Tetrahydropyrrole derivative for resisting breast cancer as well as preparation method, application and medicine thereof

Abstract

The invention discloses a tetrahydropyrrole derivative for resisting breast cancer as well as a preparation method, application and a medicine of the tetrahydropyrrole derivative. The structural formula of the tetrahydropyrrole derivative is shown as a chemical formula 1. The application refers to application of the tetrahydropyrrole derivative in preparation of anti-breast cancer drugs. The drug is an anti-breast cancer drug comprising the tetrahydropyrrole derivative or a pharmaceutically acceptable salt of the tetrahydropyrrole derivative. Experiments prove that the tetrahydropyrrole derivative has an anti-breast cancer effect and can be used as a candidate active component for preparing the anti-breast cancer medicine.

Description

Technical Field

[0001] This invention belongs to the field of synthesis and application technology of nitrogen-containing heterocyclic compounds, specifically relating to a tetrahydropyrrole derivative for anti-breast cancer, its preparation method, application and drug. Background Technology

[0002] Breast cancer is one of the most common malignant tumors in women, and drug treatment is the primary approach. With in-depth research, anti-breast cancer drugs have evolved from traditional chemotherapy drugs to targeted therapy, endocrine therapy, and novel molecular drugs, among other directions.

[0003] Nitrogen-containing heterocyclic compounds are widely found in natural products and possess a variety of pharmacological activities, including antitumor, anti-inflammatory, and antibacterial effects. According to the U.S. Food and Drug Administration (FDA), nitrogen-containing heterocyclic compounds account for approximately 82% of marketed small molecule drugs, and in 2023 alone, 13 nitrogen-containing heterocyclic drugs were approved by the FDA for the treatment of various diseases, including cancer, migraines, and postpartum depression. Therefore, nitrogen-containing heterocycles represent a class of pharmacophores with diverse pharmacological activities.

[0004] Tetrahydropyrrole is a five-membered nitrogen-containing heterocycle, often used as a base side chain in the design of new drugs. For example, the estrogen receptor antagonist lasofoxifene, used to treat hormone-dependent breast cancer, and the telomerase / telomere inhibitor Braco-19, as well as G9a and GLP histone methyltransferase inhibitor UNC0638, which have anti-triple-negative breast cancer activity, all contain a tetrahydropyrrole group. Furthermore, fusion of tetrahydropyrrole with other heterocycles also exhibits antitumor activity. For example, acetylardeemin, a compound fused with tetrahydropyrrole and an indole ring, has significant inhibitory activity against breast cancer and doxorubicin-resistant breast cancer.

[0005] Currently, a large number of marketed anti-breast cancer drugs are used in clinical practice. However, issues such as drug resistance and toxic side effects severely limit drug screening in clinical settings. Therefore, designing and developing more structurally novel anti-breast cancer drugs is of great significance.

[0006] It should be noted that the information disclosed in the background section above is only used to enhance the understanding of the background of the present invention, and therefore may include information that does not constitute prior art known to those skilled in the art. Summary of the Invention

[0007] To address the aforementioned problems in the prior art, this invention provides a tetrahydropyrrole derivative for anti-breast cancer treatment, its preparation method, application, and pharmaceutical formulation. The technical problem to be solved by this invention is achieved through the following technical solution: In a first aspect, the present invention provides a tetrahydropyrrole derivative for use in the treatment of breast cancer, the structural formula of which is shown in Chemical Formula 1:

[0008] Chemical Formula 1 R1 is selected from halogens, cycloalkyl groups with 3 to 10 carbon atoms, and alkylamine groups with 3 to 10 carbon atoms; R2 is selected from alkyl groups having 1 to 10 carbon atoms and cycloalkyl groups having 3 to 10 carbon atoms; R3 is selected from alkylamine with 1 to 10 carbon atoms, dialkylamine with 3 to 10 carbon atoms, and saturated heterocyclic alkyl with 3 to 10 carbon atoms; L is selected from 1 to 10 alkyl groups.

[0009] In one embodiment of the present invention, R1 is selected from chlorine, methylamino, isopropylamino, and cyclopropyl.

[0010] In one embodiment of the present invention, R2 is selected from methyl, ethyl, and isopropyl.

[0011] In one embodiment of the present invention, R3 is selected from tetrahydropyrrole-N-yl, dimethylamino, and diethylamino.

[0012] In one embodiment of the present invention, L is selected from methylene, ethylene, and propylene.

[0013] In one embodiment of the present invention, the tetrahydropyrrole derivative is selected from the group consisting of the following compounds:

[0014]

[0015]

[0016] .

[0017] Secondly, the present invention provides a method for preparing tetrahydropyrrole derivatives of the above-mentioned compounds TP1, TP2, TP3 and TP4, comprising the following steps: S1. Compound 1, Compound 2, 1-ethyl-3-(3-dimethylaminopropyl)carbodiimide hydrochloride and 1-hydroxybenzotriazole were added to anhydrous dichloromethane, mixed and reacted at room temperature. After the reaction was completed, intermediate 1 was obtained by separation and purification. ; S2. Add intermediate 1 to phosphorus oxychloride and react at 100-130℃. After the reaction is completed, separate and purify to obtain compound TP1. ; S3. Compound TP1, CH3-NH2 and sodium tert-butoxide are added to dioxane, and then a third-generation palladium precatalyst with BrettPhos ligand is added to form a reaction system; the reaction system is placed in a sealed tube and reacted under nitrogen protection. After the reaction is completed, compound TP2 is obtained by separation and purification. ; Compound TP1, (CH3)2CH-NH2, and sodium tert-butoxide were added to dioxane, followed by the addition of a third-generation palladium precatalyst with a BrettPhos ligand to form a reaction system. The reaction system was placed in a sealed tube and reacted under nitrogen protection. After the reaction was completed, compound TP3 was obtained by separation and purification. ; Compound TP1, cyclopropylboronic acid and K3PO4 were dissolved in toluene aqueous solution, Pd(Amphos)2Cl2 was added, and the reaction was carried out under nitrogen protection. After the reaction was completed, the compound TP4 was obtained by separation and purification. .

[0018] Thirdly, the present invention provides the application of the above-mentioned tetrahydropyrrole derivative in the preparation of anti-breast cancer drugs.

[0019] Fourthly, the present invention provides an anti-breast cancer drug comprising the above-mentioned tetrahydropyrrole derivative, or a pharmaceutically acceptable salt comprising the above-mentioned tetrahydropyrrole derivative.

[0020] In one embodiment of the present invention, the pharmaceutically acceptable salt of the tetrahydropyrrole derivative includes one of hydrochloride, sulfate, phosphate, citrate, maleate and tartrate.

[0021] Compared with the prior art, the beneficial effects of the present invention are as follows: This invention provides a tetrahydropyrrole derivative for anti-breast cancer treatment and its preparation method. Experiments have verified that this tetrahydropyrrole derivative has anti-breast cancer effects and can be used as a candidate active ingredient for preparing anti-breast cancer drugs. Compound TP4 exhibits strong inhibitory activity against doxorubicin-resistant MCF-7 / ADR cells, demonstrating high application value and promising development prospects in the preparation of drugs for treating drug-resistant breast cancer.

[0022] The present invention will be further described in detail below with reference to the accompanying drawings and embodiments. Attached Figure Description

[0023] Figure 1 This is the 1H NMR spectrum characterization of compound TP1 provided in the embodiments of the present invention; Figure 2 This is a mass spectrometry characterization of compound TP1 provided in an embodiment of the present invention; Figure 3 This is the HPLC characterization chromatogram of compound TP1 provided in the embodiments of the present invention; Figure 4 This is the proton nuclear magnetic resonance spectrum of compound TP2 provided in the embodiments of the present invention; Figure 5 This is a mass spectrometry characterization of compound TP2 provided in the embodiments of the present invention; Figure 6 This is the HPLC characterization chromatogram of compound TP2 provided in the embodiments of the present invention; Figure 7 This is the 1H NMR spectrum characterization of compound TP3 provided in the embodiments of the present invention; Figure 8 This is a mass spectrometry characterization of compound TP3 provided in an embodiment of the present invention; Figure 9 This is the HPLC characterization chromatogram of compound TP3 provided in the embodiments of the present invention; Figure 10 This is the proton NMR spectrum of compound TP4 provided in the embodiments of the present invention; Figure 11 This is a mass spectrometry characterization of compound TP4 provided in the embodiments of the present invention; Figure 12 This is an HPLC characterization chromatogram of compound TP4 provided in the embodiments of the present invention. Detailed Implementation

[0024] To further illustrate the technical means and effects adopted by the present invention to achieve the intended purpose, the following, in conjunction with the accompanying drawings and specific embodiments, provides a detailed description of a tetrahydropyrrole derivative for anti-breast cancer, its preparation method, application, and drug, as proposed in accordance with the present invention.

[0025] The foregoing and other technical contents, features, and effects of the present invention will be clearly presented in the following detailed description of specific embodiments in conjunction with the accompanying drawings. Through the description of the specific embodiments, a more in-depth and concrete understanding can be gained of the technical means and effects adopted by the present invention to achieve its intended purpose. However, the accompanying drawings are for reference and illustration only and are not intended to limit the technical solutions of the present invention.

[0026] It should be noted that, in this document, the terms "comprising," "including," or any other variations are intended to cover non-exclusive inclusion, such that an article or device that comprises a list of elements includes not only those elements but also other elements not expressly listed. Without further limitation, an element defined by the phrase "comprising one..." does not exclude the presence of other identical elements in the article or device that includes said element.

[0027] This invention provides a tetrahydropyrrole derivative for use in treating breast cancer, the structural formula of which is shown in Chemical Formula 1:

[0028] Chemical Formula 1 Wherein, R1 is selected from halogens, cycloalkyl groups with 3 to 10 carbon atoms, and alkylamino groups with 3 to 10 carbon atoms; R2 is selected from alkyl groups with 1 to 10 carbon atoms and cycloalkyl groups with 3 to 10 carbon atoms; R3 is selected from alkylamino groups with 1 to 10 carbon atoms, dialkylamino groups with 3 to 10 carbon atoms, and saturated heterocyclic alkyl groups with 3 to 10 carbon atoms; L is selected from alkyl groups with 1 to 10 carbon atoms.

[0029] In some examples, R1 is selected from chlorine, methylamino, isopropylamino, and cyclopropyl.

[0030] In some examples, R2 is selected from methyl, ethyl, and isopropyl.

[0031] In some examples, R3 is selected from tetrahydropyrrole-N-yl, dimethylamino, and diethylamino.

[0032] In some examples, L is selected from methylene, ethylene, and propylene.

[0033] In one example, the tetrahydropyrrole derivative is selected from the group consisting of the following compounds:

[0034] In one embodiment of the present invention, the preparation method of compounds TP1, TP2, TP3 and TP4 includes the following steps: S1. Compound 1 (4-methoxy-3-[3-(pyrrolidone-1-yl)propoxy]aniline), compound 2 (1-(4-methoxybenzyl)-5,5-dimethyl-2-oxopyrrole-3-carboxylic acid), 1-ethyl-3-(3-dimethylaminopropyl)carbodiimide hydrochloride (EDCI), and 1-hydroxybenzotriazole (HOBt) were added to anhydrous dichloromethane, mixed thoroughly, and reacted at room temperature. After the reaction was completed, the mixture was separated and purified by thin-layer chromatography to obtain intermediate 1. N-{4-methoxy-3-[3-(pyrrolidone-1-yl)propoxy]phenyl}-1-(4-methoxyphenyl)-5,5-dimethyl-2-oxopyrrolidone-3-carboxamide); ; S2. Intermediate 1 was added to phosphorus oxychloride and reacted at 100–130 °C. After the reaction was completed, the mixture was separated and purified by thin-layer chromatography to obtain compound TP1 (4-chloro-8-methoxy-2,2-dimethyl-7-[3-(pyrrolidin-1-yl)propoxy]-2,3-dihydro-1-yl) H -pyrrolo[3,2- c Quinoline).

[0035] ; S3. Compound TP1, CH3-NH2, and sodium tert-butoxide were added to dioxane, followed by the addition of a third-generation palladium precatalyst with a BrettPhos ligand to form a reaction system. Under nitrogen protection, the reaction system was placed in a sealed tube for reaction. After the reaction was completed, the compound TP2 (8-methoxy-) was separated and purified by thin-layer chromatography. N 2,2-Trimethyl-7-[3-(pyrrolidone-1-yl)propoxy]-2,3-dihydro-1 H -pyrrolo[3,2- c Quinoline-4-amine). The reaction formula is as follows: ; Compound TP1, (CH3)2CH-NH2, and sodium tert-butoxide were added to dioxane, followed by the addition of a third-generation palladium precatalyst with a BrettPhos ligand to form a reaction system. The reaction system was placed in a sealed tube for further reaction, and the compound TP3 was obtained after separation and purification by thin-layer chromatography monitoring. N -Isopropyl-8-methoxy-2,2-dimethyl-7-[3-(pyrrolidine-1-)propoxy]-2,3-dihydro-1 H -pyrrolo[3,2- c Quinoline-4-amine). The reaction formula is as follows: ; Compound TP1, cyclopropylboronic acid, and K3PO4 were dissolved in toluene aqueous solution, and Pd(Amphos)2Cl2 was added to carry out the reaction. After the reaction was completed, the mixture was separated and purified by thin-layer chromatography to obtain compound TP4 (4-cyclopropyl-8-methoxy-2,2-dimethyl-7-[3-(pyrrolidone-1-yl)propoxy]-2,3-dihydro-1 H -pyrrolo[3,2- c Quinoline); .

[0036] The present invention also provides the application of the above-mentioned tetrahydropyrrole derivative in the preparation of anti-breast cancer drugs.

[0037] The present invention also provides an anti-breast cancer drug, which is a hydropyrrole derivative as described in any of the above embodiments, or a pharmaceutically acceptable salt of a tetrahydropyrrole derivative as described in any of the above embodiments.

[0038] For example, pharmaceutically acceptable salts of tetrahydropyrrole derivatives include one of hydrochloride, sulfate, phosphate, citrate, maleate, and tartrate.

[0039] The solution provided by the present invention will be further described below with reference to specific embodiments.

[0040] Example 1 This embodiment provides a method for preparing tetrahydropyrrole derivatives (compounds TP1, TP2, TP3, and TP4), comprising the following steps: Step S1, the synthesis of intermediate 1, is shown in the following reaction formula: ; 5.0 g (20 mmol) of 4-methoxy-3-[3-(pyrrolidone-1-yl)propoxy]aniline (compound 1), 6.1 g (22 mmol) of 1-(4-methoxybenzyl)-5,5-dimethyl-2-oxopyrrole-3-carboxylic acid (compound 2), 5.8 g (30 mmol) of EDCI and 4.1 g (30 mmol) of HOBt were added sequentially to anhydrous dichloromethane (200 mL). After stirring, the mixture was reacted at room temperature for 2 h. After the reaction was completed, the mixture was concentrated under reduced pressure by thin-layer chromatography (TLC). The crude product was then purified by MPLC (medium-pressure liquid chromatography). MPLC was performed using a Prep-C18 column (120 g, Tianjin Bona Ager Technology Co., Ltd.), with an elution gradient of 10%–45% acetonitrile aqueous solution containing 0.1% trifluoroacetic acid, and an elution time of 10 min. Intermediate 1 was obtained with a mass of 7.4 g. Intermediate 1 was a yellowish-brown solid with a yield of 73.1%.

[0041] Physicochemical property testing of intermediate 1: Melting point (mp): 181~183℃.

[0042] Elemental analysis: C 29 H 39 The measured value of C in N3O5 is 68.30% (calculated value is 68.35%), the measured value of N is 8.21% (calculated value is 8.25%), and the measured value of H is 7.77% (calculated value is 7.71%).

[0043] Proton spectrum and mass spectrum: 1 H NMR (300 MHz, DMSO- d 6): δ 10.13 (s, 1H), 9.48 (br, 1H), 7.50 (d, J = 2.1 Hz, 1H), 7.22 (d, J = 8.7 Hz, 2H), 7.07 (dd, J = 8.7 Hz, J =2.1 Hz, 1H), 6.95 (d, J = 8.7 Hz, 1H), 6.87 (d, J = 8.4 Hz, 2H), 4.39-4.26(m, 2H), 4.03 (t, J = 6.0 Hz, 2H), 3.75 (s, 3H), 3.73 (s, 3H), 3.68-3.62 (m,3H), 3.35-3.28 (m, 2H), 3.08-3.00 (m, 2H), 2.21-2.04 (m, 6H), 1.98-1.82 (m,2H), 1.20 (s, 3H), 1.12 (s, 3H). ESI-MS [M+1] + m / z 510.3.

[0044] Step S2: Synthesis of compound TP1, the synthesis reaction formula is shown below: ; 7.1 g (14 mmol) of intermediate 1 was slowly added to POCl3 (100 mL) and reacted at 120 °C for 8 h. After the reaction was completed by TLC monitoring, the reaction system was cooled to room temperature and then concentrated under reduced pressure. The crude product was purified by MPLC (medium-pressure liquid chromatography) (Prep-C18 column, 120 g, Tianjin Bona Ager Technology Co., Ltd.; elution gradient of 10%~20% acetonitrile aqueous solution containing 0.1% formic acid, elution time 12 min) to obtain 3.4 g of compound TP1, which was a yellow solid with a yield of 62.5%.

[0045] Physicochemical property testing of compound TP1: Melting point (mp): 207~209℃.

[0046] Elemental analysis: C 21 H 28The measured value of C in ClN3O2 is 64.63% (calculated value is 64.69%), the measured value of N is 10.81% (calculated value is 10.78%), and the measured value of H is 7.31% (calculated value is 7.24%).

[0047] Proton spectrum and mass spectrum: 1 H NMR (300 MHz, CDCl3): δ 11.39 (br, 1H), 10.54 (brs, 1H), 8.12 (s, 1H), 7.76 (s, 1H), 4.26 (t, J = 6.0 Hz, 2H), 4.03 (s, 3H), 3.99-3.90(m, 2H), 3.45-3.43 (m, 2H), 3.13-3.04 (m, 4H), 2.49-2.42 (m, 2H), 2.22-2.08(m, 4H), 1.55 (s, 3H), 1.51 (s, 3H). ESI-MS [M+1] + m / z 390.1.

[0048] Step S3: Synthesis of compounds TP2, TP3, and TP4, with the following reaction formula: ; ; S31. Under nitrogen protection, 389 mg (1.0 mmol) of compound TP1, (5.0 mmol) of primary amine compound (CH3-NH2 or (CH3)2CH-NH2) and 481 mg (1.0 mmol) were added. t BuONa was added sequentially to 5 mL of dioxane, followed by 45 mg (0.05 mmol) of BrettPhos Pd G3. The reaction system was sealed and reacted at 90 °C for 1 h. After the reaction was completed by TLC monitoring, the mixture was concentrated under reduced pressure. The crude product was purified by Pre-HPLC (preparative high-performance liquid chromatography) (SunFire Prep-C18 column, 19 × 150 mm, Waters, USA; elution gradient of 15%–32% acetonitrile aqueous solution containing 0.1% trifluoroacetic acid, elution time 7 min) to obtain compounds TP2 and TP3. Compound TP2 was a brown oily liquid with a yield of 43.1%; compound TP3 was a brown oily liquid with a yield of 48.8%.

[0049] Physicochemical property testing of compound TP2: Melting point (mp): 233~235℃.

[0050] Elemental analysis: C 22 H 32 The measured value of C in N4O2 is 68.68% (calculated value is 68.72%), the measured value of N is 14.52% (calculated value is 14.57%), and the measured value of H is 8.43% (calculated value is 8.39%).

[0051] Proton spectrum and mass spectrum: 1 H NMR (300 MHz, DMSO- d 6): δ 11.31 (brs, 1H), 9.68 (br,1H), 8.35 (s, 1H), 7.62-7.58 (m, 1H), 7.43 (s, 2H), 4.15 (t, J = 5.1 Hz, 2H),3.85 (s, 3H), 3.65-3.58 (m, 2H), 3.35-3.28 (m, 2H), 3.09-3.07 (m, 2H), 3.02(s, 3H), 2.78 (s, 2H), 2.25-2.15 (m, 2H), 2.05-2.00 (m, 2H), 1.89-1.85 (m,2H), 1.41 (s, 3H), 1.39 (s, 3H). ESI-MS [M+1] + m / z 385.2.

[0052] Physicochemical property testing of compound TP3: Melting point (mp): 242~244℃.

[0053] Elemental analysis: C 24 H 36 The measured value of C in N4O2 is 69.84% (calculated value is 69.87%), the measured value of N is 13.61% (calculated value is 13.58%), and the measured value of H is 8.86% (calculated value is 8.80%).

[0054] Proton spectrum and mass spectrum: 1 H NMR (300 MHz, DMSO- d 6): δ 11.22 (brs, 1H), 9.65 (br,1H), 8.38 (s, 1H), 7.42 (s, 2H), 7.17 (d, J=8.4 Hz, 1H), 4.15-4.09 (m, 3H), 3.85 (s, 3H), 3.71-3.63 (m, 2H), 3.40-3.32 (m, 2H), 3.10-3.02 (m, 2H), 2.82(s, 2H), 2.24-2.22 (m, 2H), 2.10-2.05 (m, 2H), 1.92-1.88 (m, 2H), 1.41 (s,3H), 1.40 (s, 3H), 1.28 (s, 3H), 1.26 (s, 3H). ESI-MS [M+1] + m / z 413.2.

[0055]

[0056] Under nitrogen protection, 389 mg (1.0 mmol) of compound TP1, 258 mg (3.0 mmol) of cyclopropylboronic acid, and 636 mg (3.0 mmol) of K3PO4 were dissolved in a mixed solution of toluene (10 mL) and water (1 mL). After stirring, 35 mg (0.05 mmol) of Pd(Amphos)2Cl2 was added, and the reaction was carried out at 90 °C for 3 h. After the reaction was completed by TLC monitoring, the solution was concentrated under reduced pressure. The crude product was purified by Pre-HPLC (XBridge Prep-C18 column, 19 × 150 mm, Waters, USA; elution gradient of 5%–23% acetonitrile aqueous solution containing 0.1% formic acid, elution time 6.5 min) to obtain 167 mg of compound TP4, which was a yellow solid with a yield of 42.2%.

[0057] Physicochemical property testing of compound TP4: Melting point (mp): 226~228℃.

[0058] Elemental analysis: C 24 H 33 The measured value of C in N3O2 is 72.85% (calculated value is 72.88%), the measured value of N is 10.59% (calculated value is 10.62%), and the measured value of H is 8.48% (calculated value is 8.41%).

[0059] Proton spectrum and mass spectrum: 1 H NMR (300 MHz, DMSO- d 6): δ 8.25 (s, 2H), 7.64 (br, 1H), 7.25 (s, 1H), 7.09 (s, 1H), 4.09 (t,J = 6.3 Hz, 2H), 3.84 (s, 3H), 3.03 (s,2H), 2.68-2.57 (m, 6H), 2.02-1.96 (m, 3H), 1.79-1.72 (m, 4H), 1.39 (s, 3H), 1.38 (s, 3H), 1.08-1.04 (m, 2H), 0.96-0.93 (m, 2H). ESI-MS [M+1] + m / z 396.2.

[0060] Example 2 Compounds TP1, TP2, TP3, and TP4 prepared in Example 1 were used, with 4-hydroxytamoxifen (TAM) as a positive control. MCF-7 breast cancer cells, MDA-MB-231 triple-negative breast cancer cells, MCF-7 / ADR doxorubicin-resistant breast cancer cells, and MCF-10A non-tumorigenic breast epithelial cells were used as test cell lines. The in vitro antitumor activity of the four compounds was detected by the MTT assay. The steps included: Step 1: Using dimethyl sulfoxide (DMSO) as a solvent, compounds TP1, TP2, TP3, TP4, and 4-hydroxytamoxifen were prepared to a concentration of 0.1 mol·L⁻¹. -1 The mother liquors. That is, the mother liquors of compounds TP1, TP2, TP3, TP4 and 4-hydroxytamoxifen are obtained.

[0061] Step 2: Using Dalberg modified Eagle's (DMEM) medium, the stock solutions were serially diluted to prepare compounds TP1, TP2, TP3, TP4, and 4-hydroxytamoxifen to a drug concentration of 100 μmol·L⁻¹. -1 50 μmol·L -1 25 μmol·L -1 5 μmol·L -1 1 μmol·L -1 0.5 μmol·L -1 and 0.1 μmol·L -1 The culture medium. For example, using a culture medium with a concentration of 0.1 mol·L⁻¹. -1 The mother liquor of compound TP1 was diluted to prepare a solution containing 100 μmol·L⁻¹ of compound TP1. -1 50 μmol·L -1 25 μmol·L -1 5 μmol·L -11 μmol·L -1 0.5 μmol·L -1 and 0.1 μmol·L -1 The culture medium was prepared using the same dilution method as the stock solution of compound TP1, and will not be described further.

[0062] Step 3: Take logarithmically growing breast cancer cells MCF-7, triple-negative breast cancer cells MDA-MB-231, doxorubicin-resistant breast cancer cells MCF-7 / ADR, and non-tumorigenic breast epithelial cells MCF-10A, respectively, and suspend them in DMEM medium containing 10% fetal bovine serum to form cell suspensions. That is, obtain MCF-7 cell suspension, MDA-MB-231 cell suspension, MCF-7 / ADR cell suspension, and MCF-10A cell suspension, respectively.

[0063] Seed the cell suspension into 96-well cell culture plates (different test cell lines were seeded in different 96-well cell culture plates), adding 100 μL of cell suspension (approximately (4~10) × 10⁶ cells per well) to each well. 4 (cells). Incubate at 37°C in a 5% CO2 incubator until cells are fully adhered. Discard the original culture medium and add 200 µL of culture medium containing different concentrations of compounds TP1, TP2, TP3, TP4, and 4-hydroxytamoxifen to each well of a 96-well cell culture plate containing MCF-7 cell suspension. Similarly, 200 µL of culture medium containing different concentrations of compounds TP1, TP2, TP3, TP4, and 4-hydroxytamoxifen was added to each well of a 96-well cell culture plate containing MDA-MB-231 cell suspension; 200 µL of culture medium containing different concentrations of compounds TP1, TP2, TP3, TP4, and 4-hydroxytamoxifen was added to each well of a 96-well cell culture plate containing MCF-7 / ADR cell suspension; and 200 µL of culture medium containing different concentrations of compounds TP1, TP2, TP3, TP4, and 4-hydroxytamoxifen was added to each well of a 96-well cell culture plate containing MCF-10A cell suspension. The 96-well cell culture plates were then incubated at 37°C in a 5% CO2 incubator for 3 days. Three replicates were prepared for each concentration of each compound. Meanwhile, each 96-well cell culture plate also includes a blank control well. The blank wells do not contain the test cell lines or compounds, but only an equal amount of culture medium.

[0064] Step 4: After the culture is complete, discard the original culture medium and add 30 μL of MTT (5 g·L⁻¹) to each well. -1Cells were incubated with 100 µL of DMEM medium at 37°C in a 5% CO2 incubator for 4 h. The original culture medium was discarded, and 100 μL of DMSO was added to each well, followed by shaking for 15 min. The absorbance (OD) of each well was measured at 490 nm using a microplate reader. The inhibition rate (%) of the drug against tumor cells was calculated, and the half-maximal inhibitory concentration (IC50) was calculated using OriginPro software. 50 Values. See Table 1 for the results.

[0065] Table 1. Antitumor activity of compounds TP1, TP2, TP3, and TP4

[0066] According to the detection results in Table 1, compounds TP1, TP2, TP3, and TP4 all showed certain inhibitory activities against MCF-7 breast cancer cells, MDA-MB-231 triple-negative breast cancer cells, and doxorubicin-resistant MCF-7 / ADR breast cancer cells. 50 The values ​​are all less than 30 µmol·L -1 .

[0067] The inhibitory activity assay of breast cancer cells MCF-7 showed that, compared with the positive control drug 4-hydroxytamoxifen (TAM), the IC50 of the four compounds was significantly higher. 50 Although the value is less than 16 µmol·L -1 However, it is still lower than TAM. In the inhibitory activity assay of triple-negative breast cancer cells MDA-MB-231, compound TP1 showed inhibitory activity almost equivalent to the positive control drug, indicating the feasibility of compound TP1 as an active ingredient for treating triple-negative breast cancer. In the inhibitory activity assay of doxorubicin-resistant breast cancer cells MCF-7 / ADR, compounds TP1, TP2, TP3, and TP4 all showed excellent anti-tumor cell activity. Among them, compound TP4 had an IC50 value of [missing information]. 50 The value was 8.4 ± 1.5 μmol·L. -1 Its inhibitory activity was nearly five times that of the positive control drug 4-hydroxytamoxifen, making it a potential candidate active ingredient for the treatment of drug-resistant breast cancer. Meanwhile, compounds TP1, TP2, TP3, and TP4 showed no significant toxicity to non-tumorigenic breast epithelial cells MCF-10A, while 4-hydroxytamoxifen exhibited some toxicity.

[0068] The above results indicate that the tetrahydropyrrole derivative provided by the present invention has anti-breast cancer activity and can be used as a candidate drug for anti-breast cancer treatment.

[0069] In the description of this specification, the references to terms such as "one embodiment," "some embodiments," "example," "specific example," or "some examples," etc., indicate that a specific feature, structure, material, or characteristic described in connection with that embodiment or example is included in at least one embodiment or example of the invention. In this specification, the illustrative expressions of the above terms do not necessarily refer to the same embodiment or example. Furthermore, the specific features, structures, materials, or characteristics described may be combined in any suitable manner in one or more embodiments or examples. In addition, those skilled in the art can combine and integrate the different embodiments or examples described in this specification.

[0070] The above description, in conjunction with specific preferred embodiments, provides a further detailed explanation of the present invention. It should not be construed that the specific implementation of the present invention is limited to these descriptions. For those skilled in the art, various simple deductions or substitutions can be made without departing from the concept of the present invention, and all such modifications and substitutions should be considered within the scope of protection of the present invention.

Claims

1. A tetrahydropyrrole derivative for use in the treatment of breast cancer, characterized in that, The structural formula of the tetrahydropyrrole derivative is shown in Chemical Formula 1: Chemical Formula 1 R1 is selected from halogens, cycloalkyl groups with 3 to 10 carbon atoms, and alkylamine groups with 3 to 10 carbon atoms; R2 is selected from alkyl groups having 1 to 10 carbon atoms and cycloalkyl groups having 3 to 10 carbon atoms; R3 is selected from alkylamine with 1 to 10 carbon atoms, dialkylamine with 3 to 10 carbon atoms, and saturated heterocyclic alkyl with 3 to 10 carbon atoms; L is selected from 1 to 10 alkyl groups.

2. The tetrahydropyrrole derivative according to claim 1, characterized in that, R1 is selected from chlorine, methylamino, isopropylamino, and cyclopropyl.

3. The tetrahydropyrrole derivative according to claim 1, characterized in that, R2 is selected from methyl, ethyl, and isopropyl.

4. The tetrahydropyrrole derivative according to claim 1, characterized in that, The R3 is selected from tetrahydropyrrole-N-yl, dimethylamino, and diethylamino.

5. The tetrahydropyrrole derivative according to claim 1, characterized in that, The L is selected from methylene, ethylene, and propylene.

6. The tetrahydropyrrole derivative according to claim 1, characterized in that, The tetrahydropyrrole derivative is selected from the group consisting of the following compounds: 。 7. A method for preparing the tetrahydropyrrole derivative according to claim 6, characterized in that, Includes the following steps: S1. Compound 1, Compound 2, 1-ethyl-3-(3-dimethylaminopropyl)carbodiimide hydrochloride and 1-hydroxybenzotriazole were added to anhydrous dichloromethane, mixed and reacted at room temperature. After the reaction was completed, intermediate 1 was obtained by separation and purification. ； S2. Add intermediate 1 to phosphorus oxychloride and react at 100-130℃. After the reaction is completed, separate and purify to obtain compound TP1. ； S3. Compound TP1, CH3-NH2 and sodium tert-butoxide are added to dioxane, and then a third-generation palladium precatalyst with BrettPhos ligand is added to form a reaction system; the reaction system is placed in a sealed tube and reacted under nitrogen protection. After the reaction is completed, compound TP2 is obtained by separation and purification. ； Compound TP1, (CH3)2CH-NH2, and sodium tert-butoxide were added to dioxane, followed by the addition of a third-generation palladium precatalyst with a BrettPhos ligand to form a reaction system. The reaction system was placed in a sealed tube and reacted under nitrogen protection. After the reaction was completed, compound TP3 was obtained by separation and purification. ； Compound TP1, cyclopropylboronic acid and K3PO4 were dissolved in toluene aqueous solution, Pd(Amphos)2Cl2 was added, and the reaction was carried out under nitrogen protection. After the reaction was completed, the compound TP4 was obtained by separation and purification. 。 8. The use of the tetrahydropyrrole derivative according to any one of claims 1 to 6 in the preparation of anti-breast cancer drugs.

9. An anti-breast cancer drug, characterized in that, The anti-breast cancer drug includes a tetrahydropyrrole derivative as described in any one of claims 1 to 6, or a pharmaceutically acceptable salt comprising a tetrahydropyrrole derivative as described in any one of claims 1 to 6.

10. The anti-breast cancer drug according to claim 9, characterized in that, The pharmaceutically acceptable salts of the tetrahydropyrrole derivatives include one of the following: hydrochloride, sulfate, phosphate, citrate, maleate, and tartrate.

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