.0, 136.8, 133.5, 130.4, 130.3, 129.4, 127.8, 127.7,127.2, 126.5, 126.2, 123.4, 73.0, 22.0. HRMS (ESI) m / z: [M + H]+ calcd. forC 22 H 17 N4O5S2, 481.0635; found, 481.0641.

[0039] Compound 3ia, yield 85%, mp 262-264 °C. 1 H NMR (400 MHz, CDCl3): d 8.31 (s, 1H), 8.11 (d, J = 8.0 Hz, 2H), 8.05 (d, J = 7.6 Hz, 1H), 7.43 (t, J = 8.4 Hz, 3H), 7.29-7.23 (m, 3H), 6.99-6.96 (m, 2H), 6.47 (s, 1H), 3.89 (s, 3H), 2.50 (s, 3H). 13 C NMR (100 MHz, CDCl3): d 168.8, 165.2, 146.8, 146.3, 137.6, 137.3,135.9, 134.8, 133.9, 133.7, 132.4, 131.7, 130.2, 130.0, 129.3, 127.8, 127.5,126.0, 123.7, 72.5, 52.6, 21.9. HRMS (ESI) m / z: [M + H] + calcd. forC 24 H 20 N3O5S2, 494.0839; found, 494.0847.

[0040] Compound 3ja, yield 98%, mp 220-222 °C. 1 H NMR (400 MHz, CDCl3): d 8.10 (d, J = 8.0 Hz, 2H), 7.66 (d, J = 8.4 Hz, 1H), 7.42 (d, J = 8.0 Hz, 2H), 7.30-7.24(m, 3H), 7.04-7.02 (m, 1H), 6.93 (s, 1H), 6.88 (d, J= 8.4 Hz, 1H), 6.86 (s,1H), 6.47 (s, 1H), 3.73 (s, 3H), 2.47 (s, 3H). 13 C NMR (100 MHz, CDCl3): d 169.4, 163.1, 147.2, 146.5, 136.9, 134.1, 133.8, 131.4, 130.2, 129.9, 129.4,128.4, 127.9, 127.6, 125.5, 124.0, 121.4, 115.1, 71.8, 55.7, 21.9. HRMS (ESI)m / z: [M + H] + calcd. for C 23 H 20 N3O4S2, 466.0890; found, 466.0895.

[0041] Compound 3 Ka, yield 89%, mp 254-256 °C. 1 H NMR (400 MHz, CDCl3): d 8.09 (d, J = 8.0 Hz, 2H), 7.55 (s, 2H), 7.50 (s, 1H), 7.43 (d, J = 8.0 Hz, 2H), 7.27-7.25 (m, 3H), 7.03-7.01 (m, 1H), 6.94 (s, 1H), 6.53 (s, 1H), 2.49 (s, 3H). 13 CNMR (100 MHz, CDCl3): d 169.0, 146.8, 146.4, 138.0, 137.4, 136.1, 133.9,133.3, 132.8, 131.4, 130.2, 130.1, 129.4, 127.8, 127.6, 126.1, 123.6, 72.2,21.9. HRMS (ESI) m / z: [M + H] + calcd. for C 22 H 17 BrN3O3S2, 513.9889; found, 513.9890.

[0042] Compound 3la, yield 94%, mp 241-243 °C.1 H NMR (400 MHz, DMSO- d 6 ): d 8.14 (d, J = 8.0 Hz, 2H), 7.84 (d, J = 8.0 Hz, 1H), 7.62 (d, J = 7.6 Hz, 1H), 7.56 (d, J = 7.6 Hz, 3H), 7.44 (t, J = 8.0 Hz, 1H), 7.28-7.26 (m, 3H), 7.17 (s, 1H), 6.90-6.88 (m, 1H), 2.47 (s, 3H). 13 C NMR (100 MHz, CDCl3): d 168.7, 146.8,146.7, 140.3, 137.1, 136.8, 133.9, 131.3, 131.1, 130.2, 130.1, 129.8, 129.4,129.3, 128.7, 127.5, 125.4, 122.6, 73.1, 21.9. HRMS (ESI) m / z: [M + H] + calcd.for C 22 H 17 BrN3O3S2, 513.9889; found, 513.9889.

[0043] Compound 3ma, yield 93%, mp 240-242 °C. 1 H NMR (400 MHz, CDCl3): d 8.12 (d, J = 8.4 Hz, 2H), 7.44 (d, J = 8.0 Hz, 2H), 7.30-7.17 (m, 5H), 7.05 (d, J = 7.6Hz, 1H), 6.99-6.96 (m, 1H), 6.78 (s, 1H), 6.71 (s, 1H), 2.50 (s, 3H), 2.30(s, 3H). 13 C NMR (100 MHz, CDCl3): d168.8, 147.3, 146.5, 139.5, 137.7, 137.2,134.5, 133.3, 132.9, 131.7, 130.0, 129.8, 129.3, 128.0, 127.2, 127.0, 125.6,123.2, 72.2, 22.0, 19.7. HRMS (ESI) m / z: [M + H] + calcd. for C 23 H 20 N3O3S2,450.0941; found, 450.0950.

[0044] Compound 3ab, 90% yield, mp 292-294 °C. 1 H NMR (400 MHz, DMSO- d 6 ): d 8.14 (d, J = 7.6 Hz, 2H), 7.68 (d, J = 7.6 Hz, 1H), 7.57 (d, J = 8.4 Hz, 3H), 7.52 (t, J = 7.6 Hz, 1H), 7.46 (d, J = 8.0 Hz, 1H), 7.39 (d, J = 8.0 Hz, 1H), 7.35 (d, J = 7.6 Hz, 1H), 7.30 (s, 1H), 7.25 (t, J = 8.0 Hz, 1H), 7.22 (s, 1H), 2.47 (s, 3H). 13 C NMR (100 MHz, DMSO- d 6 ): d 169.5, 149.6, 147.0, 146.5, 137.4,135.6, 134.9, 134.0, 133.8, 132.3, 130.6, 130.0, 128.8, 128.2, 127.8, 126.7,123.6, 70.9, 21.8. HRMS (ESI) m / z: [M + H] + calcd. for C 22 H 17ClN3O3S2, 470.0394; found, 470.0402.

[0045] Compound 3ac, 98% yield, mp 232-234 °C. 1 H NMR (400 MHz, CDCl3): d 8.10 (d, J = 7.6 Hz, 2H), 7.67 (d, J = 7.6 Hz, 1H), 7.44-7.31 (m, 5H), 7.21 (d, J = 8.4Hz, 1H), 6.89 (s, 1H), 6.80 (d, J = 8.8 Hz, 1H), 6.60 (s, 1H), 6.51 (s, 1H), 3.77 (s, 3H), 2.49 (s, 3H). 13 C NMR (100 MHz, CDCl3): d 169.7, 160.6, 147.3,146.6, 138.5, 137.4, 135.9, 134.1, 133.3, 131.4, 130.2, 130.1, 129.4, 129.1,128.8, 115.8, 114.4, 109.5, 72.2, 55.5, 21.9. HRMS (ESI) m / z: [M + H] + calcd.for C 23 H 20 N3O4S2, 466.0890; found, 466.0895.

[0046] Compound 3ad, yield 88%, mp 247-249 °C. 1 H NMR (400 MHz, CDCl3): d 8.10 (d, J = 7.6 Hz, 2H), 7.68 (d, J = 7.2 Hz, 1H), 7.47-7.39 (m, 4H), 7.33-7.28 (m,2H), 6.95 (t, J = 8.4 Hz, 1H), 6.89 (s, 1H), 6.70 (d, J = 9.6 Hz, 1H), 6.61(s, 1H), 2.49 (s, 3H).13 C NMR (100 MHz, CDCl3): d 169.4, 163.1 ( J = 246.7Hz), 147.7, 146.7, 139.1 ( J = 11.4 Hz), 137.3, 135.8, 133.9, 133.4, 131.5,130.3, 130.2, 129.4, 129.3 ( J = 9.2 Hz), 128.5, 119.6 ( J = 3.3 Hz), 114.6 ( J = 22.4 Hz), 112.2 ( J = 22.5 Hz), 71.6, 21.9. HRMS (ESI) m / z: [M + H] + calcd.for C 22 H 17 FN3O3S2, 454.0690; found, 454.0690.

[0047] Compound 3ae, yield 84%, mp 276-278 °C. 1 H NMR (400 MHz, CDCl3): d 8.10 (d, J = 8.0 Hz, 2H), 7.69 (d, J = 7.2 Hz, 1H), 7.45-7.35 (m, 5H), 7.32 (s, 1H),7.20 (d, J = 8.4 Hz, 1H), 6.96 (d, J = 8.4 Hz, 1H), 6.84 (s, 1H), 6.64 (s, 1H), 2.49 (s, 3H). 13 C NMR (100 MHz, CDCl3): d 169.2, 147.3, 146.8, 137.2,135.8, 135.4, 133.8, 133.5, 132.8, 131.5, 130.5, 130.2, 130.1, 129.4, 128.1,127.7, 126.7, 125.1, 71.2, 21.9. HRMS (ESI) m / z: [M + H] +calcd. forC 22 H 17 ClN3O3S2, 470.0394; found, 470.0395.

[0048] Compound 3af, yield 97%, mp 241-243 °C. 1 H NMR (400 MHz, CDCl3): d 8.10 (d, J = 8.0 Hz, 2H), 7.67 (d, J = 7.2 Hz, 1H), 7.44-7.33 (m, 5H), 6.95 (d, J = 8.8Hz, 1H), 6.88 (s, 1H), 6.81-6.76 (m, 2H), 6.50 (s, 1H), 3.85 (s, 3H), 2.48(s, 3H). 13 C NMR (100 MHz, CDCl3): d 169.7, 158.7, 146.6, 145.0, 137.5, 135.8,134.1, 133.3, 131.4, 130.8, 130.2, 129.4, 128.8, 126.9, 124.9, 115.8, 112.4,72.1, 55.7, 21.9. HRMS (ESI) m / z: [M + H] + calcd. for C 23 H 20 N3O4S2, 466.0890; found, 466.0896.

[0049] Compound 3 ag, yield 82%, mp 246-248 ℃. 1 H NMR (400 MHz, CDCl3): d 8.10 (d, J = 7.6 Hz, 2H), 7.68 (d, J = 7.2 Hz, 1H), 7.47-7.39 (m, 4H), 7.34 (d, J = 7.6Hz, 1H), 7.05-6.91 (m, 3H), 6.86 (s, 1H), 6.59 (s, 1H), 2.49 (s, 3H). 13 C NMR (100 MHz, CDCl3): d 169.4, 161.1 ( J = 246.4 Hz), 146.7, 146.3, 137.4, 135.8,133.9, 133.7 ( J = 2.7 Hz), 133.4, 131.4, 130.4, 130.2, 129.4, 128.3, 127.4 ( J = 8.6 Hz), 125.5 ( J = 8.1 Hz), 117.0 ( J = 22.4 Hz), 114.4 ( J = 24.4 Hz),71.4, 21.9. HRMS (ESI) m / z: [M + H] + calcd. for C 22 H 17 FN3O3S2, 454.0690; found, 454.0699.

[0050] Compound 3ah, yield 89%, mp 245-247 °C. 1 H NMR (400 MHz, CDCl3): d 8.24 (d, J = 8.0 Hz, 2H), 7.76 (t, J = 7.6 Hz, 1H), 7.69-7.64 (m, 3H), 7.44 (t, J = 7.6Hz, 1H), 7.39 (t, J = 7.6 Hz, 1H), 7.33-7.31 (m, 2H), 7.26-7.23 (m, 2H), 7.03-7.00 (m, 1H), 6.92 (s, 1H), 6.61 (s, 1H). 13 C NMR (100 MHz, CDCl3): d 169.6, 146.9, 137.3, 137.1, 137.0, 135.9, 135.2, 133.5, 131.5, 130.3, 129.9,129.6, 129.4, 128.8, 128.0, 127.6, 125.6, 123.7, 72.0. HRMS (ESI) m / z: [M +H] + calcd. for C 21 H16 N3O3S2, 422.0628; found, 422.0634.

[0051] Compound 3ai, yield 85%, mp 239-241 °C. 1 H NMR (400 MHz, CDCl3): d 8.16 (d, J = 8.0 Hz, 2H), 7.68 (d, J = 7.6 Hz, 1H), 7.45-7.21 (m, 6H), 7.08 (d, J = 8.0Hz, 2H), 7.00-6.97 (m, 1H), 6.91 (s, 1H), 6.58 (s, 1H), 3.92 (s, 3H). 13 C NMR (100 MHz, CDCl3): d 169.7, 165.0, 146.9, 137.5, 137.4, 135.8, 133.3, 131.8,131.4, 130.2, 129.9, 128.9, 128.1, 127.9, 127.4, 125.7, 123.8, 114.7, 72.0,56.0. HRMS (ESI) m / z: [M + H] + calcd. for C 22 H 18 N3O4S2, 452.0733; found, 452.0735.

[0052] Compound 3aj, yield 94%, mp 243-245 °C. 1 H NMR (400 MHz, CDCl3): d 8.17 (d, J = 8.4 Hz, 2H), 7.68 (d, J = 7.6 Hz, 1H), 7.62 (d, J = 8.4 Hz, 2H), 7.45 (t, J = 7.6 Hz, 1H), 7.40 (t, J = 7.6 Hz, 1H), 7.34-7.30 (m, 2H), 7.26-7.23 (m,2H), 7.02-7.00 (m, 1H), 6.90 (s, 1H), 6.63 (s, 1H).13 C NMR (100 MHz, CDCl3): d 169.7, 146.8, 142.2, 137.1, 136.9, 135.9, 135.4, 133.6, 131.5, 130.9,130.4, 130.0, 129.9, 128.7, 128.0, 127.7, 125.6, 123.6, 72.0. HRMS (ESI) m / z:[M + H] + calcd. for C 21 H 15 ClN3O3S2, 456.0238; found, 456.0247.

[0053] Compound 3ak, yield 98%, mp 227-229 °C. 1 H NMR (400 MHz, CDCl3): d 8.14 (d, J = 8.0 Hz, 2H), 7.70 (d, J = 7.2 Hz, 1H), 7.64 (d, J = 8.0 Hz, 2H), 7.44 (t, J = 7.6 Hz, 1H), 7.39 (t, J = 7.6 Hz, 1H), 7.34-7.30 (m, 2H), 7.26-7.23 (m,2H), 7.01-7.00 (m, 1H), 6.93 (s, 1H), 6.61 (s, 1H), 1.37 (s, 9H). 13 C NMR (100MHz, CDCl3): d 169.6, 159.4, 146.9, 137.3, 137.1, 135.8, 133.9, 133.3, 131.5,130.2, 129.8, 129.2, 128.8, 127.9, 127.4, 126.6, 125.5, 123.7, 72.0, 35.6,31.0. HRMS (ESI) m / z: [M + H] + calcd. for C 25 H 24 N3O3S2, 478.1254; found, 478.1261.

[0054] Compound 3al, yield 94%, mp 223-225 °C. 1 H NMR (400 MHz, CDCl3): d 8.11 (s, 1H), 7.86 (d, J = 5.2 Hz, 1H), 7.74 (d, J = 7.6 Hz, 1H), 7.47 (t, J = 7.6 Hz, 1H), 7.41 (t, J = 7.6 Hz, 1H), 7.34-7.23 (m, 5H), 7.01-6.99 (m, 1H), 6.89 (s, 1H), 6.64 (s, 1H). 13 C NMR (100 MHz, CDCl3): d 169.5, 146.8, 137.3, 137.2,137.0, 136.7, 136.2, 135.9, 133.5, 131.5, 130.3, 130.0, 128.8, 128.2, 127.9,127.6, 125.6, 123.7, 71.9. HRMS (ESI) m / z: [M + H] + calcd. for C 19 H 14 N3O3S3,428.0192; found, 428.0195.

[0055] Compound 3am, yield 88%, mp 221-223 ℃. 1 H NMR (400 MHz, CDCl3): d 7.73 (d, J = 7.6 Hz, 1H), 7.55-7.53 (m, 2H), 7.50-7.44 (m, 4H), 7.40 (t, J = 7.6 Hz,1H), 7.29-7.26 (m, 1H), 7.19-7.13 (m, 2H), 7.06 (d, J = 6.8 Hz, 1H), 6.89 (d, J = 7.6 Hz, 1H), 6.15 (s, 1H), 5.50 (s, 1H), 5.16 (d, J = 14.0 Hz, 1H), 4.98(d,J = 14.0 Hz, 1H). 13 C NMR (100 MHz, CDCl3): d 171.8, 146.4, 137.5, 137.1,135.6, 133.3, 131.1, 130.6, 130.3, 130.1, 129.7, 129.5, 128.9, 127.5, 127.2,125.6, 123.6, 72.0, 61.1. HRMS (ESI) m / z: [M + H] + calcd. for C 22 H 18 N3O3S2,436.0784; found, 436.0795.

[0056] Example 2: In vitro antitumor activity test Cells in the logarithmic growth phase were collected and prepared into a single-cell suspension using complete culture medium containing 10% fetal bovine serum. Cells were seeded at a density of 5000 cells per well in 96-well plates, with a volume of 100 mg / well. µ L. Place the culture plates in a 37°C, 5% CO2 incubator and incubate for 24 hours, then aspirate the original culture medium. Replace the experimental groups with fresh complete culture medium containing different concentrations of the test compounds, and replace the negative control group with complete culture medium without the drug. Return the culture plates to the incubator and continue incubation for 48 hours. After the drug treatment, add 10 μL of the drug to each well. µ Incubate with L MTT solution for another 4 hours. Carefully aspirate the culture supernatant from each well and add 100 μL of MTT solution to each well. m Dissolve the crystals in L DMSO. Measure the absorbance of each well at 492 nm using a microplate reader and record the results.

[0057] Example 3: Wound healing experiment of compound 3ma Seed cells evenly into 6-well plates, approximately 5 × 10⁶ cells per well. 5 Add complete culture medium and incubate at 37°C with 5% CO2 for 24 hours until cell confluence reaches 80-90%. Use 10 µ The L-shaped pipette tip was held perpendicular to the bottom of the well plate and used to slice through the cell layer. The plate was then gently rinsed twice with PBS to remove any detached cell debris. After washing, culture medium and different concentrations of compound 3 mA were added, and images were taken under an inverted microscope, recorded as hour 0. The culture plate was then returned to the incubator, and images were taken again at 24, 36, and 48 hours. The area of ​​the scratched region at each time point was measured using ImageJ software, and cell migration rate was calculated.

[0058] Example 4: Transwell migration experiment of compound 3ma Take cells in the logarithmic growth phase and "starve" them 12 hours in advance by replacing the medium with serum-free medium. Add 600 μL of serum-free medium to the lower chamber of the Transwell chamber. µ L of complete culture medium containing 10% FBS, with different concentrations of 3mA of serum-free 200 mg / L added to the upper chamber. µ L cell suspension. The culture plate was placed in a cell culture incubator at 37°C and 5% CO2 and cultured for 24 hours. After culture, the chambers were removed, the culture medium was discarded, and the chambers were rinsed with PBS. The surface of the upper chamber membrane was gently wiped with a moistened cotton swab to remove unmigrated cells. The chambers were then fixed in 4% paraformaldehyde for 30 minutes, followed by staining with 0.1% crystal violet solution for 30 minutes. The chambers were rinsed with PBS, air-dried, and observed under a microscope. Images of migrating cells on the lower membrane surface were randomly selected from five fields of view and processed using ImageJ software.

[0059] The above description is merely a preferred embodiment of the present invention and is not intended to limit the invention. Various modifications and variations can be made to the present invention by those skilled in the art. Any modifications, equivalent substitutions, improvements, etc., made within the spirit and principles of the present invention should be included within the scope of protection of the present invention.

Claims

1. A quinazoline-fused benzothiocyanate tetracyclic compound that is a structural analog of berberine, characterized in that, Its chemical structural formula is shown in general formula 3: The general formula 3 is represented by compounds 3aa, 3ba, 3ca, 3ea, 3fa, 3ja, 3ka, 3ma, 3ad, 3af, 3aj, and 3ak. 。 2. A method for preparing a quinazoline-fused benzothiocyanate tetracyclic compound as a berberine structural analog as described in claim 1, characterized in that, The specific steps are as follows: In an organic solvent, at a certain temperature and under the action of a phosphine reagent, the benzodithiocyclopentanone compound shown in general formula 1 is reacted with the quinazoline-derived dipolar compound shown in general formula 2 for a period of time, and the compound of general formula 3 is obtained through a [4 + 3] cyclization reaction. R in general formula 1 1 And R of the general formula 2 2 and R 3 The definition is as defined in claim 1, wherein the phosphine reagent is a trialkylphosphine or a triarylphosphine.

3. The method for preparing the quinazoline-fused benzothiocyanate tetracyclic compound, which is a structural analog of berberine, according to claim 2, is characterized in that... The molar ratio of the benzodithiocyclopentanone compound shown in Formula 1 to the quinazoline-derived dipolar compound shown in Formula 2 is 1:(1-1.5); the organic solvent is a polar aprotic solvent and a haloalkanes solvent.

4. The method for preparing the quinazoline-fused benzothiocyanate tetracyclic compound, which is a structural analog of berberine, according to claim 2 or 3, is characterized in that... The molar ratio of the benzodithiocyclopentanone compound shown in Formula 1 to the quinazoline-derived dipolar compound shown in Formula 2 is 1:1; the phosphine reagent is methyldiphenylphosphine; and the organic solvent is dichloromethane.

5. The method for preparing the quinazoline-fused benzothiocyanate tetracyclic compound, which is a structural analog of berberine, according to claim 2 or 3, is characterized in that... After stirring and reacting for a period of time under certain temperature conditions, the mixture is dried under reduced pressure and separated by column chromatography to obtain the compound of general formula 3.

6. The method for preparing the quinazoline-fused benzothiocyanate tetracyclic compound as a berberine structural analog according to claim 5, characterized in that, The reaction temperature is from 25 degrees to the reflux temperature of the solvent; the reaction time is 1 to 5 hours.

7. The method for preparing the quinazoline-fused benzothiocyanate tetracyclic compound as a berberine structural analog according to claim 6, characterized in that, The reaction temperature is 35 degrees Celsius; the reaction time is 3 hours.

8. Use of the quinazoline fused benzothiocyanate compound as a berberine structural analog as described in claim 1 in the preparation of a medicament for treating triple-negative breast cancer.

9. The use according to claim 8, characterized in that, It is used to inhibit the proliferation and / or migration of triple-negative breast cancer cells.

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