Long-chain fatty acyl monoterpenoids, their preparation methods and uses in the preparation of anti-complement drugs and anti-viral pneumonia drugs

Abstract

The present invention relates to a long-chain fatty acyl monoterpenoid compound, a preparation method thereof, and uses thereof in the preparation of anti-complement drugs and anti-viral pneumonia drugs. The long-chain fatty acyl monoterpenoid compound has a chemical structure with the following general structural formula: The R group is hexadecyl or tetradecyl. Through in vitro anti-complement activity tests, the results show that the above-mentioned long-chain fatty acyl monoterpenoid compound has a strong inhibitory effect on the classical pathway of the complement system (see Table 1). (3S,5R,6S,7E)-3-tetradecanoate-5,6-epoxy-β-ionone has been confirmed through in vivo animal experiments to have a good therapeutic effect on viral pneumonia in mice induced by influenza A virus H1N1.

Description

Technical Field

[0001] The present invention relates to the field of traditional Chinese medicine pharmacy, and particularly relates to a long-chain fatty acyl monoterpenoid compound, a preparation method thereof, and uses thereof in preparing a complement inhibitor and an anti-viral pneumonia drug. Background Art

[0002] The complement system, as an important part of the body's immune system, participates in the body's specific and non-specific immune mechanisms. It plays an important role in the body's anti-microbial defense response, dissolution of immune complexes, enhancement of humoral immune response, and mediation of the damaging response of immunopathology. If the complement system is over-activated, it will not only consume a large amount of complement components, reducing the body's anti-infection ability, but also produce a large number of bioactive substances, causing a serious inflammatory response in the body, thus resulting in a pathological damage process. For example, the development process of infectious diseases such as influenza is closely related to the over-activation of complement. Research has found that the direct cell damage caused by the host's viral infection is only one of its pathogenic mechanisms, while the excessive inflammatory response and immune homeostasis imbalance caused by the host's infection and damage, and the resulting cytokine storm are the key factors leading to severe pneumonia and even death. Therefore, regulating the body's immune response to treat viral infectious pneumonia, especially in the prevention and treatment of severe pneumonia, has important clinical significance. Therefore, it is of great value and significance to search for highly effective and low-toxic complement inhibitors from traditional Chinese medicine to treat these diseases related to the over-activation of the complement system.

[0003] Fructus Trichosanthis Pericarpium is the dried and mature pericarp of the plant Trichosanthes kirilowii Maxim. or Trichosanthes rosthornii Harms of the family Cucurbitaceae. The main effects of Fructus Trichosanthis Pericarpium are clearing away heat and resolving phlegm, promoting qi circulation and relieving chest distress, and it is mainly used clinically to treat cough with yellow phlegm and chest distress and hypochondriac pain. Modern pharmacological research shows that Fructus Trichosanthis Pericarpium has various effects such as improving the cardiovascular system, relieving cough and reducing sputum, anti-inflammatory, antibacterial, and antioxidant effects. Summary of the Invention

[0004] The object of the present invention is to provide a long-chain fatty acyl monoterpenoid compound, a preparation method thereof, and uses thereof in preparing a complement inhibitor and an anti-viral pneumonia drug for a compound with anti-complement activity and the treatment of viral pneumonia.

[0005] The object of the present invention can be achieved by the following technical solutions:

[0006] When the inventor was screening the in vitro anti-complement activity of the ethanol extract of Trichosanthes kirilowii Maxim. pericarp, it was found that the ethanol extract of Trichosanthes kirilowii Maxim. pericarp had significant anti-complement activity. Therefore, an activity-guided separation method was adopted to isolate two long-chain fatty acyl monoterpenoid compounds with good anti-complement activity from Trichosanthes kirilowii Maxim. pericarp. More excitingly, during the research process, the inventor found that one of the long-chain fatty acyl monoterpenoid compounds also had good antiviral pneumonia effect.

[0007] One of the purposes of the present invention is a long-chain fatty acyl monoterpenoid compound, and the long-chain fatty acyl monoterpenoid compound has a chemical structure with the following general formula:

[0008]

[0009] The R group is hexadecyl or tetradecyl.

[0010] Furthermore, the R group is hexadecyl, and the long-chain fatty acyl monoterpenoid compound is trichosanate A.

[0011] Furthermore, the R group is tetradecyl, and the long-chain fatty acyl monoterpenoid compound is (3S,5R,6S,7E)-3-tetradecanoate-5,6-epoxy-β-ionone.

[0012] Combined Figure 1 , trichosanate A is compound 1, and (3S,5R,6S,7E)-3-tetradecanoate-5,6-epoxy-β-ionone is compound 2.

[0013] Compound 1, trichosanate A: colorless crystal; molecular formula: C 29 H 50 O 4 ; relative molecular weight: 462; 1 H NMR(400MHz,CD 3OD), δ: 7.16 (1H, d, J = 15.8 Hz, H-7), 6.22 (1H, d, J = 15.8 Hz, H-8), 4.91 (1H, m, H-3), 2.37 (1H, dd, J = 14.9, 5.5 Hz, H-4a), 2.30 (3H, s, H-10), 2.28 (2H, m, H-2'), 1.86 (1H, dd, J = 14.8, 7.6 Hz, H-4b), 1.64 (1H, dd, J = 13.3, 2.4 Hz, H-2a), 1.62 (2H, m, H-3'), 1.42 (1H, dd, J = 13.3, 9.2 Hz, H-2b), 1.29 (24H, m, H-4'–H-15'), 1.23 (3H, s, H-11), 1.19 (3H, s, H-13), 0.99 (3H, s, H-12), 0.90 (3H, t, J = 7.0 Hz, H-16'); 13 C NMR (100 MHz, CD 3 OD) δ: 200.1 (C-9), 174.9 (C-1'), 144.4 (C-7), 134.1 (C-8), 71.0 (C-6), 68.5 (C-3), 67.7 (C-5), 42.6 (C-2), 37.5 (C-4), 35.7 (C-1), 35.4 (C-2'), 33.1 (C-14'), 30.1–30.8 (C-4'–C-13'), 28.9 (C-11), 27.5 (C-10), 26.0 (C-3'), 25.7 (C-12), 23.7 (C-15'), 20.1 (C-13), 14.4 (C-16').

[0014] Compound 2, (3S,5R,6S,7E)-3-tetradecanoate-5,6-epoxy-β-ionone: colorless crystal; molecular formula: C 27 H 46 O 4 ; relative molecular weight: 434; 1 H NMR (400 MHz, CD 3OD), δ: 7.16 (1H, d, J = 15.8 Hz, H-7), 6.22 (1H, d, J = 15.8 Hz, H-8), 4.91 (1H, m, H-3), 2.37 (1H, dd, J = 14.9, 5.6 Hz, H-4a), 2.30 (3H, s, H-10), 2.28 (2H, m, H-2'), 1.86 (1H, dd, J = 14.8, 7.6 Hz, H-4b), 1.64 (1H, dd, J = 13.3, 2.4 Hz, H-2a), 1.62 (2H, m, H-3'), 1.42 (1H, dd, J = 13.3, 9.2 Hz, H-2b), 1.29 (20H, m, H-4'–H-13'), 1.23 (3H, s, H-11), 1.19 (3H, s, H-13), 0.99 (3H, s, H-12), 0.90 (3H, t, J = 7.0 Hz, H-14'); 13 C NMR (100 MHz, CD 3 OD) δ: 200.1 (C-9), 174.9 (C-1'), 144.4 (C-7), 134.1 (C-8), 71.0 (C-6), 68.5 (C-3), 67.7 (C-5), 42.6 (C-2), 37.5 (C-4), 35.7 (C-1), 35.4 (C-2'), 33.1 (C-12'), 30.1–30.8 (C-4'–C-11'), 28.9 (C-11), 27.5 (C-10), 26.0 (C-3'), 25.7 (C-12), 23.7 (C-13'), 20.1 (C-13), 14.4 (C-14').

[0015] The second object of the present invention is a method for preparing the long-chain fatty acyl monoterpenoid compound as described above, and the preparation method comprises the following steps:

[0016] After pulverizing the pericarp of Trichosanthes kirilowii Maxim., percolation extraction is carried out. After concentrating the extraction solution, it is suspended in water and extracted successively with petroleum ether, ethyl acetate, and n-butanol to obtain a petroleum ether extract, an ethyl acetate extract, and an n-butanol extract;

[0017] The petroleum ether extract is taken and separated by silica gel column chromatography, eluted with a petroleum ether-ethyl acetate gradient to obtain 5 fractions A1–A5; the fraction A2 is separated by silica gel column chromatography, eluted with a n-hexane-ethyl acetate gradient to obtain 12 fractions A2A–A2L; the fraction A2F is separated by reverse-phase ODS column chromatography, eluted with a methanol-water gradient to obtain 12 fractions A2F1–A2F12.

[0018] Fraction A2F8 was purified by semi-preparative liquid phase, and the compounds trichosanate A and (3S,5R,6S,7E)-3-tetradecanoate-5,6-epoxy-β-ionone were obtained by isocratic elution with methanol-water as the mobile phase.

[0019] Furthermore, the specific operation steps of the preparation method are as follows:

[0020] After mechanically pulverizing the pericarp of Trichosanthes kirilowii Maxim., percolation extraction was carried out with 95% (v / v) ethanol. After concentrating the extract, it was suspended in water and successively extracted with petroleum ether, ethyl acetate, and n-butanol to obtain a petroleum ether extract, an ethyl acetate extract, and an n-butanol extract;

[0021] The petroleum ether extract was separated by silica gel column chromatography and eluted with a gradient of petroleum ether-ethyl acetate (1:0 → 0:1, v / v) to obtain 5 fractions A1–A5; Fraction A2 was separated by silica gel column chromatography and eluted with a gradient of n-hexane-ethyl acetate (100:0 → 1:10, v / v) to obtain 12 fractions A2A–A2L; Fraction A2F was separated by reversed-phase ODS column chromatography and eluted with a gradient of methanol-water (30:70 → 100:0, v / v) to obtain 12 fractions A2F1–A2F12.

[0022] Fraction A2F8 was purified by semi-preparative liquid phase, and the compounds trichosanate A and (3S,5R,6S,7E)-3-tetradecanoate-5,6-epoxy-β-ionone were obtained by isocratic elution with methanol-water (97:3, v / v) as the mobile phase. Among them, the silica gel mesh number in silica gel column chromatography was 200-300 mesh.

[0023] The third object of the present invention lies in the use of a long-chain fatty acyl monoterpenoid compound as described above in the preparation of anti-complement drugs.

[0024] Furthermore, the long-chain fatty acyl monoterpenoid compound inhibits the classical pathway of the complement system.

[0025] The fourth object of the present invention lies in the use of a long-chain fatty acyl monoterpenoid compound as described above in the preparation of anti-viral pneumonia drugs.

[0026] Furthermore, the long-chain fatty acyl monoterpenoid compound reduces pulmonary pathological changes, alleviates pulmonary inflammatory reactions, and ultimately treats acute lung injury.

[0027] The present invention applies modern pharmacological research methods to test the anti-complement activity of the isolated long-chain fatty acyl monoterpenoid compounds. Two long-chain fatty acyl monoterpenoid components, namely trichosanate A and (3S,5R,6S,7E)-3-tetradecanoate-5,6-epoxy-β-ionone, were isolated from the petroleum ether extraction part of the ethanol extract of the dried ripe pericarp of the Cucurbitaceae plant Trichosanthes kirilowii Maxim. The in vitro anti-complement activity test of trichosanate A and (3S,5R,6S,7E)-3-tetradecanoate-5,6-epoxy-β-ionone showed that the above-mentioned long-chain fatty acyl monoterpenoid compounds had a strong inhibitory effect on the classical pathway of the complement system (see Table 1). The in vivo experiment of (3S,5R,6S,7E)-3-tetradecanoate-5,6-epoxy-β-ionone confirmed that it had a good therapeutic effect on murine viral pneumonia induced by influenza A virus H1N1. BRIEF DESCRIPTION OF THE DRAWINGS

[0028] Figure 1 Flow chart for the extraction and isolation of long-chain fatty acyl monoterpenoid compounds 1 and 2 from the pericarp of Trichosanthes kirilowii Maxim.

[0029] Figure 2 Effect of compound 2 on the body weight of mice with viral pneumonia

[0030] Figure 3 Effect of compound 2 on the lung index of mice with viral pneumonia

[0031] Figure 4 Effect of compound 2 on the pathological damage of the lungs of mice with viral pneumonia

[0032] Figure 5 Effect of compound 2 on the inflammatory factors in the lungs of mice with viral pneumonia DETAILED DESCRIPTION OF THE INVENTION

[0033] The present invention will be described in detail below with reference to the drawings and specific embodiments. These embodiments are implemented on the premise of the technical solution of the present invention, and detailed implementation manners and specific operation processes are given, but the protection scope of the present invention is not limited to the following embodiments.

[0034] A long-chain fatty acyl monoterpenoid compound, which has a chemical structure with the following general formula:

[0035]

[0036] The R group is hexadecyl or tetradecyl.

[0037] Example

[0038] I. Extracting and isolating long-chain fatty acyl monoterpenoid compounds 1 and 2 from Trichosanthis Fructus Peel

[0039] As Figure 1 shown, take 31.0 kg of Trichosanthis Fructus Peel medicinal materials. After mechanical pulverization, percolation extraction is carried out with 95% (v / v) ethanol. After concentrating the extraction solution, it is suspended in water and extracted successively with petroleum ether, ethyl acetate, and n-butanol to obtain petroleum ether extract, ethyl acetate extract, and n-butanol extract. Among them, the petroleum ether extract is 611 g;

[0040] Take 600 g of the petroleum ether extract and separate it by silica gel column chromatography, eluting with a gradient of petroleum ether - ethyl acetate (1:0 → 0:1, v / v) to obtain 5 fractions A1–A5; Fraction A2 is separated by silica gel column chromatography, eluting with a gradient of n-hexane - ethyl acetate (100:0 → 1:10, v / v) to obtain 12 fractions A2A–A2L; Fraction A2F is separated by reverse-phase ODS column chromatography, eluting with a gradient of methanol - water (30:70 → 100:0, v / v) to obtain 12 fractions A2F1–A2F12.

[0041] Fraction A2F8 is purified by semi-preparative liquid phase, eluting isocratically with methanol - water (97:3, v / v) as the mobile phase to obtain compound trichosanate A and (3S,5R,6S,7E)-3-tetradecanoate-5,6-epoxy-β-ionone.

[0042] Among them, the silica gel mesh number in silica gel column chromatography is 200 - 300 mesh.

[0043] Combined with Figure 1 , trichosanate A is compound 1, and (3S,5R,6S,7E)-3-tetradecanoate-5,6-epoxy-β-ionone is compound 2.

[0044] Compound 1, trichosanate A: colorless crystal; molecular formula: C 29 H 50 O 4 ; relative molecular weight: 462; 1 H NMR (400 MHz, CD 3OD), δ: 7.16 (1H, d, J = 15.8 Hz, H-7), 6.22 (1H, d, J = 15.8 Hz, H-8), 4.91 (1H, m, H-3), 2.37 (1H, dd, J = 14.9, 5.5 Hz, H-4a), 2.30 (3H, s, H-10), 2.28 (2H, m, H-2'), 1.86 (1H, dd, J = 14.8, 7.6 Hz, H-4b), 1.64 (1H, dd, J = 13.3, 2.4 Hz, H-2a), 1.62 (2H, m, H-3'), 1.42 (1H, dd, J = 13.3, 9.2 Hz, H-2b), 1.29 (24H, m, H-4'–H-15'), 1.23 (3H, s, H-11), 1.19 (3H, s, H-13), 0.99 (3H, s, H-12), 0.90 (3H, t, J = 7.0 Hz, H-16'); 13 C NMR (100 MHz, CD 3 OD) δ: 200.1 (C-9), 174.9 (C-1'), 144.4 (C-7), 134.1 (C-8), 71.0 (C-6), 68.5 (C-3), 67.7 (C-5), 42.6 (C-2), 37.5 (C-4), 35.7 (C-1), 35.4 (C-2'), 33.1 (C-14'), 30.1–30.8 (C-4'–C-13'), 28.9 (C-11), 27.5 (C-10), 26.0 (C-3'), 25.7 (C-12), 23.7 (C-15'), 20.1 (C-13), 14.4 (C-16').

[0045] Compound 2, (3S,5R,6S,7E)-3-tetradecanoate-5,6-epoxy-β-ionone: colorless crystal; molecular formula: C 27 H 46 O 4 ; relative molecular weight: 434; 1 H NMR (400 MHz, CD 3OD), δ: 7.16 (1H, d, J = 15.8 Hz, H-7), 6.22 (1H, d, J = 15.8 Hz, H-8), 4.91 (1H, m, H-3), 2.37 (1H, dd, J = 14.9, 5.6 Hz, H-4a), 2.30 (3H, s, H-10), 2.28 (2H, m, H-2'), 1.86 (1H, dd, J = 14.8, 7.6 Hz, H-4b), 1.64 (1H, dd, J = 13.3, 2.4 Hz, H-2a), 1.62 (2H, m, H-3'), 1.42 (1H, dd, J = 13.3, 9.2 Hz, H-2b), 1.29 (20H, m, H-4'–H-13'), 1.23 (3H, s, H-11), 1.19 (3H, s, H-13), 0.99 (3H, s, H-12), 0.90 (3H, t, J = 7.0 Hz, H-14'); 13 C NMR (100 MHz, CD 3 OD) δ: 200.1 (C-9), 174.9 (C-1'), 144.4 (C-7), 134.1 (C-8), 71.0 (C-6), 68.5 (C-3), 67.7 (C-5), 42.6 (C-2), 37.5 (C-4), 35.7 (C-1), 35.4 (C-2'), 33.1 (C-12'), 30.1–30.8 (C-4'–C-11'), 28.9 (C-11), 27.5 (C-10), 26.0 (C-3'), 25.7 (C-12), 23.7 (C-13'), 20.1 (C-13), 14.4 (C-14').

[0046] II. In vitro anti-complement classical pathway test

[0047] Take 0.1 mL of complement prepared from guinea pig serum and add it to barbital buffer solution (BBS) to prepare a 1:10 (v / v) solution. Then, serially dilute it with BBS to 1:20 (v / v), 1:40 (v / v), 1:80 (v / v), 1:160 (v / v), 1:320 (v / v), 1:640 (v / v), and 1:1280 (v / v) solutions. Take 0.1 mL of 1:1000 (v / v) hemolysin, complement at each concentration, and 2% sheep red blood cells (SRBC), and dissolve them in 0.3 mL of BBS. Mix well, place in a water bath at 37 °C for 30 min, then put it into a low-temperature high-speed centrifuge and centrifuge at 5000 rpm and 4 °C for 10 min. Take 0.2 mL of the supernatant from each tube and place it in a 96-well plate, and measure the absorbance at 405 nm. At the same time, set up a complete hemolysis group (0.1 mL of 2% SRBC dissolved in 0.5 mL of distilled water) in the experiment. Take the absorbance of the distilled water lysed blood vessel as the complete hemolysis standard and calculate the hemolysis rate. Plot the complement dilution on the X-axis and the hemolysis percentage on the Y-axis. Select the lowest complement concentration that reaches a similar high hemolysis rate as the critical complement concentration required to ensure normal hemolysis of the system. Mix the complement at the critical concentration with the test samples at different concentrations, add an appropriate amount of BBS, hemolysin, and 2% SRBC, place in a water bath at 37 °C for 30 min, then put it into a low-temperature high-speed centrifuge, centrifuge at 5000 rpm and 4 °C for 10 min, and then take 0.2 mL of the supernatant from each tube and place it in a 96-well plate, and measure the absorbance at 405 nm. At the same time, set up a test sample control group, a complement group, and a complete hemolysis group in the experiment. After subtracting the absorbance value of the corresponding test sample control group from the absorbance value of the test sample, calculate the hemolysis rate. Plot the test sample concentration on the X-axis and the hemolysis inhibition rate on the Y-axis, and calculate the concentration of the test sample required to inhibit 50% hemolysis (CH 50 ). The results are shown in Table 1.

[0048] Table 1. Inhibitory effects of Compounds 1 and 2 on the classical pathway of the complement system (Mean±SD, n = 3)

[0049]

[0050] Determined by in vitro anti-complement activity assay, the results show that the long-chain fatty acyl monoterpenoid compounds 1 and 2 extracted and isolated in this example have strong inhibitory effects on the classical pathway of the complement system, and the minimum test sample concentrations required for 50% hemolysis are 432.9 μg / mL and 206.9 μg / mL, respectively.

[0051] III. Pharmacodynamic experiment of Compound 2 in the treatment of viral pneumonia

[0052] Thirty-two male BALB / c mice, weighing 14–15 g, were randomly divided into 4 groups according to body weight: a normal control group (abbreviated as Normal), a model group (abbreviated as Model), a compound 2 group (abbreviated as C2), and a positive drug group (the positive drug was oseltamivir, abbreviated as Oseltamivir), with 8 mice in each group.

[0053] All experimental animals were anesthetized with isoflurane gas and then infected by nasal drip with 30 μL of 2LD 50 H1N1 virus solution. In the normal group, 30 μL of 1640 medium was used for nasal drip infection as a control, and the other 3 groups were infected with H1N1 virus solution. Two hours after infection, intragastric administration was carried out. The administration dose of the compound 2 group was 40 mg / kg, which served as a control for the normal and virus-infected groups. The positive drug group was given oseltamivir at a dose of 20 mg / kg, once a day for four consecutive days, and the body weight of the mice was recorded daily. Four days after the mice were infected with the virus, their body weights were measured, blood was taken by cutting the eyeball, the whole lungs were cut off, the bloodstains were blotted dry with filter paper, and the weights were recorded. The upper lobe of the right lung was placed in 10% formalin for pathological evaluation of the mouse lung tissue. The right lung was rinsed clean with normal saline and stored in a -80°C refrigerator. When the frozen lung tissue was to be used, it was homogenized with pre-cooled PBS using a homogenizer, the homogenate was collected and centrifuged, the supernatant was aliquoted and stored at -80°C for later use. It was used for BCA quantification and measurement of indexes such as lung tissue inflammatory factors.

[0054] (1) Effect of compound 2 on the body weight of H1N1 virus-infected mice

[0055] Body weight change is a macroscopic index to evaluate whether a drug has a protective effect on mice with viral pneumonia. The body weight change rate was obtained by dividing the daily body weight of the mice by the body weight on the day of infection, and the body weight change curve was plotted using the daily body weight change rate of the mice. The results showed that the body weight of the mice in the model group decreased the most, and the body weight decrease in each drug-administered group was significantly less than that in the model group. The results are as Figure 2 shown.

[0056] (2) Effect of compound 2 on the lung index of H1N1 virus-infected mice

[0057] The lung index is the ratio of the wet weight of the mouse lung to the body weight. The larger the ratio, the more severe the lung lesion, which is also an important indicator of viral pneumonia. The specific calculation process is as follows: On the fourth day after virus infection, the animals were weighed, blood was taken sufficiently, the lungs were dissected and removed intact, and the surface blood was blotted dry with filter paper and then weighed. This was used as the wet weight of the lungs, and the lung index was calculated. Lung index = wet weight of lungs (mg) / body weight (g).

[0058] The results showed that compared with the model group, the lung index of the mice in the normal group was significantly lower (p < 0.001); after drug administration, the lung indexes of the C2 group and the positive drug group were significantly lower than that of the model group (p < 0.001, p < 0.001), as Figure 3 shown.

[0059] (3) Effect of Compound 2 on the pathological changes in the lungs of mice infected with H1N1 virus

[0060] Take the lung tissue samples in the fixing solution, embed them in paraffin to form tissue blocks with a volume of 1 cm × 1 cm × 1 cm, cut them into paraffin sections with a width of 5 μm and perform HE staining. Finally, scan the sections with an Olympus VS200 whole slide scanner.

[0061] The pathological examination results showed that in the normal group, the alveolar contours were clear, the structure was complete, there was no bleeding, and there was basically no inflammation; the pathological sections of the model group showed that the alveolar walls were significantly thickened, the alveoli were atrophied and deformed, and a large number of neutrophils and lymphomononuclear cells infiltrated the lung interstitium, with severe inflammation. Both the C2 group and the positive drug group could significantly improve the pathological damage of the lungs. The alveolar contours were relatively clear, the structure was relatively complete, and the infiltration of inflammatory cells was significantly reduced, as Figure 4 shown.

[0062] (4) Effect of Compound 2 on the content of inflammatory factors in the lungs of mice infected with H1N1 virus

[0063] Take the supernatant of the lung tissue homogenate, detect the protein content by the BCA method according to the instruction manual, and determine the contents of MCP-1, IL-6 and TNF-α in the lung tissues of mice in each group in the experiment by the ELISA method according to the kit instruction manual.

[0064] Compared with the model group, the levels of MCP-1, IL-6, and TNF-α in the lung homogenate of mice in the normal group were significantly lower than those in the model group (p < 0.001, p < 0.001, p < 0.001); after administration, the levels of MCP-1, IL-6, and TNF-α in the lung homogenate of the C2 group were significantly lower than those in the model group (p < 0.01, p < 0.001, p < 0.01), and the levels of MCP-1, IL-6, and TNF-α in the lung homogenate of the positive drug group were significantly lower than those in the model group (p < 0.001, p < 0.001, p < 0.05). The results are as Figure 5 shown.

[0065] The above is only a preferred embodiment of the present invention, and it is not a limitation of the present invention in other forms. Any person skilled in the art may use the disclosed technical content to make changes or modifications into equivalent embodiments with equivalent changes. However, any simple modification, equivalent change and modification made to the above embodiments based on the technical essence of the present invention without departing from the technical solution content of the present invention still fall within the protection scope of the technical solution of the present invention.

Claims

1. A preparation method of long-chain fatty acyl monoterpenoid compounds, characterized in that, the preparation method comprises the following steps: After pulverizing the pericarpium trichosanthis, percolation extraction is carried out with 95% v / v ethanol. After concentrating the extract, it is suspended in water and successively extracted with petroleum ether, ethyl acetate, and n-butanol to obtain a petroleum ether extract, an ethyl acetate extract, and an n-butanol extract; Take the petroleum ether extract and separate it by silica gel column chromatography, eluting with a petroleum ether - ethyl acetate 1:0 → 0:1, v / v gradient to obtain 5 fractions A1–A5; Fraction A2 is separated by silica gel column chromatography, eluting with a n-hexane - ethyl acetate 100:0 → 1:10, v / v gradient to obtain 12 fractions A2A–A2L; Fraction A2F is separated by reverse-phase ODS column chromatography, eluting with a methanol - water 30:70 → 100:0, v / v gradient to obtain 12 fractions A2F1–A2F12; Fraction A2F8 is purified by semi-preparative liquid phase, eluting isocratically with methanol - water 97:3, v / v as the mobile phase to obtain the compounds trichosanate A and (3S,5R,6S,7E)-3-tetradecanoate-5,6-epoxy-β-ionone; Compound 1, trichosanate A: colorless crystal; molecular formula: C 29 H 50 O 4 ; relative molecular weight: 462; 1 H NMR (400 MHz, CD 3 OD), δ: 7.16 (1H, d, J = 15.8 Hz, H-7), 6.22 (1H, d, J = 15.8 Hz, H-8), 4.91 (1H, m, H-3), 2.37 (1H, dd, J = 14.9, 5.5 Hz, H-4a), 2.30 (3H, s, H-10), 2.28 (2H, m, H-2'), 1.86 (1H, dd, J = 14.8, 7.6 Hz, H-4b), 1.64 (1H, dd, J = 13.3, 2.4 Hz, H-2a), 1.62 (2H, m, H-3'), 1.42 (1H, dd, J = 13.3, 9.2 Hz, H-2b), 1.29 (24H, m, H-4'–H-15'), 1.23 (3H, s, H-11), 1.19 (3H, s, H-13), 0.99 (3H, s, H-12), 0.90 (3H, t, J = 7.0 Hz, H-16'); 13 C NMR (100 MHz, CD 3 OD) δ: 200.1 (C-9), 174.9 (C-1'), 144.4 (C-7), 134.1 (C-8), 71.0 (C-6), 68.5 (C-3), 67.7 (C-5), 42.6 (C-2), 37.5 (C-4), 35.7 (C-1), 35.4 (C-2'), 33.1 (C-14'), 30.1–30.8 (C-4'–C-13'), 28.9 (C-11), 27.5 (C-10), 26.0 (C-3'), 25.7 (C-12), 23.7 (C-15'), 20.1 (C-13), 14.4 (C-16'); Compound 2, (3S,5R,6S,7E)-3-tetradecanoate-5,6-epoxy-β-ionone: colorless crystal; molecular formula: C 27 H 46 O 4 ; relative molecular weight: 434; 1 H NMR (400 MHz, CD 3 OD), δ: 7.16 (1H, d, J = 15.8 Hz, H-7), 6.22 (1H, d, J = 15.8 Hz, H-8), 4.91 (1H, m, H-3), 2.37 (1H, dd, J = 14.9, 5.6 Hz, H-4a), 2.30 (3H, s, H-10), 2.28 (2H, m, H-2'), 1.86 (1H, dd, J = 14.8, 7.6 Hz, H-4b), 1.64 (1H, dd, J = 13.3, 2.4 Hz, H-2a), 1.62 (2H, m, H-3'), 1.42 (1H, dd, J = 13.3, 9.2 Hz, H-2b), 1.29 (20H, m, H-4'–H-13'), 1.23 (3H, s, H-11), 1.19 (3H, s, H-13), 0.99 (3H, s, H-12), 0.90 (3H, t, J = 7.0 Hz, H-14'); 13 C NMR (100 MHz, CD 3 OD) δ: 200.1 (C-9), 174.9 (C-1'), 144.4 (C-7), 134.1 (C-8), 71.0 (C-6), 68.5 (C-3), 67.7 (C-5), 42.6 (C-2), 37.5 (C-4), 35.7 (C-1), 35.4 (C-2'), 33.1 (C-12'), 30.1–30.8 (C-4'–C-11'), 28.9 (C-11), 27.5 (C-10), 26.0 (C-3'), 25.7 (C-12), 23.7 (C-13'), 20.1 (C-13), 14.4 (C-14').

2. Use of the long-chain fatty acyl monoterpenoid compound according to claim 1 in the preparation of anti-complement drugs.

3. Use of the long-chain fatty acyl monoterpenoid compound according to claim 1 in the preparation of anti-viral pneumonia drugs.

4. Use of a long-chain fatty acyl monoterpenoid compound according to claim 3 in the preparation of anti-viral pneumonia drugs, characterized in that, the long-chain fatty acyl monoterpenoid compound alleviates pulmonary pathological changes and relieves pulmonary inflammatory reactions.

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