Estrogen-degrading agent derivatives, and methods of making and using the same

Abstract

The present application belongs to the field of pharmaceutical chemistry, and relates to the use of a compound shown in formula (I) or a solvate thereof and a pharmaceutically acceptable salt thereof for preparing a drug for treating breast cancer. The compound has the characteristics of significantly improving the stability, solubility and dissolution of the drug and the metabolizability in vivo, has ideal pharmacokinetic characteristics and a good drug development prospect.

Description

Technical Field

[0001] The invention belongs to the field of medicinal chemistry, specifically relating to a Girdestrant derivative or its salt or its solvate, its preparation method, a pharmaceutical composition comprising the derivative, and its pharmaceutical uses. Background Technology

[0002] Breast cancer is the most common malignant tumor among women worldwide, with estrogen receptor (ER)α-positive breast cancer accounting for approximately 70%. To date, the primary intervention for treating ERα-positive breast cancer is traditional endocrine therapy, with treatment drugs including aromatase inhibitors (AIs), selective estrogen receptor modulators (SERMs), and selective estrogen receptor degraders (SERDs). Among the selective estrogen receptor degraders, fulvestrant has proven to be effective and well-tolerated in clinical practice. However, fulvestrant has significant limitations, such as easy resistance, low receptor affinity, and the need for parenteral administration. These limitations have spurred the development of novel orally administered SERDs. Elacestrant is an orally administered SERD that received FDA approval on January 27, 2023, for the treatment of ER-positive, HER2-negative, estrogen receptor 1 (ESR1)-mutant postmenopausal advanced or metastatic breast cancer.

[0003] Giredestrant (GDC-9545; RG-6171), chemical name: It is a small molecule drug developed by Genentech, and is an ER degrader. Currently, this drug is in Phase III clinical trials for the treatment of breast cancer and metastatic breast cancer. The structural formula of its tartrate salt in clinical development is as follows:

[0004] ,

[0005] WO2019245974, WO2022005943, US11873305, US2024174678, US9980947, US2018235945, US2016175289, US2021353601, CN115551513, and US2022002304 disclose reports on synthetic methods, compounds, compositions, and uses.

[0006] Giredestrant is another non-steroidal SERD drug with potent anticancer activity. Upon binding to the estrogen receptor, it causes conformational changes, inhibiting transcriptional activity and leading to receptor degradation, thereby suppressing the growth and survival of ER-positive breast cancer cells. In vitro studies have shown that among SERD drugs, giredestrant is the most potent ER antagonist, with a half-maximal inhibitory concentration (IC50) of 0.05 nmol in MCF-7 cells, and it induces 100% estrogen receptor degradation in MCF-7 cells. In PDX studies, daily administration of a low dose (1 mg / kg) of giredestrant to experimental animals resulted in tumor regression.

[0007] Giredestrant's physicochemical characteristics include metabolic resistance due to its difluoropropanol structure and high lipophilicity. Its extremely low water solubility (less than 0.02 mg / ml) presents the greatest challenge in achieving adequate bioavailability. Furthermore, regarding its API stability, literature reports that giredestrant powder should be stored at -20°C for up to 3 years or at 4°C for up to 2 years, with lower temperatures preferred. Giredestrant solutions have a much shorter stability period; some sources suggest storage at -80°C for 6 months or at -20°C for 1 month, always keeping the container sealed and avoiding direct sunlight and high temperatures.

[0008] The discovery of new derivatives of active ingredients or their new polymorphs offers opportunities to improve their properties, increasing the possibilities available to formulation experts when developing new drug forms, drugs with specific release profiles, or drugs with specific solubilities. For example, WO2019245974 discloses Girdestrant tartrate, and some of its crystalline forms, and studies have found that the B form exhibits unique mechanical behavior, but makes it very difficult to process via mechanical compression. Therefore, mechanical compression applied during the tableting process of conventional pharmaceutical compositions often leads to partial decomposition, color change, and agglomeration of the API. One way to avoid harmful compressive forces applied during tableting is to directly fill the API or the pharmaceutical composition containing it into capsules, but this exhibits an uneven particle size distribution, making it difficult to produce tablets and capsules on an industrial scale.

[0009] Based on these considerations, there remains a need for new Girdestrant derivatives and new polymorphs with further improved physical and / or chemical properties. Therefore, the inventors of this application believe it is worthwhile to explore new pharmaceutical polymorphs of Girdestrant esters with good chemical purity and improved stability properties, which may further improve the properties of Girdestrant in finished pharmaceutical products, especially improving oral bioavailability and long-term storage stability, as well as simplifying the manufacturing process. Summary of the Invention

[0010] This invention relates to specific derivatives of Girdestrant, and to the preparation of derivative structures with improved drug-like properties, aiming to at least partially address the technical problems existing in the prior art. To this end, one aspect of the invention provides a Girdestrant derivative of formula (I) or a solvate thereof, and pharmaceutically acceptable salts thereof:

[0011] ,

[0012] Where: R is selected from

[0013] -CH2OC(O)R1;

[0014] -C 1-6 Alkyl groups, wherein the alkyl groups are optionally irradiated by 1-3 R groups. a Group substitution;

[0015] -CH2-m-dioxacyclopentenyl, optionally via 1-2 R... a Group substitution;

[0016] -C 6-10 Aryl, -C 6-10 Heteroaryl, wherein the aryl-heteroaryl group is optionally etherified by 1-3 R... a Group substitution;

[0017] R1 is selected from C 1-6 Alkyl, C 6-10 Aryl, C 6-10 heteroaryl and C 3-6 cycloalkyl; the alkyl, aryl, heteroaryl and cycloalkyl groups are optionally derived from 1 to 3 R... a Group substitution;

[0018] R a Selected independently from C 1-6 Alkyl, OC 1-6 Alkyl, oxo, and halogen.

[0019] It has been found that the light and heat stability and hydrophilicity of Giredestrant can be enhanced by reacting Giredestrant or some of its salts with suitable reagents to generate certain amides at the secondary amine sites of the molecule.

[0020] The Giredestrant derivatives produced by the process of this invention are superior to Giredestrant and its well-known salts in several respects. First, the tested amides are more resistant to light and heat in solution. Second, due to their more suitable lipophilicity, these derivatives should have higher bioavailability when administered orally or via membrane routes. These derivatives are expected to have a more suitable octanol / water partition coefficient.

[0021] These and other advantages will become more apparent upon consideration of the following description of the invention. The present invention relates to derivatives of Girdestrant that are metabolized in the human body into bioavailable forms of the drug.

[0022] Another aspect of the invention provides a compound of preferred formula I:

[0023] ,

[0024] .

[0025] The salts of the compounds of formula (I) described by the applicant, which have amino functionalities, can form with any pharmaceutically acceptable inorganic or organic anion. Exemplary inorganic anions include chlorides, bromides, nitrates, carbonates, bicarbonates, phosphates, monohydrophosphates, dihydrophosphates, etc. Exemplary organic anions include formates, acetates, lactates, propionates, butyrates, isobutyrates, palmitates, glutamates, maleates, malonates, benzoates, succinates, fumarates, salicylates, citrates, tartrates, methanesulfonates, benzenesulfonates, etc.

[0026] In another aspect of the invention, a pharmaceutical composition is provided. According to embodiments of the invention, the pharmaceutical composition comprises crystals of the aforementioned Girdestrant derivative and a pharmaceutically acceptable carrier. The carrier in the pharmaceutical composition is "acceptable," meaning it is compatible with (and preferably, capable of stabilizing) the active ingredient of the composition and is not harmful to the treated subject. The compounds and pharmaceutically acceptable excipients of the invention are generally formulated into dosage forms suitable for administration to a patient via the desired route of administration. Suitable dosage forms for oral administration include tablets, capsules, pills, powders, syrups, suspensions, solutions, emulsions, and granules. Suitable pharmaceutically acceptable excipients will vary depending on the specific dosage form selected. Furthermore, suitable pharmaceutically acceptable excipients can be selected for the specific function they may perform in the composition. For example, certain pharmaceutically acceptable excipients can be selected to facilitate the production of a uniform dosage form. Certain pharmaceutically acceptable excipients can be selected because they are capable of stabilizing one or more compounds of the invention in the pharmaceutical composition. Certain pharmaceutically acceptable excipients can be selected because they can improve patient compliance. Suitable pharmaceutically acceptable excipients include binders, lubricants, slippers, disintegrants, granulators, coating agents, wetting agents, solvents, solubilizers, suspending agents, flavoring agents, flavor masking agents, anti-caking agents, humectants, chelating agents, plasticizers, viscosity modifiers, antioxidants, preservatives, stabilizers, surfactants, emulsifiers, and buffers. The pharmaceutical compositions of the present invention are prepared using techniques and methods known to those skilled in the art, such as those described in Mack Publishing Company, Remington.

[0027] Oral dosage forms may include carriers in the form of diluents or fillers. Suitable diluents and fillers generally include lactose, sucrose, glucose, sorbitol, mannitol, cellulose or cellulose derivatives such as microcrystalline cellulose, starch, calcium sulfate, and calcium hydrogen phosphate. Liquid dosage forms typically consist of a suspension or solution of a compound or salt in a liquid carrier, such as ethanol, olive oil, glycerin, glucose syrup, or water. When the composition is in tablet or lozenge form, any pharmaceutical carrier conventionally used for preparing solid dosage forms can be used, such as magnesium stearate, kaolin, talc, gelatin, arabinocyanurate, stearic acid, starch, lactose, and sucrose. When the composition is in capsule form, any conventional encapsulation is suitable. When the composition is in the form of soft-shell capsules (e.g., gelatin), any pharmaceutical carrier conventionally used for preparing dispersions or suspensions can be incorporated, such as aqueous gels or oils. Formulations can be appropriately formulated to provide controlled / sustained release of the active compound. Oral solid dosage forms may further include excipients in the form of binders. Suitable binders include starch, gelatin, alginate, sodium alginate, astragalus, acacia, guar gum, povidone, and cellulose or cellulose derivatives, such as microcrystalline cellulose. Oral solid dosage forms may also include excipients in the form of disintegrants. Suitable disintegrants include crospovidone, sodium glycolate starch, croscarmellose, alginate, and sodium carboxymethyl cellulose. Oral solid dosage forms may further include excipients in the form of lubricants. Suitable lubricants include stearic acid, magnesium stearate, calcium stearate, and talc.

[0028] Giredestrant itself, when stored as an API and mixed with various pharmaceutical excipients and carriers, is highly susceptible to producing genotoxic impurities with the following structure (referred to as impurity A elsewhere in this article). Therefore, the storage conditions for Giredestrant are extremely stringent, such as storage at 2-8°C and not exceeding 12 months.

[0029] .

[0030] The Girdestrant derivative described in this invention is used to manufacture a drug for treating breast cancer.

[0031] In another aspect of the present invention, the use of the compound of formula (I) of this application, or a solvate thereof, or a salt thereof, in the preparation of a medicament for treating breast cancer is provided.

[0032] Another aspect of this application provides the use of a pharmaceutical composition containing a compound of formula (I) of this application or a solvate thereof or a salt thereof in the preparation of a medicament for treating breast cancer.

[0033] The key points of this invention are:

[0034] Primarily, this invention relates to the introduction of suitable amide ester derivatives or pharmaceutically acceptable salts thereof at the secondary amine position of Giredestrant. By using these derivatives to replace all or part of the Giredestrant salt in pharmaceutical formulations, significant improvements are made in the temperature and light stability, solubility and dissolution rate of the drug, as well as its metabolizability in vivo. Attached Figure Description

[0035] Figure 1 This is a comparison chart of the dissolution curves of compound 1 capsules, compound 6 capsules, and compound 8 capsules compared to Girdestrant tartrate capsules. Detailed Implementation

[0036] The technical solution of the present invention will now be clearly and completely described with reference to the accompanying drawings. Obviously, the described embodiments are only some, not all, of the embodiments of the present invention. Based on the embodiments of the present invention, all other embodiments obtained by those skilled in the art without creative effort are within the scope of protection of the present invention.

[0037] The present invention will be further explained below with reference to specific embodiments.

[0038] The following embodiments can further describe the present invention; however, these embodiments should not be construed as limiting the scope of the present invention.

[0039] Implementation Step 1: Synthesis of Compound 1:

[0040] .

[0041] Step 1: Add Giredestrant (5.2 g, 1.0 eq) and 50 mL DMSO to a 100 mL three-necked flask and stir to dissolve. Add side chain 1 (3.1 g, 1.05 eq) at room temperature and stir to react for 2 h. Monitor the reaction by HPLC until it ends. Add 100 mL dichloromethane and stir to disperse. Cool to 10-15 °C. Wash three times with water (50 mL x 3). Dry the dichloromethane layer and concentrate under reduced pressure. Purify by column chromatography to obtain 5.2 g of compound 1, yield: 76.6%.

[0042] Step 2: Add Giredestrant (5.2 g, 1.0 eq) and 100 mL DCM to a 100 mL three-necked flask and stir. Add side chain 1 (3.1 g, 1.05 eq) at room temperature and stir overnight. Monitor the reaction by HPLC until it ends. Cool to 10-15 °C, wash three times with water (50 mL x 3), dry the dichloromethane layer and concentrate under reduced pressure. Purify by column chromatography to obtain 4.2 g of compound 1, yield: 61.9%.

[0043] 1H NMR (400MHz, d-DMSO, ppm): δ 10.46(s,1H),7.39 (d,1H), 7.19 (d,1H),7.01~6.93 (m,2H), 6.11(d,2H),5.16(s,1H),4.86(s,2H),4.52~4.38 (m,2H),4.16(s,1H), 4.05~4.03 (m,1H), 3.80~3.74(m,3H), 3.63~3.42(m,2H), 3.20~3.10 (m,1H),2.96~2.92 (m,3H), 2.82~2.71 (m,1H), 2.64~2.58 (m,3H), 1.81~1.68 (m,2H), 1.14(d,3H), 0.92(s,3H).

[0044] Example 2: Synthesis of Compound 2:

[0045]

[0046] Step 1: Add Giredestrant (5.2 g, 1.0 eq) and 50 mL DMSO to a 100 mL three-necked flask and stir to dissolve. Add side chain 2 (3.2 g, 1.05 eq) at room temperature and stir to react for 2 h. Monitor the reaction by HPLC until it ends. Add 100 mL dichloromethane and stir to disperse. Cool to 10-15 °C. Wash three times with 50 mL x 3 water. Dry the dichloromethane layer and concentrate under reduced pressure. Purify by column chromatography to obtain 4.7 g of compound 2, yield: 68.6%.

[0047] Step 2: Add Giredestrant (5.2 g, 1.0 eq) and 50 mL tetrahydrofuran to a 100 mL three-necked flask and stir. Add side chain 2 (3.2 g, 1.05 eq) at room temperature and stir for 2 h. Monitor the reaction by HPLC until it ends. Add 100 mL dichloromethane, stir and disperse, then cool to 10-15 °C. Wash three times with water (50 mL x 3). Dry the dichloromethane layer and concentrate under reduced pressure. Purify by column chromatography to obtain 3.5 g of compound 2, yield: 51.1%.

[0048] 1H NMR (400MHz,dDMSO,ppm): δ 10.46(s,1H),7.62~7.41(m,5H),7.37(d,1H),7.20(d,1H),7.01~6.94(m,2H),6.13(d,2H),5.33(s,2H),5.17(s,1H), 4.52~4.39(m,2H),4.14(s,1H), 4.05~4.03(m,1H), 3.80~3.74(m,3H), 3.63~3.42(m,2H), 3.20~3.10 (m,1H), 2.96~2.92 (m,3H), 2.82~2.71 (m,1H), 2.64~2.58 (m,3H), 1.81~1.69 (m,2H),1.14 (d,3H).

[0049] Example 3: Synthesis of Compound 3:

[0050]

[0051] Step 1: Add Giredestrant (5.2 g, 1.0 eq) and 50 mL DMSO to a 100 mL three-necked flask and stir to dissolve. Add side chain 3 (2.7 g, 1.1 eq) at room temperature and stir to react for 2 h. Monitor the reaction by HPLC until it ends. Add 100 mL dichloromethane and stir to disperse. Cool to 10-15 °C. Wash three times with water (50 mL x 3). Dry the dichloromethane layer and concentrate under reduced pressure. Purify by column chromatography to obtain 5.4 g of compound 3, yield: 86.2%.

[0052] Step 2: Add Giredestrant (5.2 g, 1.0 eq) and 50 mL of acetone to a 100 mL three-necked flask and stir. Add side chain 3 (2.7 g, 1.1 eq) at room temperature, heat to 40-50 °C and stir for 4 h. Monitor the reaction by HPLC until it ends. Add 100 mL of dichloromethane and stir to disperse. Cool to 10-15 °C and wash three times with 50 mL x 3 of water. Dry the dichloromethane layer and concentrate under reduced pressure. Purify by column chromatography to obtain 4.5 g of compound 3, yield: 71.8%.

[0053] 1H NMR (400MHz,dDMSO,ppm): δ 10.46(s,1H),7.54~7.41(m,5H),7.38(d,1H),7.17(d,1H),7.01~6.93(m,2H),6.11(d,2H), 5.15(s,1H), 4.52~4.37(m,2H),4.18(s,1H)4.05~4.03(m,1H), 3.80~3.76(m,3H), 3.63~3.47(m,2H), 3.20~3.16 (m,1H), 2.96~2.92 (m,3H), 2.82~2.71 (m,1H), 2.64~2.58 (m,3H), 1.81~1.68 (m,2H), 1.16(d,3H).

[0054] Example 4: Synthesis of Compound 4

[0055]

[0056] Step 1: Add Giredestrant (5.2 g, 1.0 eq) and 50 mL DMSO to a 100 mL three-necked flask and stir until dissolved. Add side chain 4 (2.6 g, 1.1 eq) at room temperature and stir for 2 h. Monitor the reaction by HPLC until it ends. Add 100 mL dichloromethane and stir to disperse. Cool to 10-15 °C and wash three times with 50 mL x 3 water. Dry the dichloromethane layer and concentrate under reduced pressure. Purify by column chromatography to obtain 5.4 g of compound 4, yield: 86.7%.

[0057] Step 2: Add Giredestrant (5.2 g, 1.0 eq) and 50 mL N,N-dimethylformamide to a 100 mL three-necked flask and stir. Add side chain 4 (2.6 g, 1.1 eq) at room temperature and stir for 4 h. Monitor the reaction by HPLC until it ends. Add 100 mL dichloromethane and stir to disperse. Cool to 10-15 °C and wash three times with 50 mL x 3 water. Dry the dichloromethane layer and concentrate under reduced pressure. Purify by column chromatography to obtain 4.7 g of compound 4, yield: 75.5%.

[0058] 1H NMR (400MHz,dDMSO,ppm): δ 10.46(s,1H),7.38(d,1H),7.19(d,1H),7.00~6.93(m,2H),6.13(d,2H), 5.15(s,1H), 4.50~4.39(m,2H),4.19(s,1H) 4.05~4.03(m,1H), 3.92(t,2H),3.80~3.76(m,3H), 3.63~3.47(m,2H), 3.20~3.16 (m,1H), 2.96~2.92(m,3H), 2.82~2.71 (m,1H), 2.64~2.58 (m,3H), 1.81~1.68 (m,2H), 1.41~1.27(m,4H),1.16(d,3H),0.92(t,3H).

[0059] Example 5: Synthesis of Compound 5

[0060]

[0061] Step 1: Add Giredestrant (5.2 g, 1.0 eq) and 50 mL DMSO to a 100 mL three-necked flask and stir to dissolve. Add side chain 5 (2.9 g, 1.1 eq) at room temperature and stir to react for 2 h. Monitor the reaction by HPLC until it ends. Add 100 mL dichloromethane and stir to disperse. Cool to 10-15 °C and wash three times with 50 mL x 3 water. Dry the dichloromethane layer and concentrate under reduced pressure. Purify by column chromatography to obtain 4.2 g of compound 5, yield: 64.5%.

[0062] Step 2: Add Giredestrant (5.2 g, 1.0 eq) and 50 mL N,N-dimethylformamide to a 100 mL three-necked flask and stir. Add side chain 5 (2.9 g, 1.1 eq) at room temperature and stir for 4 h. Monitor the reaction by HPLC until it ends. Add 100 mL dichloromethane and stir to disperse. Cool to 10-15 °C and wash three times with 50 mL x 3 water. Dry the dichloromethane layer and concentrate under reduced pressure. Purify by column chromatography to obtain 3.6 g of compound 5, yield: 55.3%.

[0063] 1H NMR (400MHz,dDMSO,ppm): δ 10.46(s,1H),7.38(d,1H),7.17(d,1H),7.01~6.93(m,2H),6.11(d,2H), 5.30(s,2H),5.15(s,1H), 4.52~4.37(m,2H),4.18(s,1H) 4.05~4.03(m,1H), 3.80~3.76(m,3H), 3.63~3.47(m,2H), 3.20~3.16 (m,1H), 2.96~2.92(m,3H), 2.82~2.71 (m,1H), 2.64~2.58 (m,3H), 2.37(t,2H),1.81~1.62(m,4H), 1.16(d,3H),0.97(t,3H).

[0064] Example 6: Synthesis of Compound 6

[0065]

[0066] Step 1: Add Giredestrant (5.2 g, 1.0 eq) and 50 mL DMSO to a 100 mL three-necked flask and stir to dissolve. Add side chain 6 (2.8 g, 1.1 eq) at room temperature and stir to react for 2 h. Monitor the reaction by HPLC until it ends. Add 100 mL dichloromethane and stir to disperse. Cool to 10-15 °C and wash three times with 50 mL x 3 water. Dry the dichloromethane layer and concentrate under reduced pressure. Purify by column chromatography to obtain 4.0 g of compound 6, yield: 62.8%.

[0067] Step 2: Add Giredestrant (5.2 g, 1.0 eq) and 100 mL of dichloromethane to a 100 mL three-necked flask and stir. Add side chain 6 (2.8 g, 1.1 eq) at room temperature and stir for 4 h. Monitor the reaction by HPLC until it ends. Cool to 10-15 °C, wash three times with water (50 mL x 3), dry the dichloromethane layer and concentrate under reduced pressure. Purify by column chromatography to obtain 3.4 g of compound 6, yield: 53.4%.

[0068] 1H NMR (400MHz,dDMSO,ppm): δ 10.44(s,1H),7.40(d,1H),7.19(d,1H),7.03~6.95(m,2H),6.14(d,2H), 5.32(s,2H),5.16(s,1H), 4.51~4.38(m,2H),4.17(s,1H) 4.05~4.03(m,1H), 3.80~3.76(m,3H), 3.63~3.47(m,2H), 3.20~3.16 (m,1H), 2.96~2.92(m,3H), 2.82~2.71 (m,1H), 2.64~2.58 (m,3H), 2.39(q,2H),1.81~1.62(m,2H), 1.18(d,3H),0.96(t,3H).

[0069] Example 7: Synthesis of Compound 7

[0070]

[0071] Step 1: At room temperature, add Giredestrant (10.45 g, 20 mmol), dry dichloromethane (100 mL), and triethylamine (6.07 g, 60 mmol) to a 500 mL three-necked flask, and stir to disperse. Cool to -5 to 0 °C, add side chain 7 (5.39 g, 25 mmol), and control the temperature below 20 °C. After the addition is complete, react at room temperature (20 to 30 °C) for 12 h. After the reaction was completed, the temperature was lowered to 0-10℃, and the mixture was quenched in water (200mL). The phases were separated, and the aqueous phase was extracted with dichloromethane (100mL x 2). The organic phases were combined and washed successively with 10% sodium carbonate (200mL x 3) and saturated brine (200mL x 3). The mixture was dried over anhydrous sodium sulfate, filtered, and the mother liquor was concentrated under reduced pressure and purified by column chromatography (PE / EA = 4 / 1 to PE / EA = 1 / 2) to obtain compound 7 (7.46g, 53.2%).

[0072] Step 2: At room temperature, Giredestrant (10.45 g, 20 mmol), dried tetrahydrofuran (100 mL), and triethylamine (6.07 g, 60 mmol) were added to a 500 mL three-necked flask and stirred until dissolved. The temperature was lowered to -5 to 0 °C, and side chain 7 (5.39 g, 25 mmol) was added, maintaining the temperature below 20 °C. After the addition was complete, the temperature was raised to 50 to 55 °C and reacted for 3 h. After the reaction was completed, the temperature was lowered to 0 to 10 °C, quenched in water (300 mL), and extracted with dichloromethane (200 mL x 2). The organic phases were combined and washed successively with 10% sodium carbonate (300 mL x 2) and saturated brine (300 mL x 2). The mixture was dried over anhydrous sodium sulfate, filtered, and the mother liquor was concentrated under reduced pressure and purified by column chromatography (PE / EA = 4 / 1 to PE / EA = 1 / 2) to obtain compound 7 (9.58 g, 68.3%).

[0073] Step 3: At room temperature, add Giredestrant (10.45 g, 20 mmol), dry tetrahydrofuran (200 mL), and N,N-diisopropylethylamine (7.76 g, 60 mmol) to a 500 mL three-necked flask, and stir until dissolved. Cool to -5 to 0 °C, add side chain 7 (5.39 g, 25 mmol), and control the temperature below 20 °C. After the addition is complete, heat to reflux (65 to 70 °C) and react for 3 hours. After the reaction was completed, the temperature was lowered to 0-10℃, quenched in water (300mL), and extracted with dichloromethane (200mL x 2). The organic phases were combined and washed successively with 10% sodium carbonate (300mL x 2) and saturated brine (300mL x 2). The mixture was dried over anhydrous sodium sulfate, filtered, and the mother liquor was concentrated under reduced pressure and purified by column chromatography (PE / EA = 4 / 1 to PE / EA = 1 / 2) to give compound 7 (9.86g, 70.3%).

[0074] Step 4: At room temperature, add Giredestrant (10.45 g, 20 mmol), dried N,N-dimethylformamide (100 mL), and potassium carbonate powder (8.29 g, 60 mmol) to a 500 mL three-necked flask, and stir to disperse. Cool to -5 to 0 °C, add side chain 7 (5.39 g, 25 mmol), and control the temperature below 20 °C. After the addition is complete, react at room temperature (20 to 30 °C) for 12 h. After the reaction is complete, cool to 0 to 10 °C, quench in water (300 mL), extract with dichloromethane (200 mL x 2), combine the organic phases, wash with saturated brine (300 mL x 2), dry with anhydrous sodium sulfate, filter, concentrate the mother liquor under reduced pressure, and purify by column chromatography (PE / EA = 4 / 1 to PE / EA = 1 / 2) to obtain compound 7 (6.85 g, 48.8%).

[0075] 1H-NMR (400MHz, d-DMSO, ppm): δ10.31 (s, 1H); 7.99-7.87 (m, 2H); 7.40 (d, J=7.53Hz, 1H); 7.23 (d, J =8.03Hz,1H);7.20-7.11(m,2H);7.07-6.86(m,2H);6.70(d,J=6.78Hz,1H);6.34(s,2H);6.10(d, J=12.03Hz, 2H); 5.19-5.10 (m, 1H); 5.05 (s, 1H); 4.66-4.33 (m, 2H); 3.73-3.57 (m, 3H); 3.47-3.40 (m, 2H); 3 .17-3.02 (m,1H); 2.82-2.74 (m,3H); 2.66-2.53 (m,2H); 2.52-2.46 (m,2H); 1.84-1.50 (m,2H); 1.06 (d,3H).

[0076] Example 8: Synthesis of Compound 8

[0077]

[0078] Step 1: At room temperature, add Giredestrant (10.45 g, 20 mmol), dry dichloromethane (100 mL), and triethylamine (6.07 g, 60 mmol) to a 500 mL three-necked flask, and stir to disperse. Cool to -5 to 0 °C, add side chain 8 (5.39 g, 25 mmol), and control the temperature below 20 °C. After the addition is complete, react at room temperature (20 to 30 °C) for 12 h. After the reaction was completed, the temperature was lowered to 0-10℃, and the mixture was quenched in water (200mL). The phases were separated, and the aqueous phase was extracted with dichloromethane (100mL x 2). The organic phases were combined and washed successively with 10% sodium carbonate (200mL x 3) and saturated brine (200mL x 3). The mixture was dried over anhydrous sodium sulfate, filtered, and the mother liquor was concentrated under reduced pressure and purified by column chromatography (PE / EA = 4 / 1 to PE / EA = 1 / 2) to obtain compound 8 (7.82g, 55.8%).

[0079] Step 2: At room temperature, Giredestrant (10.45 g, 20 mmol), dried tetrahydrofuran (100 mL), and triethylamine (6.07 g, 60 mmol) were added to a 500 mL three-necked flask and stirred until dissolved. The temperature was lowered to -5 to 0 °C, and side chain 8 (5.39 g, 25 mmol) was added, with the temperature controlled below 20 °C. After the addition was complete, the temperature was raised to 50 to 55 °C and reacted for 3 h. After the reaction was completed, the temperature was lowered to 0 to 10 °C, quenched in water (300 mL), and extracted with dichloromethane (200 mL x 2). The organic phases were combined and washed successively with 10% sodium carbonate (300 mL x 2) and saturated brine (300 mL x 2). The mixture was dried over anhydrous sodium sulfate, filtered, and the mother liquor was concentrated under reduced pressure and purified by column chromatography (PE / EA = 4 / 1 to PE / EA = 1 / 2) to obtain compound 8 (10.10 g, 72.0%).

[0080] 1 H-NMR (400MHz, d-DMSO, ppm): δ10.40 (s, 1H); 8.05-7.92 (m, 1H); 7.55-7.45 (m, 1H); 7.38 (d, J=7.38Hz, 1H); 7.25 (d, J=7.98Hz, 1H); 7.20-7.11 (m, 2H); 7.08-6.83 (m, 2H); 6.75 (d, J=6.75Hz, 1H); 6.34 (s, 2H); 6.08 (d, J=12.00Hz, 2H); 5.23-5.13 (m, 1H); 5.00 (s, 1H); 4.70-4.38 (m, 2H); 3.75-3.53 (m, 3H); 3.48-3.35 (m, 2H); 3 .20-3.06 (m,1H); 2.85-2.71 (m,3H); 2.68-2.56 (m,2H); 2.50-2.43 (m,2H); 1.88-1.55 (m,2H); 1.11 (m,3H).

[0081] Example 9: Synthesis of Compound 9

[0082]

[0083] Step 1: At room temperature, add Giredestrant (10.45 g, 20 mmol), dry dichloromethane (200 mL), and triethylamine (6.07 g, 60 mmol) to a 500 mL three-necked flask, and stir to disperse. Cool to -5 to 0 °C, add side chain 9 (5.82 g, 25 mmol), and control the temperature below 20 °C. After the addition is complete, heat to reflux (35 to 40 °C) and react for 6 hours. After the reaction was completed, the temperature was lowered to 0-10℃, and the mixture was quenched in water (200mL). The phases were separated, and the aqueous phase was extracted with dichloromethane (100mL x 2). The organic phases were combined and washed successively with 10% sodium carbonate (200mL x 3) and saturated brine (200mL x 3). The mixture was dried over anhydrous sodium sulfate, filtered, and the mother liquor was concentrated under reduced pressure and purified by column chromatography (PE / EA = 4 / 1 to PE / EA = 1 / 2) to obtain compound 9 (8.67g, 60.3%).

[0084] Step 2: At room temperature, Giredestrant (10.45 g, 20 mmol), dried tetrahydrofuran (200 mL), and triethylamine (6.07 g, 60 mmol) were added to a 500 mL three-necked flask and stirred until dissolved. The temperature was lowered to -5 to 0 °C, and side chain 9 (5.82 g, 25 mmol) was added, maintaining the temperature below 20 °C. After the addition was complete, the temperature was raised to reflux (65 to 70 °C) and reacted for 3 h. After the reaction was completed, the temperature was lowered to 0 to 10 °C, quenched in water (300 mL), and extracted with dichloromethane (200 mL x 3). The organic phases were combined and washed successively with 10% sodium carbonate (300 mL x 2) and saturated brine (300 mL x 2). The mixture was dried over anhydrous sodium sulfate, filtered, and the mother liquor was concentrated under reduced pressure and purified by column chromatography (PE / EA = 4 / 1 to PE / EA = 1 / 2) to obtain compound 9 (10.78 g, 75.0%).

[0085] 1H-NMR (400MHz, d-DMSO, ppm): δ10.45 (s, 1H); 7.43 (d, J=7.45Hz, 1H); 7.30 (d, J=8.10Hz, 1H); 7.25-7.16 (m, 2H); 7.13-6.88 (m, 2H); 6.80 (d, J=6.80Hz, 1H); 6.75-6.65 (m, 2H); 6.39 (s, 2H); 6.13 (d, J=12.26Hz, 2H); 5.28-5.18 (m, 1H); 5.05 (s, 1H); 4.75-4.43 (m, 2H); 3.80-3.53 (m, 3H); 3.53-3.40 (m, 2H); 3 .25-3.11 (m,1H); 2.90-2.76 (m,3H); 2.73-2.61 (m,2H); 2.55-2.48 (m,2H); 1.93-1.58 (m,2H); 1.16 (m,3H).

[0086] Example 10: Synthesis of Compound 10

[0087]

[0088] Step 1: At room temperature, add Giredestrant (10.45 g, 20 mmol), dry dichloromethane (300 mL), and triethylamine (6.07 g, 60 mmol) to a 500 mL three-necked flask, and stir to disperse. Cool to -5 to 0 °C, add side chain 10 (6.12 g, 25 mmol), and control the temperature below 20 °C. After the addition is complete, heat to reflux (35 to 40 °C) and react for 6 hours. After the reaction was completed, the temperature was lowered to 0-10℃, and the mixture was quenched in water (200mL). The phases separated, and the aqueous phase was extracted with dichloromethane (200mL). The organic phases were combined and washed successively with 10% sodium carbonate (300mL x 3) and saturated brine (300mL x 3). The mixture was dried over anhydrous sodium sulfate, filtered, and the mother liquor was concentrated under reduced pressure and purified by column chromatography (PE / EA = 4 / 1 to PE / EA = 1 / 2) to obtain compound 10 (8.48g, 57.8%).

[0089] Step 2: At room temperature, Giredestrant (10.45 g, 20 mmol), dried DMF (100 mL), and triethylamine (6.07 g, 60 mmol) were added to a 500 mL three-necked flask and stirred until dissolved. The temperature was lowered to -5 to 0 °C, and side chain 10 (6.12 g, 25 mmol) was added, maintaining the temperature below 20 °C. After the addition was complete, the reaction was allowed to proceed at room temperature (20 to 30 °C) for 12 h. After the reaction was complete, the temperature was lowered to 0 to 10 °C, quenched in water (300 mL), and extracted with dichloromethane (200 mL x 3). The organic phases were combined and washed successively with 10% sodium carbonate (300 mL x 2) and saturated brine (300 mL x 2). The mixture was dried over anhydrous sodium sulfate, filtered, and the mother liquor was concentrated under reduced pressure and purified by column chromatography (PE / EA = 4 / 1 to PE / EA = 1 / 2) to obtain compound 10 (8.80 g, 69.9%).

[0090] 1 H-NMR (400MHz, d-DMSO, ppm): δ10.47 (s, 1H); 7.48 (d, J=7.48Hz, 1H); 7.35 (d, J=8.08Hz, 1H); 7.20-7.11 (m, 2H); 7.05-6.91 (m, 2H); 6.80 (d, J=6.81Hz, 1H); 6.39 (s, 2H); 6.25-6.16 (m, 2H); 6.10 (d, J=12.10Hz, 2H); 5.30-5.20 (m, 1H); 5.09 (s, 1H); 4.79-4.45 (m, 2H); 3.90-3.62 (m, 3H); 3.61-3.48 (m, 2H); 3.35 (s ,3H); 3.27-3.15 (m,1H); 2.93-2.80 (m,3H); 2.75-2.63 (m,2H); 2.59-2.51 (m,2H); 1.94-1.60 (m,2H); 1.12 (m,3H).

[0091] Example 11: Synthesis of Compound 11

[0092]

[0093] Step 1: At room temperature, Giredestrant (10.45 g, 20 mmol), dried tetrahydrofuran (300 mL), and triethylamine (6.07 g, 60 mmol) were added to a 500 mL three-necked flask and stirred to disperse them. The temperature was lowered to -5 to 0 °C, and side chain 11 (6.12 g, 25 mmol) was added, with the temperature controlled below 20 °C. After the addition was complete, the temperature was raised to 50 to 55 °C and reacted for 3 h. After the reaction was completed, the temperature was lowered to 0 to 10 °C, quenched in water (200 mL), extracted with dichloromethane (200 mL x 3), and the organic phases were combined. The mixture was washed successively with 10% sodium carbonate (300 mL x 3) and saturated brine (300 mL x 3), dried over anhydrous sodium sulfate, filtered, and the mother liquor was concentrated under reduced pressure and purified by column chromatography (PE / EA = 4 / 1 to PE / EA = 1 / 2) to obtain compound 11 (10.10 g, 68.8%).

[0094] Step 2: At room temperature, add Giredestrant (10.45 g, 20 mmol), dry DMF (100 mL), and N,N-diisopropylethylamine (7.76 g, 60 mmol) to a 500 mL three-necked flask, and stir until dissolved. Cool to -5 to 0 °C, add side chain 11 (6.12 g, 25 mmol), and maintain the temperature below 20 °C. After the addition is complete, raise the temperature to 70 to 80 °C and react for 3 hours. After the reaction was completed, the temperature was lowered to 0-10℃, quenched in water (300mL), extracted with dichloromethane (200mL x 3), the organic phases were combined, washed successively with 10% sodium carbonate (300mL x 2) and saturated brine (300mL x 2), dried over anhydrous sodium sulfate, filtered, the mother liquor was concentrated under reduced pressure, and purified by column chromatography (PE / EA = 4 / 1 to PE / EA = 1 / 2) to obtain compound 11 (10.43g, 71.0%).

[0095] 1H-NMR (400MHz, d-DMSO, ppm): δ10.37 (s, 1H); 7.38 (d, J=7.40Hz, 1H); 7.25 (d, J=7.99Hz, 1H); 6.95-6.81 (m, 2H); 6.78-6.73 (m, 2H); 6.70 (d, J=6.70Hz, 1H); 6.60-6.53 (m, 1H); 6.45-6.37 (m, 1H); 6.29 (s, 2H); 6.00 (d, J=12.00Hz, 2H); 5.20-5.10 (m, 1H) ); 4.99 (s, 1H); 4.69-4.35 (m, 2H); 3.80-3.52 (m, 3H); 3.51-3.38 (m, 2H); 3.30 (s, 3H); 3.17-3.05 (m,1H); 2.83-2.70 (m,3H); 2.65-2.53 (m,2H); 2.49-2.41 (m,2H); 1.84-1.50 (m,2H); 1.02 (m,3H).

[0096] Example 12: Synthesis of Compound 12

[0097]

[0098] Step 1: At room temperature, add Giredestrant (10.45 g, 20 mmol), dry tetrahydrofuran (100 mL), and N,N-diisopropylethylamine (7.76 g, 60 mmol) to a 500 mL three-necked flask, and stir to disperse. Cool to -5 to 0 °C, add side chain 12 (6.12 g, 25 mmol), and control the temperature below 20 °C. After the addition is complete, raise the temperature to 50 to 55 °C and react for 3 h. After the reaction was completed, the temperature was lowered to 0-10℃, quenched in water (200mL), extracted with dichloromethane (200mL x 3), the organic phases were combined, washed successively with 10% sodium carbonate (300mL x 3) and saturated brine (300mL x 3), dried over anhydrous sodium sulfate, filtered, the mother liquor was concentrated under reduced pressure, and purified by column chromatography (PE / EA = 4 / 1 to PE / EA = 1 / 2) to obtain compound 12 (9.97g, 67.9%).

[0099] Step 2: At room temperature, add Giredestrant (10.45 g, 20 mmol), dry acetonitrile (300 mL), and N,N-diisopropylethylamine (7.76 g, 60 mmol) to a 500 mL three-necked flask, and stir until dissolved. Cool to -5 to 0 °C, add side chain 12 (6.12 g, 25 mmol), and maintain the temperature below 20 °C. After the addition is complete, heat to reflux (75 to 80 °C) and react for 3 hours. After the reaction was completed, the temperature was lowered to 0-10℃, quenched in water (300mL), extracted with dichloromethane (200mL x 3), the organic phases were combined, washed successively with 10% sodium carbonate (300mL x 3) and saturated brine (300mL x 3), dried over anhydrous sodium sulfate, filtered, the mother liquor was concentrated under reduced pressure, and purified by column chromatography (PE / EA = 4 / 1 to PE / EA = 1 / 2) to give compound 12 (7.32g, 49.9%).

[0100] 1 H-NMR (400MHz, d-DMSO, ppm): δ10.33 (s, 1H); 7.65 (d, J=7.55Hz, 1H); 7.35-7.47 (m, 2H); 7.15-7.01 (m, 2H); 6.90 (d, J=6.80Hz, 1H); 6.80-6.73 (m, 2H); 6.65-6.57 (m, 1H); 6.49 (s, 2H); 6.20 (d, J=11.97Hz, 2H); 5.40-5.30 (m, 1H); 5.19 (s, 1H); 4.89-4.55 (m, 2H); 4.00-3.72 (m, 3H); 3.71-3.58 (m, 2H); 3.50 (s ,3H); 3.37-3.25 (m,1H); 3.03-2.90 (m,3H); 2.85-2.73 (m,2H); 2.69-2.61 (m,2H); 2.04-1.70 (m,2H); 1.22 (m,3H).

[0101] Table 1. Structural formulas and mass spectrometry data of compounds 1-12

[0102] ,

[0103] ,

[0104] .

[0105] Example 13: Stability Test of Influencing Factors

[0106] Table 2 Related Material Analysis Methods

[0107] ,

[0108] .

[0109] 2. Influencing Factors Experiment

[0110] The compound from the examples, along with Girdestrant, were packaged in pharmaceutical-grade low-density polyethylene bags and a polyester / aluminum / polyethylene pharmaceutical composite film. Under influencing conditions, a 30-day test was conducted using light exposure (total fluorescent lamp illuminance not less than 1.2 × 10⁶ Lux·hr, and near-ultraviolet lamp energy not less than 200 W·hr / m²), high temperature (60°C), and high humidity (25°C, RH: 90% ± 5). The test results are as follows:

[0111] Table 3. Results of the experiment on influencing factors

[0112] ,

[0113] .

[0114] The data in the table show that Giredestrant produces highly genotoxic impurity A after 30 days of exposure to light, high temperature, and high humidity. However, the crystal form of the Giredestrant derivative remains stable under these conditions. This indicates that the derivative provided by this invention has good stability and is suitable for routine storage of the active pharmaceutical ingredient.

[0115] Example 14: Solubility Test

[0116] The water solubility of the compounds was measured by suspending them in phosphate-buffered saline or an equivalent pH 7 buffer, sonicating (30 min), and slurrying (2 h, 1000 rpm) with excess solids (typically 5–10 mg). The suspension was then sonicated again for 30 min and stirred again at 1000 rpm for at least 12 h. Approximately 300 μL of sample was transferred to a rotary filter. Each sample was rotated at 16 K rpm for 10 min using a 0.45 μm PVDF rotary filter. A portion of the filtrate was transferred and diluted with acetonitrile:isopropanol (70:30 v / v). The obtained samples were analyzed by HPLC, and the results are as follows:

[0117] Table 4 Solubility Test Results

[0118] .

[0119] The results showed that the solubility of compounds 1, 6 and 8 was significantly improved, and the solubility of the other compounds was improved accordingly, which was unexpected before the salt formation.

[0120] Example 15: Comparative Example: Preparation of Girdestrant Tartaric Acid Capsules

[0121] Table 5 Comparative Example Prescriptions

[0122] .

[0123] 2. Preparation process

[0124] Mix the prescribed amount of Girdestrant tartaric acid with microcrystalline cellulose for 10 minutes and pass through a 200-mesh sieve;

[0125] Add the prescribed amount of lactose monohydrate and sodium carboxymethyl cellulose to step 1), mix for 15 minutes, and pass through a 100-mesh sieve;

[0126] Add the prescribed amount of talcum powder to step 2) and mix for 20 minutes.

[0127] Simply insert capsule number 3.

[0128] Example 16: Preparation of the inventive compound capsule

[0129] The formulation and preparation process are the same as in the comparative examples. The applicant selected specific representative compounds 1, 6 and 8 for testing. Their dosages are equivalent to 30 mg / capsule of Giredestrant, and the formulation dosages of the capsules are 390.4 mg, 366.3 mg and 403.6 mg, respectively.

[0130] Example 17: Dissolution Test

[0131] Take capsules of Compound 1, Compound 6, and Compound 8, and Girdestrant tartrate capsules, and perform dissolution testing according to the method outlined in the 2025 edition of the Pharmacopoeia of the People's Republic of China, Part IV, General Chapter 0931, Method II. Use 900 mL of 0.1 mol / L hydrochloric acid solution (or 900 mL of pH 4.5 acetate buffer, or 900 mL of pH 6.8 phosphate buffer containing 0.5% SDS, or 900 mL of purified water containing 0.5% SDS) as the dissolution medium, at a rotation speed of 50 r·min⁻¹. At specified time points (5, 10, 15, 30, 45, 60, 90, and 120 min), collect 7 mL of the solution (simultaneously adding 7 mL of isothermal medium), and filter through a 0.45 μm aqueous filter membrane. Accurately measure 1 mL of the filtrate, place it in a 10 mL volumetric flask, dilute to the mark with 0.1 mol·L⁻¹ hydrochloric acid solution, and mix well to obtain the sample solution. Take the above control solution and sample solution respectively, and measure the absorbance at 365 nm using a UV-Vis spectrophotometer [Pharmacopoeia of the People's Republic of China 2025 Edition, Part IV, General Chapter 0401]. Calculate the dissolution rate and plot the dissolution curve, as shown below. Figure 1 As shown.

[0132] The results showed that the dissolution rate and dissolution rate of compound 1 capsules, compound 6 capsules, and compound 8 capsules were significantly higher than those of Girdestrant tartrate capsules.

[0133] Example 18: Metabolic stability test

[0134] The applicant selected compounds 1, 6, and 8 as representatives, and incubated solutions of these compounds at certain concentrations with human, rat, and mouse liver microsomes in vitro. The percentage of remaining compounds was then determined using LC-MS / MS to compare the metabolic stability of compounds 1, 6, and 8 in human, rat, and mouse liver microsomes. The experimental results are shown in the table below:

[0135] Table 6. Comparison of metabolic stability of compounds 1, 6 and 8 in liver microsomes of humans, rats and mice.

[0136] .

[0137] The results showed that compounds 1, 6, and 8 could rapidly release bioactive giredestrants from liver microsomes in humans, rats, and mice. Specifically, the metabolic bioavailability (MF%) of compound 1 in human liver microsomes was 6.56%. The metabolism of compound 1 in rat and mouse liver microsomes, as well as compounds 6 and 8 in the same three types of liver microsomes, was very rapid, making accurate data impossible to obtain. Further analysis revealed that the rapid metabolic breakdown of these compounds is due to the readily metabolizable groups used in their formation.

[0138] Implementation Step 19: Experimental Study on In Vitro Cytotoxicity of Normal Human Hepatocytes

[0139] 1. Experimental Materials

[0140] 1.1 Cells: LO2 cells, a human hepatitis cell line

[0141] 1.2 Drug: Compound 1 of this invention is selected as a typical representative (HPLC purity 99.1%, because compound 1 has the best water solubility and crystal structure, and is the most representative).

[0142] 1.3 Reagents and Instruments: Modified RPMI-1640 culture medium, penicillin-streptomycin solution, 0.25% trypsin-EDTA, fetal bovine serum, MTT, dimethyl sulfoxide, CO-150 carbon monoxide incubator, SW-CJ-2F medical clean bench, CKX-41-32 inverted microscope, CU600 electric thermostatic water bath, RT-2100C enzyme-linked immunosorbent assay (ELISA) analyzer.

[0143] 2 Experimental Methods

[0144] 2.1 Reagent Preparation

[0145] 2.1.1 Preparation of MTT: Weigh 0.25g of MTT using a precision balance and place it in a 50mL volumetric flask. Add an appropriate amount of PBS, incubate in a 50-60℃ water bath, and shake well to dissolve completely. Add PBS to the mark to prepare a 5mg / kg solution. Filter the solution through a 0.22μm microporous membrane for sterilization, aliquot, and store in a refrigerator at 4℃ protected from light.

[0146] 2.1.2 Preparation of cell cryopreservation solution: The cell cryopreservation solution was prepared by mixing 20% ​​serum, 10% DMSO and 70% 1640 medium evenly and storing at -20℃.

[0147] 2.1.3 Preparation of Compound 1: Prepare a stock solution of the drug using DMSO, and then dilute it with culture medium to the concentration of the drug to be used. The final concentration of DMSO should be controlled at ≤0.1%.

[0148] 2.2 LO2 Cell Culture: Normal human LO2 cells were placed in a 25cm² cell culture flask, and approximately 4–5 mL of RPMI-1640 culture medium containing 10% FBS was added. The flasks were then incubated at 37°C in a 5% CO2 saturated humidity incubator. The culture medium was changed every 2 days, and cell growth was observed daily. Cells were passaged or cryopreserved after reaching 80% confluence. Cells from passages 5–7 were used for formal experiments.

[0149] 2.3 Grouping and administration: The experiment was divided into a normal cell control group and different concentrations of the drug administration group of the present invention based on the preliminary experimental results. The concentrations were 4.0, 8.0, 16.0, 32.0, 64.0, 128, 256, 512, 800 and 1000 μmol / L.

[0150] 2.4 Hepatocyte MTT assay: Logarithmic growth phase LO2 cells were prepared into a cell suspension of 5.0 × 10³ cells / mL and seeded into 96-well plates. A normal control group and treatment groups were established using different concentrations of compound 1 (4.0, 8.0, 16.0, 32.0, 64.0, 128, 256, 512, 800, and 1000 μmol / mL). After 24 h of culture, the culture medium was aspirated, and the cells were washed 2–3 times with PBS. Different concentrations of the compound of the present invention were added, with 8 replicates per concentration. The plates were incubated at 37°C in a 5% CO₂ incubator. After 24, 48, and 72 h of culture, MTT solution was added at each time point, and the plates were incubated in the dark for 4 h. After 4 h, the supernatant was aspirated, and 150 LDMSO was added to each well to dissolve the thiazolyl blue crystals. The plates were gently shaken to ensure uniform dissolution. The absorbance of each well was measured at 490 nm using a microplate reader. The absorbance value per well reflects the number of cells, and the two are directly proportional. The experiment was repeated three times. Cell viability was calculated as follows: Cell viability (%) = Absorbance of each group (OD490) × 100 / Absorbance of the control group (OD490)

[0151] 2.5 Statistical Analysis: All data are expressed as mean ± standard deviation and were processed using SPSS 17.0 statistical software. t-tests were performed for statistical analysis; paired t-tests were used for self-comparisons, and unpaired t-tests were used for inter-group comparisons. The significance level was P < 0.05.

[0152] 3. Experimental Results: The effects of different concentrations of compound 1 on cell growth at different time periods are shown in the table below:

[0153] Table 7 Effect of compound 1 on LO2 cell growth (x ± s, n = 8)

[0154] .

[0155] Experimental data showed that 24 h after administration, the OD values ​​of different concentrations of compound 1 were not statistically significant compared with the normal group, and the cell survival rate was similar to that of the normal group. 48 h after administration, the OD values ​​of all groups were higher than those after 24 h, indicating normal cell growth. Cells in different concentrations of phlorizin administration showed normal growth, with a survival rate greater than 90%, which was not statistically significant compared with the normal group. 72 h after administration, the OD values ​​of all groups showed an increasing trend, but the rate of increase decreased, indicating that the cells were still in the growth stage, although the growth rate was somewhat inhibited. Different concentrations of phlorizin administration had no inhibitory effect on cells, and the OD values ​​were not statistically significant compared with the normal group. The cell survival rate was greater than 90%. Therefore, it can be considered that compound 1, in the range of 4.0–1000 mol / L, has little inhibitory effect on cell growth and is non-toxic to cells.

[0156] Finally, it should be noted that the above embodiments are only used to illustrate the technical solutions of the present invention, and not to limit them. Although the present invention has been described in detail with reference to the foregoing embodiments, those skilled in the art should understand that modifications can still be made to the technical solutions described in the foregoing embodiments, or equivalent substitutions can be made to some or all of the technical features therein. Such modifications or substitutions do not cause the essence of the corresponding technical solutions to deviate from the scope of the technical solutions of the embodiments of the present invention.

Claims

1. A compound of the following formula or a pharmaceutically acceptable salt thereof, characterized in that... It has the following structure: 。 2. Use of the compound of claim 1 or a pharmaceutically acceptable salt thereof in the preparation of a medicament for treating breast cancer.

3. A pharmaceutical composition, characterized in that, It contains the compound as described in claim 1 or a pharmaceutically acceptable salt thereof, and a pharmaceutically acceptable carrier.

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