Furanocoumarin compounds and use thereof in the preparation of drugs for treating osteoporosis

Abstract

The application discloses a furan coumarin compound and application thereof in preparation of a drug for treating osteoporosis, and belongs to the technical field of drug research and development. 50 The furan coumarin compound has an IC of 37.86 nM, has no obvious cytotoxicity and systemic toxicity, can inhibit a key path of RANKL-induced osteoclast differentiation, and can be used for preparing the drug for treating osteoporosis.

Description

Technical Field

[0001] This invention belongs to the field of drug development technology, and in particular relates to a class of furanocoumarin compounds and their application in the preparation of drugs for treating osteoporosis. Background Technology

[0002] Osteoporosis (OP) is the most common metabolic disease of the skeletal system, characterized by decreased bone mineral density and deterioration of bone microstructure, leading to an increased risk of fragility fractures. With the accelerating aging of the global population, the incidence of osteoporosis continues to rise, and osteoporotic fractures—especially hip and spinal fractures—are associated with significant morbidity, mortality, and a heavy socioeconomic burden. Currently, drug treatment for osteoporosis mainly relies on anti-resorption agents and bone-forming agents. Although these therapies are effective in reducing fracture risk, their long-term use is limited by safety concerns, single-target mechanisms of action, and poor patient compliance. Therefore, developing alternative treatment strategies with higher safety and multi-target efficacy remains an important goal. Plant-derived bioactive compounds have long been considered an important resource for drug development due to their good efficacy and safety profile. Among them, isoimperatorin (ISO), a linear furanocoumarin mainly found in medicinal plants such as Salvia miltiorrhiza and Angelica sinensis, has attracted increasing attention due to its various pharmacological activities, including anti-inflammatory, antioxidant, and immunomodulatory effects. Previous studies have shown that isoimperatorin can inhibit osteoclast differentiation and function, reduce excessive bone resorption, and prevent bone loss due to estrogen deficiency. However, isoimperatorin is mainly metabolized by the liver, and studies have shown that long-term high-dose administration may produce hepatotoxic effects. Isoimperatorin is a strong inhibitor of P450 subtype 1a2 (Cyp1a2) and P450 subtype 2C (CYP2C), which may affect the metabolism of various clinical drugs (caffeine, omeprazole, clomipramine) (Wang Dujun et al., Effects of imperatorin and isoimperatorin on mouse liver CYP450, 2016). Moreover, most existing studies involve oral administration, but isoimperatorin has a complex metabolism in vivo, with 32 metabolites detected. Its bioavailability and pharmacokinetic characteristics still need to be optimized. Summary of the Invention

[0003] To address the aforementioned technical problems, this invention proposes a class of furanocoumarin compounds and their application in the preparation of drugs for treating osteoporosis.

[0004] To achieve the above objectives, the present invention provides the following technical solution: This invention provides a class of furanocoumarin compounds, the general structural formulas of which are shown in Formula I and Formula II: In Equation I, R1 is selected from , , , , , , , , , , , , , , or ; In Equation II, R2 is selected from , , , , , , , , or .

[0005] This invention utilizes furanocoumarins as the parent nucleus for structural modification, resulting in a series of novel furanocoumarin compounds, IC 50 It reaches 37.86 nM, has no obvious cytotoxicity or systemic toxicity, and can inhibit the key pathway of RANKL-induced osteoclast differentiation, which can be used to prepare drugs for the treatment of osteoporosis.

[0006] Furthermore, the furanocoumarin compounds are selected from the following A1-A groups. 16 B1-B 10 Any compound: , , , , , , , , , , , , , , , , , , , , , , , , or .

[0007] This invention also provides a method for preparing the above-mentioned furanocoumarin compounds. When the furanocoumarin compound has the structural formula of Formula I, the method includes the following steps: dissolving compound 1 and potassium carbonate in N,N-dimethylformamide, adding a first bromoalkyl compound, and reacting at 80°C for 8 hours to obtain the furanocoumarin compound shown in Formula I, wherein the structural formula of compound 1 is... The synthetic route is as follows ; When the furanocoumarin compound has the structural formula of Formula II, the process includes the following steps: dissolving compound 2 in anhydrous dichloromethane, adding boron tribromide under nitrogen protection, and reacting to obtain compound 3; dissolving potassium carbonate and compound 3 in N,N-dimethylformamide, adding a second bromoalkyl compound, and reacting at 80°C for 8 hours to obtain the furanocoumarin compound shown in Formula II, wherein the structural formula of compound 2 is... The structural formula of compound 3 is as follows: The synthetic route is as follows .

[0008] This invention uses a nucleophilic substitution reaction to prepare furanocoumarin compounds. The synthetic route is simple, the conditions are mild (80℃, 8h), and the yield is moderate (7.9%~80%).

[0009] Furthermore, the equivalent ratio of compound 1 to the first bromoalkyl compound is 1:1.1.

[0010] Further, the first bromoalkyl compound is selected from bromomethylcyclopentane, methyl bromoacetate, 4-methoxybenzyl bromide, bromopropene, 2-bromomethyl-6-methylpyridine, 4-bromomethyltetrahydropyran, 3-bromomethylpyridine hydrobromide, 3-bromomethylbenzaldehyde, (1-bromomethyl)-3-methylbenzene, 2-bromoethylacetate, ethyl bromoacetate, 4-(bromomethyl)piperidine-1-carboxylic acid tert-butyl ester, 2-bromo-N,N-diethylethylamine hydrobromide, ethyl 6-bromohexanoate, (R)-5-bromomethyl-2-pyrrolidone, or ethyl 5-bromopentanoate.

[0011] Furthermore, the equivalent ratio of compound 3 to the second bromoalkyl compound is 1:1.1.

[0012] Further, the second bromoalkyl compound is selected from bromomethylcyclopropane, methyl bromoacetate, bromopropene, methyl 4-bromomethylbiphenyl-2-carboxylate, methyl 3-bromomethylbenzoate, 3-bromomethylbenzaldehyde, (1-bromomethyl)-3-methylbenzene, 2-bromoethylacetate, 8-bromomethylquinoline, or tert-butyl 4-(bromomethyl)piperidine-1-carboxylate.

[0013] The present invention also provides the use of the above-mentioned furanocoumarin compounds or their pharmaceutically acceptable salts or solvates in the preparation of drugs for treating osteoporosis.

[0014] The present invention also provides a medicament for treating osteoporosis, comprising the above-mentioned furanocoumarin compounds or their pharmaceutically acceptable salts or solvates.

[0015] Compared with the prior art, the present invention has the following advantages and technical effects: Compared to ISO, the furanocoumarin compounds provided by this invention have lower toxicity, stronger activity, and better stability.

[0016] Compared to sodium alendronate (Alen-Na), the furanocoumarin compounds provided in this invention, as plant-derived compounds, have better biocompatibility and multi-target regulatory advantages.

[0017] Compared with existing anti-osteoporosis drugs, the furanocoumarin compounds provided by this invention avoid the safety issues of long-term use and provide a new choice of structural types. Attached Figure Description

[0018] The accompanying drawings, which form part of this invention, are used to provide a further understanding of the invention. The illustrative embodiments of the invention and their descriptions are used to explain the invention and do not constitute an undue limitation of the invention. In the drawings: Figure 1 To test Example 1, the expression of proto-oncogene protein (c-Fos) mRNA (A) and tumor necrosis factor α (TNF-α) mRNA (B) after different compounds were used to treat macrophage colony-stimulating factor (M-CSF) and nuclear factor-κB receptor activator ligand (RANKL) induced BMDMs.

[0019] Figure 2 For example 1, compound B 10 (A) Cell viability of BMDMs after treatment with compound Isoimperatorin (ISO) (B).

[0020] Figure 3 In Test Example 1, BMDMs were induced by M-CSF and RANKL, via compound B 10 The staining status of TRAP after treatment (A), area statistics (B) and its inhibition ability change curve (C); the staining status of TRAP after ISO treatment (D), area statistics (E) and its inhibition ability change curve (F), where the scale bar in A is 100 μm and the scale bar in D is 100 μm.

[0021] Figure 4 In Test Example 1, RANKL-induced BMDMs were processed via compound B 10 After treatment, the expression of MMP9 (A), Dc-stamp (B), and NFATc1 (C) mRNAs was observed.

[0022] Figure 5 For example 1, the staining of F-actin ring structure (A), area statistics (B), and the number of cell nuclei in osteoclasts after different treatments of RANKL-induced BMDMs (C) are shown.

[0023] Figure 6 The images show the Micro-CT three-dimensional reconstruction images (A) of the femur of ovariectomized (OVX) mice in test example 2, as well as the determination of bone structure parameters: BV / TV (B), BS / BV (C), and Tb.Th (D).

[0024] Figure 7 In Test Example 2, the levels of serum and biochemical indicators ALT (A), AST (B), BUN (C), CRP (D), CK-MB (E), cTNI (F), and E2 (G) in mice of each experimental group were measured. Detailed Implementation

[0025] Various exemplary embodiments of the present invention will now be described in detail. This detailed description should not be considered as a limitation of the present invention, but rather as a more detailed description of certain aspects, features, and embodiments of the present invention.

[0026] It should be understood that the terminology used in this invention is merely for describing particular embodiments and is not intended to limit the invention. Furthermore, with respect to numerical ranges in this invention, it should be understood that each intermediate value between the upper and lower limits of the range is also specifically disclosed. Every smaller range between any stated value or intermediate value within a stated range, and any other stated value or intermediate value within said range, is also included in this invention. The upper and lower limits of these smaller ranges may be independently included or excluded from the range.

[0027] Unless otherwise stated, all technical and scientific terms used herein have the same meaning as commonly understood by one of ordinary skill in the art. While only preferred methods and materials have been described herein, any methods and materials similar or equivalent to those described herein may be used in the implementation or testing of this invention. All references to this specification are incorporated by way of citation to disclose and describe methods and / or materials associated with those references. In the event of any conflict with any incorporated reference, the content of this specification shall prevail.

[0028] Various modifications and variations can be made to the specific embodiments described in this specification without departing from the scope or spirit of the invention, as will be apparent to those skilled in the art. Other embodiments derived from this specification will also be apparent to those skilled in the art. This specification and embodiments are merely exemplary.

[0029] The terms “include,” “including,” “have,” “contain,” etc., used in this article are all open-ended terms, meaning that they include but are not limited to.

[0030] Unless otherwise specified, the room temperature in this invention is 25±2℃.

[0031] All raw materials and reagents used in this invention were purchased commercially. For example, C57 / BL6 mice were purchased from Zhuhai Beston Technology Co., Ltd. Mouse bone marrow-derived macrophages (BMDMs) were extracted from C57 / BL6 mice. The specific extraction method involved euthanizing the mice, separating the femur and tibia under aseptic conditions, and removing attached soft tissue. The bone marrow cavity was rinsed with sterile buffer to obtain a bone marrow cell suspension. After erythrocyte lysis and filtration to remove impurities, the cells were seeded into a culture system containing macrophage colony-stimulating factor (M-CSF) to induce differentiation. The culture medium was changed periodically during the culture process, and mature adherent BMDMs were obtained after approximately 6–7 days.

[0032] It should be noted that any aspects not described in detail in this invention are conventional practices in the field and are not the focus of this invention.

[0033] The technical solution of the present invention will be further illustrated by the following embodiments.

[0034] Example 1 The furanocoumarin compounds prepared in this embodiment are 9-(cyclopentylmethoxy)-7H-furo[3,2- g]chromen-7-one (9-(cyclopentylmethoxy)-7H-furano[3,2-g]chromen-7-one, A1), the synthetic route is as follows: The specific preparation process was as follows: Bromomethylcyclopentane (44.4 mg, 0.25 mmol, 1.0 equiv) was added to a 2 mL DMF (N,N-dimethylformamide) mixture of compound 1 (50.0 mg, 0.25 mmol, 1.0 equiv) and K₂CO₃ (136.7 mg, 1.0 mmol, 4.0 equiv), and the reaction was carried out at 80 °C for 8 h. After the reaction was completed by TLC monitoring, water (5 mL) was added, followed by dilution with ethyl acetate (15 mL). The organic layer was separated, and the aqueous phase was extracted with ethyl acetate (3 × 15 mL). The combined organic phases were washed with brine, dried over anhydrous Na₂SO₄, filtered, and concentrated under reduced pressure. The residue was purified by silica gel chromatography to give 30.3 mg of compound A1 as a white solid, with a yield of 42.7%. 1 H NMR (400 MHz, CDCl3) δ 7.76 (d, J= 9.6 Hz, 1H), 7.69 (d, J = 2.4 Hz, 1H), 7.35 (s, 1H), 6.81 (d, J = 2.4 Hz, 1H), 6.37 (d, J =9.6 Hz, 1H), 4.35 (d, J = 7.2 Hz, 2H), 2.51 – 2.39 (m, 1H), 1.93 – 1.84 (m,2H), 1.68 – 1.63 (m, 2H), 1.61 – 1.56 (m, 2H), 1.51 – 1.42 (m, 2H). 13 C NMR (101 MHz, CDCl3) δ 160.8, 148.5, 146.8, 144.5, 143.7, 132.4, 126.1, 116.7,114.9, 113.1, 106.8, 78.4, 40.0, 29.4 (2×C), 25.6 (2×C). ESI-HRMS m / z:calculated for C 17 H 17 O4 + [M + H] + , 285.1121; found, 285.1120. Example 2 The furanocoumarin compounds prepared in this embodiment are methyl 2-((7-oxo-7H-furo[3,2-g] chromen-9-yl)oxy)acetate (2-((7-oxo-7H-furano[3,2-g]chromene-9-yl)oxy)methyl acetate, A2), its structure is ; The specific preparation process is as follows: methyl bromoacetate is used instead of methyl bromocyclopentane, and other conditions remain unchanged. The operation of Example 1 is repeated to obtain 38.0 mg of A2, which is a white solid with a yield of 56.1%. 1 H NMR (400 MHz, DMSO- d 6) δ8.11 (d, J = 9.6 Hz, 1H), 8.07 (d, J = 2.4 Hz, 1H), 7.63 (s, 1H), 7.05 (d, J = 2.4Hz, 1H), 6.41 (d, J= 9.6 Hz, 1H), 5.16 (s, 2H), 3.66 (s, 3H). 13 C NMR (101 MHz, DMSO- d 6) δ 169.6, 160.0, 148.4, 146.4, 145.7, 142.1, 130.6, 126.3, 116.8,114.7, 114.3, 107.5, 68.9, 52.4. ESI-HRMS m / z: calculated for C 14 H 11 O6 + [M + H] + , 275.0550; found, 275.0551. Example 3 The furanocoumarin compounds prepared in this embodiment are 9 - ((4-methoxybenzyl)oxy)-7H-furo[3, 2-g]chromen-7-one (9-((4-methoxybenzyl)oxy)-7H-furano[3,2-g]chromene-7-one, A3), its structural formula is ; The specific preparation process is as follows: 4-methoxybenzyl bromide is used instead of bromomethylcyclopentane, and other conditions remain unchanged. The operation of Example 1 is repeated to obtain 30.6 mg of A3, which is a white solid with a yield of 38.4%. 1 H NMR (400 MHz, DMSO- d 6) δ 8.15 – 8.11 (m, 2H), 7.67 (s, 1H), 7.44 – 7.40 (m, 2H), 7.09 (d, J = 2.4 Hz,1H), 6.94 – 6.89 (m, 2H), 6.43 (d, J = 9.6 Hz, 1H), 5.39 (s, 2H), 3.73 (s, 3H). 13 C NMR (101 MHz, DMSO- d 6) δ 159.8, 159.3, 147.9, 147.5, 145.3, 143.0, 130.5,130.1 (2×C), 128.6, 125.7, 116.4, 114.3, 114.2, 113.8 (2×C), 107.1, 74.4,55.1. ESI-HRMS m / z: calculated for C19 H 15 O5 + [M + H] + , 323.0914; found, 323.0911. Example 4 The furanocoumarin compounds prepared in this embodiment are 9-(allyloxy)-7H-furo[3,2-g]chromen- 7-one (9-Allyloxy-7H-furano[3,2-g]chromene-7-one, A4), its structural formula is ; The specific preparation process is as follows: bromopropene is used instead of bromomethylcyclopentane, and other conditions remain unchanged. The operation of Example 1 is repeated to obtain 41.4 mg of A4, which is a white solid with a yield of 69.2%. 1 H NMR (400 MHz, DMSO- d 6) δ 8.14(d, J = 14.0 Hz, 1H), 8.13 (d, J = 2.4 Hz, 1H), 7.69 (s, 1H), 7.10 (d, J = 2.0 Hz, 1H), 6.44 (d, J = 10.0 Hz, 1H), 6.16 – 6.03 (m, 1H), 5.45 – 5.37 (m, 1H), 5.27– 5.21 (m, 1H), 4.95 (t, J = 1.6 Hz, 1H), 4.93 (t, J = 1.6 Hz, 1H). 13 C NMR (101MHz, DMSO- d 6) δ 159.8, 147.9, 147.4, 145.3, 142.9, 133.7, 130.4, 125.8, 118.6,116.4, 114.3, 114.2, 107.1, 73.7. ESI-HRMS m / z: calculated for C 14 H 11 O4 + [M + H] + , 243.0652; found, 243.0641. Example 5 The furanocoumarin compounds prepared in this embodiment are 9-((6-methylpyridin-2-yl)methoxy)- 7H-furo[3,2-g]chromen-7-one(9-((6-methylpyridin-2-yl)methoxy)-7H-furano[3,2-g]chromene-7-one, A5), its structural formula is ; The specific preparation process is as follows: 2-bromomethyl-6-methylpyridine is used instead of bromomethylcyclopentane, and other conditions remain unchanged. The operation of Example 1 is repeated to obtain 60.7 mg of A5, which is a white solid with a yield of 80.0%. 1 H NMR (400 MHz, DMSO- d 6) δ 8.16 (d, J = 9.6 Hz, 1H), 8.13 (d, J = 2.4 Hz, 1H), 7.76 (t, J = 7.6 Hz,1H), 7.71 (s, 1H), 7.51 (d, J = 7.6 Hz, 1H), 7.22 (d, J = 7.6 Hz, 1H), 7.11 (d, J =2.4 Hz, 1H), 6.45 (d, J = 9.6 Hz, 1H), 5.50 (s, 2H), 2.44 (s, 3H). 13 C NMR (101MHz, DMSO- d 6) δ 159.7, 157.4, 155.7, 148.0, 147.2, 145.3, 142.8, 137.4, 130.7,125.9, 122.6, 118.8, 116.5, 114.5, 114.3, 107.1, 75.4, 23.8. ESI-HRMS m / z:calculated for C 18 H 14 O4N + [M + H] + , 308.0917; found, 308.0926. Example 6 The furanocoumarin compounds prepared in this embodiment are 9-((tetrahydro-2H-pyran-4-yl) methoxy)-7H-furo[3,2-g]chromen-7-one (9-((tetrahydro-2H-pyran-4-yl)methoxy)-7H-furano[3,2-g]chromene-7-one, A6), its structure is as follows: ; The specific preparation process is as follows: 4-bromomethyltetrahydropyran was used instead of bromomethylcyclopentane, and other conditions remained unchanged. The operation of Example 1 was repeated to obtain 42.3 mg of A6, which was a white solid with a yield of 57.0%. 1 H NMR (400 MHz, DMSO- d 6) δ 8.14 (d, J = 12.0 Hz, 1H), 8.13 (s, 1H), 7.68 (s, 1H), 7.10 (d, J = 2.4 Hz, 1H), 6.43 (d, J = 9.6 Hz, 1H), 4.25 (d, J = 6.4 Hz, 2H), 3.93 – 3.84 (m, 2H), 3.34 (dd, J = 11.6, 2.4 Hz, 1H), 3.33 (d, J = 2.4 Hz, 1H), 2.09 –1.97 (m, 1H), 1.81 – 1.72 (m, 2H), 1.45 – 1.31 (m, 2H). 13 C NMR (101 MHz, DMSO- d 6) δ 159.8,147.9, 147.3, 145.3, 142.8, 131.2, 125.8, 116.5, 114.2, 114.1, 107.1, 78.0,66.6 (2×C), 35.4, 29.0 (2×C). ESI-HRMS m / z: calculated for C 17 H 17 O5 + [M + H] + ,301.1071; found, 301.1072. Example 7 The furanocoumarin compounds prepared in this embodiment are 9-(pyridin-3-ylmethoxy)-7H-furo[3, 2-g]chromen-7-one (9-((pyridin-3-yl)methoxy)-7H-furano[3,2-g]chromene-7-one, A7), its structural formula is ; The specific preparation process is as follows: 3-bromomethylpyridine hydrobromide was used instead of bromomethylcyclopentane, and other conditions remained unchanged. The operation of Example 1 was repeated to obtain 28.4 mg of A7, which was a white solid with a yield of 30.7%. 1H NMR (400 MHz, DMSO- d 6) δ 8.69 (d, J = 2.0Hz, 1H), 8.54 (dd, J = 4.8, 1.6Hz, 1H), 8.15 – 8.11(m, 2H), 7.93 (dt, J = 7.6, 2.0 Hz, 1H), 7.70 (s, 1H), 7.41 (ddd, J = 8.0, 4.8,2.0Hz, 1H), 7.09 (d, J = 2.4Hz, 1H), 6.44 (d, J = 9.6 Hz, 1H), 5.50 (s, 2H). 13 CNMR (101 MHz, DMSO- d 6) δ 159.8, 149.7, 149.5, 148.1, 147.5, 145.4, 143.1,136.2, 132.4, 130.4, 125.8, 123.7, 116.5, 114.8, 114.3, 107.3, 72.5. ESI-HRMSm / z: calculated for C 17 H 12 O4N + [M + H] + , 294.0761; found, 294.0742. Example 8 The furanocoumarin compounds prepared in this embodiment are 3-(((7-oxo-7H-furo[3,2-g]chromen-9- yl)oxy)methyl)benzaldehyde (3-(((7-oxo-7H-furano[3,2-g]chromen-9-yl)oxy)methyl)benzaldehyde, A8), its structure is ; The specific preparation process is as follows: 3-bromomethylbenzaldehyde is used instead of bromomethylcyclopentane, and other conditions remain unchanged. The operation of Example 1 is repeated to obtain 12.7 mg of A8, which is a white solid with a yield of 16.1%. 1 H NMR (400 MHz, DMSO- d 6)δ 10.02 (s, 1H), 8.18 – 8.10 (m, 2H), 8.06 (t, J = 2.0Hz, 1H), 7.87 (ddt, J=13.6, 7.6, 1.6Hz, 2H), 7.70 (s, 1H), 7.63 (t, J = 7.6 Hz, 1H), 7.10 (d, J = 2.0Hz, 1H), 6.44 (d, J = 9.6 Hz, 1H), 5.56 (s, 2H). 13 C NMR (101 MHz, DMSO- d 6) δ193.1, 159.8, 148.1, 147.5, 145.4, 143.1, 138.1, 136.4, 134.1, 130.5, 129.6,129.5, 128.8, 125.9, 116.6, 114.8, 114.4, 107.3, 74.1. ESI-HRMS m / z:calculated for C 19 H 13 O5 + [M + H] + , 321.0758; found, 321.0755. Example 9 The furanocoumarin compounds prepared in this embodiment are 9-((3-methylbenzyl)oxy)-7H-furo[3, 2-g]chromen-7-one (9-((3-methylbenzyl)oxy)-7H-furano[3,2-g]chromene-7-one, A9), its structure is as follows: ; The specific preparation process is as follows: (1-bromomethyl)-3-methylbenzene is used instead of bromomethylcyclopentane, and other conditions remain unchanged. The operation of Example 1 is repeated to obtain 52.7 mg of A9, which is a white solid with a yield of 69.7%. 1 H NMR (400 MHz, DMSO- d 6) δ 8.15 – 8.11 (m, 2H), 7.67 (s, 1H), 7.34 (s, 1H), 7.30 – 7.22 (m,2H), 7.13 (d, J = 8.0Hz, 1H), 7.09 (d, J = 2.0 Hz, 1H), 6.43 (d, J = 9.6 Hz, 1H), 5.41 (s, 2H), 2.29 (s, 3H). 13 C NMR (101 MHz, DMSO- d6) δ 159.9, 148.0, 147.6,145.4, 143.1, 137.6, 136.7, 130.7, 129.0, 128.9, 128.4, 125.9, 125.3, 116.5,114.5, 114.3, 107.2, 74.8, 21.1. ESI-HRMS m / z: calculated for C 19 H 15 O4 + [M + H] + , 307.0965; found, 307.0966. Example 10 The furanocoumarin compounds prepared in this embodiment are 2-((7-oxo-7H-furo[3,2-g]chromen-9- yl)oxy)ethyl acetate (2-((7-oxo-7H-furano[3,2-g]chromen-9-yl)oxy)ethyl acetate, A 10 Its structure is ; The specific preparation process was as follows: 2-bromoethyl acetate (72.6 mg, 0.44 mmol, 1.1 equiv) was added to a 3 mL DMF mixture of compound 1 (80.0 mg, 0.40 mmol, 1.0 equiv) and K₂CO₃ (218.8 mg, 1.58 mmol, 4.0 equiv), and the mixture was reacted at 80 °C for 8 h. After the reaction was completed by TLC monitoring, water (5 mL) was added, followed by dilution with ethyl acetate (15 mL). The organic layer was separated, and the aqueous phase was extracted with ethyl acetate (3 × 15 mL). The combined organic phases were washed with brine, dried over anhydrous Na₂SO₄, filtered, and concentrated under reduced pressure. The residue was purified by silica gel chromatography to obtain 67.4 mg of compound A. 10 It is a white solid with a yield of 59.0%. 1 H NMR (400 MHz, DMSO- d 6) δ 8.15 (d, J = 9.6 Hz, 1H), 8.13 (d, J = 2.2 Hz, 1H), 7.70 (s, 1H), 7.10 (d, J = 2.0 Hz, 1H), 6.44 (d, J = 9.6Hz, 1H), 4.62 – 4.57 (m, 2H), 4.37 – 4.32 (m, 2H), 1.98 (s, 3H). 13 C NMR (101MHz, DMSO-d 6) δ 170.4, 159.8, 148.1, 147.5, 145.4, 143.0, 130.6, 125.9,116.6, 114.6, 114.3, 107.3, 71.3, 63.2, 20.7. ESI-HRMS m / z: calculated forC 15 H 13 O6 + [M + H] + , 289.0707; found, 289.0704. Example 11 The furanocoumarin compounds prepared in this embodiment are ethyl 2-((7-oxo-7H-furo[3,2-g] chromen-9-yl)oxy)acetate (2-((7-oxo-7H-furano[3,2-g]chromen-9-yl)oxy)ethyl acetate, A 11 Its structure is ; The specific preparation process is as follows: Ethyl bromoethyl is used instead of bromomethylcyclopentane, and other conditions remain unchanged. The operation of Example 10 is repeated to obtain 59.3 mg of A. 11 It is a white solid with a yield of 51.9%. 1 H NMR (400 MHz, DMSO- d 6) δ8.14 (d, J = 9.6 Hz, 1H), 8.09 (d, J = 2.2 Hz, 1H), 7.65 (s, 1H), 7.08 (d, J = 2.0Hz, 1H), 6.44 (d, J = 9.6 Hz, 1H), 5.17 (s, 2H), 4.15 (q, J = 7.2 Hz, 2H), 1.16(t, J = 7.2 Hz, 3H). 13 C NMR (101 MHz, DMSO- d 6) δ 168.7, 159.7, 148.0, 146.0,145.4, 141.7, 130.3, 126.0, 116.5, 114.4, 113.8, 107.1, 68.6, 60.8, 14.1.ESI-HRMS m / z: calculated for C 15 H 13 O6 + [M + H]+ , 289.0707; found, 289.0704. Example 12 The furanocoumarin compounds prepared in this embodiment are tert-butyl 4-(((7-oxo-7H-furo[3,2- g]chromen-9-yl)oxy)methyl)piperidine-1-carboxylate (4-(((7-oxo-7H-furano[3,2-g]chromen-9-yl)oxy)methyl)piperidine-1-carboxylic acid tert-butyl ester, A 12 Its structure is ; The specific preparation process is as follows: 4-(bromomethyl)piperidine-1-carboxylic acid tert-butyl ester was used instead of bromomethylcyclopentane, and other conditions remained unchanged. The operation of Example 10 was repeated to obtain 100.7 mg of A. 12 It is a white solid with a yield of 63.7%. 1 H NMR (400 MHz, DMSO-) d 6) δ 8.14 – 8.09 (m, 2H), 7.66 (s, 1H), 7.08 (d, J = 2.4 Hz, 1H), 6.42 (d, J = 9.6 Hz, 1H), 4.23 (d, J = 6.4 Hz, 2H), 4.03 – 3.91 (m, 2H), 2.74 (s, 2H), 1.99 – 1.91 (m, 1H), 1.83 (dd, J = 12.8, 3.6 Hz, 2H), 1.38 (s,9H), 1.26 – 1.14 (m, 2H). 13 C NMR (101 MHz, DMSO- d 6) δ 159.9, 154.0, 148.0,147.4, 145.4, 142.9, 131.2, 125.9, 116.6, 114.3, 114.2, 107.2, 78.7, 77.8,43.3 (2×C), 42.7 (2×C), 36.3, 28.2 (3×C). ESI-HRMS m / z: calculated forC 22 H 26 NO6 + [M + H] + , 400.1754; found, 400.1739. Example 13 The furanocoumarin compounds prepared in this embodiment are 9-(2-(diethylamino)ethoxy)-7H-furo[3,2-g]chromen-7-one (9-(2-(diethylamino)ethoxy)-7H-furano[3,2-g]chromone-7-one, A 13 Its structure is ; The specific preparation process is as follows: 2-bromo-N,N-diethylethylamine hydrobromide was used instead of bromomethylcyclopentane, and other conditions remained unchanged. The operation of Example 10 was repeated to obtain 20.3 mg of A. 13 It is a pale yellow solid with a yield of 13.4%. 1 H NMR (400MHz, DMSO- d 6) δ 8.14 – 8.09 (m, 2H), 7.66 (s, 1H), 7.08 (d, J = 2.0 Hz, 1H), 6.42 (d, J = 9.6 Hz, 1H), 4.48 (t, J = 6.0 Hz, 2H), 2.90 (t, J = 6.0 Hz, 2H), 2.58(q, J = 7.2 Hz, 4H), 0.92 (t, J = 7.2 Hz, 6H). 13 C NMR (101 MHz, DMSO- d 6) δ 160.3,148.3, 147.6, 145.8, 143.2, 131.4, 126.2, 116.8, 114.6, 114.4, 107.6, 71.6,52.5, 47.4 (2×C), 11.9 (2×C). ESI-HRMS m / z: calculated for C 17 H 20 NO4 + [M + H] + , 302.1387; found, 302.1395. Example 14 The furanocoumarin compounds prepared in this embodiment are ethyl 6-((7-oxo-7H-furo[3,2-g] chromen-9-yl)oxy)hexanoate (Ethyl 6-((7-oxo-7H-furano[3,2-g]chromen-9-yl)oxy)hexanoate, A 14 Its structure is ; The specific preparation process is as follows: Ethyl 6-bromohexanoate was used instead of methyl bromocyclopentane, and other conditions remained unchanged. The operation of Example 10 was repeated to obtain 94.0 mg of A. 14It is a yellow oily substance with a yield of 68.9%. 1 H NMR (400 MHz, DMSO- d 6)δ 8.12 (d, J = 8.0 Hz, 1H), 8.09 (s, 1H), 7.66 (s, 1H), 7.08 (s, 1H), 6.41 (d, J = 9.6 Hz, 1H), 4.39 – 4.34 (m, 2H), 4.04 – 3.99 (m, 2H), 2.29 (t, J = 7.2 Hz, 2H), 1.74 (q, J = 7.2 Hz, 2H), 1.58 (p, J = 7.2 Hz, 2H), 1.48 (p, J = 8.4 Hz, 2H), 1.14 (td, J = 7.2, 1.2 Hz, 3H). 13 C NMR (101 MHz, DMSO- d 6) δ 173.0, 159.9, 147.9,147.5, 145.4, 143.0, 131.1, 125.9, 116.6, 114.3, 114.2, 107.2, 73.4, 59.8,33.6, 29.3, 24.9, 24.3, 14.2. ESI-HRMS m / z: calculated for C 19 H 21 O6 + [M + H] + ,345.1333; found, 345.1326. Example 15 The furanocoumarin compounds prepared in this embodiment are 5-(((7-oxo-7H-furo[3,2-g]chromen-9- yl)oxy)methyl)pyrrolidin-2-one (5-(((7-oxo-7H-furano[3,2-g]chromen-9-yl)oxy)methyl)pyrrolidone-2-one, A) 15 Its structure is ; The specific preparation process is as follows: (R)-5-bromomethyl-2-pyrrolidone was used instead of bromomethylcyclopentane, and other conditions remained unchanged. The operation of Example 10 was repeated to obtain 22.7 mg of A. 15 It is a yellow solid with a yield of 19.2%. 1 H NMR (400MHz, DMSO-d 6) δ 8.17 – 8.11 (m, 2H), 7.75 (s, 1H), 7.69 (s, 1H), 7.10 (d, J =2.0 Hz, 1H), 6.43 (d, J = 9.6 Hz, 1H), 4.32 (h, J = 4.8 Hz, 2H), 3.92 (dt, J = 8.4,4.4 Hz, 1H), 2.40 – 2.32 (m, 1H), 2.27 – 2.19 (m, 1H), 2.18 – 2.12 (m, 1H), 2.10 – 2.04 (m, 1H). 13 C NMR (101 MHz, DMSO- d 6) δ 177.1, 159.8, 148.0, 147.3,145.4, 142.8, 131.1, 125.9, 116.6, 114.4, 114.3, 107.3, 75.9, 53.2, 29.6,23.0. ESI-HRMS m / z: calculated for C 16 H 14 NO5 + [M + H] + , 300.0867; found, 300.0879. Example 16 The furanocoumarin compounds prepared in this embodiment are ethyl 5-((7-oxo-7H-furo[3,2-g] chromen-9-yl)oxy)pentanoate Ethyl (5-((7-oxo-7H-furano[3,2-g]chromen-9-yl)oxy)valerate, A 16 Its structure is ; The specific preparation process is as follows: Ethyl 5-bromopentanoate was used instead of methyl bromocyclopentane, and other conditions remained unchanged. The operation of Example 1 was repeated to obtain 20.0 mg of A. 16 It is a colorless oily substance with a yield of 24.5%. 1 H NMR (400 MHz, DMSO- d 6) δ 8.15 (d, J = 9.6 Hz, 1H), 8.12 (d, J = 2.4 Hz, 1H), 7.69 (s, 1H), 7.10 (d, J= 2.2Hz, 1H), 6.44 (d, J = 9.6 Hz, 1H), 4.42 – 4.36 (m, 2H), 4.04 (q, J = 7.2 Hz, 2H),2.42 – 2.35 (m, 2H), 1.80 – 1.72 (m, 4H), 1.16 (t, J = 7.2 Hz, 3H). 13 C NMR (101MHz, DMSO- d 6) δ 172.8, 159.8, 147.9, 147.4, 145.3, 142.9, 130.9, 125.8, 116.5,114.2, 114.1, 107.1, 73.0, 59.7, 33.1, 28.9, 21.0, 14.1. ESI-HRMS m / z:calculated for C 18 H 19 O6 + [M + H] + , 331.1176; found, 331.1175. Example 17 This embodiment prepared a compound. 4-hydroxy-7H-furo[3,2-g]chromen-7-one (4-hydroxy-7H-furano[3,2-g]benzopyran-7-one, compound 3), the synthetic route is as follows: ; The specific preparation process was as follows: Under a nitrogen atmosphere, compound 2 (4.85 g, 22.4 mmol, 1.0 equiv) was dissolved in 50 mL of anhydrous dichloromethane (CH2Cl2), and a 2 M BBr3 dichloromethane solution (33.7 mL, 67.3 mmol, 3.0 equiv) was added dropwise at 0 °C. The reaction mixture was stirred at this temperature for 1 hour, then heated to room temperature and stirred for another 3 hours. After confirming the completion of the reaction by TLC, the mixture was poured into an ice-water bath in a fume hood. The resulting precipitate was allowed to stand for 30 minutes, then collected by filtration and dried to give 4.5 g of compound 3, which was a yellow solid with a yield of 99%. 1 H NMR (400 MHz, CD3OD) δ 8.32(d, J = 9.6 Hz, 1H), 7.70 (d, J = 2.4 Hz, 1H), 7.05 (dd, J= 2.4, 1.2 Hz, 1H), 7.02(s, 1H), 6.22 (d, J = 9.6 Hz, 1H). 13 C NMR (101 MHz, CD3OD) δ 163.8, 159.3, 154.4,149.4, 146.0, 141.6, 114.1, 111.6, 105.5, 105.0, 92.3. ESI-HRMS m / z:calculated for C 11 H7O4 + [M + H] + , 203.0266; found, 203.0335. Example 18 The furanocoumarin compounds prepared in this embodiment are 4-(cyclopropylmethoxy)-7H-furo[3,2- g]chromen-7-one (4-(cyclopropylmethoxy)-7H-furano[3,2-g]benzopyran-7-one, B1), its synthetic route is as follows: ; The specific preparation process was as follows: Bromomethylcyclopropane (54.0 mg, 0.44 mmol, 1.1 equiv) was added to a 2 mL DMF mixture of compound 3 (80.0 mg, 0.39 mmol, 1.0 equiv) and K₂CO₃ (218.8 mg, 1.58 mmol, 4.0 equiv), and the mixture was reacted at 80 °C for 8 h. After the reaction was completed by TLC monitoring, water (5 mL) was added, followed by dilution with ethyl acetate (15 mL). The organic layer was separated, and the aqueous phase was extracted with ethyl acetate (3 × 15 mL). The combined organic phases were washed with brine, dried over anhydrous Na₂SO₄, filtered, and concentrated under reduced pressure. The residue was purified by silica gel chromatography to give 26.2 mg of compound B1 as a white solid, with a yield of 28.1%. 1 H NMR (400 MHz, DMSO- d 6) δ 8.21 (dd, J = 9.6, 0.8 Hz, 1H), 8.02 (d, J = 2.4 Hz, 1H), 7.34 (t, J = 0.8 Hz, 1H), 7.27 (dd, J = 2.4, 1.0 Hz, 1H), 6.33 (d, J = 9.6 Hz, 1H), 4.29 (d, J= 7.2 Hz, 2H), 1.31 – 1.23 (m, 1H), 0.60 –0.54 (m, 2H), 0.41 – 0.33 (m, 2H). 13 C NMR (101 MHz, DMSO- d 6) δ 160.3, 157.6,152.2, 148.8, 146.1, 139.7, 113.8, 112.6, 106.7, 105.6, 93.6, 77.4, 10.8, 3.1(2×C). ESI-HRMS m / z: calculated for C 15 H 13 O4 + [M + H] + , 257.0808; found,257.0806. Example 19 The furanocoumarin compounds prepared in this embodiment are methyl 2-((7-oxo-7H-furo[3,2-g] chromen-4-yl)oxy)acetate (2-((7-oxo-7H-furano[3,2-g]chromen-4-yl)oxy)methyl acetate, B2), its structure is ; The specific preparation process is as follows: methyl bromoacetate is used instead of methyl bromocyclopentane, and other conditions remain unchanged. The operation of Example 18 is repeated to obtain 8.6 mg of B2, which is a white solid with a yield of 7.9%. 1 H NMR (400 MHz, DMSO- d 6) δ 8.31(dd, J = 9.6, 0.8 Hz, 1H), 8.05 (d, J = 2.4 Hz, 1H), 7.44 (t, J = 0.8 Hz, 1H), 7.23(dd, J = 2.4, 1.2 Hz, 1H), 6.39 (d, J = 9.6 Hz, 1H), 5.22 (s, 2H), 3.70 (s, 3H). 13 C NMR (101 MHz, DMSO- d6) δ 169.2, 160.2, 157.5, 152.0, 147.8, 146.7, 139.8,113.7, 113.0, 107.0, 104.8, 94.6, 68.9, 52.2. ESI-HRMS m / z: calculated forC 14 H 11 O6 + [M + H] + , 321.0758; found, 321.0756. Example 20 The furanocoumarin compounds prepared in this embodiment are 4-(allyloxy)-7H-furo[3,2-g]chromen- 7-one (4-Allyloxy-7H-furano[3,2-g]chromene-7-one, B3), its structure is as follows: ; The specific preparation process is as follows: bromopropene is used instead of bromomethylcyclopentane, and other conditions remain unchanged. The operation of Example 18 is repeated to obtain 57.7 mg of B3, which is a white solid with a yield of 60.1%. 1 H NMR (400 MHz, DMSO- d 6) δ 8.19(d, J = 9.6 Hz, 1H), 8.02 (d, J = 2.4 Hz, 1H), 7.39 – 7.27 (m, 2H), 6.32 (d, J =10.0 Hz, 1H), 6.19 – 6.08 (m, 1H), 5.47 (dd, J = 17.2, 1.6Hz, 1H), 5.30 (dd, J =10.8, 1.6 Hz, 1H), 5.05 – 5.00 (m, 2H). 13 C NMR (101 MHz, DMSO- d 6) δ 160.2,157.6, 152.2, 148.3, 146.2, 139.6, 133.5, 118.3, 113.5, 112.6, 106.5, 105.6,93.7, 73.0. ESI-HRMS m / z: calculated for C 14 H 11 O4 + [M + H] + , 243.0652; found,243.0641. Example 21 The furanocoumarin compounds prepared in this embodiment are methyl 4'-(((7-oxo-7H-furo[3,2-g] chromen-4-yl)oxy)methyl)-[1,1'-biphenyl]-2-carboxylate (4'-(((7-oxo-7H-furano[3,2-g]chromen-4-yl)oxy)methyl)-[1,1'-biphenyl]-2-carboxylic acid methyl ester, B4), its structure is ; The specific preparation process is as follows: methyl 4-bromomethylbiphenyl-2-carboxylic acid ester is used instead of bromomethylcyclopentane, and other conditions remain unchanged. The operation of Example 18 is repeated to obtain 126.3 mg of B4, which is a white solid with a yield of 74.8%. 1 H NMR (400 MHz, DMSO- d 6) δ 8.19 (d, J = 10.0 Hz, 1H), 8.08 (d, J = 2.4 Hz, 1H), 7.73 (d, J = 7.6Hz, 1H), 7.62 (t, J = 7.6 Hz, 1H), 7.58 (s, 1H), 7.56 (s, 1H), 7.51 – 7.47 (m,1H), 7.46 – 7.42 (m, 2H), 7.40 (s, 1H), 7.33 (s, 1H), 7.31 (s, 1H), 6.33 (d, J = 10.0 Hz, 1H), 5.62 (s, 2H), 3.55 (s, 3H). 13 C NMR (101 MHz, DMSO- d 6) δ 168.6,160.2, 157.7, 152.2, 148.4, 146.3, 140.9, 140.6, 139.7, 135.6, 131.7, 130.9,130.6, 129.5, 128.4 (2×C), 128.1 (2×C), 127.7, 113.7, 112.7, 106.7, 105.8,93.9, 73.9, 52.0. ESI-HRMS m / z: calculated for C 26 H 19 O6 + [M + H] + , 427.1176;found, 427.1172. Example 22 The furanocoumarin compounds prepared in this embodiment are methyl 3-(((7-oxo-7H-furo[3,2-g] chromen-4-yl)oxy)methyl)benzoate (3-(((7-oxo-7H-furano[3,2-g]chromen-4-yl)oxy)methyl)benzoate, B5), its structure is as follows ; The specific preparation process is as follows: methyl 3-bromomethylbenzoate is used instead of bromomethylcyclopentane, and other conditions remain unchanged. The operation of Example 18 is repeated to obtain 101.6 mg of B5, which is a white solid with a yield of 73.2%. 1 H NMR (400 MHz, DMSO- d 6) δ 8.16 (d, J = 9.6 Hz, 1H), 8.13 (s, 1H), 8.07 (d, J = 2.4 Hz, 1H), 7.95 (d, J = 9.6 Hz, 1H), 7.84 (d, J = 6.4 Hz, 1H), 7.59 (d, J = 7.6 Hz, 1H), 7.44 (d, J = 2.0Hz, 1H), 7.41 (s, 1H), 6.32 (s, 1H), 5.66 (s, 2H), 3.86 (s, 3H). 13 C NMR (101MHz, DMSO- d 6) δ 166.1, 160.2, 157.7, 152.2, 148.3, 146.3, 139.5, 137.5,132.9, 130.0, 129.3, 129.1, 128.7, 113.6, 112.8, 106.5, 105.8, 94.0, 73.4,52.4. ESI-HRMS m / z: calculated for C 20 H 15 O6 + [M + H] + , 351.0863; found, 351.0862. Example 23 The furanocoumarin compounds prepared in this embodiment are 3-(((7-oxo-7H-furo[3,2-g]chromen-4- yl)oxy)methyl)benzaldehyde (3-(((7-oxo-7H-furano[3,2-g]chromen-4-yl)oxy)methyl)benzaldehyde, B6), its structure is ; Replacing bromomethylbenzaldehyde with bromomethylcyclopentane, and keeping other conditions unchanged, the operation of Example 18 was repeated to obtain 59.2 mg of B6, which was a yellow solid, with a yield of 46.7%. 1 H NMR (400 MHz, DMSO- d 6) δ 10.04 (s, 1H), 8.18 (d, J = 9.6 Hz, 1H), 8.10 – 8.04 (m, 2H), 7.94 – 7.85 (m, 2H), 7.66 (t, J =7.6 Hz, 1H), 7.44 (d, J = 2.0 Hz, 1H), 7.39 (s, 1H), 6.31 (d, J = 9.6 Hz, 1H), 5.67 (s, 2H). 13 C NMR (101 MHz, DMSO- d 6) δ 193.2, 160.2, 157.7, 152.2, 148.3,146.3, 139.6, 137.9, 136.5, 134.0, 129.7, 129.6, 128.7, 113.6, 112.8, 106.5,105.8, 94.0, 73.3. ESI-HRMS m / z: calculated for C 19 H 13 O5 + [M + H] + , 321.0758;found, 321.0756. Example 24 The furanocoumarin compounds prepared in this embodiment are 4-((3-methylbenzyl)oxy)-7H-furo[3, 2-g]chromen-7-one (4-((3-methylbenzyl)oxy)-7H-furano[3,2-g]benzopyran-7-one, B7), its structure is as follows: ; The specific preparation process is as follows: (1-bromomethyl)-3-methylbenzene is used instead of bromomethylcyclopentane, and other conditions remain unchanged. The operation of Example 18 is repeated to obtain 58.0 mg of B7, which is a white solid with a yield of 47.8%. 1 H NMR (400 MHz, DMSO- d 6) δ 8.12 (d, J = 10.0 Hz, 1H), 8.04 (d, J= 2.4 Hz, 1H), 7.40 (s, 1H), 7.36 (s, 1H), 7.33 (s, 1H), 7.30-7.26 (m, 2H), 7.17 (d, J = 8.0 Hz, 1H), 6.30(d, J = 9.6 Hz, 1H), 5.50 (s, 2H), 2.31 (s, 3H). 13 C NMR (101 MHz, DMSO- d 6) δ160.2, 157.7, 152.2, 148.5, 146.2, 139.6, 137.9, 136.5, 129.1, 128.7, 128.6,125.3, 113.6, 112.7, 106.6, 105.8, 93.8, 74.2, 21.1. ESI-HRMS m / z: calculated for C 19 H 15 O4 + [M + H] + , 307.0965; found, 307.0965. Example 25 The furanocoumarin compounds prepared in this embodiment are 2-((7-oxo-7H-furo[3,2-g]chromen-4- yl)oxy)ethyl acetate (4-(2-acetoxyethoxy)-7H-furano[3,2-g]chromene-7-one, B8), its structure is as follows: ; The specific preparation process is as follows: 2-bromoethyl acetate is used instead of bromomethylcyclopentane, and other conditions remain unchanged. The operation of Example 18 is repeated to obtain 41.6 mg of B8, which is a white solid with a yield of 36.4%. 1 H NMR (400 MHz, DMSO- d 6)δ 8.17 (d, J = 10.4 Hz, 1H), 8.05 (d, J = 2.4 Hz, 1H), 7.39 (s, 1H), 7.30 (dd, J =2.4, 1.2 Hz, 1H), 6.36 (d, J = 9.6 Hz, 1H), 4.69 – 4.62 (m, 2H), 4.43 – 4.37 (m, 2H), 2.03 (s, 3H). 13 C NMR (101 MHz, DMSO-d 6) δ 170.5, 160.2, 157.6, 152.1,148.3, 146.4, 139.4, 113.7, 112.8, 106.6, 105.3, 94.0, 71.1, 62.9, 20.8. ESI-HRMS m / z: calculated for C 15 H 13 O6 + [M + H] + , 289.0707; found, 289.0706. Example 26 The furanocoumarin compounds prepared in this embodiment are 4-(quinolin-8-ylmethoxy)-7H-furo[3, 2-g]chromen-7-one (4-(quinoline-8-methoxy)-7H-furano[3,2-g]benzopyran-7-one, B9), its structure is as follows: ; The specific preparation process is as follows: 8-bromomethylquinoline was used instead of bromomethylcyclopentane, and other conditions remained unchanged. The operation of Example 18 was repeated to obtain 62.3 mg of B9, which was a white solid with a yield of 45.8%. 1 H NMR (600 MHz, CDCl3) δ8.95 (dd, J = 4.2, 1.8 Hz, 1H), 8.23 ​​(dd, J = 8.4, 1.8 Hz, 1H), 8.21 (d, J = 9.6Hz, 1H), 7.89 (d, J = 7.2 Hz, 1H), 7.87 (d, J = 8.4 Hz, 1H), 7.59 (t, J = 7.8 Hz, 1H), 7.55 (d, J = 2.4 Hz, 1H), 7.49 (dd, J = 8.4, 4.2 Hz, 1H), 7.19 (s, 1H), 7.06(d, J = 2.4 Hz, 1H), 6.23 (d, J = 9.6 Hz, 1H), 6.19 (s, 2H). 13C NMR (151 MHz, CDCl3) δ 161.3, 158.2, 152.6, 150.0, 149.1, 145.7, 144.9, 139.6, 136.4,134.5, 128.4, 128.2, 128.1, 126.3, 121.6, 114.2, 112.6, 107.4, 105.3, 94.4,71.4. ESI-HRMS m / z: calculated for C 21 H 14 NO4 + [M + H] + , 344.0917; found, 344.0918. Example 27 The furanocoumarin compounds prepared in this embodiment are tert-Butyl 4-(((7-oxo-7H-furo[3,2- g]chromen-4-yl)oxy)methyl)piperidine-1-carboxylate (4-(((7-oxo-7H-furano[3,2-g]chromen-4-yl)oxy)methyl)piperidine-1-carboxylic acid tert-butyl ester, B 10 Its structure is ; The specific preparation process is as follows: 4-(bromomethyl)piperidine-1-carboxylic acid tert-butyl ester was used instead of bromomethylcyclopentane, and other conditions remained unchanged. The operation of Example 18 was repeated to obtain 93.8 mg of B. 10 It is a white solid with a yield of 59.3%. 1 H NMR (400MHz, DMSO- d 6) δ 8.17 (d, J = 10.0 Hz, 1H), 8.02 (d, J = 2.4 Hz, 1H), 7.32 (s,1H), 7.30 (dd, J = 2.4, 1.2 Hz, 1H), 6.31 (d, J = 10.0 Hz, 1H), 4.34 (d, J = 6.4Hz, 2H), 4.00 (d, J = 12.8 Hz, 2H), 2.75 (s, 2H), 2.06 – 1.95 (m, 1H), 1.85 –1.77 (m, 2H), 1.39 (s, 9H), 1.28 – 1.20 (m, 2H). 13 C NMR (101 MHz, DMSO- d6) δ160.2, 157.7, 154.0, 152.2, 148.8, 146.1, 139.5, 113.0, 112.5, 106.0, 105.8,93.4, 78.6, 76.6, 43.7 (2×C), 42.8 (2×C), 36.3, 28.2 (3×C). ESI-HRMS m / z:calculated for C 22 H 26 NO6 + [M + H] + , 400.1755; found, 400.1773. Test Example 1: In vitro efficacy evaluation of furanocoumarin compounds 1. Cell culture Mouse bone marrow-derived macrophages (BMDMs) were cultured in α-MEM medium (Gibco, catalog number: C12571500BT) supplemented with 10% (v / v) FBS (Gibco, catalog number: 10270-106), 100 U / mL penicillin, and 100 μg / mL streptomycin (Gibco, catalog number: 15140122). The cells were cultured in a constant-temperature cell culture incubator at 37°C (atmosphere: 95% O2 and 5% CO2, all by volume percentage; all gas percentages mentioned below refer to volume percentages), with the culture medium being routinely refreshed every 48 hours.

[0035] 2. Detection of the effects of compounds on c-Fos mRNA and TNF-α mRNA expression (1) Seeding plate: BMDMs cells were seeded at a rate of 1×10⁻⁶. 6 Cells / well were seeded at a density of 1 mL culture medium per well in 12-well plates and cultured according to standard procedures until the cells adhered. Specific experimental groups were set as follows: negative control group, RANKL group, and compound treatment group.

[0036] (2) RANKL induction: Except for the negative control, all other groups were given 50 ng / mL M-CSF and 100 ng / mL RANKL to induce BMDMs cells to differentiate into osteoclasts. The drug-treated group was given 10 μM of the compound and induced differentiation for 5 days.

[0037] (3) RNA extraction: Discard the original cell culture medium, wash the cells twice with PBS, add 100 μL of RL from the RNA extraction kit to each well for lysis; collect the cell lysate and extract Total RNA according to the kit instructions (Vazyme, catalog number: RC112), and measure its concentration with an ultra-micro UV-Vis spectrophotometer. (4) Reverse transcription reaction A 20 μL reverse transcription system was established, including 4 μL of 5× reaction buffer, X μL of total RNA (500 ng), and μL of DEPC-H2O (16-X). After being placed in 8-tube sets, the tubes were labeled and placed in a polymerase chain reaction (PCR) amplification instrument at 50 °C for 15 min and 85 °C for 5 s for reverse transcription. (5) Quantitative fluorescence Add 80 μL DEPC-H2O to the transcribed cDNA to a final volume of 100 μL, centrifuge to mix, and store at -20℃. Multiply the sample number by the following ratio: (SYBR Green fluorescent dye and its reaction mixture, purchased from TransGen Biotech Co., Ltd., model AQ601-02-V2) 10 μL + DEPC-H2O 10 μL + primer 0.5 μL, and transfer 18 μL / well to a 96-well plate. Add 2 μL cDNA to make a 20 μL system, centrifuge at 1200 rpm for 1 min to mix. Place the sample plate in a CFX Connect Real-Time PCR system for detection, cycling 39 times at 95℃ for 2 min; 95℃ for 20 s; 57℃ for 20 s; 72℃ for 20 s; 95℃ for 1 min; 55℃ for 30 s; 95℃ for 30 s. The program s is used for detection; (6) Analysis Use 2 ‑ΔΔCt The experimental results were analyzed using the following formulas: △Ct target gene = Ct target gene - Ct internal reference gene; △△Ct target gene = △Ct experimental group target gene - △Ct control group target gene. 2 ‑ΔΔCt This indicates the fold increase in the expression of the target gene in the experimental group compared to the control group. The primers are shown in Table 1.

[0038] Table 1 Primer Sequences The results are as follows Figure 1 As shown, in the c-Fos mRNA expression assay, compared with the RANKL-induced group, various compounds (such as A) showed increased expression. 16 B5, B7, B 10 Treatment with these compounds all reduced c-Fos mRNA expression levels, suggesting that they inhibit RANKL-induced early osteoclast differentiation. In TNF-α mRNA expression detection, some compounds (such as A5, A6, A9, and A...) showed lower expression levels. 13 B7, B8, B10 Compound B (e.g., [specific compounds]) showed inhibitory effects on RANKL-induced TNF-α expression, indicating potential activity in regulating osteoclast-related inflammatory signaling. A comprehensive comparison of the inhibitory effects of different compounds on c-Fos and TNF-α mRNA expression revealed that compound B... 10 Compound B exhibits a significant inhibitory effect on the aforementioned genes, demonstrating superior overall activity characteristics. Therefore, compound B was selected. 10 As a representative compound, it will be used for subsequent in vitro and in vivo efficacy studies.

[0039] 3. Cell viability detection BMDMs cells at 1×10 6 Cells / well were seeded at a density of approximately 80% in 96-well plates. Then, different concentrations of compound B were added. 10 (0, 0.01, 0.1, 0.5, 1, 5, 10, 15 μM) or ISO (0, 0.01, 0.1, 0.5, 1, 5, 10, 15 μM), with 3 replicates per group and a blank control group. After culturing for 24 h, 10 μL of CCK8 solution was added to each well, and the cells were incubated for another 2 h at 37℃ in a 5% CO2 incubator. The absorbance (OD) value was measured at 450 nm using a microplate reader, and the cell inhibition rate was calculated.

[0040] The results are as follows Figure 2 As shown, compound B 10 No significant toxicity was observed. Figure 2 (A), while ISO showed significant toxicity to BMDMs cells at a concentration of 5 μM ( Figure 2 (B). Experimental results show that, compared with ISO, compound B of the present invention... 10 It is safer.

[0041] 4. TRAP staining experiment (1) Osteoclast induction and differentiation and experimental grouping: BMDMs cells were divided into groups of 1×10 6 Cells were seeded at a density of cells / well in 24-well plates and cultured in α-MEM medium. The following groups were established: negative control, RANKL-induced group, and RANKL+B group. 10 Group B and RANKL+ISO group. Compound B is among them. 10 Cells were treated with ISO at concentrations of 10, 20, 30, 35, 40, 45, and 50 nM. To promote osteoclast maturation, 50 ng / mL M-CSF and 100 ng / mL RANKL were added to the culture medium. The drug-containing medium was changed every 48 hours until a large number of well-defined, multinucleated osteoclasts were observed under a microscope.

[0042] (2) Staining: The differentiation of osteoclasts was assessed using the TRAP staining kit according to the manufacturer's instructions. The original cell culture medium was removed, and the cells were washed three times with PBS and fixed with 4% paraformaldehyde for 15 minutes. The TRAP staining solution was prepared according to the manufacturer's instructions. Then, 200 μL of the staining solution was added to a 24-well plate, and the cells were incubated at 37°C in the dark for 20 minutes. The staining solution was removed and the cells were washed three times. The cells were observed and statistically analyzed under a bright field microscope.

[0043] The results are as follows Figure 4 As shown, compound B 10 It can significantly reduce the area of ​​osteoclasts ( Figure 3 (A), its half-maximal inhibitory concentration (IC50) for inhibiting osteoclast formation 50 The value is 37.86 nM. Figure 3 (C), while ISO's IC 50 Value greater than 50 nM ( Figure 3 (F). Experimental results show that, compared with ISO, compound B... 10 It exhibits stronger osteoclastogenesis inhibitory ability. The IC50 value was determined based on TRAP staining analysis. 50 Value, subsequent experiments used compound B at a concentration of 40 nM. 10 To conduct research.

[0044] 5. Cellular mRNA extraction (1) Seeding plate: BMDMs cells were seeded at a rate of 1×10⁻⁶. 6 Cells / well were seeded at a density of 1 mL culture medium per well in 12-well plates and cultured according to standard protocols until cell adhesion was achieved. Specific experimental groups were set as follows: negative control group, RANKL group, Alen-Na group (1 μM), ISO group (1 μM), and RANKL+B group. 10 Group (40 nM).

[0045] (2) RANKL induction: Except for the negative control, all other groups were given 50 ng / mL M-CSF and 100 ng / mL RANKL to induce BMDMs cells to differentiate into osteoclasts. The drug-treated groups were given their respective concentrations of B... 10 Differentiation was induced for 5 days, with the culture medium changed once during this period, and M-CSF, RANKL, and B were reintroduced. 10 .

[0046] (3) RNA extraction: Discard the original cell culture medium, wash the cells twice with PBS, add 100 μL of RL from the RNA extraction kit to each well for lysis; collect the cell lysate and extract Total RNA according to the kit instructions (Vazyme, catalog number: RC112), and measure its concentration with an ultra-micro UV-Vis spectrophotometer. (4) Reverse transcription reaction A 20 μL reverse transcription system was established, including 4 μL of 5×Buffer, X μL of Total RNA (500 ng), and μL of DEPC-H2O (16-X). After being placed in 8-tube sets, the tubes were labeled and placed in a PCR instrument at 50 ℃ for 15 min and 85 ℃ for 5 s for reverse transcription. (5) Quantitative fluorescence Add 80 μL DEPC-H2O to the transcribed cDNA to a final volume of 100 μL, centrifuge to mix, and store at -20℃. Take 18 μL of the mixture (SYBR 10 μL + DEPC-H2O 10 μL + primer 0.5 μL) multiplied by the number of samples and transfer it to a 96-well plate. Add 2 μL of cDNA to make a 20 μL mixture, centrifuge at 1200 rpm for 1 min to mix. Place the sample plate in a CFX Connect Real-Time System for detection, cycling 39 times (95 ℃, 2 min; 95 ℃, 20 s; 57 ℃, 20 s; 72 ℃, 20 s), followed by a cycle of 95 ℃, 1 min; 55 ℃, 30 s; 95 ℃, 30 s. (6) Analysis Use 2 ‑ΔΔCt The experimental results were analyzed using the following formulas: △Ct target gene = Ct target gene - Ct internal reference gene; △△Ct target gene = △Ct experimental group target gene - △Ct control group target gene. 2 ‑ΔΔCt This indicates the fold increase in the expression of the target gene in the experimental group compared to the control group. The primers are shown in Table 1.

[0047] The results are as follows Figure 4 As shown, MMP9 in RANKL-induced BMDMs ( Figure 4 (A), Dc-stamp ( Figure 4 (B), NFATC1 ( Figure 4 The expression of mRNA in compound C was significantly increased, while that of compound B was significantly increased. 10 After treatment, the expression levels of the above genes were significantly reduced, indicating that compound B... 10 It can significantly inhibit the formation of osteoclasts.

[0048] 6. F-actin staining experiment (1) Seeding plate: Same as BMDMs cells at 1×10 6 Cells / well were seeded at a density of 1 / 24 wells and cultured according to standard methods until cell adhesion. Specific experimental groups were set as follows: negative control group, RANKL group, Alen-Na group, ISO group, and B group. 10 Group (40 nM).

[0049] (2) RANKL induction: Except for the Control group, all other groups were given 50 ng / mL M-CSF and 100 ng / mL RANKL to induce BMDMs cells to differentiate into osteoclasts. The drug-treated groups were given Alen-Na, ISO, and B, respectively. 10 Differentiation was induced for 5 days, with the culture medium changed once during this period, and M-CSF, RANKL, and B were reintroduced. 10 .

[0050] (3) Staining: F-actin in cells was detected using phalloidin labeled with an actin tracker, following the manufacturer's instructions. The original cell culture medium was removed, and the cells were fixed with 0.1% Triton X-100 and washed with PBS. The staining agent was diluted 1:50 and incubated with the cells for 60 minutes. After washing again, the cells were observed under a fluorescence microscope.

[0051] The results are as follows Figure 5 As shown, compared with the control group, the actin rings in the RANKL group exhibited larger ring structures and a significantly increased number of cell nuclei within the rings; while in B... 10 In the treatment group, the area of ​​the actin ring was significantly reduced, and the number of cell nuclei within the ring was also significantly decreased. These results indicate that compound B... 10 It can significantly inhibit the formation of osteoclasts.

[0052] Test Example 2 Compound B 10 In vivo efficacy evaluation 1. Ovariectomized (OVX) mouse model ① Animal groups: Sham (8 animals), OVX (8 animals), Alen-Na (10mg / kg) (8 animals), B 10 -L (5mg / kg) (8 animals), B 10 -M (10mg / kg) (8 animals), B 10 -H (20mg / kg) (8 animals).

[0053] ② Modeling method: C57 / BL6 mice, female, weighing 18-22 g, were acclimatized for 7 days, and their general condition, including diet, body weight, and mental state, was observed. After acclimatization, an osteoporosis model was established by ovariectomy, followed by 6 weeks of gavage administration.

[0054] ③ Animal administration: using compound B 10 Low (5 mg / kg), medium (10 mg / kg), and high (20 mg / kg) dose groups were administered via gavage. The positive control group received alendronate sodium (Alen-Na, 10 mg / kg), while the control group (Sham) and the model group (OVX) were administered an equal volume of physiological saline via gavage. The treatment groups received gavage once daily for 4 weeks starting from the beginning of the formal experiment.

[0055] ④ Animal sampling: Mice were injected intraperitoneally with 0.2 mL of 0.6% sodium pentobarbital, and blood was collected from the orbital cavity. The mice died from spinal dislocation. The mouse serum was collected and stored at -80℃, and the femur was fixed in paraformaldehyde.

[0056] 2. In vivo detection 2.1. Micro-CT Detection of Mouse Femoral Parameters Femoral tissue was placed on the scanning platform of the Micro-CT imaging system. Scanning parameters were set as follows: voltage 90 kV, current 180 μA, field of view (FOV) 40 mm, and a 360° spiral scan mode for 4.5 min to acquire high-resolution tomographic images. After scanning, the sample was reconstructed in three dimensions using Caliper analyze software to remove non-bone tissue residues (such as muscle). The reconstructed 3D data was imported into the Measure module of Caliperanalyze to measure parameters such as bone mineral density, bone volume / tissue volume (BV / TV), trabecular bone thickness (Tb.Th), and bone surface area to bone volume ratio (BS / BV). These values ​​were recorded for subsequent analysis.

[0057] The results are as follows Figure 6 As shown, BV / TV after oophorectomy Figure 6 (A) and Tb. Th ( Figure 6 C) decreased significantly, while BS / BV ( Figure 6 The level of compound B was significantly increased; while the level of compound B was significantly increased. 10 After treatment, all the above changes were significantly reversed. The results indicate that compound B... 10 It can slow the progression of osteoporosis in the body.

[0058] 2.2. Biochemical Indicator Detection Collect blood supernatant: Collect blood into a 1.5 mL EP tube, centrifuge at 1000 rpm for 10 minutes, and use the supernatant for ELISA detection of biochemical indicators.

[0059] The secretion of alanine aminotransferase (ALT), aspartate aminotransferase (AST), C-reactive protein (CRP), and estradiol (E2) in the supernatant was detected using an enzyme-linked immunosorbent assay (ELISA) kit (Shanghai Jianglai Biotechnology Co., Ltd., catalog number: E2: JL18734-48T, ALT: JL12668, AST: JL13992, CRP: JL13196). The procedure was performed according to the kit's instructions, as follows: (1) Reagent preparation ① Remove from the refrigerator and allow to equilibrate to room temperature for 20 minutes. ② Dilute the washing buffer (20×) with double-distilled water to 1× to prepare the required washing buffer. ③ Add the standard diluent to one bottle of standard according to the volume indicated on the standard label and incubate at room temperature for 15 minutes. ④ Take six clean 1.5 mL centrifuge tubes, pre-add 500 μL of standard diluent to each tube, and perform serial dilutions of the standard to obtain seven standard concentrations: 50, 25, 12.5, 6.25, 3.12, 1.56, and 0.78 pg / mL. Finally, add the diluted standard to the wells of the pre-coated plate sequentially, adding the standard diluent directly as the 0 pg / mL concentration, for a total of eight standard concentrations. ⑤ Add 300 μL to each well, and repeat the washing process after approximately 15-30 seconds. Wash the plate five times in total, and then pat it dry on paper.

[0060] (2) Operation steps ① Calculate the number of pre-coated strips required for one experiment, and take out the required strips and place them in a 96-well frame.

[0061] ② Add the sample or standard of different concentrations to the corresponding well at a rate of 50 μL / well, seal the reaction well with a sealing film (transparent), and incubate at room temperature for 60 min.

[0062] ③ Wash the board 5 times, and pat it dry on thick absorbent paper on the last wash.

[0063] ④ Add 50 μL of biotinylated antibody (a secondary antibody that specifically binds to the target indicators (ALT, AST, CRP, E2) and is chemically linked to biotin) from the ELISA kit purchased from Shanghai Jianglai Biotechnology Co., Ltd. Seal the reaction wells with a sealing film (clear) and incubate at room temperature for 60 minutes.

[0064] ⑤ Wash the board 5 times, and pat it dry on thick absorbent paper on the last wash.

[0065] ⑥ Add the enzyme conjugate working solution (horseradish peroxidase (HRP) labeled streptavidin (HRP), diluted according to the ELISA kit instructions) from the ELISA kit purchased from Shanghai Jianglai Biotechnology Co., Ltd., at a volume of 100 μL / well. Seal the reaction wells with white sealing film and incubate at room temperature in the dark for 30 minutes.

[0066] ⑦ Wash the board 5 times, and pat it dry on thick absorbent paper on the last wash.

[0067] ⑧ Add 100 μL of TMB colorimetric reagent solution per well, seal the reaction wells with white sealing film, and incubate at room temperature in the dark for 20 minutes.

[0068] ⑨ Add the stop solution from the ELISA kit purchased from Shanghai Jianglai Biotechnology Co., Ltd., at a volume of 50 μL / well. After mixing, immediately measure the expression levels of ALT, AST, CRP, and E2.

[0069] The secretion of blood urea nitrogen (BUN) and creatine kinase isoenzyme MB (CK-MB) in the supernatant was detected using an enzyme-linked immunosorbent assay (ELISA) kit (Shanghai Keabob Biotechnology Co., Ltd., catalog number: BUN: CB10533-Mu, CK-MB: CB12417-Mu). The procedure was performed according to the kit's instructions, as follows: (1) Operation steps ① Remove the required strips from the aluminum foil bag after equilibration at room temperature for 20 minutes. Seal the remaining strips in a self-sealing bag and store them at 4℃.

[0070] ② Set up standard wells and sample wells, and add 50 μL of standard at different concentrations to each standard well.

[0071] ③ Add 50 μL of the sample to be tested to the sample well, and do not add any to the blank well.

[0072] ④ Except for the blank wells, add 100 μL of horseradish peroxidase (HRP) labeled detection antibody to each of the standard and sample wells, seal the reaction wells with sealing film, and incubate at 37°C in a water bath or incubator for 60 min.

[0073] ⑤ Discard the liquid, pat dry on absorbent paper, fill each well with washing solution (350 μL), let stand for 1 min, then discard the washing solution, pat dry on absorbent paper, and repeat this washing process 5 times (or a plate washer can be used).

[0074] ⑥ Add 50 μL each of substrate A and B from the ELISA kit purchased from Shanghai Keabob Biotechnology Co., Ltd. to each well, and incubate at 37°C in the dark for 15 min.

[0075] ⑦ Add 50 μL of stop solution to each well, mix well, and immediately measure the expression levels of BUN and CK-MB.

[0076] The secretion of cardiac troponin I (cTNI) in the supernatant was detected using an enzyme-linked immunosorbent assay (ELISA) kit (Beijing Solarbio Science & Technology Co., Ltd., catalog number: cTNI: SEKM-0153). The procedure was performed according to the kit's instructions, as follows: (1) Reagent preparation ① Remove the kit and test samples from the refrigerator 20-30 minutes before the experiment and allow them to equilibrate to room temperature. If crystals appear in the 20× concentrated wash buffer, place it in a 37 ℃ water bath until completely dissolved. ② Dilute the 20× concentrated wash buffer to 1× wash buffer with double-distilled water or deionized water according to the required volume for the experiment. Store any unused concentrated wash buffer at 4 ℃. Before opening the tube, ensure that all lyophilized standards are at the bottom. Add 0.92 mL of SR1 (the highest concentration in the standard curve) standard / sample diluent to reconstitute the standards (final concentration of 40000 pg / mL). Let it stand at room temperature for 10-30 minutes and then gently mix. Subsequently, use a 2-fold gradient method to prepare standard solutions of 40000, 20000, 10000, 5000, 2500, 1250, and 625 pg / mL, where 40000 pg / mL is the highest concentration of the standard curve. The SR1 standard / sample diluent is used as well 0 (0 pg / mL). Any unused portion of the reconstituted standard stock solution should be discarded, or aliquoted into single-use portions and stored at -20 to -80 °C. ④ Dilute 100× biotinylated antibody concentrate to 1× working solution using the appropriate diluent according to the required experimental volume. Mix thoroughly and use within 30 min. ⑤ Dilute 100× enzyme conjugate concentrate to 1× working solution using the appropriate diluent according to the required experimental volume. Mix thoroughly and use within 30 min. ⑥ Add 300 μL of washing buffer to each well, let stand for 15–30 s, then discard the buffer. Repeat the washing process 5 times, patting dry on absorbent paper after the last wash.

[0077] (2) Operation steps ① Calculate the number of pre-coated strips required for one experiment, take out the required strips and put them into a 96-well plate frame, and seal the remaining strips and put them back into the aluminum foil bag for storage.

[0078] ② Add 300 μL of 1× washing solution to each well, let stand for 30 seconds, then discard the solution and pat dry on absorbent paper. Repeat twice.

[0079] ③ Set up standard wells, 0 wells and sample wells respectively: add 100 μL of standard at different concentrations to the standard wells; add 100 μL of standard / sample diluent to the 0 wells; add 100 μL of the sample to be tested to the sample wells.

[0080] ④ Seal the plate with a new sealing film and incubate at room temperature (25±2 ℃) with shaking for 120 min.

[0081] ⑤ Wash the plate 4 times: Add 300 μL of washing solution to each well, discard the solution, and pat dry on absorbent paper.

[0082] ⑥ Add 100 μL of biotinylated detection antibody working solution to each well, seal the plate with a new sealing film, and incubate at room temperature with shaking for 60 min.

[0083] ⑦ Wash the plate 4 times: Add 300 μL of washing solution to each well, discard the solution, and pat dry on absorbent paper.

[0084] ⑧ Add 100 μL of enzyme conjugate working solution to each well, seal the plate with a new sealing membrane, and incubate at room temperature with shaking for 30 min.

[0085] ⑨ Wash 5 times: Add 300 μL of washing solution to each well, discard the solution and pat dry on absorbent paper.

[0086] ⑩ Add 100 μL of TMB chromogenic substrate to each well and incubate at room temperature in the dark for 5-30 min; then add 50 μL of the stop solution from the ELISA kit purchased from Beijing Solarbio Science & Technology Co., Ltd. to each well to terminate the reaction, mix well and immediately measure the expression level of cTNI.

[0087] The results are as follows Figure 7 As shown, ALT ( Figure 7 (A), AST ( Figure 7 in B), BUN ( Figure 7 C), CRP ( Figure 7 Medium D), CK-MB ( Figure 7 (E), cTNI ( Figure 7 The expression of F in the middle was not affected by compound B. 10 Effects. However, through compound B 10 After treatment, the expression level of estradiol (E2) increased. Figure 7 (G). This result indicates that compound B 10 It may play a role in the treatment of osteoporosis by increasing estrogen expression levels.

[0088] The above are merely preferred embodiments of the present invention, but the scope of protection of the present invention is not limited thereto. Any variations or substitutions that can be easily conceived by those skilled in the art within the scope of the technology disclosed in the present invention should be included within the scope of protection of the present invention.

Claims

1. A class of furanocoumarin compounds, characterized in that, The general structural formulas are shown in Formula I and Formula II: In Equation I, R1 is selected from , , , , , , , , , , , , , , or ; In Equation II, R2 is selected from , , , , , , , , or .

2. The furanocoumarin compound according to claim 1, characterized in that, Selected from A1-A below 16 B1-B 10 Any compound: , , , , , , , , , , , , , , , , , , , , , , , , or .

3. A method for preparing a furanocoumarin compound according to any one of claims 1-2, characterized in that, When the furanocoumarin compound has the structural formula of Formula I, the process includes the following steps: dissolving compound 1 and potassium carbonate in N,N-dimethylformamide, adding a first bromoalkyl compound, and reacting at 80°C for 8 hours to obtain the furanocoumarin compound shown in Formula I, wherein the structural formula of compound 1 is... ; When the furanocoumarin compound has the structural formula of Formula II, the following steps are included: dissolving potassium carbonate and compound 3 in N,N-dimethylformamide, adding a second bromoalkyl compound, and reacting at 80°C for 8 hours to obtain the furanocoumarin compound shown in Formula II, wherein the structural formula of compound 3 is... .

4. The method for preparing furanocoumarin compounds according to claim 3, characterized in that, The equivalent ratio of compound 1 to the first bromoalkyl compound is 1:1.

1.

5. The method for preparing furanocoumarin compounds according to claim 4, characterized in that, The first bromoalkyl compound is selected from bromomethylcyclopentane, methyl bromoacetate, 4-methoxybenzyl bromide, bromopropene, 2-bromomethyl-6-methylpyridine, 4-bromomethyltetrahydropyran, 3-bromomethylpyridine hydrobromide, 3-bromomethylbenzaldehyde, (1-bromomethyl)-3-methylbenzene, 2-bromoethylacetate, ethyl bromoacetate, tert-butyl 4-(bromomethyl)piperidine-1-carboxylate, 2-bromo-N,N-diethylethylamine hydrobromide, ethyl 6-bromohexanoate, (R)-5-bromomethyl-2-pyrrolidone, or ethyl 5-bromopentanoate.

6. The method for preparing furanocoumarin compounds according to claim 1, characterized in that, The equivalent ratio of compound 3 to the second bromoalkyl compound is 1:1.

1.

7. The method for preparing furanocoumarin compounds according to claim 6, characterized in that, The preparation method of compound 3 is as follows: compound 2 is dissolved in anhydrous dichloromethane, boron tribromide is added under nitrogen protection, and compound 3 is obtained after the reaction; the structural formula of compound 2 is as follows. .

8. The method for preparing furanocoumarin compounds according to claim 6, characterized in that, The second bromoalkyl compound is selected from bromomethylcyclopropane, methyl bromoacetate, bromopropene, methyl 4-bromomethylbiphenyl-2-carboxylic acid, methyl 3-bromomethylbenzoate, 3-bromomethylbenzaldehyde, (1-bromomethyl)-3-methylbenzene, 2-bromoethylacetate, 8-bromomethylquinoline, or tert-butyl 4-(bromomethyl)piperidine-1-carboxylate.

9. The use of a furanocoumarin compound as described in any one of claims 1-2 or a pharmaceutically acceptable salt or solvate thereof in the preparation of a medicament for treating osteoporosis.

10. A drug for treating osteoporosis, characterized in that, Includes the furanocoumarin compounds as described in any one of claims 1-2, or their pharmaceutically acceptable salts or solvates.

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