Application of small molecule compound with 3-(methylthio)-2-phenyl-3a, 8a-dihydrofuro [2, 3-b] benzofuran structure in preparation of acute lung injury resisting medicine

Abstract

The invention discloses an application of a small molecule compound with a 3-(methylthio)-2-phenyl-3a, 8a-dihydrofuro [2, 3-b] benzofuran structure in preparation of a medicine for resisting acute lung injury, and the structure of the small molecule compound with the 3-(methylthio)-2-phenyl-3a, 8a-dihydrofuro [2, 3-b] benzofuran structure is shown as a formula (I). In the formula (I), R is H, halogen, C1-C3 alkyl and C1-C3 alkoxy. Cell experiments prove that the compound provided by the invention has a remarkable inhibition effect on expression up-regulation of various inflammatory factors of an LPS-induced macrophage system RAW264.7, and has no obvious cytotoxicity. In conclusion, the small molecule compound with the 3-(methylthio)-2-phenyl-3a, 8a-dihydrofuro [2, 3-b] benzofuran structure has a relieving effect on the acute lung injury, and can be used as a novel candidate therapeutic drug for the acute lung injury.

Description

Technical Field

[0001] This invention belongs to the field of pharmaceutical technology, specifically relating to the application of a small molecule compound having a 3-(methylthio)-2-phenyl-3a,8a-dihydrofurano[2,3-b]benzofuran structure in the preparation of drugs for treating acute lung injury. Background Technology

[0002] Acute lung injury (ALI) is a syndrome caused by the excessive release of inflammatory factors (such as interleukin-1β, IL-6, and tumor necrosis factor-α, TNF-α) due to various reasons, leading to damage to alveolar epithelial cells and capillary endothelial cells, and subsequently causing diffuse interstitial and alveolar edema, resulting in acute hypoxic respiratory failure. When the lungs encounter external stimuli, such as infection, trauma, or exposure to toxic substances, an inflammatory response in the lungs is rapidly triggered. An appropriate inflammatory response is a self-protective mechanism initiated by the body to clear pathogens and damaged cells and promote tissue repair. However, an excessive inflammatory response will disrupt lung homeostasis, damage alveolar epithelial cells and capillary endothelial cells, causing decreased lung compliance and impaired oxygenation, leading to dyspnea. At the same time, the damaged alveolar epithelial cells and vascular endothelial cells will release inflammatory factors, further aggravating the pulmonary inflammatory response, ultimately forming a vicious cycle of inflammation, which can eventually lead to acute respiratory distress syndrome and even death. Therefore, in the treatment of acute lung injury, it is necessary to closely monitor and control the development of the inflammatory response in order to alleviate the patient's symptoms and promote recovery.

[0003] In the clinical treatment of acute lung injury, anti-inflammatory drugs are most commonly used, such as glucocorticoids (e.g., methylprednisolone, prednisolone acetate). While glucocorticoids have strong anti-inflammatory effects, long-term use can lead to side effects such as lower extremity edema, weight gain, elevated blood sugar, and gastrointestinal ulcers. These side effects not only affect patients' quality of life but may also trigger other complications, increasing the difficulty of treatment. Therefore, developing new anti-inflammatory drugs can better meet the treatment needs of patients with acute lung injury. Summary of the Invention

[0004] The purpose of this application is to address the aforementioned shortcomings of the prior art by providing a novel use for preparing an active small molecule compound with the structure 3-(methylthio)-2-phenyl-3a,8a-dihydrofurano[2,3-b]benzofuran in the treatment of acute lung injury, thus providing a new drug candidate for the treatment of acute lung injury. Whole animal experiments have confirmed that compounds with this structure have a significant therapeutic effect on lipopolysaccharide (LPS)-induced acute lung injury.

[0005] To achieve the above objectives, the technical solution adopted by the present invention to solve its technical problem is as follows:

[0006] The application of a small molecule compound having the 3-(methylthio)-2-phenyl-3a,8a-dihydrofurano[2,3-b]benzofuran structure in the preparation of drugs for treating acute lung injury, wherein the structure of the small molecule compound having the 3-(methylthio)-2-phenyl-3a,8a-dihydrofurano[2,3-b]benzofuran structure is as follows:

[0007] ;

[0008] Or, the pharmaceutically acceptable salts or eutectics, deuterated compounds, solvates, or enantiomers corresponding to the above formula;

[0009] Wherein, R is H, halogen, C1~C3 alkyl, or C1~C3 alkoxy.

[0010] Furthermore, the structural formula of the small molecule compound is as follows:

[0011] ;

[0012] Wherein, R is H, halogen, C1~C3 alkyl, or C1~C3 alkoxy.

[0013] Furthermore, the structure of the R substituent is H, Me, Br, or OMe (methoxy).

[0014] More specifically, the small molecule compound is selected from the following structures:

[0015] ;

[0016] Furthermore, the small molecule compound is a compound represented by formula Q3.

[0017] A small molecule that can effectively alleviate lung inflammation caused by various reasons such as infection and trauma, including the above-mentioned compounds and their pharmaceutically acceptable salts or cocrystals, deuterated products, solvates, and enantiomers.

[0018] Furthermore, the drug is an anti-acute lung injury drug that inhibits the expression of inflammatory factors in lung tissue.

[0019] Furthermore, the inflammatory factors include one or more of IL-1α, IL-1β, and TNF-α.

[0020] Furthermore, the acute lung injury is an acute lung injury caused by one or more of bacteria, trauma, or exposure to toxic substances. Even further, the acute lung injury is an acute lung injury caused by bacteria or hydrochloric acid.

[0021] Furthermore, the acute lung injury is lipopolysaccharide-induced acute lung injury.

[0022] Furthermore, the active ingredient in the drug is a small molecule compound having the structure 3-(methylthio)-2-phenyl-3a,8a-dihydrofurano[2,3-b]benzofuran; the drug also includes a pharmaceutically acceptable carrier or excipient. Even further, the dosage form of the drug is a tablet, injection, capsule, oral liquid, or drop pill.

[0023] Furthermore, the drug is one that can be administered via nebulization.

[0024] The beneficial effects of this invention are as follows:

[0025] The active small molecule compounds in this invention can effectively alleviate lung inflammation caused by various reasons such as infection and trauma.

[0026] The four compounds provided by this invention were demonstrated in cell experiments to significantly inhibit the upregulation of various inflammatory factors in the LPS-induced macrophage line RAW264.7, without significant cytotoxicity. Furthermore, compound Q3, selected through cell biology, demonstrated good anti-inflammatory and therapeutic effects on an animal model of acute lung injury induced by LPS intrapulmonary nebulization. In summary, these small molecule compounds with the structure 3-(methylthio)-2-phenyl-3a,8a-dihydrofurano[2,3-b]benzofuran exhibit alleviating effects on acute lung injury and can serve as novel candidate drugs for the treatment of acute lung injury.

[0027] It should be understood that the above general description and the following detailed description are exemplary and explanatory only, and do not limit this application. Attached Figure Description

[0028] Figure 1 The 1H NMR spectrum of 3-(methylthio)-2-phenyl-3a,8a-dihydrofuran[2,3-b]benzofuran given in Example Q1 is shown.

[0029] Figure 2 The 1H NMR spectrum of compound Q2 in Example 1 is shown.

[0030] Figure 3 The 1H NMR spectrum of 5-bromo-3-(methylthio)-2-phenyl-3a,8a-dihydrofuran[2,3-b]benzofuran is given for compound Q3 in Example.

[0031] Figure 4The 1H NMR spectrum of compound Q4 in Example 1 is shown.

[0032] Figure 5 The image shows the carbon NMR spectrum of 3-(methylthio)-2-phenyl-3a,8a-dihydrofuran[2,3-b]benzofuran given in Example Q1.

[0033] Figure 6 The carbon NMR spectrum of 5-methyl-3-(methylthio)-2-phenyl-3a,8a-dihydrofuran[2,3-b]benzofuran is given for compound Q2 in Example.

[0034] Figure 7 The carbon NMR spectrum of 5-bromo-3-(methylthio)-2-phenyl-3a,8a-dihydrofuran[2,3-b]benzofuran is given for compound Q3 in Example.

[0035] Figure 8 The carbon NMR spectrum of 5-methoxy-3-(methylthio)-2-phenyl-3a,8a-dihydrofuran[2,3-b]benzofuran is given for compound Q4 in Example.

[0036] Figure 9 The effect of compounds Q1-Q4 on LPS-induced expression of inflammatory factors in RAW264.7 (##P<0.01 vs Ctrl group; *P<0.05; **P<0.01 vs LPS group, N=3).

[0037] Figure 10 The effect of the preferred compound Q3 on LPS-induced acute lung injury in mice (##P<0.01 vs Ctrl group; *P<0.05; **P<0.01 vs LPS group, N=6). Detailed Implementation

[0038] The present invention will be further described below with reference to embodiments and accompanying drawings. The following embodiments are merely illustrative of the present invention and are not intended to limit the present invention in any way.

[0039] Synthesis of compounds Q1-Q4: Direct Access to 3-Thioether-SubstitutedDihydrofuro[2,3-b]benzofurans via Tandem Reactions of Sulfur Ylides and 2-Nitrobenzofurans. J Org Chem. 2023; 88(15): 10810-10817.

[0040] Example 1 3-(methylthio)-2-phenyl-3a,8a-dihydrofuran[2,3-b]benzofuran Q1

[0041]

[0042] The hydrogen NMR spectrum and carbon NMR spectrum are shown below. Figure 1 and Figure 5 The corresponding NMR and mass spectrometry data are as follows:

[0043] 1 H NMR (400 MHz, CDCl3) δ 7.99 - 7.96 (m, 2H), 7.54 (d, J = 7.2 Hz,1H), 7.40 - 7.33 (m, 3H), 7.20 (td, J = 8.0, 0.8 Hz, 1H), 6.96 (td, J = 7.6,0.8 Hz, 1H), 6.92 (d, J = 8.0 Hz, 1H), 6.76 (d, J = 7.2 Hz, 1H), 4.75 (d, J =7.2 Hz, 1H), 2.27 (s, 3H). 13 C NMR (100 MHz, CDCl3) δ 157.9, 153.7, 129.6,129.5, 129.0, 128.2, 127.8, 126.6, 124.8, 121.6, 109.9, 109.2, 105.7, 54.2,17.3; ESI-MS: m / z = 283.0790 [M+H] + .

[0044] Example 2 5-Methyl-3-(methylthio)-2-phenyl-3a,8a-dihydrofuran[2,3-b]benzofuran Q2

[0045]

[0046] The hydrogen NMR spectrum and carbon NMR spectrum are shown below. Figure 2 and Figure 6 The corresponding NMR and mass spectrometry data are as follows: 1HNMR (400 MHz, CDCl3) δ 8.08 - 7.81 (m, 2H), 7.42 - 7.31 (m, 3H), 7.13 (d, J =2.4 Hz, 1H), 6.83 (d, J = 8.8 Hz, 1H), 6.74 (d, J = 7.6 Hz, 1H), 4.72 (d, J =7.2 Hz, 2H), 3.80 (s, 3H), 2.27 (s, 3H). 13 C NMR (100 MHz, CDCl3) δ 154.9,153.9, 151.9, 129.6, 129.5, 128.2, 127.8, 127.6, 113.7, 111.1, 109.8, 109.5,105.3, 56.2, 54.5, 29.9, 17.3; ESI-MS: m / z = 297.0940 [M+H] + .

[0047] Example 3 5-Bromo-3-(methylthio)-2-phenyl-3a,8a-dihydrofurano[2,3-b]benzofuran Q3

[0048]

[0049] The hydrogen NMR spectrum and carbon NMR spectrum are shown below. Figure 3 and Figure 7 The corresponding NMR and mass spectrometry data are as follows:

[0050] 1 H NMR (400 MHz, CDCl3) δ 7.99 - 7.95 (m, 2H), 7.63 (d, J = 2.0 Hz,1H), 7.42 - 7.34 (m, 3H), 7.30 (dd, J = 8.4, 2.0 Hz, 1H), 6.79 (d, J = 8.4Hz, 1H), 6.75 (d, J = 7.2 Hz, 1H), 4.73 (d, J = 7.2 Hz, 1H), 2.27 (s, 3H). 13CNMR (100 MHz, CDCl3) δ 157.1, 154.1, 131.8, 129.7, 129.4, 129.1, 128.3,127.8, 127.7, 113.5, 111.5, 109.5, 105.1, 54.1, 17.3; ESI-MS: m / z = 382.9716[M+Na] + .

[0051] Example 4 5-Methoxy-3-(methylthio)-2-phenyl-3a,8a-dihydrofuran[2,3-b]benzofuran Q4

[0052]

[0053] The hydrogen NMR spectrum and carbon NMR spectrum are shown below. Figure 4 and Figure 8 The corresponding NMR and mass spectrometry data are as follows:

[0054] 1 H NMR (400 MHz, CDCl3) δ 8.08 - 7.81 (m, 2H), 7.42 - 7.31 (m, 3H), 7.13 (d, J = 2.4 Hz, 1H), 6.83 (d, J = 8.8 Hz, 1H), 6.74 (d, J = 7.6 Hz, 1H), 4.72 (d, J = 7.2 Hz, 2H), 3.80 (s, 3H), 2.27 (s, 3H). 13 C NMR (100 MHz, CDCl3)δ 154.9, 153.9, 151.9, 129.6, 129.5, 128.2, 127.8, 127.6, 113.7, 111.1,109.8, 109.5, 105.3, 56.2, 54.5, 29.9, 17.3; ESI-MS: m / z = 313.0893 [M+H] + .

[0055] Example 5: Cytotoxic effects of the compound on the macrophage cell line RAW264.7

[0056] The macrophage cell line RAW264.7 was adjusted to 2×10 3Cells were seeded into 96-well plates at the specified density, with 9 groups per plate: one control group (containing only the drug solvent dimethyl sulfoxide) and eight drug-treated groups (containing 1, 5, 10, 50, 100, 500, 1000, and 5000 nM of compounds from the Q1-Q4 series, respectively). Each group had 5 replicates. The culture medium was complete (DMEM + 10% FBS + 1% antibiotics), and the cells were cultured for 24, 48, and 72 hours, respectively. At the corresponding time points, 10 μl of CCK-8 reagent was added to each well, and the cells were incubated in the dark for 1 hour. After incubation, the cells were shaken thoroughly on a microplate reader, and the absorbance at 450 nm was measured (the higher the number of viable cells, the higher the absorbance). The IC50 values ​​of each compound for cytotoxicity against RAW264.7 cells are shown in Table 1.

[0057] Table 1. Cytotoxic effects of the compounds on the RAW264.7 macrophage cell line.

[0058]

[0059] The test results showed that the cytotoxicity of compounds Q1-Q4 against RAW264.7 macrophages was all above 2000 nM, exhibiting good low toxicity characteristics. We will further explore the anti-inflammatory effects of compounds Q1-Q4 at non-cytotoxic doses.

[0060] Example 6: Effects of the compound on LPS-induced expression of inflammatory factors in RAW264.7

[0061] The compound from Example 5, which showed no cytotoxicity to the RAW264.7 cell line, was selected at a dose of 500 nM. The inhibitory effect of this dose on LPS-induced inflammatory cytokine expression was then examined. RAW264.7 cells were seeded in 12-well plates and grouped as follows: PBS + solvent, LPS + solvent, LPS + Q1 500 nM, LPS + Q2 500 nM, LPS + Q3 500 nM, and LPS + Q4 500 nM. Each group had three replicates. When the cells reached 70% confluence, they were starved for 8 hours in serum-free DMEM medium. Then, 500 nM of the solvent or compounds Q1-Q4 were added for 1 hour each, followed by incubation in PBS or 100 ng / ml LPS (Sigma L3129) for 12 hours. Cells were then collected for RNA extraction.

[0062] Commercially available kits (Novizan RC201, R302, and Q312) were used for RNA extraction, reverse transcription, and real-time quantitative PCR. Instructions for use were provided in the respective product manuals. Primers for IL-1α, IL-1β, IL-6, and TNF-α used in the quantitative PCR are shown in Table 2.

[0063] Table 2 Primer sequences for quantitative PCR of IL-1α, IL-1β, IL-6 and TNF-α

[0064]

[0065] The results showed that compounds Q1-Q4, possessing the structure 3-(methylthio)-2-phenyl-3a,8a-dihydrofurano[2,3-b]benzofuran, effectively inhibited the LPS-induced increase in the expression levels of IL-1α, IL-1β, and TNF-α in RAW264.7 cells, with Q3 showing the most significant inhibitory effect at the same dose. Figure 9 ).

[0066] Example 7: The inhibitory effect of the preferred compound Q3 on LPS-induced acute lung injury in mice.

[0067] Establishment of an acute lung injury model: C57 / BL-6 mice were divided into a control group (N=6) and an LPS-induced group (N=18). Mice were anesthetized with isoflurane inhalation and fixed in a supine position on a mouse restraint board. The pharynx was irradiated with a fiber optic cable, and the tongue was gently pulled outwards with forceps; tracheal contractions were observed through the mouth. At a dose of 10 μl / 10g of mouse body weight, the appropriate volume of LPS solution (final LPS concentration 4 mg / ml; the control group received an equal volume of physiological saline) was drawn using a quantitative nebulizer. The nebulizer was slowly inserted into the trachea to nebulize the corresponding volume of LPS solution. After instilling the LPS solution (or physiological saline), the mice were held upright for approximately 5 minutes, then laid supine in their cages until they awoke.

[0068] Intratracheal nebulization administration: Mice in the LPS-induced group were randomly divided into a model group, a compound Q3 0.3 mg / ml group, and a compound Q3 0.9 mg / ml group, with 6 mice in each group. The preferred candidate compound Q3 was administered via intratracheal nebulization at 2 and 6 hours after LPS instillation, at doses of 0.3 and 0.9 mg / ml, 10 μl / 10g mouse body weight. The control group and the model group received only the same volume of solvent.

[0069] Mouse tissue collection: Mice were sacrificed 24 hours after LPS administration. 1 ml of 4% paraformaldehyde was injected intratracheally, and the entire lung was immersed in the 4% paraformaldehyde solution. One week later, the lungs were embedded in paraffin, sectioned, and stained with hematoxylin and eosin (HE) to observe inflammatory cell infiltration. The standard Smith scoring method was used to semi-quantitatively score pulmonary edema, alveolar and interstitial inflammation, alveolar and interstitial hemorrhage, atelectasis, and hyaline membrane formation.

[0070] Results: The test results showed that LPS could induce a large number of inflammatory cells to enter the lung tissue, leading to alveolar edema, rupture, and other damage. In contrast, intratracheal nebulization of compound Q3 at concentrations of 0.3 and 0.9 mg / ml dose-dependently inhibited LPS-induced infiltration of inflammatory cells in the lungs and alveolar damage. Figure 10 (A) Semi-quantitative scoring of the degree of alveolar damage, such as... Figure 10 As shown in B.

[0071] The above descriptions are merely some preferred embodiments of the present invention, used to help illustrate the invention. The scope of protection of the present invention is not limited to the above embodiments; all technical solutions falling within the scope of the present invention's concept are within its protection.

Claims

1. The application of a small molecule compound having a 3-(methylthio)-2-phenyl-3a,8a-dihydrofurano[2,3-b]benzofuran structure in the preparation of drugs for treating acute lung injury, characterized in that, The structure of the small molecule compound having the 3-(methylthio)-2-phenyl-3a,8a-dihydrofurano[2,3-b]benzofuran structure is shown in formula (I): （I）； Or, the pharmaceutically acceptable salts or eutectics, deuterated compounds, solvates, or enantiomers corresponding to the above formula; Wherein, R is H, halogen, C1~C3 alkyl, or C1~C3 alkoxy.

2. The application of the small molecule compound having the 3-(methylthio)-2-phenyl-3a,8a-dihydrofurano[2,3-b]benzofuran structure according to claim 1 in the preparation of drugs for treating acute lung injury, characterized in that, The structure of the R substituent is H, Me, Br or -OMe.

3. The application of the small molecule compound having the 3-(methylthio)-2-phenyl-3a,8a-dihydrofurano[2,3-b]benzofuran structure according to claim 1 in the preparation of drugs for treating acute lung injury, characterized in that, The structure of the small molecule compound having the 3-(methylthio)-2-phenyl-3a,8a-dihydrofurano[2,3-b]benzofuran structure is as follows: 。 4. The application of the small molecule compound having the 3-(methylthio)-2-phenyl-3a,8a-dihydrofurano[2,3-b]benzofuran structure according to claim 1 in the preparation of drugs for treating acute lung injury, characterized in that, The structure of the small molecule compound having the 3-(methylthio)-2-phenyl-3a,8a-dihydrofurano[2,3-b]benzofuran structure is as follows: , , or .

5. The application of the small molecule compound having the 3-(methylthio)-2-phenyl-3a,8a-dihydrofurano[2,3-b]benzofuran structure according to claim 1 in the preparation of drugs for treating acute lung injury, characterized in that, The drug is an anti-acute lung injury drug that inhibits the expression of inflammatory factors in lung tissue.

6. The application of the small molecule compound having the 3-(methylthio)-2-phenyl-3a,8a-dihydrofurano[2,3-b]benzofuran structure according to claim 5 in the preparation of drugs for treating acute lung injury, characterized in that, The inflammatory factors include one or more of IL-1α, IL-1β, and TNF-α.

7. The application of the small molecule compound having the 3-(methylthio)-2-phenyl-3a,8a-dihydrofurano[2,3-b]benzofuran structure according to claim 1 in the preparation of drugs for treating acute lung injury, characterized in that, The acute lung injury mentioned above is an acute lung injury caused by one or more of the following: bacteria, trauma, or exposure to toxic substances.

8. The application of the small molecule compound having the 3-(methylthio)-2-phenyl-3a,8a-dihydrofurano[2,3-b]benzofuran structure according to claim 1 in the preparation of drugs for treating acute lung injury, characterized in that, The acute lung injury mentioned is lipopolysaccharide-induced acute lung injury.

9. The application of the small molecule compound having the 3-(methylthio)-2-phenyl-3a,8a-dihydrofurano[2,3-b]benzofuran structure according to claim 1 in the preparation of drugs for treating acute lung injury, characterized in that, The active ingredient in the drug is a small molecule compound having the structure 3-(methylthio)-2-phenyl-3a,8a-dihydrofurano[2,3-b]benzofuran; the drug also includes a pharmaceutically acceptable carrier or adjuvant.

10. The application of the small molecule compound having the 3-(methylthio)-2-phenyl-3a,8a-dihydrofurano[2,3-b]benzofuran structure according to claim 1 in the preparation of drugs for treating acute lung injury, characterized in that, The drug is one that can be administered via nebulization.

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