Hydroxyl cinnamic acid derivative as well as preparation method and application thereof

Abstract

The invention belongs to the technical field of new application of medicines, and particularly relates to a hydroxycinnamic acid derivative as well as a preparation method and application thereof. According to the method, an LPS tracheal instillation induced ARDS model is combined with a hydroxycinnamic acid derivative for treatment, alveolar lavage fluid cell count is measured, and pathological characteristics and inflammation gene expression level of lung tissue are detected; results show that the hydroxycinnamic acid derivative can significantly reduce lung cell infiltration, alleviate lung tissue damage and reduce inflammatory factor expression level.

Description

[0001] This invention application is a subsequent application of an invention patent with an application date of 2024-03-08, an application number of 2024102721361, and a patent name of "Application of KS-3 butyrate or a pharmaceutically acceptable salt thereof in the preparation of a drug for the treatment of acute respiratory distress syndrome." Technical Field

[0002] The invention belongs to the technical field of medicines, and in particular relates to hydroxycinnamic acid derivatives and preparation methods and applications thereof. Background Art

[0003] Acute respiratory distress syndrome (ARDS) is a common respiratory critical illness, with clinical features of bilateral chest opacity and severe hypoxemia caused by non-cardiogenic pulmonary edema. The etiology of ARDS is complex, and its most common risk factor is pneumonia, such as pneumonia caused by infection with SARS-CoV, H5N1 avian influenza virus, H7N9 avian influenza virus, or SARS-CoV-2.

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

[0005] Therefore, it is of great significance to develop a drug that can treat ARDS. Summary of the invention

[0006] In view of the above problems, the purpose of the present invention is to provide a hydroxycinnamic acid derivative, which can be used in drugs for alleviating acute respiratory distress syndrome, and can alleviate acute respiratory distress syndrome by effectively reducing lung cell infiltration, reducing lung tissue damage and reducing the expression level of inflammatory factors.

[0007] In order to achieve the above object, the present invention can adopt the following technical solutions:

[0008] In one aspect, the present 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 independently selected from the second alkyl group, the second alkyl group has a carbon chain number of ≤5; and / or the first alkyl group has a carbon chain number of ≤15; and / or the alkoxy group has a carbon chain number of ≤5.

[0014] Preferably, the 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, which 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 carbon chain number of the first alkyl group is ≤5; or R2 is selected from the alkoxy group, R1 is selected from the first alkyl group, and the carbon chain number of the first alkyl group is ≤3; the preparation method of the hydroxycinnamic acid derivative comprises: mixing a compound represented by formula I, a compound represented by formula II, a compound represented by formula III and TsCl to obtain a 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 carbon chain number of the first alkyl group is ≥6; or R2 is selected from the alkoxy group, R1 is selected from the first alkyl group, and the carbon chain number of the first alkyl group is ≥4; the preparation method of the hydroxycinnamic acid derivative comprises: mixing the compound represented by formula I and the compound represented by formula II, then adding an organic base catalyst, and then adding a first condensing agent to react to obtain the hydroxycinnamic acid derivative;

[0020] (c) In the hydroxycinnamic acid derivative, R2 is selected from -H, R1 is selected from or -C=CCOOR6; the preparation method of the hydroxycinnamic acid derivative comprises: mixing the compound represented by formula I, the compound represented by formula II, a hydroxyl activator, a catalyst and a second condensing agent to obtain the hydroxycinnamic acid derivative;

[0021] Wherein, the structural formulas of the compound represented by formula I, the compound represented by formula II and the compound represented by formula III are as follows:

[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, which comprises the hydroxycinnamic acid derivative of the present invention or the pharmaceutical composition of the present invention.

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

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

[0027] (i) Use of a hydroxycinnamic acid derivative or a pharmaceutical composition or methyl hydroxycinnamate in the preparation of a pharmaceutical preparation for reducing lung cell infiltration;

[0028] (ii) Use of a hydroxycinnamic acid derivative or a pharmaceutical composition in the preparation of a pharmaceutical preparation for alleviating pathological damage to lung tissue;

[0029] (iii) Use of a hydroxycinnamic acid derivative or a pharmaceutical composition or methyl hydroxycinnamate in the preparation of a pharmaceutical preparation for alleviating lung inflammation.

[0030] Preferably, the use of the above-mentioned hydroxycinnamic acid derivatives or methyl hydroxycinnamate in the preparation of a drug for reducing lung inflammation includes: the use of hydroxycinnamic acid derivatives or methyl hydroxycinnamate in the preparation of a pharmaceutical preparation for reducing the mRNA expression level 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 application includes one or more combinations of the following applications:

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

[0033]

[0034] (b) Use of the second compound in the preparation of a pharmaceutical preparation for alleviating pathological damage to lung tissue, wherein the first compound is selected from the following compounds:

[0035]

[0036] (c) Use of a third compound in the preparation of a pharmaceutical preparation for reducing 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) Use of a fourth compound in the preparation of a pharmaceutical preparation for reducing the mRNA expression level of the pulmonary inflammatory factor HO-1, wherein the fourth compound is selected from the following compounds:

[0039]

[0040] (e) Use of the fifth compound in the preparation of a pharmaceutical preparation for reducing 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 preparation for reducing the mRNA expression level of the lung 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 preparation for reducing the mRNA expression level of the pulmonary inflammatory factor IL-1β, wherein the seventh compound is selected from any one of the following compounds:

[0045]

[0046]

[0047] Preferably, in the above application, the dosage form of the drug may include oral preparation, aerosol preparation, powder, tablet, paste preparation or injection.

[0048] The beneficial effects of the present invention include at least: the present invention adopts LPS tracheal instillation to induce ARDS model combined with hydroxycinnamic acid derivative treatment, and measures the cell count of alveolar lavage fluid, histopathology and inflammatory gene expression level; the results show that hydroxycinnamic acid derivatives can significantly reduce lung cell infiltration, alleviate lung tissue damage and reduce the expression level of inflammatory factors. BRIEF DESCRIPTION OF THE DRAWINGS

[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 The effect of 6 administrations of hydroxycinnamic acid and its derivatives on the 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 The effect of 4 administrations of hydroxycinnamic acid and its derivatives on the lung cell infiltration of 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 The effect of 4 administrations of hydroxycinnamic acid and its derivatives on lung tissue damage in ARDS mice induced by LPS (upper figure, 100×, scale bar=100μm; lower figure, 400×, scale bar=100μm);

[0053] Figure 5 The effect of hydroxycinnamic acid derivatives on the mRNA level of IL-6, an inflammatory factor in the lung tissue of ARDS mice induced by LPS (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 inflammatory factor TNF-α in lung tissue of ARDS mice induced by LPS (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 IL-1β, an inflammatory factor in the lung tissue of ARDS mice induced by LPS (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 ARDS mice induced by LPS (data are expressed as mean ± standard deviation, N = 4, *P < 0.05; **P < 0.01; ***P < 0.001);

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

[0058] Fig.10 The effect of 4 administrations of hydroxycinnamic acid and its derivatives on the lung cell infiltration of LPS-induced ARDS mice (data are expressed as mean ± standard deviation, N = 6, *P < 0.05; **P < 0.01; ***P < 0.001);

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

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

[0061] Fig.13 The effect of hydroxycinnamic acid derivatives on the mRNA level of IL-1β, an inflammatory factor in the lung tissue of ARDS mice induced by LPS (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 unit of the bar chart value is 10 5 / mL, for example, if the bar graph value is 34.26, it means 34.26*10 5 / mL; Figure 5 , Figure 6 , Figure 7 , Fig. 9 , Fig.10 , Fig.11 and Fig.12 In the figure, the data are normalized and the vertical axis shows the multiple relationship. DETAILED DESCRIPTION

[0063] The examples are provided to better illustrate the present invention, but the present invention is not limited to the examples. Therefore, those skilled in the art may make non-essential improvements and adjustments to the implementation scheme according to the above invention content, which still fall within the protection scope of the present invention.

[0064] The terms used herein are only used to describe specific embodiments and are not intended to limit the present disclosure. Unless the context has a significantly different meaning, expressions in the singular include expressions in the plural. As used herein, it should be understood that terms such as "include", "have", "include" are intended to indicate the presence of features, numbers, operations, materials, or combinations. The terms of the present invention are disclosed in the specification, and are not intended to exclude the possibility that one or more other features, numbers, operations, materials, or combinations thereof may exist or may be added. As used herein, " / " may be interpreted as "and" or "or", depending on the circumstances.

[0065] In the present invention, the terms "first" and "second" have no special meanings and only refer to substances with the same function in different environments.

[0066] The embodiment of the present invention provides a hydroxycinnamic acid derivative, and the structural formula of the hydroxycinnamic acid derivative is as follows:

[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 independently selected from the second alkyl group, the second alkyl group has a carbon chain number of ≤5; and / or the first alkyl group has a carbon chain number of ≤15; and / or the alkoxy group has a carbon chain number of ≤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; in addition, R1COO- and R2 are located at different positions. In addition, R4, R5 and R6 are independently selected from the second alkyl group, which means that R4, R5 and R6 can select 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 cereals, fruits, and vegetables. It has been widely studied because of its anti-inflammatory, antioxidant, antibacterial, anti-tumor and anti-platelet aggregation effects. However, since the main functional group of antioxidant activity is phenolic hydroxyl, it has pharmacokinetic problems of low bioavailability, low stability and rapid metabolism. In addition, due to the dissociation effect of the carboxylic acid group, it is difficult to penetrate the cell membrane, further limiting its biological activity. Therefore, it is necessary to structurally modify hydroxycinnamic acid to increase its hydrophobicity so that it can penetrate the cell membrane, prolong the drug effect, improve bioavailability, and increase biological activity. It is of great significance to acylate and esterify the phenolic hydroxyl group and carboxylic acid group of the hydroxycinnamic acid molecule, respectively, which is an effective way to perform structural modification. Therefore, the hydroxycinnamic acid derivatives in the present invention also improve the bioavailability of hydroxycinnamic acid or hydroxycinnamic acid.

[0074] In some specific examples, the hydroxycinnamic acid derivative is selected from any one of the following compounds:

[0075]

[0076]

[0077] The present invention also provides a method for preparing the hydroxycinnamic acid derivative of the present 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 carbon chain number of the first alkyl group is ≤5; or R2 is selected from alkoxy, R1 is selected from the first alkyl group, and the carbon chain number of the first alkyl group is ≤3; the preparation method of the hydroxycinnamic acid derivative comprises: mixing the compound represented by formula I, the compound represented by formula II, the compound represented by 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 carbon chain number 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 carbon chain number 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 the first alkyl group, and the carbon chain number of the first alkyl group is ≥6; or R2 is selected from alkoxy, R1 is selected from the first alkyl group, and the carbon chain number of the first alkyl group is ≥4; the preparation method of the hydroxycinnamic acid derivative comprises: mixing the compound represented by formula I and the compound represented by formula II, then adding an organic base catalyst, and then adding a first condensing agent to react 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) R2 is selected from -H, R1 is selected from the first alkyl group, and the carbon chain number of the first alkyl group is ≥6, such as compounds A4, A5 and A6; (ii) R2 is selected from alkoxy, R1 is selected from the first alkyl group, and the carbon chain number of the first alkyl group is ≥4, such as compound B2; in addition, the above-mentioned organic base catalyst and the first condensing agent are well known in the art, such as the organic base catalyst can be pyridine The condensing agent may be DCC (dicyclohexylcarbodiimide);

[0080] (c) In the hydroxycinnamic acid derivative, R2 is selected from -H, R1 is selected from or -C=CCOOR6; the preparation method of the hydroxycinnamic acid derivative comprises: mixing the compound represented by formula I, the compound represented by formula II, a hydroxy activator, a catalyst and a second condensing agent 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, respectively: (i) R2 is selected from -H, R1 is selected from (ii) R2 is selected from -H, R1 is selected from -C=CCOOR6, such as compound A7; in addition, the above-mentioned hydroxy activator, catalyst and second condensing agent are all well known in the art, for example, the hydroxy activator can be DIEPA (N,N-diisopropylethylamine), the catalyst can be DMAP (4-dimethylaminopyridine), and the second condensing agent can be EDCI (1-ethyl-(3-dimethylaminopropyl)carbodiimide hydrochloride);

[0081] Wherein, the structural formulas of the compound represented by formula I, the compound represented by formula II and the compound represented by formula III are as follows:

[0082]

[0083] In addition, it should be understood that in the above-mentioned preparation method of the hydroxycinnamic acid derivative, the corresponding R1, R2 and R3 in the raw material are consistent with the corresponding R1, R2 and R3 in the hydroxycinnamic acid derivative.

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

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

[0086] The embodiment of the present invention further provides a pharmaceutical preparation for treating acute respiratory distress syndrome, which comprises the hydroxycinnamic acid derivative of the present invention or the pharmaceutical composition of the present invention.

[0087] It should be noted that the hydroxycinnamic acid derivatives of the present invention or the above-mentioned composition can also be added with excipients to prepare pharmaceutical preparations; the excipients can be selected according to the dosage form of the pharmaceutical preparation; the dosage form of the drug can include oral preparations, nebulizers, powders, tablets, pastes or injections.

[0088] The embodiment of the present invention also provides a use of the hydroxycinnamic acid derivative of the present invention or the pharmaceutical composition of the present invention or methyl hydroxycinnamate in the preparation of a pharmaceutical preparation for treating acute respiratory distress syndrome. The structural formula of methyl hydroxycinnamate (also referred to as compound A1, the same below) 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 has the effect of reducing lung inflammation of hydroxycinnamic acid and butyric acid. In addition, through drug screening, it was found that hydroxycinnamic acid derivatives can significantly reduce the cell infiltration of ARDS mice. The results of low-dose drug verification showed that after four oral administrations of hydroxycinnamic acid derivatives to KM mice, LPS was instilled into the trachea. Compared with LPS-induced ARDS mice, the cell infiltration of alveolar lavage fluid, pathological tissue damage, and the expression of inflammatory factors in lung tissue were all reduced. According to the above experimental results, it can be known that hydroxycinnamic acid derivatives can relieve ARDS by reducing lung inflammation and oxidative damage, and the effect is better than methyl hydroxycinnamic acid (methyl hydroxycinnamic acid can relieve ARDS by reducing lung inflammation and oxidative damage).

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

[0091] (i) Use of hydroxycinnamic acid derivatives or pharmaceutical compositions or methyl hydroxycinnamate in the preparation of pharmaceutical preparations for reducing lung cell infiltration; it should be noted that lung cell infiltration is one of the causes of acute respiratory distress syndrome. It can be seen from the present invention that hydroxycinnamic acid derivatives can effectively reduce lung cell infiltration and thus reduce acute respiratory distress syndrome; specifically, the number of lung cell infiltrations is an important indicator of the lung cell infiltration situation. It can be seen from the present invention that hydroxycinnamic acid derivatives can reduce the number of lung cell infiltrations and thus reduce lung cell infiltration and thus reduce acute respiratory distress syndrome;

[0092] (ii) Use of a hydroxycinnamic acid derivative or a pharmaceutical composition in the preparation of a pharmaceutical preparation for alleviating pathological damage to lung tissue; it should be noted that pathological damage to lung tissue is one of the causes of acute respiratory distress syndrome. It can be seen from the verification of the present invention that hydroxycinnamic acid derivatives can significantly reduce pathological damage to lung tissue and thus alleviate acute respiratory distress syndrome;

[0093] (iii) Use of hydroxycinnamic acid derivatives or pharmaceutical compositions or methyl hydroxycinnamate in the preparation of pharmaceutical preparations for alleviating lung inflammation; it should be noted that lung inflammation is one of the causes of acute respiratory distress syndrome. It can be seen from the verification of the present invention that 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 for reducing lung inflammation includes: the use of hydroxycinnamic acid derivatives or methyl hydroxycinnamate in the preparation of pharmaceutical preparations for reducing 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. Through verification of the present invention, it can be known that hydroxycinnamic acid derivatives can significantly reduce the mRNA expression level 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 then alleviating acute respiratory distress syndrome.

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

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

[0098]

[0099] (b) Use of the second compound in the preparation of a pharmaceutical preparation for alleviating pathological damage to lung tissue, wherein the first compound is selected from the following compounds:

[0100]

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

[0102]

[0103]

[0104] (d) Use of a fourth compound in the preparation of a pharmaceutical preparation for reducing the mRNA expression level of the pulmonary inflammatory factor HO-1, wherein the fourth compound is selected from the following compounds:

[0105]

[0106] (e) Use of the fifth compound in the preparation of a pharmaceutical preparation for reducing 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 preparation for reducing the mRNA expression level of the lung 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 preparation for reducing the mRNA expression level of the pulmonary inflammatory factor IL-1β, wherein the seventh compound is selected from any one of the following compounds:

[0112]

[0113]

[0114] In some specific embodiments, the above-mentioned application, the dosage form of the drug may include oral agent, atomizer, powder, tablet, paste dosage form or injection. It should be noted that the above-mentioned application of the drug can be a drug in different preparation forms, and the preparation of drugs in different preparation forms is well known in the art; in addition, the specific drug dosage form can be selected according to clinical needs.

[0115] In order to better understand the present invention, the content of the present invention is further explained below in conjunction with specific examples, but the content of the present invention is not limited to the following examples.

[0116] 1. 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] Its synthetic route is as follows:

[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 Compound A, Compound B, Compound C and Compound D in the preparation of Compound A2

[0124]

[0125] The specific preparation steps are as follows:

[0126] (a) Compound A (2.0 g), Compound B (1.026 mL) and Compound D (2.146 g) were added to a test tube, and the mixture was stirred evenly after the addition was completed. Compound C (2.798 mL) was then added to obtain a reaction mixture, and the reaction mixture was heated to 60° C. and stirred for 50 minutes;

[0127] (b) After the TLC plate showed that the reaction was completed, 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, and dried with anhydrous MgSO4 for 2 hours after washing. After drying, the product was purified by column chromatography with PE / EA (volume ratio 6:1) to obtain 2.5 g of white solid, which was compound A2, with a yield of 89.6%.

[0128] The compound A2 prepared above was subjected to hydrogen nuclear magnetic resonance ( 1 HNMR) detection, the results are as follows:

[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 referred to as Compound A3, hereinafter the same)

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

[0132]

[0133] Its synthetic route is as follows:

[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 Compounds corresponding to Compound A, Compound B, Compound C and Compound D in the preparation of Compound A3

[0137]

[0138] The specific preparation steps are as follows:

[0139] (a) Compound A (2.0 g), Compound B (1.221 mL) and Compound D (2.146 g) were added to a test tube, and the mixture was stirred evenly after the addition was completed. Compound C (2.798 mL) was then added to obtain a reaction mixture, and the reaction mixture was heated to 60° C. and stirred for 50 minutes;

[0140] (b) After the TLC plate showed that the reaction was completed, 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, it was dried with anhydrous MgSO4 for 2 hours. After drying, the product was purified by column chromatography with PE / EA (volume ratio 6:1) to obtain 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 hydrogen nuclear magnetic resonance ( 1 HNMR) detection, the results are as follows:

[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, the same below)

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

[0145]

[0146] Its synthetic route is as follows:

[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 Compounds corresponding to Compound A, Compound B, Compound C and Compound D in the preparation of Compound A4

[0150]

[0151] The specific preparation steps are as follows:

[0152] (a) Compound A (0.5 g) and Compound B (0.526 mL) were added to a test tube at 0°C. After the addition was complete, the mixture was stirred evenly. Compound C (0.340 mL) was added dropwise. Compound D (0.867 g) was then dissolved in DCM and added dropwise to the reaction system. After the addition was complete, the reaction system was heated to room temperature and stirred for 24 hours.

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

[0154] The compound A4 prepared above was subjected to hydrogen nuclear magnetic resonance ( 1 HNMR) detection, the results are as follows:

[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] Its synthetic route is as follows:

[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 Compounds corresponding to Compound A, Compound B, Compound C and Compound D in the preparation of Compound A5

[0164]

[0165] The specific preparation steps are as follows:

[0166] (a) Compound A (2.0 g) and Compound B (2.386 mL) were added to a test tube at 0°C. After the addition was complete, the mixture was stirred evenly. Compound C (1.360 mL) was added dropwise. Compound D (3.468 g) was dissolved in DCM and added dropwise to the reaction system. After the addition was complete, the reaction system was heated to room temperature and stirred for 24 hours.

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

[0168] The compound A5 prepared above was subjected to hydrogen nuclear magnetic resonance ( 1 HNMR) detection, the results are as follows:

[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] Its synthetic route is as follows:

[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 Compounds corresponding to Compound A, Compound B, Compound C and Compound D in the preparation of Compound A6

[0178]

[0179] The specific preparation steps are as follows:

[0180] (a) Compound A (1.0 g) and Compound B (2.040 g) were added to a test tube at 0°C. After the addition was completed, the mixture was stirred evenly. Compound C (0.681 mL) was added dropwise. Compound D (1.739 g) was then dissolved in DCM and added dropwise to the reaction system. After the addition was completed, the temperature of the reaction system was raised to room temperature and stirred for 24 hours.

[0181] (b) After the TLC plate showed that the reaction was completed, the mixture was filtered, poured into 50 mL of water, extracted twice with 50 mL of dichloromethane, and then the organic layer was washed with 50 mL of saturated NaHCO3 solution, and dried with anhydrous Na2SO4 for 2 hours. After drying, the product was purified by column chromatography with PE / EA (volume ratio 10:1) to obtain 0.9 g of a white solid, namely compound A6, with a yield of 38.4%.

[0182] The compound A6 prepared above was subjected to hydrogen nuclear magnetic resonance ( 1 HNMR) detection, the results are as follows:

[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 referred to as Compound A7, hereinafter the same)

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

[0187]

[0188] Its synthetic route is as follows:

[0189]

[0190] The specific preparation steps are as follows, and 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 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, and 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, and stirring was continued overnight;

[0196] (b) After the TLC plate showed that the reaction was completed, 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 and allowed to stand for stratification. The lower organic layer was collected and the operation was repeated once. The mixture was dried over 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 a white solid, i.e., compound A7, with a yield of 6.1%.

[0197] The compound A7 prepared above was subjected to hydrogen nuclear magnetic resonance ( 1 HNMR) detection, the results are as follows:

[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 referred to as Compound A8, hereinafter the same)

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

[0202]

[0203] Its synthetic route is as follows:

[0204]

[0205] The specific preparation steps are as follows, and 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 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, and 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, and stirring was continued overnight;

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

[0212] The compound A8 prepared above was subjected to hydrogen nuclear magnetic resonance ( 1 HNMR) detection, the results are as follows:

[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, the same below)

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

[0217]

[0218] Its synthetic route is as follows:

[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 Compounds corresponding to Compound A, Compound B, Compound C and Compound D in the preparation of Compound B1

[0222]

[0223]

[0224] The specific preparation steps are as follows:

[0225] (a) Compound A (0.9 g) and compound B (0.432 mL) were added to a test tube at 0°C. After the addition was complete, the mixture was stirred evenly. C (1.077 mL) was added dropwise. D (0.825 g) was then dissolved in DCM and added dropwise to the reaction system. After the addition was complete, the reaction system was heated to room temperature and stirred for 24 hours.

[0226] (b) After the TLC plate showed that the reaction was completed, the mixture was filtered, poured into 50 mL of water, extracted twice with 50 mL of dichloromethane, and then the organic layer was washed with 50 mL of saturated NaHCO3 solution. After washing, the organic layer was dried with anhydrous Na2SO4 for 2 hours. After drying, the product was purified by column chromatography with PE / EA (volume ratio 15:1) to obtain 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 hydrogen nuclear magnetic resonance ( 1 HNMR) detection, the results are as follows:

[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 referred to as Compound B2, the same below)

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

[0232]

[0233] Its synthetic route is as follows:

[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 Compounds corresponding to Compound A, Compound B, Compound C and Compound D in the preparation of Compound B2

[0237]

[0238] The specific preparation steps are as follows:

[0239] (a) Compound A (2.6 g) and compound B (1.493 mL) were added to a test tube at 0°C. After the addition was complete, the mixture was stirred evenly. C (1.105 mL) was added dropwise. D (2.83 g) was then dissolved in DCM and added dropwise to the reaction system. After the addition was complete, the reaction system was heated to room temperature and stirred for 24 hours.

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

[0241] The compound B2 prepared above was subjected to hydrogen nuclear magnetic resonance ( 1 HNMR) detection, the results are as follows:

[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] 2. Animal Clinical Trials

[0245] In the following experiments, 8-week-old KM male mice were selected and maintained at room temperature at 23±1°C and humidity at 50±60%, with a 12-hour light-dark cycle and free access to clean food and water.

[0246] In the following experiments, the alveolar lavage fluid cell count was performed as follows: mix the EP tube containing BALF (alveolar lavage fluid), draw 10 μL of BALF, drop it into the cell counting plate, insert the cell counting plate into the whole cell automatic counter, observe and record the readings.

[0247] In the following experiments, tissue damage analysis was performed according to the following method: the collected lung tissue samples were fixed with freshly prepared 4% paraformaldehyde and then stained with hematoxylin and eosin (H&E); specifically, the paraffin sections of the lung tissue were baked in an oven at 65°C for at least 1.5 hours to ensure that the paraffin on the tissue sections was fully melted; then, the sections were placed in xylene twice to remove the melted paraffin and dehydrated in graded ethanol. The sections were stained with hematoxylin and eosin in turn and then dehydrated 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 magnifications of 100 times and 400 times, respectively.

[0248] In the following experiments, the mRNA level of inflammatory factors in lung tissue was detected according to the following method:

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

[0250] ① After homogenizing the lung tissue, add 750 μL of Trizol reagent and place it on ice for 10 min. Then, repeatedly blow and beat it into a uniform suspension. Suck it into a 1.5 mL RNase-free centrifuge tube, add 150 μL of chloroform, mix well and let it stand for 5 min.

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

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

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

[0254] ⑤ Continue centrifugation at 4°C and 12,000 rpm / min for 5 min. Repeat the previous step and generally wash 2-3 times. After washing, place in a fume hood to dry. When the white precipitate at the bottom of the centrifuge tube becomes transparent, add 20 μL of DEPC water, which is the total RNA of the lung tissue.

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

[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 RNA to be tested and gently drop it onto the measuring probe to measure the concentration, record the measurement results, and observe the OD260 / 280 ratio, which should be 1.8-2.0.

[0258] (3) Reverse transcription of RNA into cDNA (TaKaRa)

[0259] A 20 μL transcription system in the TaKaRa reverse transcription kit was selected for the experiment; the RNA content required for transcription was calculated, specifically including: adding 4 μL 5x PrimeScript RT Master Mix and the required volume of RNA to a 200 μL EP tube, adding water to make up the volume to 16 μL, shaking and mixing, and then centrifuging to the bottom; setting the reaction conditions on the PCR instrument: 37°C, 15 min; 85°C, 5 s; 4°C termination.

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

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

[0262] Table 10 Primer sequences

[0263]

[0264] ② Prepare sterile and enzyme-free pipette tips and 8-well tubes or 96-well plates for PCR;

[0265] ③Prepare the PCR reaction solution in a 0℃ metal bath; then configure the mixing tube according to the instructions based on the genes used in this experiment;

[0266] ④After the sample is loaded, observe whether there is any error in the loading, and then use a centrifuge to centrifuge the 8-tube strip or 96-well plate;

[0267] ⑤Finally, set the reaction program for the 7500fast instrument according to the instruction manual. After the setting is completed, run the sample on the machine;

[0268] ⑥Export data and calculate 2 -ΔΔCT Make bar graphs and perform statistical analysis.

[0269] In the following experiments, data were expressed as mean ± standard deviation (SEM), and the differences between the two groups were evaluated using independent sample T test. All graphs were created by GraphPad Prism 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. They were gavaged with 200 μL of corresponding drugs (30 mM) on days 0, 2, 4, and 6, respectively. The LPS group was gavaged with corn oil. On the 6th day, 50 μL LPS (0.2 g / L) was instilled into the trachea. 72 h after LPS tracheal instillation, BALF (alveolar lavage fluid) was collected for cell counting.

[0272] The cell counting results are as follows Figure 1 As shown, the results showed that compared with the LPS group, the total number of cells in the A2+LPS group was significantly reduced; and other drug treatments did not reduce the total number of cells in BALF; the above indicated that A2 could alleviate the lung cell infiltration caused by LPS.

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

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

[0275] The cell counting results are 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 indicate that A2 can alleviate the lung cell infiltration caused by LPS.

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

[0277] Twenty-four KM mice were divided into LPS group, A1+LPS group, and A2+LPS group. They were gavaged with 100 μL of the corresponding drug (15 mM) on days 0, 2, 4, and 6, respectively. The LPS group was gavaged with corn oil. On the 6th day, 50 μL of LPS (0.2 g / L) was instilled into the trachea. 72 h after LPS tracheal instillation, lung tissues and BALF (alveolar lavage fluid) were collected for cell counting, tissue damage, and inflammatory factor detection.

[0278] The cell counting results are as follows Figure 3 As shown in the figure, compared with 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 shows that A2 can still effectively alleviate the lung cell infiltration caused by ARDS with a reduced dosage, and its effect is better than that of A1.

[0279] Tissue damage 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 in the A1+LPS group after treatment; and compared with the pathological damage of the lungs in the LPS group, it was alleviated in the A2+LPS group. Therefore, the supplementation of A2 can reduce the pathological damage of the lung tissue of mice caused by LPS, and the effect is better than that of A1.

[0280] The mRNA levels of inflammatory factors interleukin-6 (IL-6), tumor necrosis factor-α (TNF-α), interleukin-1β (IL-1β), heme oxygenase 1 (HO-1), and myeloperoxidase (MPO) were shown in Figure 2 . Figure 5 , Figure 6 , Figure 7 , Figure 8 and Fig. 9 As shown in the results, 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 the 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, it is shown that the supplementation of A2 can significantly reduce the mRNA expression of inflammatory factors in the lung tissue of mice induced by LPS, and the therapeutic effect is better than that of A1 (p<0.05*, p<0.01**, p<0.001***).

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

[0282] A total of 66 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. They were given 100 μL of corresponding drugs (15 mM) by gavage on days -3, -2, -1, and 1, respectively. The Control group and LPS group were given corn oil by gavage. On day 0, 50 μL of LPS (0.2 g / L) was instilled into the trachea. 72 hours after tracheal instillation of LPS, lung tissues and BALF (bone alveolar lavage fluid) were collected for cell counting, tissue damage and inflammatory factor detection.

[0283] The cell counting results are as follows Fig.10 As shown in the figure, compared with 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 shows that A5 can effectively alleviate the lung cell infiltration caused by ARDS, and its effect is better than A1.

[0284] The mRNA levels of inflammatory factors interleukin-6 (IL-6), tumor necrosis factor-α (TNF-α), and interleukin-1β (IL-1β) were Fig.11 , Fig.12 and Fig.13 The results showed that the hydroxycinnamic acid derivatives can down-regulate 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 derivatives of the present invention have a relieving effect on LPS-induced ARDS mice. Through drug screening, it was found that the hydroxycinnamic acid derivatives of the present invention can significantly reduce the cell infiltration of ARDS mice, and its effect was further verified by administering 6 times and 4 times. It was found that supplementing the hydroxycinnamic acid derivatives of the present invention can reduce the lung cell infiltration, increased lung inflammation and lung tissue pathological damage caused by LPS, and its effect is better than 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 solution of the present invention rather than to limit it. Although the present invention has been described in detail with reference to the preferred embodiments, those skilled in the art should understand that the technical solution of the present invention can be modified or replaced by equivalents without departing from the purpose and scope of the technical solution of the present invention, which should be covered by the scope of the claims of the present invention.

Claims

1. The structural formula of hydroxycinnamic acid derivatives is shown below. R3 is -COOR4; R2 is selected from -H or alkoxy; R1 is selected from the first alkyl group, or -C=CCOOR6; in, n is an integer, 1≤n≤5; and / or R4, R5 and R6 are independently selected from a second alkyl group, the carbon chain number of the second alkyl group is ≤5; and / or the number of carbon chains in the first alkyl group is ≤15; and / or the number of carbon chains in the alkoxy group is ≤5.

2. The hydroxycinnamic acid derivative according to claim 1, characterized in that The hydroxycinnamic acid derivative is selected from any one of the following compounds:

3. The method for preparing the hydroxycinnamic acid derivative according to claim 1 or 2, characterized in that: Choose one of the following methods: (a) In the hydroxycinnamic acid derivative, R2 is selected from -H, R1 is selected from the first alkyl group, and the carbon chain number of the first alkyl group is ≤5; or R2 is selected from the alkoxy group, R1 is selected from the first alkyl group, and the carbon chain number of the first alkyl group is ≤3; The preparation method of the hydroxycinnamic acid derivative comprises: mixing a compound represented by formula I, a compound represented by formula II, a compound represented by formula III and TsCl to obtain a hydroxycinnamic acid derivative; (b) in the hydroxycinnamic acid derivative, R2 is selected from -H, R1 is selected from the first alkyl group, and the carbon chain number of the first alkyl group is ≥ 6; or R2 is selected from the alkoxy group, R1 is selected from the first alkyl group, and the carbon chain number of the first alkyl group is ≥ 4; The preparation method of the hydroxycinnamic acid derivative comprises: mixing the compound represented by formula I and the compound represented by formula II, then adding an organic base catalyst, and finally adding a condensation agent to react to obtain the hydroxycinnamic acid derivative; (c) In the hydroxycinnamic acid derivative, R2 is selected from -H, R1 is selected from or -C=CCOOR6; The preparation method of the hydroxycinnamic acid derivative comprises: mixing and reacting a compound represented by formula I, a compound represented by formula II, a hydroxyl activator, a catalyst and a condensation agent to obtain the hydroxycinnamic acid derivative; Wherein, the structural formulas of the compound represented by formula I, the compound represented by formula II and the compound represented by formula III are as follows:

4. A pharmaceutical composition for treating acute respiratory distress syndrome, characterized in that: The invention comprises the hydroxycinnamic acid derivative according to claim 1 or 2.

5. A pharmaceutical preparation for treating acute respiratory distress syndrome, characterized in that: The invention comprises the hydroxycinnamic acid derivative according to claim 1 or 2 or the pharmaceutical composition according to claim 4.

6. Use of the hydroxycinnamic acid derivative according to claim 1 or 2, or the pharmaceutical composition according to claim 4, or methyl hydroxycinnamate in the preparation of a pharmaceutical preparation for treating acute respiratory distress syndrome, wherein the structural formula of methyl hydroxycinnamate is:

7. The use according to claim 6, characterized in that: Applications include one or a combination of the following: (i) Use of a hydroxycinnamic acid derivative or a pharmaceutical composition or hydroxycinnamic acid in the preparation of a pharmaceutical preparation for reducing lung cell infiltration; (ii) Use of a hydroxycinnamic acid derivative or a pharmaceutical composition in the preparation of a pharmaceutical preparation for alleviating pathological damage to lung tissue; (iii) Use of a hydroxycinnamic acid derivative or a pharmaceutical composition or methyl hydroxycinnamate in the preparation of a pharmaceutical preparation for alleviating lung inflammation.

8. The use according to claim 7, characterized in that: The application of hydroxycinnamic acid derivatives or methyl hydroxycinnamate in the preparation of drugs for reducing lung inflammation includes: the application of hydroxycinnamic acid derivatives or methyl hydroxycinnamate in the preparation of pharmaceutical preparations for reducing 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.

9. The use according to claim 8, characterized in that: Applications include one or a combination of the following: (a) Use of a first compound in the preparation of a pharmaceutical preparation for reducing lung cell infiltration, wherein the first compound is selected from any one of the following compounds: (b) Use of the second compound in the preparation of a pharmaceutical preparation for alleviating pathological damage to lung tissue, wherein the first compound is selected from the following compounds: (c) Use of a third compound in the preparation of a pharmaceutical preparation for reducing the mRNA expression level of the pulmonary inflammatory factor IL-6, wherein the third compound is selected from any one of the following compounds: (d) Use of a fourth compound in the preparation of a pharmaceutical preparation for reducing the mRNA expression level of the pulmonary inflammatory factor HO-1, wherein the fourth compound is selected from the following compounds: (e) Use of the fifth compound in the preparation of a pharmaceutical preparation for reducing the mRNA expression level of the pulmonary inflammatory factor MPO, wherein the fifth compound is selected from the following compounds: (f) Use of the sixth compound in the preparation of a pharmaceutical preparation for reducing the mRNA expression level of the lung inflammatory factor TNF-α, wherein the sixth compound is selected from any one of the following compounds: (g) Use of the seventh compound in the preparation of a pharmaceutical preparation for reducing the mRNA expression level of the pulmonary inflammatory factor IL-1β, wherein the seventh compound is selected from any one of the following compounds:

10. The use according to any one of claims 6 to 9, characterized in that The dosage forms of the pharmaceutical preparation include oral preparation, aerosol, powder, tablet, paste or injection.

Images