Hydroxycinnamic acid derivatives, process for their preparation and use

Abstract

The application belongs to the technical field of new application of medicines, and particularly relates to hydroxycinnamic acid derivatives, a preparation method and application thereof. The ARDS model is induced by LPS tracheal instillation combined with hydroxycinnamic acid derivatives, the alveolar lavage fluid cell count is determined, and the pathological characteristics and the expression level of inflammatory genes of lung tissues are detected; the results show that the hydroxycinnamic acid derivatives can significantly reduce lung cell infiltration, alleviate lung tissue damage and reduce the expression level of inflammatory factors.

Description

[0001] This invention application is a follow-up application to the invention patent filed on March 8, 2024, with application number 2024102721361 and patent title "The use of KS-3 butyrate or its pharmaceutically acceptable salt in the preparation of a medicament for treating acute respiratory distress syndrome". Technical Field

[0002] This invention belongs to the field of pharmaceutical technology, specifically relating to hydroxycinnamic acid derivatives, their preparation methods, and applications. Background Technology

[0003] Acute respiratory distress syndrome (ARDS) is a common respiratory critical illness, clinically characterized by bilateral opaque chest X-rays accompanied by severe hypoxemia caused by non-cardiac pulmonary edema. The etiology of ARDS is complex, with pneumonia being the most common risk factor, such as pneumonia caused by infection with SARS-CoV, H5N1 avian influenza virus, H7N9 avian influenza virus, or SARS-CoV-2.

[0004] Current treatments for ARDS primarily rely on mechanical ventilation (such as low tidal volume mechanical ventilation). Other potential treatments are controversial or have uncertain efficacy. For example, the efficacy of hormone therapy is uncertain and controversial; and tyrosine kinase inhibitors and kinins that specifically target the extracellular matrix and its receptors have no clear efficacy.

[0005] Therefore, developing a drug that can treat ARDS is of great significance. Summary of the Invention

[0006] To address the above problems, the present invention aims to provide a hydroxycinnamic acid derivative that can be used in drugs to alleviate acute respiratory distress syndrome (ARDS). This derivative can effectively reduce lung cell infiltration, reduce lung tissue damage, and lower the expression level of inflammatory factors, thereby alleviating ARDS.

[0007] To achieve the above objectives, the present invention can adopt the following technical solutions:

[0008] In one aspect, this invention provides a hydroxycinnamic acid derivative, the structural formula of which is shown below.

[0009]

[0010] R3 is -COOR4;

[0011] R2 is selected from -H or alkoxy;

[0012] R1 is selected from the first alkyl group, Or -C = CCOOR6;

[0013] Wherein, n is an integer, 1≤n≤5; and / or R4, R5 and R6 are each independently selected from the second alkyl group, the number of carbon chains of the second alkyl group is ≤5; and / or the number of carbon chains of the first alkyl group is ≤15; and / or the number of carbon chains in the alkoxy group is ≤5.

[0014] Preferably, the above-mentioned hydroxycinnamic acid derivative is selected from any one of the following compounds:

[0015]

[0016]

[0017] Another aspect of the present invention provides a method for preparing the hydroxycinnamic acid derivative of the present invention, wherein the method is selected from one of the following methods:

[0018] (a) In the hydroxycinnamic acid derivative, R2 is selected from -H, R1 is selected from the first alkyl group, and the number of carbon chains of the first alkyl group is ≤5; or R2 is selected from alkoxy, R1 is selected from the first alkyl group, and the number of carbon chains of the first alkyl group is ≤3; the preparation method of the hydroxycinnamic acid derivative includes: mixing and reacting the compound shown in Formula I, the compound shown in Formula II, the compound shown in Formula III and TsCl to obtain the hydroxycinnamic acid derivative.

[0019] (b) In the hydroxycinnamic acid derivative, R2 is selected from -H, R1 is selected from the first alkyl group, and the first alkyl group has ≥6 carbon chains; or R2 is selected from the alkoxy group, R1 is selected from the first alkyl group, and the first alkyl group has ≥4 carbon chains; the preparation method of the hydroxycinnamic acid derivative includes: mixing the compound shown in Formula I and the compound shown in Formula II, then adding an organic base catalyst, and then adding a first condensing agent to react and obtain the hydroxycinnamic acid derivative;

[0020] (c) In hydroxycinnamic acid derivatives, R2 is selected from -H, and R1 is selected from... Or -C=CCOOR6; The preparation method of hydroxycinnamic acid derivatives includes: mixing and reacting the compound shown in Formula I, the compound shown in Formula II, a hydroxyl activator, a catalyst, and a second condensing agent to obtain hydroxycinnamic acid derivatives;

[0021] The structural formulas of compounds I, II, and III are shown below:

[0022]

[0023] In another aspect, the present invention provides a pharmaceutical composition for treating acute respiratory distress syndrome, comprising the hydroxycinnamic acid derivative of the present invention.

[0024] In another aspect, the present invention provides a pharmaceutical preparation for treating acute respiratory distress syndrome, comprising the hydroxycinnamic acid derivative of the present invention or the pharmaceutical composition of the present invention.

[0025] In another aspect, this invention provides the use of the hydroxycinnamic acid derivative of this invention, or the pharmaceutical composition of this invention, or methyl hydroxycinnamate in the preparation of a pharmaceutical formulation for treating acute respiratory distress syndrome, wherein the structural formula of methyl hydroxycinnamate is:

[0026] Preferably, the above applications include one or more combinations of the following applications:

[0027] (i) Use of hydroxycinnamic acid derivatives or pharmaceutical compositions or methyl hydroxycinnamic acid in the preparation of pharmaceutical formulations that reduce lung cell infiltration;

[0028] (ii) Use of hydroxycinnamic acid derivatives or pharmaceutical compositions in the preparation of pharmaceutical formulations that alleviate pathological damage to lung tissues;

[0029] (iii) Use of hydroxycinnamic acid derivatives or pharmaceutical compositions or methyl hydroxycinnamic acid in the preparation of pharmaceutical formulations for reducing lung inflammation.

[0030] Preferably, the use of the above-mentioned hydroxycinnamic acid derivative or methyl hydroxycinnamate in the preparation of a drug for reducing lung inflammation includes: the use of the hydroxycinnamic acid derivative or methyl hydroxycinnamate in the preparation of a drug formulation that reduces the mRNA expression levels of lung inflammatory factors IL-6 and / or HO-1 and / or MPO and / or TNF-α and / or IL-1β.

[0031] Preferably, the above applications include one or more combinations of the following applications:

[0032] (a) Use of the first compound in the preparation of a pharmaceutical formulation that reduces lung cell infiltration, wherein the first compound is selected from any of the following compounds:

[0033]

[0034] (b) The use of the second compound in the preparation of a pharmaceutical formulation that alleviates pathological damage to lung tissue, wherein the first compound is selected from the following compounds:

[0035]

[0036] (c) Use of the third compound in the preparation of a pharmaceutical formulation that reduces the mRNA expression level of the pulmonary inflammatory factor IL-6, wherein the third compound is selected from any one of the following compounds:

[0037]

[0038] (d) The use of the fourth compound in the preparation of a pharmaceutical formulation that reduces the mRNA expression level of the pulmonary inflammatory factor HO-1, wherein the fourth compound is selected from the following compounds:

[0039]

[0040] (e) The use of the fifth compound in the preparation of pharmaceutical formulations that reduce the mRNA expression level of the pulmonary inflammatory factor MPO, wherein the fifth compound is selected from the following compounds:

[0041]

[0042] (f) Use of the sixth compound in the preparation of a pharmaceutical formulation that reduces the mRNA expression level of the pulmonary inflammatory factor TNF-α, wherein the sixth compound is selected from any one of the following compounds:

[0043]

[0044] (g) Use of the seventh compound in the preparation of a pharmaceutical formulation that reduces the mRNA expression level of the pulmonary inflammatory factor IL-1β, wherein the seventh compound is selected from any of the following compounds:

[0045]

[0046]

[0047] Preferably, in the above applications, the dosage form of the drug may include oral dosage form, nebulizer, powder, tablet, ointment, or injection.

[0048] The beneficial effects of this invention include at least the following: the invention uses an LPS tracheal infusion-induced ARDS model combined with treatment with hydroxycinnamic acid derivatives to measure the cell count, histopathology, and inflammatory gene expression levels in bronchoalveolar lavage fluid; the results show that hydroxycinnamic acid derivatives can significantly reduce lung cell infiltration, alleviate lung tissue damage, and reduce the expression levels of inflammatory factors. Attached Figure Description

[0049] Figure 1 Effects of small molecule drugs on lung cell infiltration in LPS-induced ARDS mice (data are expressed as mean ± standard deviation, N = 6, *P < 0.05; **P < 0.01; ***P < 0.001);

[0050] Figure 2 Effects of six administrations of hydroxycinnamic acid and its derivatives on lung cell infiltration in LPS-induced ARDS mice (data are expressed as mean ± standard deviation, N = 6, *P < 0.05; **P < 0.01; ***P < 0.001);

[0051] Figure 3 Effects of four administrations of hydroxycinnamic acid and its derivatives on lung cell infiltration in LPS-induced ARDS mice (data are expressed as mean ± standard deviation, N = 6, *P < 0.05; **P < 0.01; ***P < 0.001);

[0052] Figure 4 Effects of four administrations of hydroxycinnamic acid and its derivatives on LPS-induced ARDS lung tissue damage in mice (top figure, 100×, scale bar = 100 μm; bottom figure, 400×, scale bar = 100 μm);

[0053] Figure 5 The effect of hydroxycinnamic acid derivatives on the mRNA level of IL-6, an inflammatory factor in the lung tissue of LPS-induced ARDS mice (data are expressed as mean ± standard deviation, N = 4, *P < 0.05; **P < 0.01; ***P < 0.001);

[0054] Figure 6 The effect of hydroxycinnamic acid derivatives on the mRNA level of TNF-α, an inflammatory factor in the lung tissue of LPS-induced ARDS mice (data are expressed as mean ± standard deviation, N = 4, *P < 0.05; **P < 0.01; ***P < 0.001);

[0055] Figure 7 The effect of hydroxycinnamic acid derivatives on the mRNA level of the inflammatory factor IL-1β in the lung tissue of LPS-induced ARDS mice (data are expressed as mean ± standard deviation, N = 4, *P < 0.05; **P < 0.01; ***P < 0.001);

[0056] Figure 8 The effect of hydroxycinnamic acid derivatives on the mRNA level of HO-1, an inflammatory factor in the lung tissue of LPS-induced ARDS mice (data are expressed as mean ± standard deviation, N = 4, *P < 0.05; **P < 0.01; ***P < 0.001);

[0057] Figure 9 The effect of hydroxycinnamic acid derivatives on the mRNA level of MPO, an inflammatory factor in the lung tissue of LPS-induced ARDS mice (data are expressed as mean ± standard deviation, N = 4, *P < 0.05; **P < 0.01; ***P < 0.001);

[0058] Figure 10 Effects of four administrations of hydroxycinnamic acid and its derivatives on lung cell infiltration in LPS-induced ARDS mice (data are expressed as mean ± standard deviation, N = 6, *P < 0.05; **P < 0.01; ***P < 0.001);

[0059] Figure 11 The effect of hydroxycinnamic acid derivatives on the mRNA level of IL-6, an inflammatory factor in the lung tissue of LPS-induced ARDS mice (data are expressed as mean ± standard deviation, N = 4, *P < 0.05; **P < 0.01; ***P < 0.001);

[0060] Figure 12 The effect of hydroxycinnamic acid derivatives on the mRNA level of TNF-α, an inflammatory factor in the lung tissue of LPS-induced ARDS mice (data are expressed as mean ± standard deviation, N = 4, *P < 0.05; **P < 0.01; ***P < 0.001);

[0061] Figure 13 The effect of hydroxycinnamic acid derivatives on the mRNA level of the inflammatory factor IL-1β in the lung tissue of LPS-induced ARDS mice (data are expressed as mean ± standard deviation, N = 4, *P < 0.05; **P < 0.01; ***P < 0.001);

[0062] in, Figure 1 , Figure 2 Figure 3 The units for the values ​​in the medium bar chart are 10. 5 / mL, for example, if the bar chart value is marked as 34.26, it means 34.26 * 10 5 / mL; Figure 5 , Figure 6 , Figure 7 , Figure 9 , Figure 10 , Figure 11 and Figure 12 In the diagram, the data has been normalized, and the vertical axis shows the multiple relationships. Detailed Implementation

[0063] The embodiments described are provided to better illustrate the present invention, but are not intended to limit the scope of the invention to the embodiments described. Therefore, non-essential improvements and adjustments made to the embodiments by those skilled in the art based on the above description are still within the scope of protection of the present invention.

[0064] The terminology used herein is for the purpose of describing particular embodiments only and is not intended to limit this disclosure. Singular expressions include plural expressions unless they have a distinct meaning in the context. As used herein, it should be understood that terms such as “comprising,” “having,” “including,” are intended to indicate the presence of a feature, number, operation, material, or combination thereof. The terminology of the invention is disclosed in the specification and is not intended to exclude the possibility that one or more other features, numbers, operations, materials, or combinations thereof may be present or added. As used herein, “ / ” may be interpreted as “and” or “or,” depending on the context.

[0065] In this invention, the terms "first" and "second" have no special meaning and refer only to substances that have the same effect in different environments.

[0066] This invention provides a hydroxycinnamic acid derivative, the structural formula of which is shown below.

[0067]

[0068] R3 is -COOR4;

[0069] R2 is selected from -H or alkoxy;

[0070] R1 is selected from the first alkyl group, Or -C = CCOOR6;

[0071] Wherein, n is an integer, 1≤n≤5; and / or R4, R5 and R6 are each independently selected from the second alkyl group, the number of carbon chains of the second alkyl group is ≤5; and / or the number of carbon chains of the first alkyl group is ≤15; and / or the number of carbon chains in the alkoxy group is ≤5.

[0072] It should be understood that R1COO- can be located at the ortho, para, and meta positions of R2, and R2 can also be located at the ortho, para, and meta positions of R2; furthermore, R1COO- and R2 are located at different sites. Additionally, R4, R5, and R6 being independently selected from the second alkyl group means that R4, R5, and R6 can be alkyl groups with the same or different carbon chain numbers.

[0073] It should be noted that hydroxycinnamic acid is a plant phenolic acid widely found in grains, fruits, and vegetables. It has been extensively studied for its anti-inflammatory, antioxidant, antibacterial, antitumor, and antiplatelet aggregation effects. However, because the main functional group responsible for its antioxidant activity is the phenolic hydroxyl group, it suffers from low bioavailability, low stability, and rapid metabolism, leading to pharmacokinetic problems. Furthermore, the dissociation effect of the carboxylic acid group makes it difficult to penetrate cell membranes, further limiting its biological activity. Therefore, structural modification of hydroxycinnamic acid to increase its hydrophobicity, enabling it to penetrate cell membranes, prolonging its duration of action, improving bioavailability, and enhancing its biological activity is of great significance. Acylation and esterification of the phenolic hydroxyl and carboxylic acid groups in the hydroxycinnamic acid molecule are effective methods for structural modification. Therefore, the hydroxycinnamic acid derivative in this invention also improves the bioavailability of hydroxycinnamic acid.

[0074] In some specific examples, the above-mentioned hydroxycinnamic acid derivatives are selected from any of the following compounds:

[0075]

[0076]

[0077] This invention also provides a method for preparing the hydroxycinnamic acid derivative of this invention, which is selected from one of the following methods:

[0078] (a) In the hydroxycinnamic acid derivative, R2 is selected from -H, R1 is selected from the first alkyl group, and the number of carbon chains of the first alkyl group is ≤5; or R2 is selected from alkoxy, R1 is selected from the first alkyl group, and the number of carbon chains of the first alkyl group is ≤3; the preparation method of the hydroxycinnamic acid derivative includes: mixing and reacting the compound shown in Formula I, the compound shown in Formula II, the compound shown in Formula III and TsCl to obtain the hydroxycinnamic acid derivative; specifically, it should be understood that the above preparation method is applicable to the hydroxycinnamic acid derivatives in the above two cases, namely: (i) hydroxycinnamic acid derivatives in which R2 is selected from -H, R1 is selected from the first alkyl group, and the number of carbon chains of the first alkyl group is ≤5, such as compounds A2 and A3; (i) hydroxycinnamic acid derivatives in which R2 is selected from alkoxy, R1 is selected from the first alkyl group, and the number of carbon chains of the first alkyl group is ≤3, such as compound B1;

[0079] (b) In the hydroxycinnamic acid derivative, R2 is selected from -H, R1 is selected from a first alkyl group, and the first alkyl group has a carbon chain number ≥6; or R2 is selected from an alkoxy group, R1 is selected from a first alkyl group, and the first alkyl group has a carbon chain number ≥4; the preparation method of the hydroxycinnamic acid derivative includes: mixing the compound shown in Formula I and the compound shown in Formula II, then adding an organic base catalyst, and then adding a first condensing agent to react and obtain the hydroxycinnamic acid derivative; specifically, it should be understood that the above preparation method is applicable to the hydroxycinnamic acid derivatives in the above two cases, namely: (i) hydroxycinnamic acid derivatives in which R2 is selected from -H, R1 is selected from a first alkyl group, and the first alkyl group has a carbon chain number ≥6, such as compounds A4, A5 and A6; (ii) hydroxycinnamic acid derivatives in which R2 is selected from an alkoxy group, R1 is selected from a first alkyl group, and the first alkyl group has a carbon chain number ≥4, such as compound B2; in addition, the above organic base catalyst and the first condensing agent are known in the art, such as pyridine can be selected as the organic base catalyst. The condensing agent can be DCC (dicyclohexylcarbodiimide);

[0080] (c) In hydroxycinnamic acid derivatives, R2 is selected from -H, and R1 is selected from... Or -C=CCOOR6; The preparation method of hydroxycinnamic acid derivatives includes: mixing and reacting the compound shown in Formula I, the compound shown in Formula II, a hydroxyl activator, a catalyst, and a second condensing agent to obtain hydroxycinnamic acid derivatives; Specifically, it should be understood that the above preparation method is applicable to hydroxycinnamic acid derivatives under the above two conditions, namely: (i) R2 is selected from -H, R1 is selected from (ii) hydroxycinnamic acid derivatives, such as compound A8; (ii) R2 is selected from -H, and R1 is selected from -C=CCOOR6, such as compound A7; in addition, the above-mentioned hydroxyl activator, catalyst and second condensing agent are all known in the art, such as DIEPA (N,N-diisopropylethylamine) as the hydroxyl activator, DMAP (4-dimethylaminopyridine) as the catalyst, and EDCI (1-ethyl-(3-dimethylaminopropyl)carbodiimide hydrochloride) as the second condensing agent;

[0081] The structural formulas of compounds I, II, and III are shown below:

[0082]

[0083] In addition, it should be understood that in the above-mentioned preparation method of hydroxycinnamic acid derivatives, the corresponding R1, R2 and R3 in the raw materials are the same as the R1, R2 and R3 in the corresponding hydroxycinnamic acid derivatives.

[0084] This invention also provides a pharmaceutical composition for treating acute respiratory distress syndrome, comprising the hydroxycinnamic acid derivative of this invention.

[0085] It should be noted that the hydroxycinnamic acid derivatives in this invention can be combined with other compounds used to treat acute respiratory distress syndrome to prepare a pharmaceutical composition to increase efficacy. The compounds used to treat acute respiratory distress syndrome are drugs known in the art, such as β2 receptor agonists, phosphodiesterase 4 inhibitors, or nikethamide.

[0086] This invention also provides a pharmaceutical preparation for treating acute respiratory distress syndrome, comprising the hydroxycinnamic acid derivative or the pharmaceutical composition of this invention.

[0087] It should be noted that the hydroxycinnamic acid derivatives of this invention or the above-mentioned compositions can also be prepared into pharmaceutical formulations by adding excipients; the excipients can be selected according to the dosage form of the pharmaceutical formulation; the dosage form of the drug can include oral dosage, nebulized dosage, powder, tablet, ointment or injection.

[0088] This invention also provides the use of the hydroxycinnamic acid derivative of this invention, or the pharmaceutical composition of this invention, or methyl hydroxycinnamate in the preparation of a pharmaceutical formulation for treating acute respiratory distress syndrome. The structural formula of methyl hydroxycinnamate (also known as compound A1, hereinafter the same) is:

[0089] It should be noted that the hydroxycinnamic acid derivative (compound A2) is a derivative synthesized from a natural small molecule compound (methyl 4-hydroxycinnamate) and a short-chain fatty acid (butyric acid). Theoretically, it possesses the lung-inflammation-reducing effects of both hydroxycinnamic acid and butyric acid. Furthermore, drug screening revealed that the hydroxycinnamic acid derivative significantly reduced cell infiltration in ARDS mice. Low-dose validation showed that after four oral administrations of the hydroxycinnamic acid derivative to KM mice followed by intratracheal infusion of LPS, compared to LPS-induced ARDS mice, the cell infiltration in bronchoalveolar lavage fluid, pathological tissue damage, and expression of inflammatory factors in lung tissue were all reduced. Based on these experimental results, it can be concluded that the hydroxycinnamic acid derivative can alleviate ARDS by reducing lung inflammation and oxidative damage, and its effect is superior to that of methyl hydroxycinnamate (methyl hydroxycinnamate can alleviate ARDS by reducing lung inflammation and oxidative damage).

[0090] In some specific examples, the above applications include one or more combinations of the following applications:

[0091] (i) The use of hydroxycinnamic acid derivatives or pharmaceutical compositions or methyl hydroxycinnamate in the preparation of pharmaceutical formulations for reducing pulmonary cell infiltration; it should be noted that pulmonary cell infiltration is one of the causes of acute respiratory distress syndrome. As verified by the present invention, hydroxycinnamic acid derivatives can effectively reduce pulmonary cell infiltration, thereby alleviating acute respiratory distress syndrome; specifically, the number of pulmonary cell infiltrations is an important indicator reflecting the status of pulmonary cell infiltration. As verified by the present invention, hydroxycinnamic acid derivatives can reduce the number of pulmonary cell infiltrations, thereby reducing pulmonary cell infiltration and thus alleviating acute respiratory distress syndrome;

[0092] (ii) Application of hydroxycinnamic acid derivatives or pharmaceutical compositions in the preparation of pharmaceutical formulations that alleviate pathological damage to lung tissues; It should be noted that pathological damage to lung tissues is one of the causes of acute respiratory distress syndrome. As verified by the present invention, hydroxycinnamic acid derivatives can significantly alleviate pathological damage to lung tissues, thereby alleviating acute respiratory distress syndrome.

[0093] (iii) The use of hydroxycinnamic acid derivatives or pharmaceutical compositions or methyl hydroxycinnamate in the preparation of pharmaceutical formulations for reducing lung inflammation; it should be noted that lung inflammation is one of the causes of acute respiratory distress syndrome. As verified by the present invention, hydroxycinnamic acid derivatives can significantly reduce lung inflammation and thus alleviate acute respiratory distress syndrome.

[0094] In some specific examples, the use of the above-mentioned hydroxycinnamic acid derivatives or methyl hydroxycinnamate in the preparation of drugs that reduce lung inflammation includes: the use of hydroxycinnamic acid derivatives or methyl hydroxycinnamate in the preparation of pharmaceutical formulations that reduce the mRNA expression levels of lung inflammatory factors IL-6 and / or HO-1 and / or MPO and / or TNF-α and / or IL-1β.

[0095] It should be noted that the level of inflammatory factors is an important indicator of lung inflammation. As verified by this invention, hydroxycinnamic acid derivatives can significantly reduce the mRNA expression levels of lung inflammatory factors IL-6 and / or HO-1 and / or MPO and / or TNF-α and / or IL-1β, thereby alleviating lung inflammation and thus alleviating acute respiratory distress syndrome.

[0096] In some specific examples, the above applications include one or more combinations of the following applications:

[0097] (a) Use of the first compound in the preparation of a pharmaceutical formulation that reduces lung cell infiltration, wherein the first compound is selected from any of the following compounds:

[0098]

[0099] (b) The use of the second compound in the preparation of a pharmaceutical formulation that alleviates pathological damage to lung tissue, wherein the first compound is selected from the following compounds:

[0100]

[0101] (c) Use of the third compound in the preparation of a pharmaceutical formulation that reduces the mRNA expression level of the pulmonary inflammatory factor IL-6, wherein the third compound is selected from any of the following compounds:

[0102]

[0103]

[0104] (d) The use of the fourth compound in the preparation of a pharmaceutical formulation that reduces the mRNA expression level of the pulmonary inflammatory factor HO-1, wherein the fourth compound is selected from the following compounds:

[0105]

[0106] (e) The use of the fifth compound in the preparation of pharmaceutical formulations that reduce the mRNA expression level of the pulmonary inflammatory factor MPO, wherein the fifth compound is selected from the following compounds:

[0107]

[0108] (f) Use of the sixth compound in the preparation of a pharmaceutical formulation that reduces the mRNA expression level of the pulmonary inflammatory factor TNF-α, wherein the sixth compound is selected from any one of the following compounds:

[0109]

[0110]

[0111] (g) Use of the seventh compound in the preparation of a pharmaceutical formulation that reduces the mRNA expression level of the pulmonary inflammatory factor IL-1β, wherein the seventh compound is selected from any of the following compounds:

[0112]

[0113]

[0114] In some specific embodiments, the dosage form of the drug in the above applications may include oral dosage forms, nebulizers, powders, tablets, ointments, or injections. It should be noted that the drug used in the above applications can be a drug in different formulations, and the preparation of drugs in different formulations is well known in the art; furthermore, the specific drug dosage form can be selected according to clinical needs.

[0115] To better understand the present invention, specific examples are provided below to further illustrate the content of the present invention, but the content of the present invention is not limited to the examples below.

[0116] I. Preparation and Characterization of Hydroxycinnamic Acid Derivatives

[0117] Example 1: Preparation and Characterization of Hydroxycinnamic Acid Derivative A2 (also referred to as Compound A2, hereinafter the same)

[0118] The structural formula of compound A2 is shown below:

[0119]

[0120] The synthetic route is shown below:

[0121]

[0122] The specific preparation steps are as follows. The compounds A, B, C and D in the preparation steps correspond to the compounds in the synthetic route as shown in Table 1 below.

[0123] Table 1. Compounds corresponding to compounds A, B, C, and D in the preparation of compound A2.

[0124]

[0125] The specific preparation steps are as follows:

[0126] (a) Add compound A (2.0 g), compound B (1.026 mL) and compound D (2.146 g) to a test tube. After the addition is complete, stir well. Then add compound C (2.798 mL) to obtain the reaction mixture. Raise the reaction mixture to 60 °C and keep stirring for 50 minutes.

[0127] (b) After the reaction was complete as shown by TLC, the reaction mixture was cooled to room temperature, poured into 20 mL of water, and extracted twice with 20 mL of dichloromethane. The organic layer was then washed with 20 mL of saturated NaHCO3 solution, and dried over anhydrous MgSO4 for 2 hours. After drying, the product was purified by PE / EA (6:1 v / v) column chromatography to give 2.5 g of a white solid, which was compound A2, with a yield of 89.6%.

[0128] The compound A2 prepared above was subjected to proton nuclear magnetic resonance (NMR) spectroscopy. 1 The results of HNMR detection are shown below:

[0129] A2 (CDCl3, 600MHz) δ (ppm) 7.67 (d, J = 16.0Hz, 1H), 7.56–7.50 (m, 2H), 7.15–7.08 (m, 2H), 6.39 (d, J=16.0Hz,1H),3.80(s,3H),2.55(t,J=7.4Hz,2H),1.79(h,J=7.4Hz,2H),1.05(t,J=7.4Hz,3H).

[0130] Example 2: Preparation and characterization of hydroxycinnamic acid derivative A3 (also known as compound A3, hereinafter the same).

[0131] The structural formula of compound A3 is shown below:

[0132]

[0133] The synthetic route is shown below:

[0134]

[0135] The specific preparation steps are as follows. The compounds A, B, C and D in the preparation steps correspond to the compounds in the synthetic route as shown in Table 2 below.

[0136] Table 2 shows the compounds corresponding to compounds A, B, C, and D in the preparation of compound A3.

[0137]

[0138] The specific preparation steps are as follows:

[0139] (a) Add compound A (2.0 g), compound B (1.221 mL) and compound D (2.146 g) to a test tube. After the addition is complete, stir well. Then add compound C (2.798 mL) to obtain the reaction mixture. Raise the reaction mixture to 60 °C and keep stirring for 50 minutes.

[0140] (b) After the reaction was complete as shown by TLC spotting, the reaction mixture was cooled to room temperature, poured into 20 mL of water, extracted twice with 20 mL of dichloromethane, and then the organic layer was washed with 20 mL of saturated NaHCO3 solution. After washing, the mixture was dried with anhydrous MgSO4 for 2 hours. After drying, the product was purified by PE / EA (6:1 v / v) column chromatography to give 1.7 g of white solid, which was compound A3, with a yield of 57.6%.

[0141] The compound A3 prepared above was subjected to proton NMR spectroscopy. 1 The results of HNMR detection are shown below:

[0142] A3 (CDCl3, 600MHz) δ (ppm) 7.67 (d, J = 16.0 Hz, 1H), 7.56–7.49 (m, 2H), 7.14–7.06 (m, 2H), 6.39 (d, J = 16.0 Hz,1H),3.80(s,3H),2.56(t,J=7.5Hz,2H),1.80–1.68(m,2H),1.51–1.39(m,2H),0.97(t,J=7.3Hz,3H).

[0143] Example 3: Preparation and Characterization of Hydroxycinnamic Acid Derivative A4 (also referred to as Compound A4, hereinafter the same)

[0144] The structural formula of compound A4 is shown below:

[0145]

[0146] The synthetic route is shown below:

[0147]

[0148] The specific preparation steps are as follows, and the compounds A, B, C and D in the preparation steps correspond to the compounds in the synthetic route as shown in Table 3 below;

[0149] Table 3 shows the compounds corresponding to compounds A, B, C, and D in the preparation of compound A4.

[0150]

[0151] The specific preparation steps are as follows:

[0152] (a) Add compound A (0.5 g) and compound B (0.526 mL) to a test tube at 0 °C. After the addition is complete, stir well. Add compound C (0.340 mL) dropwise. Then dissolve compound D (0.867 g) in DCM and add it dropwise to the reaction system. After the addition is complete, raise the temperature of the reaction system to room temperature and keep stirring for 24 hours.

[0153] (b) After the reaction was complete as shown by TLC spotting, the mixture was filtered, poured into 50 mL of water, extracted twice with 50 mL of dichloromethane, and then washed with 50 mL of saturated NaHCO3 solution. After washing, the mixture was dried with anhydrous Na2SO4 for 2 hours. After drying, the product was purified by column chromatography using PE / EA (volume ratio 15:1) to give 0.29 g of white solid, which is compound A4, with a yield of 37.3%.

[0154] The compound A4 prepared above was subjected to proton nuclear magnetic resonance (NMR) spectroscopy. 1 The results of HNMR detection are shown below:

[0155] A4(CDCl3,600MHz)δ(ppm)7.67(d,J=16.0Hz,1H),7.54(d,J=8.5Hz,2H),7.13–7.09

[0156] (m,2H),6.40(d,J=16.0Hz,1H),3.81(s,3H),2.56(t,J=7.5Hz,2H),1.79–1.72(m,2H),1.43–1.36(m,4H),0.96–0.90(m,3H).

[0157] Example 4: Preparation and Characterization of Hydroxycinnamic Acid Derivative A5 (also referred to as Compound A5, hereinafter the same)

[0158] The structural formula of compound A5 is shown below:

[0159]

[0160] The synthetic route is shown below:

[0161]

[0162] The specific preparation steps are as follows, and the compounds A, B, C and D in the preparation steps correspond to the compounds in the synthetic route as shown in Table 4 below;

[0163] Table 4 shows the compounds corresponding to compounds A, B, C, and D in the preparation of compound A5.

[0164]

[0165] The specific preparation steps are as follows:

[0166] (a) Add compound A (2.0 g) and compound B (2.386 mL) to a test tube at 0 °C. After the addition is complete, stir well. Add compound C (1.360 mL) dropwise. Then dissolve compound D (3.468 g) in DCM and add it dropwise to the reaction system. After the addition is complete, raise the temperature of the reaction system to room temperature and keep stirring for 24 hours.

[0167] (b) After the reaction was complete as shown by TLC, the mixture was filtered and poured into 50 mL of water. It was extracted twice with 50 mL of dichloromethane, and then the organic layer was washed with 50 mL of saturated NaHCO3 solution. After washing, the mixture was dried over anhydrous Na2SO4 for 2 hours. After drying, the product was purified by column chromatography using PE / EA (10:1 v / v) to give 2.6 g of a white solid, which was compound A5, with a yield of 79.7%.

[0168] The compound A5 prepared above was subjected to proton nuclear magnetic resonance (NMR) spectroscopy. 1 The results of HNMR detection are shown below:

[0169] A5(CDCl3,600MHz)δ(ppm)7.67(d,J=16.0Hz,1H),7.56–7.51(m,2H),7.13–7.09(m,

[0170] 2H),6.40(d,J=16.0Hz,1H),3.81(s,3H),2.56(t,J=7.5Hz,2H),1.75(p,J=7.5H z,2H),1.44–1.38(m,2H),1.33(ddd,J=7.1,4.4,3.1Hz,4H),0.93–0.87(m,3H).

[0171] Example 5: Preparation and Characterization of Hydroxycinnamic Acid Derivative A6 (also referred to as Compound A6, hereinafter the same)

[0172] The structural formula of compound A6 is shown below:

[0173]

[0174] The synthetic route is shown below:

[0175]

[0176] The specific preparation steps are as follows, and the compounds A, B, C and D in the preparation steps correspond to the compounds in the synthetic route as shown in Table 5 below;

[0177] Table 5 shows the compounds corresponding to compounds A, B, C, and D in the preparation of compound A6.

[0178]

[0179] The specific preparation steps are as follows:

[0180] (a) Add compound A (1.0 g) and compound B (2.040 g) to a test tube at 0 °C. After the addition is complete, stir well. Add compound C (0.681 mL) dropwise. Then dissolve compound D (1.739 g) in DCM and add it dropwise to the reaction system. After the addition is complete, raise the temperature of the reaction system to room temperature and keep stirring for 24 hours.

[0181] (b) After the reaction was complete as shown by TLC, the mixture was filtered and poured into 50 mL of water. It was extracted twice with 50 mL of dichloromethane, and then the organic layer was washed with 50 mL of saturated NaHCO3 solution. After washing, the mixture was dried with anhydrous Na2SO4 for 2 hours. After drying, the product was purified by column chromatography using PE / EA (10:1 v / v) to give 0.9 g of a white solid, which was compound A6, with a yield of 38.4%.

[0182] The compound A6 prepared above was subjected to proton nuclear magnetic resonance (NMR) spectroscopy. 1 The results of HNMR detection are shown below:

[0183] A6(CDCl3,600MHz)δ(ppm)7.67(d,J=16.0Hz,1H),7.55–7.52(m,2H),7.13–7.09(m,

[0184] 2H), 6.39 (d, J = 16.0Hz, 1H), 3.81 (s, 3H), 2.56 (t, J = 7.5Hz, 2H), 1.63 (q, J = 7.4Hz, 2H), 1.26 (d, J = 3.4Hz, 22H), 0.88 (s, 3H).

[0185] Example 6 Preparation and characterization of hydroxycinnamic acid derivative A7 (also known as compound A7, hereinafter the same)

[0186] The structural formula of compound A7 is shown below:

[0187]

[0188] The synthetic route is shown below:

[0189]

[0190] The specific preparation steps are as follows. The compounds A, B, C, D and E in the preparation steps correspond to the compounds in the synthetic route as shown in Table 7 below.

[0191] Table 6 shows the compounds corresponding to compounds A to E in the preparation of compound A7.

[0192]

[0193]

[0194] The specific preparation steps are as follows:

[0195] (a) At room temperature, compound B (1.607 g) was added to dichloromethane, then compound A (2.0 g), compound C (1.892 mL), and compound D (0.274 g) were added to the solution, and finally compound E (2.157 g) was added. The mixture was stirred overnight.

[0196] (b) After the reaction was complete as shown by TLC spotting, 20 mL of ammonium chloride and 20 mL of water were added to the reaction mixture and stirred for about 5 minutes. The mixture was then poured into a separatory funnel, allowed to stand for separation, and the lower organic layer was collected. This process was repeated once, and the mixture was dried with Na2SO4 for 2 hours. After drying, the product was purified by column chromatography using PE / EA (volume ratio 4:1) to obtain 0.2 g of white solid, which was compound A7, with a yield of 6.1%.

[0197] The compound A7 prepared above was subjected to proton NMR spectroscopy. 1 The results of HNMR detection are shown below:

[0198] A7(CDCl3,600MHz)δ(ppm)7.68(d,J=16.0Hz,1H),7.59–7.55(m,2H),7.19(d,J=8.6

[0199] Hz, 2H), 7.05 (s, 2H), 6.41 (d, J = 16.0Hz, 1H), 3.86 (s, 3H), 3.81 (s, 3H).

[0200] Example 7 Preparation and characterization of hydroxycinnamic acid derivative A8 (also known as compound A8, hereinafter the same)

[0201] The structural formula of compound A8 is shown below:

[0202]

[0203] The synthetic route is shown below:

[0204]

[0205] The specific preparation steps are as follows. The compounds A, B, C, D and E in the preparation steps correspond to the compounds in the synthetic route as shown in Table 6 below.

[0206] Table 7 shows the compounds corresponding to compounds A to E in the preparation of compound A8.

[0207]

[0208]

[0209] The specific preparation steps are as follows:

[0210] (a) At room temperature, compound B (1.780 g) was added to dichloromethane, then compound A (2.0 g), compound C (1.892 mL), and compound D (0.274 g) were added to the solution, and finally compound E (2.157 g) was added. The mixture was stirred overnight.

[0211] (b) After the reaction was complete as shown by TLC, 20 mL of ammonium chloride and 20 mL of water were added to the reaction mixture, and the mixture was stirred for about 5 minutes. The mixture was then poured into a separatory funnel, allowed to stand for separation, and the lower organic layer was collected. This process was repeated once, and the mixture was dried with Na₂SO₄ for 2 hours. After drying, the product was purified by column chromatography using PE / EA (4:1 v / v) to give 2.6 g of a white solid, which was compound A8, with a yield of 76%.

[0212] The compound A8 prepared above was subjected to proton nuclear magnetic resonance (NMR) spectroscopy. 1 The results of HNMR detection are shown below:

[0213] A8(CDCl3,600MHz)δ(ppm)7.68(d,J=16.0Hz,1H),7.56–7.52(m,2H),7.18–7.14(m,

[0214] 2H), 6.57 (d, J = 0.7Hz, 1H), 6.40 (d, J = 16.0Hz, 1H), 5.91 (q, J = 1.0Hz, 1H), 3.81 (s, 3H), 3.73 (s, 3H), 3.46 (d, J = 1.0Hz, 2H).

[0215] Example 8: Preparation and Characterization of Hydroxycinnamic Acid Derivative B1 (also referred to as Compound B1, hereinafter the same)

[0216] The structural formula of compound B1 is shown below:

[0217]

[0218] The synthetic route is shown below:

[0219]

[0220] The specific preparation steps are as follows, and the compounds A, B, C and D in the preparation steps correspond to the compounds in the synthetic route as shown in Table 8 below;

[0221] Table 8 shows the compounds corresponding to compounds A, B, C, and D in the preparation of compound B1.

[0222]

[0223]

[0224] The specific preparation steps are as follows:

[0225] (a) Add compound A (0.9 g) and compound B (0.432 mL) to a test tube at 0 °C. After the addition is complete, stir well. Add C (1.077 mL) dropwise. Then dissolve D (0.825 g) in DCM and add it dropwise to the reaction system. After the addition is complete, raise the temperature of the reaction system to room temperature and keep stirring for 24 hours.

[0226] (b) After the reaction was complete as shown by TLC, the mixture was filtered and poured into 50 mL of water. It was extracted twice with 50 mL of dichloromethane, and then the organic layer was washed with 50 mL of saturated NaHCO3 solution. After washing, the mixture was dried with anhydrous Na2SO4 for 2 hours. After drying, the product was purified by column chromatography using PE / EA (15:1 v / v) to give 0.5 g of a white solid, which was compound B1, with a yield of 41.5%.

[0227] The compound B1 prepared above was subjected to proton NMR spectroscopy. 1 The results of HNMR detection are shown below:

[0228] B1(CDCl3,600MHz)δ(ppm)7.65(d,J=16.0Hz,1H),7.13–7.08(m,2H),7.04(d,J=8.1

[0229] Hz, 1H), 6.38 (d, J = 16.0Hz, 1H), 3.85 (s, 3H), 3.81 (s, 3H), 2.57 (t, J = 7.4Hz, 2H), 1.80 (h, J = 7.4Hz, 2H), 1.05 (t, J = 7.4Hz, 3H).

[0230] Example 9: Preparation and Characterization of Hydroxycinnamic Acid Derivative B2 (also known as Compound B2, hereinafter the same)

[0231] The structural formula of compound B2 is shown below:

[0232]

[0233] The synthetic route is shown below:

[0234]

[0235] The specific preparation steps are as follows, and the compounds A, B, C and D in the preparation steps correspond to the compounds in the synthetic route as shown in Table 9 below;

[0236] Table 9 shows the compounds corresponding to compounds A, B, C, and D in the preparation of compound B2.

[0237]

[0238] The specific preparation steps are as follows:

[0239] (a) Add compound A (2.6 g) and compound B (1.493 mL) to a test tube at 0 °C. After the addition is complete, stir well. Add C (1.105 mL) dropwise. Then dissolve D (2.83 g) in DCM and add it dropwise to the reaction system. After the addition is complete, raise the temperature of the reaction system to room temperature and keep stirring for 24 hours.

[0240] (b) After the reaction was complete as shown by TLC, the mixture was filtered and poured into 50 mL of water. It was extracted twice with 50 mL of dichloromethane, and then the organic layer was washed with 50 mL of saturated NaHCO3 solution. After washing, the mixture was dried with anhydrous Na2SO4 for 2 hours. After drying, the product was purified by column chromatography using PE / EA (15:1 v / v) to give 1.12 g of a white solid, which was compound B2, with a yield of 21.8%.

[0241] The compound B2 prepared above was subjected to proton nuclear magnetic resonance (NMR) spectroscopy. 1 The results of HNMR detection are shown below:

[0242] B2(CDCl3,600MHz)δ(ppm)7.65(d,J=16.0Hz,1H),7.13–7.08(m,2H),7.04(d,J=8.1

[0243] Hz,1H),6.38(d,J=16.0Hz,1H),3.85(s,3H),3.81(s,3H),2.58(t,J=7.5Hz,2H),1.66–1.61(m,2H),1.50–1.42(m,2H),0.97(t,J=7.4Hz,3H).

[0244] II. Animal Clinical Trials

[0245] In the following experiment, 8-week-old male KM mice were selected and kept at a room temperature of 23±1℃ and a humidity of 50±60%. The light-dark cycle was 12 hours, and they were provided with clean food and water at will.

[0246] In the following experiments, the cell counting of bronchoalveolar lavage fluid was performed as follows: Mix the EP tube containing BALF (bronchoalveolar lavage fluid), draw 10 μL of BALF, drop it into a cell counting chamber, insert the cell counting chamber into an automated whole-cell counter, observe the readings and record them.

[0247] In the following experiments, tissue damage analysis was performed as follows: Collected lung tissue samples were fixed with freshly prepared 4% paraformaldehyde and then stained with hematoxylin and eosin (H&E). Specifically, paraffin sections of lung tissue were baked in an oven at 65°C for at least 1.5 hours to ensure the paraffin on the tissue sections was fully melted. The sections were then placed twice in xylene to remove the melted paraffin and dehydrated in a gradient of ethanol. The sections were stained sequentially with hematoxylin and eosin, followed by dehydration with ethanol. After staining, the sections were clarified twice in xylene and fixed with neutral resin. The stained sections were observed under an optical microscope at 100x and 400x magnification, respectively.

[0248] In the following experiments, the levels of inflammatory factor mRNA in lung tissue were detected using the following methods:

[0249] (1) Extraction of total RNA (Trizol method)

[0250] ① After homogenizing lung tissue, add 750 μL of Trizol reagent, place on ice and let stand for 10 min, then repeatedly pipette to form a uniform suspension. Aspirate into a 1.5 mL RNase-free centrifuge tube, add 150 μL of chloroform, mix well and let stand for 5 min.

[0251] ② After standing, centrifuge at 12000 rpm / min for 15 min at 4℃, and use a pipette to transfer 300 μL of the supernatant into another new 1.5 mL RNase-free centrifuge tube;

[0252] ③ Add 300 μL of isopropanol to a new RNasefree centrifuge tube, mix well, and place in a -20℃ refrigerator for 20 min;

[0253] ④ After standing, centrifuge at 4℃ and 12000rpm / min for 10min, discard the upper liquid in the centrifuge tube, and add pre-cooled 75% ethanol for cleaning.

[0254] ⑤ Continue centrifuging at 4℃ and 12000rpm / min for 5 minutes. After the centrifuge tube is cleaned, repeat the previous step. Generally, wash 2-3 times. After cleaning, place the tube in a fume hood to dry. Once the white precipitate at the bottom of the centrifuge tube is completely transparent, add 20μL of DEPC water. This is the total RNA from the lung tissue.

[0255] (2) Detection of RNA concentration, purity, and integrity

[0256] ① Turn on the Nanodrop-2000 instrument and use a pipette to draw DEPC water to clean and calibrate the instrument;

[0257] ②Take 2 μL of the RNA to be tested and gently add it to the measuring probe to determine the concentration. Record the results and observe the ratio of OD260 / 280, which should be between 1.8 and 2.0.

[0258] (3) RNA is reverse transcribed into cDNA (TaKaRa).

[0259] The transcription system of 20 μL from the TaKaRa reverse transcription kit was selected for the experiment; the amount of RNA required for transcription was calculated, specifically as follows: 4 μL of 5x PrimeScript RT Master Mix and the required volume of RNA were added to a 200 μL LEP tube, water was added to make up to a volume of 16 μL, the mixture was vortexed and centrifuged to the bottom; the reaction conditions on the PCR instrument were set as follows: 37℃, 15 min; 85℃, 5 s; 4℃ to terminate.

[0260] (4) Real-Time PCR (TaKaRa)

[0261] ① Design primer sequences (Table 10);

[0262] Table 10 Primer Sequences

[0263]

[0264] ② Prepare sterile, enzyme-free pipette tips and PCR-specific 8-tube or 96-well plates;

[0265] ③ Prepare the PCR reaction solution on a 0℃ metal bath; then, according to the gene used in this experiment, prepare the mixed tubes according to the instructions.

[0266] ④ After the sample is added, check for any errors in the sample loading. Then, use a centrifuge to centrifuge the 8-tube or 96-well plate.

[0267] ⑤ Finally, set the reaction program for the 7500fast instrument according to the instruction manual, and then run the sample on the instrument.

[0268] ⑥ Export the data and calculate 2 -ΔΔCT The values ​​were analyzed, and a bar chart was created. Statistical analysis was then performed.

[0269] In the following experiments, data are expressed as mean ± standard deviation (SEM). The independent samples t-test was used to assess the differences between the two groups. All graphs were created using GraphPadPrism 8.0.

[0270] (I) Drug Screening

[0271] Forty-two KM mice were divided into LPS group, B1+LPS group, A1+LPS group, A2+LPS group, and A3+LPS group. On days 0, 2, 4, and 6, the mice were administered 200 μL of the corresponding drug (30 mM) by gavage. The LPS group was administered corn oil by gavage. On day 6, 50 μL LPS (0.2 g / L) was instilled into the trachea. 72 h after LPS instillation, BALF (bronchoalveolar lavage fluid) was collected for cell counting.

[0272] Cell count test results as follows Figure 1 As shown, the results indicate that the total number of cells in the A2+LPS group was significantly reduced compared to the LPS group; and other drug treatments did not reduce the total number of cells in BALF; this suggests that A2 can alleviate lung cell infiltration caused by LPS.

[0273] (II) Verification of the A2 effect of compound

[0274] Twenty-four KM mice were divided into three groups: LPS group, A1+LPS group, and A2+LPS group. 200 μL of the corresponding drug (30 mM) was administered by gavage on days 0, 2, 4, 6, 8, and 10, respectively. The LPS group was administered corn oil by gavage. On day 10, 50 μL of LPS (0.2 g / L) was instilled into the trachea. 72 hours after the LPS instillation, BALF (bronchoalveolar lavage fluid) was collected for cell counting.

[0275] Cell counting results as follows Figure 2 As shown, compared with the LPS group, the total number of cells in the A2+LPS group was significantly reduced; and A1 treatment did not reduce the total number of cells in the BALF of ARDS mice; the above data further illustrate that A2 can alleviate lung cell infiltration caused by LPS.

[0276] (III) Validation of the low-dose effect of compound A2

[0277] Twenty-four KM mice were divided into three groups: LPS group, A1+LPS group, and A2+LPS group. On days 0, 2, 4, and 6, the mice were administered 100 μL of the corresponding drug (15 mM) by gavage. The LPS group was administered corn oil by gavage. On day 6, 50 μL of LPS (0.2 g / L) was instilled into the trachea. 72 hours after the LPS instillation, lung tissue and BALF (bronchoalveolar lavage fluid) were collected for cell counting, tissue damage, and inflammatory factor detection.

[0278] Cell counting results as follows Figure 3 As shown, compared to the LPS group, the total number of cells in the A2+LPS group was significantly reduced; and A1 treatment did not significantly reduce the total number of cells in the BALF of ARDS mice. This indicates that reducing the dosage of A2 can still effectively alleviate lung cell infiltration caused by ARDS, and is superior to A1.

[0279] Tissue damage conditions such as Figure 4 As shown in the figure, compared with the LPS group, no significant changes were found in the lung tissue of mice treated with A1+LPS; however, the pathological damage in the lungs was alleviated in the A2+LPS group compared with the LPS group. Therefore, A2 supplementation can reduce the pathological damage in the lung tissue of mice induced by LPS, and the effect is better than that of A1.

[0280] The mRNA levels of inflammatory cytokines interleukin-6 (IL-6), tumor necrosis factor-α (TNF-α), interleukin-1β (IL-1β), heme oxygenase-1 (HO-1), and myeloperoxidase (MPO) are as follows: Figure 5 , Figure 6 , Figure 7 , Figure 8 and Figure 9 As shown, the results indicated that the mRNA expression levels of IL-6, HO-1, and MPO in the lung tissue of mice in the A2+LPS group were significantly lower than those in LPS-treated mice, and the mRNA expression levels of IL-6, TNF-α, IL-1β, HO-1, and MPO in the lung tissue of mice in the A2+LPS group were significantly lower than those in the A1 group. Therefore, this demonstrates that A2 supplementation can significantly alleviate LPS-induced inflammatory factor mRNA expression in mouse lung tissue, and its efficacy is superior to that of A1 (p < 0.05*, p < 0.01**, p < 0.001***).

[0281] (iv) Verification of the effect of hydroxycinnamic acid derivatives

[0282] Sixty-six KM mice were divided into Control group, LPS group, A1+LPS group, A3+LPS group, A4+LPS group, A5+LPS group, A6+LPS group, A7+LPS group, A8+LPS group, B1+LPS group, and B2+LPS group. On days -3, -2, -1, and 1, 100 μL of the corresponding drug (15 mM) was administered by gavage. The Control group and LPS group were administered corn oil by gavage. On day 0, 50 μL of LPS (0.2 g / L) was instilled into the trachea. 72 h after LPS instillation, lung tissue and BALF (bronchoalveolar lavage fluid) were collected for cell counting, tissue damage, and inflammatory factor detection.

[0283] Cell counting results as follows Figure 10 As shown, compared to the LPS group, the total number of cells in the A5+LPS group was significantly reduced; and A1 treatment did not significantly reduce the total number of cells in the BALF of ARDS mice. This indicates that A5 can effectively alleviate lung cell infiltration caused by ARDS, and is superior to A1.

[0284] The mRNA levels of inflammatory cytokines interleukin-6 (IL-6), tumor necrosis factor-α (TNF-α), and IL-1β (IL-1β) are as follows: Figure 11 , Figure 12 and Figure 13 As shown in the figure. The results showed that all hydroxycinnamic acid derivatives could downregulate the mRNA levels of IL-6, TNF-α, and IL-1β, indicating that they have excellent anti-inflammatory activity. (p < 0.05*, p < 0.01**, p < 0.001***).

[0285] In summary, the hydroxycinnamic acid derivative of this invention has a mitigating effect on LPS-induced ARDS in mice. Drug screening revealed that the hydroxycinnamic acid derivative of this invention significantly reduced cell infiltration in ARDS mice. Further verification of its effect through 6-dose and 4-dose administration showed that supplementing with the hydroxycinnamic acid derivative of this invention can reduce LPS-induced lung cell infiltration, increased lung inflammation, and pathological damage to lung tissue, and its effect is superior to that of methyl hydroxycinnamate. This beneficial effect may be achieved by inhibiting the expression of inflammatory factors.

[0286] 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 it. Although the present invention has been described in detail with reference to preferred embodiments, those skilled in the art should understand that modifications or equivalent substitutions can be made to the technical solutions of the present invention without departing from the spirit and scope of the technical solutions of the present invention, and all such modifications or substitutions should be covered within the scope of the claims of the present invention.

Claims

1. Application of hydroxycinnamic acid derivatives in the preparation of pharmaceutical formulations for the treatment of acute respiratory distress syndrome, wherein the hydroxycinnamic acid derivatives are selected from the following compounds: .

2. Application of hydroxycinnamic acid derivatives in the preparation of pharmaceutical formulations that reduce lung cell infiltration or lung tissue pathological damage, wherein the hydroxycinnamic acid derivatives are selected from the following compounds: .

3. Application of hydroxycinnamic acid derivatives in the preparation of pharmaceutical formulations to reduce lung cell infiltration; the hydroxycinnamic acid derivatives are selected from the following compounds: .

4. The application according to any one of claims 1 to 3, characterized in that, Pharmaceutical formulations can be in oral, nebulized, or injectable forms.

5. The application according to any one of claims 1 to 3, characterized in that, Pharmaceutical preparations can be in the form of powders, tablets, or ointments.

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