Compounds containing a 3-alkynylpyrrole structure and uses thereof

Abstract

The application discloses a compound containing a 3-alkynyl pyrrole structure and application thereof. The compound containing the 3-alkynyl pyrrole structure provided by the application is confirmed to have a down-regulation effect on inflammatory factors secreted by LPS-induced primary alveolar macrophages through cell experiments, and has no obvious cytotoxicity. The application provides a new treatment strategy for clinical acute lung injury (ALI) treatment.

Description

TECHNICAL FIELD

[0001] The present application belongs to the field of chemical medicine, and relates to a 3-alkynyl pyrrole structure and application. BACKGROUND

[0002] Pulmonary macrophages (PM) are resident cells in the lung, widely distributed in alveoli and bronchi, lung interstitium, pulmonary vessels and pleural cavity, and play a key role in the regulation of inflammatory response. They have physiological characteristics such as adhesion, deformation, migration, chemotaxis, phagocytosis and secretion, and are the first line of defense for the lung. They can phagocytose and kill pathogens such as bacteria and viruses, thereby protecting the lung from exogenous pathogenic invasion. When the lung is infected or damaged, PMs rapidly migrate to the inflammatory site and release inflammatory mediators such as interleukin (IL)-1β, IL-6, tumor necrosis factor (TNF-α) and the like, causing a typical inflammatory response. These mediators can cause vasodilation and increased permeability, thereby helping to clear pathogens and damaged cells. However, excessive inflammation can cause tissue damage, leading to acute lung injury and even type 1 respiratory failure and death. Since PM plays a key role in the regulation of inflammatory response, the development of inhibitors of macrophage inflammatory factor secretion is of great significance for the treatment of acute lung injury (ALI) in clinic.

[0003] The traditional treatment methods for acute lung injury mainly rely on supportive treatments such as mechanical ventilation, fluid management, and treatments for specific causes (such as anti-infection treatment). However, these methods cannot completely solve the pathophysiological process of acute lung injury. By inhibiting the secretion of inflammatory factors by PM, the progress of the inflammatory response can be controlled at the source, thereby providing strong support for the development of new treatment strategies. SUMMARY

[0004] The present application aims to solve the above problems in the prior art, and provides a new medical use of 3-alkynyl pyrrole structure-containing compounds. The 3-alkynyl pyrrole structure small molecule compounds have been confirmed by cell experiments to have a significant inhibitory effect on the secretion of inflammatory factors by pulmonary macrophages induced by lipopolysaccharide (LPS), and have no obvious cytotoxicity. These compounds and their pharmaceutical compositions can be used for the prevention or treatment of pulmonary inflammatory diseases such as acute lung injury and acute respiratory distress syndrome.

[0005] To achieve the above object, the technical scheme adopted by the present application to solve its technical problems is:

[0006] The application of a compound containing a 3-alkynylpyrrole structure in the preparation of an inhibitor of alveolar macrophage inflammatory factor secretion; the general structural formula of this type of compound is:

[0007]

[0008] The R substituent is one of -H, -OCH3, -CN, -Ph, and -COOCH3.

[0009] Furthermore, the compound is used to prepare a drug for the prevention and / or treatment of acute lung injury.

[0010] Furthermore, the compound is used to prepare drugs for the prevention and / or treatment of inflammatory lung diseases.

[0011] More specifically, the compounds are selected from the following structures:

[0012]

[0013] A drug that can effectively inhibit the secretion of PM inflammatory factors includes the above-mentioned compounds or their pharmaceutically acceptable salts or cocrystals, deuterated compounds, solvates, and enantiomers.

[0014] The inhibitor or drug may be in the form of tablets, pills, powders, capsules, injections, oral liquids, ointments, or creams, and all dosage forms can be prepared according to conventional methods in the pharmaceutical field. The administration route may be oral, injection, or topical.

[0015] The inhibitor or drug comprises an active ingredient containing a 3-alkynylpyrrole structure and medically acceptable pharmaceutical excipients. The excipients include pharmaceutically conventional diluents, excipients, fillers, binders, humectants, disintegrants, absorption enhancers, surfactants, adsorbents, lubricants, etc.

[0016] The beneficial effects of this invention are as follows:

[0017] This invention discloses a small molecule compound containing a 3-alkynylpyrrole structure, which, through cell experiments, has been shown to significantly inhibit the secretion of inflammatory factors by lipopolysaccharide (LPS)-induced alveolar macrophages, without significant cytotoxicity. Therefore, these compounds or their pharmaceutical compositions hold promise for the prevention or treatment of inflammatory lung diseases, such as acute lung injury and acute respiratory distress syndrome.

[0018] It should be understood that the above general description and the following detailed description are exemplary and explanatory only, and do not limit this application. Attached Figure Description

[0019] The accompanying drawings, which are incorporated in and form part of this specification, illustrate embodiments consistent with this application and, together with the specification, serve to explain the principles of this application.

[0020] Figure 1 It is the diethyl-3-(phenylethynyl)-1 given in compound H1 of example. H -Pyrrole-2,4-dicarboxylate 1H NMR spectrum.

[0021] Figure 2 The compound H2 in the examples is diethyl 3-((4-methoxyphenyl)ethynyl)-1 H -Pyrrole-2,4-dicarboxylate 1H NMR spectrum.

[0022] Figure 3 It is the diethyl 3-((4-cyanophenyl)ethynyl)-1 given by compound H3 in the examples. H -Pyrrole-2,4-dicarboxylate 1H NMR spectrum.

[0023] Figure 4 It is the diethyl 3-([1,1'-biphenyl]-4-ylethynyl)-1 given by compound H4 in example. H -Pyrrole-2,4-dicarboxylate 1H NMR spectrum.

[0024] Figure 5 It is the diethyl 3-((4-(methoxycarbonyl)phenyl)ethynyl)-1 given by compound H5 in example. H -Pyrrole-2,4-dicarboxylate 1H NMR spectrum.

[0025] Figure 6 It is the diethyl-3-(phenylethynyl)-1 given in compound H1 of example. H 2,4-pyrrole-2,4-dicarboxylate NMR carbon spectrum.

[0026] Figure 7 The compound H2 in the examples is diethyl 3-((4-methoxyphenyl)ethynyl)-1 H 2,4-pyrrole-2,4-dicarboxylate NMR carbon spectrum.

[0027] Figure 8 It is the diethyl 3-((4-cyanophenyl)ethynyl)-1 given by compound H3 in the examples. H 2,4-pyrrole-2,4-dicarboxylate NMR carbon spectrum.

[0028] Figure 9 It is the diethyl 3-([1,1'-biphenyl]-4-ylethynyl)-1 given by compound H4 in example. H 2,4-pyrrole-2,4-dicarboxylate NMR carbon spectrum.

[0029] Figure 10 It is the diethyl 3-((4-(methoxycarbonyl)phenyl)ethynyl)-1 given by compound H5 in example. H 2,4-pyrrole-2,4-dicarboxylate NMR carbon spectrum.

[0030] Figure 11 ELISA assay was used to detect the effects of H1-H5 series compounds on LPS-induced secretion of inflammatory factors in primary PM cells. Detailed Implementation

[0031] The present invention will be further described below with reference to embodiments and accompanying drawings. The following embodiments are merely illustrative and not intended to limit the invention in any way. The preparation methods of the small molecule compounds H1-H5 containing the 3-alkynylpyrrole structure involved in the embodiments of the present invention can be found in CN113248418A. Their 1H NMR and 1C NMR spectra are as follows: Figures 1-10 As shown.

[0032] Example 1: Diethyl-3-(phenylethynyl)-1 H -Pyrrole-2,4-dicarboxylate H1

[0033]

[0034] 1 H NMR (400 MHz, CDCl3) δ 9.80 (s, 1H), 7.58 - 7.54 (m, 3H), 7.37 -7.33 (m, 3H), 4.42 (q, J = 7.2 Hz, 2H), 4.35 (q, J = 7.2 Hz, 2H), 1.43 (t, J = 7.2Hz, 3H), 1.38 (t, J = 7.2 Hz, 3H). 13 C NMR (100 MHz, CDCl3) δ 163.3, 160.6,131.6, 128.3, 127.0, 125.7, 123.8, 119.5, 111.0, 96.2, 82.1, 61.3, 60.3,14.43, 14.41; ESI-MS: m / z = 312.1245 [M+H] + .

[0035] Example 2: Diethyl-3-((4-methoxyphenyl)ethynyl)-1 H -Pyrrole-2,4-dicarboxylate H2

[0036]

[0037] 1 H NMR (400 MHz, CDCl3) δ 9.71 (s, 1H), 7.55 - 7.50 (m, 3H), 6.88 (d, J = 8.4 Hz, 2H), 4.41 (q, J = 7.2 Hz, 2H), 4.36 (q, J = 7.2 Hz, 2H), 3.83 (s, 3H), 1.43 (t, J = 7.2 Hz, 3H), 1.38 (t, J = 7.2 Hz, 3H). 13 C NMR (100 MHz, CDCl3) δ163.3, 160.6, 159.7, 133.1, 126.9, 125.4, 115.9, 114.0, 111.4, 110.4, 96.4,80.8, 61.2, 60.3, 55.3, 14.5, 14.4; ESI-MS: m / z = 342.1331 [M+H] + .

[0038] Example 3: Diethyl-3-((4-cyanophenyl)ethynyl)-1 H -pyrrole-2,4-dicarboxylate H3

[0039]

[0040] 1 H NMR (400 MHz, CDCl3) δ 9.72 (s, 1H), 7.64 (s, 4H), 7.56 (d, J = 3.2Hz, 1H), 4.42 (q, J = 7.2 Hz, 2H), 4.35 (q, J = 7.2 Hz, 2H), 1.44 (t, J = 7.2 Hz, 3H), 1.38 (t, J = 7.2 Hz, 3H). 13C NMR (100 MHz, CDCl3) δ 126.9, 160.1, 132.0,132.0, 128.7, 126.9, 126.2, 119.6, 118.6, 111.4, 110.0, 94.2, 86.6, 61.4,60.4, 14.44, 14.43; ESI-MS: m / z = 337.1179 [M+H] + .

[0041] Example 4: Diethyl-3-([1,1'-biphenyl]-4-ylethynyl)-1 H -Pyrrole-2,4-dicarboxylate H4

[0042]

[0043] 1 H NMR (400 MHz, CDCl3) δ 9.71 (s, 1H), 7.65 - 7.55 (m, 7H), 7.45 (t, J = 7.6 Hz, 2H), 7.36 (t, J = 7.2 Hz, 1H), 4.43 (q, J = 7.2 Hz, 2H), 4.37 (q, J =7.2 Hz, 2H), 1.45 (t, J = 7.2 Hz, 3H), 1.40 (t, J = 7.2 Hz, 3H). 13 C NMR (100 MHz, CDCl3) δ 163.3, 160.6, 141.0, 140.4, 132.0, 128.9, 127.6, 127.0, 127.0,125.7, 122.7, 119.5, 111.0, 96.1, 82.8, 61.3, 60.3, 14.48, 14.47; ESI-MS: m / z= 388.1540 [M+H] + .

[0044] Example 5: Diethyl-3-((4-(methoxycarbonyl)phenyl)ethynyl)-1 H -Pyrrole-2,4-dicarboxylate H5

[0045]

[0046] 1H NMR (400 MHz, CDCl3) δ 9.73 (s, 1H), 8.02 (d, J = 8.4 Hz, 2H), 7.63(d, J = 8.4 Hz, 2H), 7.56 (d, J = 3.2 Hz, 1H), 4.42 (q, J = 7.2 Hz, 2H), 4.36 (q, J =7.2 Hz, 2H), 3.93 (s, 3H), 1.42 (t, J = 7.2 Hz, 3H), 1.38 (t, J = 7.2 Hz, 3H). 13 C NMR (100 MHz, CDCl3) δ 166.7, 163.1, 160.3, 131.5, 129.5, 129.5,128.5, 126.9, 126.1, 119.6, 110.4, 95.2, 85.2, 61.4, 60.4, 52.3, 14.44,14.41; ESI-MS: m / z = 370.1300 [M+H] + .

[0047] Example 6: Evaluation of the cytotoxicity of the H1-H5 series compounds and their inhibitory effect on LPS-induced secretion of inflammatory factors in primary mouse PM.

[0048] Experimental animals: C57BL / 6J mice, male, 8 weeks old. They were housed in an SPF environment at a temperature of 20-25℃, relative humidity of 30-70%, 12-hour light / dark cycle, and with free access to water and food.

[0049] The specific experiment is as follows:

[0050] Assay 1: Detection of the cytotoxicity of H1-H5 series compounds against primary mouse PM using the CCK-8 assay.

[0051] (1) Isolation and culture of primary mouse PM

[0052] C57BL / 6J mice were euthanized by cervical dislocation and sterilized with 70% alcohol for 3 minutes. The mice were then placed in a 10cm sterile culture dish. Under aseptic conditions, a small incision was made in the skin at the midline of the neck using ophthalmic forceps to separate the muscles around the trachea and expose it. A sterile ligature was placed below the trachea, and a flat needle was inserted into the trachea to ligate it. 1ml of serum-free high-glucose culture medium was injected into the lungs. After three aspirations, bronchoalveolar lavage fluid was collected. This was repeated three times, and 2.5-3ml was collected. Cell pellet was collected at 1000rpm for 5 minutes and washed once with PBS. Finally, cells were reselected with PM culture medium (high-glucose culture medium containing 20% ​​fetal bovine serum), counted, and the cell concentration was adjusted to 5*102. 5 / ml was used for plating. After static incubation in an incubator for 3-4 hours, the supernatant was discarded, and the culture wells were repeatedly rinsed 3 times with HANK fluid to remove non-adhesive cells. At this point, the adherent cells were PM.

[0053] (2) Detection of the cytotoxicity of H1-H5 series compounds to mouse primary PM by CCK-8 assay

[0054] Primary mouse PM cells were seeded onto 96-well plates using the method described above. Each plate contained six groups: one control group (containing only dimethyl sulfoxide as the drug solvent) and five drug-treated groups (containing 0.3125, 0.625, 1.25, 2.5, and 5 μM of compounds H1-H5, respectively, with dimethyl sulfoxide as the drug solvent). Each group had five replicates. The culture medium was PM growth medium, and the cells were cultured for 24, 48, and 72 hours, with the medium changed every other day. At the corresponding time points, 10 μL of CCK-8 reagent was added to each well, and the cells were incubated in the dark for 1 hour. The cells were then removed, shaken thoroughly on a microplate reader, and the absorbance at 450 nm was measured (the higher the number of viable cells, the higher the absorbance). The IC50 values ​​of each compound for the toxicity to primary mouse PM cells were determined. 50 As shown in Table 1.

[0055] Table 1. IC50 of the cytotoxic effects of compounds H1-H5 on primary PM cells in mice 50

[0056] Examples Assay 1 : Cytotoxicity (nM) H1 1526.78±19.23 H2 2142.91±16.91 H3 1852.15±18.58 H4 2145.58±18.24 H5 2654.95±12.49

[0057] Small molecule compounds H1-H5 containing 3-alkynylpyrrole structure showed no significant cytotoxicity to primary mouse PM cells.

[0058] Assay 2: The effects of H1-H5 series compounds on the function of LPS-induced secretion of inflammatory factors in primary mouse PM were detected by ELISA.

[0059] The above-mentioned primary PM mice were administered at a rate of 5 × 10⁶. 6Cells were seeded in 24-well plates and starved for 8 hours in serum-free high-glucose medium after cell attachment. A gradient concentration of candidate compounds was added, with the final dose determined according to Test 1, using six non-cytotoxic drug doses: 5, 10, 50, 100, 500, and 1000 nM. After 30 min of treatment, 1 μg / ml of LPS was added for 24 h of stimulation. Cell supernatant was then collected for ELISA testing of IL-1β, IL-6, and TNF-α secretion levels (ELISA kit from Linke Biotechnology). Results showed that LPS-induced levels of TNF-α, IL-6, and IL-1β in the mouse primary PM supernatant were significantly higher than in the control group. Compounds H1-H5 series dose-dependently reduced the LPS-induced increase in TNF-α, IL-6, and IL-1β levels. Figure 11 In the figure, #, P<0.05 vs control group; *, P<0.05; **, P<0.01 vs LPS group, N=3).

[0060] The above descriptions are merely some preferred embodiments of the present invention, used to help illustrate the invention. The scope of protection of the present invention is not limited to the above embodiments; all technical solutions falling within the scope of the present invention's concept are within its protection.

Claims

1. The application of compounds containing a 3-alkynylpyrrole structure in the preparation of inhibitors of alveolar macrophage inflammatory factor secretion, characterized in that, The structural formula of the compound containing the 3-alkynylpyrrole structure is shown below: ； The R is one of -H, -OCH3, -CN, -Ph, and -COOCH3.

2. The application as described in claim 1, characterized in that, The compound is used to prepare drugs for the prevention and / or treatment of acute lung injury.

3. The application as described in claim 1, characterized in that, The compound is used to prepare drugs for the prevention and / or treatment of inflammatory lung diseases.

4. The application as described in any one of claims 1-3, characterized in that, The compound is any one of the following formulas: 。 5. The application according to any one of claims 1-3, characterized in that: The inhibitor or drug is in the form of tablets, pills, powders, capsules, injections, oral liquids, ointments, or creams, and is administered orally, by injection, or topically.

6. The application according to any one of claims 1-3, characterized in that: The inhibitor or drug comprises an active ingredient being the compound containing a 3-alkynylpyrrole structure and a medically acceptable pharmaceutical excipient.

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