Ferulic acid-sesquiterpene lactone hybrid molecule as well as pharmaceutical composition, preparation method and application thereof

Abstract

The invention discloses a ferulic acid-sesquiterpene lactone hybrid molecule as well as a pharmaceutical composition, a preparation method and application thereof. The structural general formula of the ferulic acid-sesquiterpene lactone hybrid molecule is as shown in formula I: # imgabs0 #, wherein R1 is hydrogen or deuterium, R2 is # imgabs1, and R3 represents an amine compound; the compound is simple to prepare and high in purity, and can be used for preparing a medicine for treating acute lung injury or preparing an auxiliary medicine for treating acute lung injury.

Description

Technical Field

[0001] The present invention belongs to the field of pharmaceutical technology, and specifically relates to a ferulic acid-sesquiterpene lactone hybrid molecule, a preparation method and a pharmaceutical composition for treating acute lung injury or assisting in the treatment of acute lung injury, in particular, the use of such pharmaceutical compounds and compositions in the treatment of bleomycin-induced acute lung injury. Background Art

[0002] Inflammation is a basic innate immune response to disturbances in tissues (such as allergens, pathogens, damaged cells, infection or tissue damage). Many diseases are caused by infection and inflammation, including bacterial infection, atherosclerosis, diabetes, obesity, neurodegenerative diseases and cancer. Among the many diseases associated with inflammation, acute lung injury (ALI) is a respiratory disease with a high mortality rate caused by pneumonia, pulmonary contusion, severe sepsis, gastroesophageal reflux, shock, blood transfusion, drug poisoning and acute pancreatitis. In recent years, a series of studies have revealed that blocking the inflammatory response is considered to be an effective strategy for the treatment of acute lung injury (ALI). Although the clinical treatment of acute lung injury (ALI) has made great progress, the mortality rate of acute lung injury (ALI) remains high due to the lack of effective drugs. Therefore, it is very necessary to find new and effective drugs for the treatment of acute lung injury (ALI). For this purpose, this patent designed and synthesized a series of ferulic acid-sesquiterpene lactone hybrid molecules, and evaluated their anti-inflammatory activity and ability to improve bleomycin (BLM)-induced acute lung injury (Formula I, Compound IV). Summary of the invention

[0003] In order to solve the above technical problems, the present invention provides a ferulic acid-sesquiterpene lactone hybrid molecule and a pharmaceutical composition, a preparation method and an application thereof.

[0004] The technical solution adopted by the present invention is: a ferulic acid-sesquiterpene lactone hybrid molecule, the general structural formula of which is the structure shown in Formula I:

[0005] Where: R1 is hydrogen or deuterium, R2 is R3 represents an amine compound.

[0006] Preferably, when R3 is an amine compound, it represents any one of alkylamine, arylamine, dialkylamine, diarylamine, cyclic amine or heterocyclic amine.

[0007] Preferably, R3 represents any one functional group or a combination of multiple functional groups of alkylthiol, aromatic thiol, alkoxy or aromatic oxy.

[0008] Preferably, the pharmaceutically acceptable salt is a salt of a compound of formula I and any of the following inorganic or organic acids: hydrofluoric acid, hydrochloric acid, hydrobromic acid, hydroiodic acid, sulfuric acid, phosphoric acid, nitric acid, phosphorous acid, sulfurous acid, carbonic acid, boric acid, phosphomolybdic acid, selenious acid, methanesulfonic acid, substituted methanesulfonic acid, phenylsulfonic acid, substituted phenylsulfonic acid, fumaric acid, citric acid, maleic acid, tartaric acid, oxalic acid, D-malic acid, L-malic acid, DL-malic acid, L-lactic acid, D-lactic acid, DL-lactic acid, formic acid, substituted formic acid, acetic acid, propionic acid, butyric acid, valeric acid, oleic acid, lauric acid, p-toluenesulfonic acid, 1-naphthalenesulfonic acid, 2-naphthalenesulfonic acid, phthalic acid, malonic acid, succinic acid, glycolic acid, mercaptoic acid, glycine, sarcosine, sulfonic acid, nicotinic acid, methylpyridine acid, isonicotinic acid, dichloroacetic acid, benzoic acid or substituted benzoic acid.

[0009] A pharmaceutical composition comprising the ferulic acid-sesquiterpene lactone hybrid molecule according to claim 1 and a pharmaceutically acceptable pharmaceutical carrier, excipient or other drug for treating acute lung injury.

[0010] A pharmaceutical composition, wherein the pharmaceutical carrier or excipient is one or more solid, semisolid, liquid preparations and pharmaceutical product adjuvants, and the pharmaceutical composition is administered in two forms: injection or oral administration. The injection is via intravenous injection or intramuscular injection, and the oral dosage form is tablets or capsules.

[0011] The pharmaceutical composition is used in drugs for treating acute lung injury, especially for bleomycin-induced lung injury.

[0012] A method for preparing a ferulic acid-sesquiterpene lactone hybrid molecule, the synthetic route is as follows:

[0013] S1. Ferulic acid and TBDMSCl were added to a round-bottom flask, followed by CH2Cl2 and 30 mmol DIPEA for reaction at room temperature for 4 h, wherein the molar ratio of ferulic acid, TBDMSCl and DIPEA was 1:2.5:3. The crude product after concentration was dissolved in 100 mL of ethyl acetate, washed, dried and distilled under reduced pressure to obtain a yellow oily initial product. The crude product and K2CO3 were then added to a mixture of THF and H2O at room temperature for sufficient reaction for 1 h, and then extracted, washed and dried to obtain compound 7.

[0014] S2. At 0°C and nitrogen, SOCl2 and DMF were added to the toluene solution of compound 7 for sufficient reaction. After the reaction mixture was stirred at 80°C for 1 hour, the excess SOCl2 was removed under vacuum to obtain the acid chloride 8 for standby use. Subsequently, DIPEA was added to the anhydrous CH2Cl2 solution of Michelia lactone at 0°C and nitrogen for reaction, wherein the millimolar ratio of Michelia lactone to DIPEA was 1:2. Then, the anhydrous CH2Cl2 solution of acid chloride 8 was added dropwise at this temperature and stirred at room temperature for 6 hours. The reaction solution was quenched with a saturated NH4Cl aqueous solution. After three extractions, washing, drying, filtering and concentration, a crude oil was obtained. The crude product was purified to obtain a colorless oily compound 9 for standby use.

[0015] S3, respectively select compound 3, compound 9 and compound 11, then mix any of the above compounds with K2CO3 and Me2NH·HCl respectively, add them to dry DCM, filter the solid after sufficient reaction, concentrate the filtered solution under reduced pressure, dissolve it in CH2Cl2, wash, dry, and concentrate under reduced pressure again, then elute and purify to obtain the corresponding compound 10, compound 12 or compound 13 for standby use;

[0016] S4. At 0°C, TBAF was added to a THF solution of compound 9 or compound 10, and the reaction mixture was stirred at room temperature for 40 min, the reaction was quenched with a saturated ammonium chloride solution, and extracted with ethyl acetate three times. The three organic layers were combined, washed, dried, and concentrated to obtain a crude product. The crude product was purified to obtain the corresponding compound 1 or compound 2;

[0017] S5, adding citric acid to the ethyl acetate solution of compound 2 or compound 12, and then stirring, filtering and washing the reaction mixture to obtain the corresponding compound 5 or compound 6;

[0018] S6. At room temperature, DPPA was added to a THF solution of compound 13 and DBU, wherein the millimolar ratio of compound 13 to DBU was 1:2. After sufficient reaction, the organic layer was removed under vacuum to obtain a yellow oily substance. The yellow oily substance was then dissolved in an EtOAc / H2O solution at room temperature, wherein the volume ratio of EtOAc to H2O was 1:1. The compound 14 was then purified on a silica gel column after extraction, washing, and drying.

[0019] S7. Add PPh3 / H2O solution to the THF solution of compound 14, stir the reaction mixture at room temperature overnight, remove the organic layer under vacuum to obtain a yellow oily crude product, and purify the crude product on a silica gel column to obtain compound 15;

[0020] S8. Add compound 17 to the ferulic acid and DCE solution, stir the reaction mixture at 80°C until compound 17 is completely consumed, and purify the reaction mixture by silica gel column chromatography to obtain compound 16. Then, add compound 16 to the DMF solution of compound 15 at room temperature, stir the reaction mixture at room temperature until compound 16 is completely consumed, quench the reaction with brine, extract with ethyl acetate three times, wash, dry, filter, concentrate, and purify on a silica gel column to obtain compound 4.

[0021] Preferably, the washing in step S1 is performed by mixing 1M HCl and brine in a volume ratio of 1:1, the washing in S2 and S3 is performed by brine, the drying in the above steps is performed by anhydrous Na2SO4, and the purification is performed by column chromatography on a silica gel column.

[0022] Preferably, the compound 17 is allylated and the compound 11 is a PTL allylic oxidation product.

[0023] The beneficial effects of the present invention are as follows: the present invention has simple preparation and high purity, and the hybrid of ferulic acid and sesquiterpene lactone can not only inhibit the inflammatory response of RAW 264.7 macrophages induced by LPS by passivating the NF-kB pathway, but also effectively alleviate the acute lung injury induced by lipopolysaccharide, and can be used for preparing drugs for treating acute lung injury or preparing auxiliary drugs for treating acute lung injury. BRIEF DESCRIPTION OF THE DRAWINGS

[0024] Figure 1 It is ferulic acid, PTL and MCL and their derivatives;

[0025] Figure 2 is the hydrogen spectrum of compound 7;

[0026] Figure 3 is the hydrogen spectrum of compound 9;

[0027] Figure 4 is the carbon spectrum of compound 9;

[0028] Figure 5 is the hydrogen spectrum of compound 1;

[0029] Figure 6 is the carbon spectrum of compound 1;

[0030] Figure 7 is the hydrogen spectrum of compound 10;

[0031] Figure 8 is the carbon spectrum of compound 10;

[0032] Fig. 9 is the hydrogen spectrum of compound 2;

[0033] Fig.10 is the carbon spectrum of compound 2;

[0034] Fig.11 is the hydrogen spectrum of compound 5;

[0035] Fig.12 is the carbon spectrum of compound 5;

[0036] Fig.13 is the hydrogen spectrum of compound 3;

[0037] Fig.14 is the hydrogen spectrum of compound 12;

[0038] Fig.15 is the carbon spectrum of compound 12;

[0039] Fig.16 is the hydrogen spectrum of compound 6;

[0040] Fig.17 is the carbon spectrum of compound 6;

[0041] Fig.18 is the hydrogen spectrum of compound 13;

[0042] Fig.19 is the hydrogen spectrum of compound 14;

[0043] Fig. 20 is the carbon spectrum of compound 14;

[0044] Fig.21 is the hydrogen spectrum of compound 15;

[0045] Fig. 22 is the carbon spectrum of compound 15;

[0046] Fig.23 is the hydrogen spectrum of compound 16;

[0047] Fig.24 is the carbon spectrum of compound 16;

[0048] Fig.25 is the hydrogen spectrum of compound 4;

[0049] Fig.26 is the carbon spectrum of compound 4;

[0050] Fig. 27 This is the synthetic route of compound 1;

[0051] Fig.28 This is the synthetic route of compound 5;

[0052] Fig.29 This is the synthetic route of compound 6;

[0053] Fig.30 This is the synthetic route of compound 4;

[0054] Fig.31 Anti-inflammatory effects of compounds 1-6 in RAW264.7 cells;

[0055] Fig.32 Pro-inflammatory protein levels of compound 6 in RAW264.7 cells;

[0056] Fig.33 Effects of compound 6 on body weight and survival rate after bleomycin-induced acute lung injury in mice;

[0057] Fig.34 The protective effect of compound 6 on ALI mice was observed pathologically. DETAILED DESCRIPTION

[0058] In order to understand the present invention, the present invention is further described below with reference to embodiments.

[0059] Example 1

[0060] like Fig. 27 As shown, the preparation of compound 7:

[0061] Ferulic acid (10 mmol) and TBDMSCl (25 mmol) were added to a round-bottom flask, followed by CH2Cl2 (50 mL) and DIPEA (30 mmol), and the reaction was carried out at room temperature for 4 h. The crude product after the reaction solution was concentrated was dissolved in ethyl acetate (100 mL). The organic phase was washed with 1M HCl and brine, and then dried over anhydrous Na2SO4. The organic layer was distilled under reduced pressure to obtain a yellow oil. The crude product and K2CO3 (1.3 g) were then added to THF (20 mL) / H2O (2.5 mL) at room temperature. The reaction mixture was stirred at room temperature for 1 h and then diluted with water (50 mL). The reaction mixture was extracted with ethyl acetate (100 mL). The combined organic layers were washed with 1M HCl (50 mL) and brine (50 mL), and dried over anhydrous Na2SO4. The organic layer was concentrated under vacuum to obtain a white solid. Yield: 97%. Melting point: 182-185°C. 1 H NMR(400MHz, CDCl3)δ=7.73(d,J=15.9Hz,1H),7.07–7.04(m,2H),6.86(d,J =8.7Hz,1H),6.31(d,J=15.9Hz,1H),3.85(s,3H),1.00(s,9H),0.18(s,6H).

[0062] like Fig. 27 As shown, the preparation of compound 9:

[0063] SOCl2 (2 mL) and DMF (one drop) were added to a solution of compound 7 (5 mmol) in toluene (2 mL) at 0°C under nitrogen. The reaction mixture was stirred at 80°C for 1 h, and then the excess SOCl2 was removed under vacuum to give the acid chloride 8.

[0064] DIPEA (5 mmol) was added to a solution of Michelia lactone (2.5 mmol) in anhydrous CH2Cl2 (5 mL) under nitrogen and 0°C. Then, a solution of acyl chloride 8 in anhydrous CH2Cl2 (2 mL) was added dropwise at this temperature and stirred at room temperature for 6 h. The reaction solution was quenched with saturated NH4Cl aqueous solution and extracted with CH2Cl2 (3×15 mL). The combined organic layer was washed with saturated brine, dried over anhydrous Na2SO4, filtered and concentrated to obtain a crude oil, which was purified on a silica gel column (petroleum ether: ethyl acetate = 10:1) to obtain a colorless oil 9. Yield: 30%. 1 H NMR (400MHz, CDCl3): δ=7.66(d,J=15.9Hz,1H),7.08–7.03(m,2H),6.82(d,J=8.1Hz,1H ),6.27–6.20(m,2H),5.48(d,J=3.1Hz,1H),3.88–3.83(m,4H),3.15(d,J=10.1Hz,1H), 2.75–2.70(m,1H),2.63–2.57(m,1H),2.51–2.46(m,1H),2.28(s,3H),2.23–2.09(m,1H ),2.04–1.95(m,1H),1.73(s,3H),1.62(s,1H),1.59(s,3H),0.98(s,9H),0.16(s,6H). 13 C NMR (100MHz, CDCl3): δ=170.4,166.6,151.1,147.1,144.7,139.4,131.6,129.9,128.6,122.3,120.9,118.8,117. 4,110.8,88.4,83.1,57.3,55.4,49.9,36.6,34.9,30.6,25.9,25.6,24.2,18.5,18.4,-4.7.HRMS(ESI):m / z[M+H] + Calculate for C 31 H 43 O6Si:539.2823; found:539.2821.

[0065] like Fig. 27 As shown, the preparation of compound 1:

[0066] To a solution of compound 9 (0.9 mmol) in THF (3 mL) was added TBAF (1.8 mmol) at 0°C. The reaction mixture was then stirred at room temperature for 40 min, quenched with saturated ammonium chloride solution, and extracted with ethyl acetate (3×15 mL). The combined organic layer was washed with saturated brine, dried over Na2SO4, and concentrated to give a crude product. The crude product was purified on a silica gel column (petroleum ether: ethyl acetate = 3:1) to give compound 1 as a yellow oil. Yield: 76%; 1 H NMR (400MHz, CDCl3): δ = 7.66 (d, J = 15.9Hz, 1H), 7.09–7.07 (m, 2H), 6.89 (d, J = 8.7Hz, 1H), 6.2 7-6.21(m,2H),5.49(d,J=3.1Hz,1H),3.92(s,3H),3.86(t,J=10.1Hz,1H),3.15(d,J=10.2Hz, 1H),2.75–2.70(m,1H),2.63–2.58(m,1H),2.51–2.45(m,1H),2.28(s,3H),2.12–2.09(m,1H) ,2.04–1.95(m,1H),1.72(d,J=1.8Hz,3H),1.59(s,3H),1.43–1.33(m,1H),1.28–1.21(m,1H). 13 C NMR (100MHz, CDCl3): δ=170.4,166.6,147.7,146.7,144.7,139.5,131.6,129.9,127.3,123.1,118.8,11 7.0,114.6,109.6,88.5,83.1,57.3,55.9,49.9,36.7,35.0,30.6,25.9,24.2,18.5.HRMS(ESI):m / z[M+H] + Calculate for C 25 H 29 O6:425.1959; found:425.1957.

[0067] Example 2

[0068] Attached Fig.28 The synthetic route of compound 5

[0069] As attached Fig.28 As shown, the preparation of compound 10:

[0070] Compound 9 (0.6 mmol), K2CO3 (9 mmol) and Me2NH·HCl (4.8 mmol) were added to dry DCM (5 mL) and stirred at room temperature for 5 hours. The solid in the mixture was filtered off and the resulting solution was concentrated under reduced pressure. The residue was dissolved in CH2Cl2 and then washed with water. The combined organic layer was dried over Na2SO4 and concentrated under reduced pressure. The crude product was purified by column chromatography using ethyl acetate-petroleum ether as eluent to obtain the desired product 10 as a yellow oil. Yield: 86%; 1 H NMR (400MHz, CDCl3): δ = 7.66 (d, J = 15.9Hz, 1H), 7.09–7.01 (m, 2H), 6.81 (d, J = 8.1 Hz,1H),6.24(d,J=15.9Hz,1H),3.87–3.80(m,4H),3.07(d,J=9.0Hz,1H),2.74(dd ,J=12.9,4.8Hz,1H),2.64–2.54(m,2H),2.50–2.35(m,2H),2.26(s,6H),2.19–1. 89(m,5H),1.70(s,3H),1.58(s,3H),1.37–1.13(m,2H),0.98(s,9H),0.16(s,6H). 13 C NMR (100MHz, CDCl3): δ=177.5,166.6,151.1,147.1,144.7,131.9,129.9,128.7,122.1,120.9,117.5,111.1,88.6,82. 5,58.5,56.9,55.4,51.5,46.0,45.9,44.94,36.8,35.4,30.4,27.4,25.7,23.9,18.6,18.5,-4.6.HRMS(ESI):m / z[M+H] + Calcd for Chemical Formula:C 33 H 50 NO6Si:584.3402; found:584.3399.

[0071] As attached Fig.28 As shown, the preparation of compound 2:

[0072] TBAF was added to a solution of compound 10 (0.6 mmol) in THF (3 mL) at 0°C. The reaction mixture was then stirred at room temperature for 40 min, quenched with saturated ammonium chloride solution, and extracted with ethyl acetate (3×15 mL). The combined organic layer was washed with saturated brine, dried over anhydrous Na2SO4, and concentrated to give a crude product, which was purified on a silica gel column (CH2Cl2:MeOH=20:1) to give compound 2 as a yellow solid. Yield: 64%. mp: 139-141°C. 1 H NMR (400MHz, CDCl3): δ = 7.64 (d, J = 15.9Hz, 1H), 7.06 (d, J = 8.6Hz, 2H), 6.86 (d ,J=7.7Hz,1H),6.22(d,J=15.9Hz,1H),3.90(s,3H),3.84(t,J=10.1Hz,1H),3 .05(d,J=10.0Hz,1H),2.74(dd,J=13.0,4.9Hz,1H),2.67–2.53(m,2H),2.50– 2.35(m,2H),2.27–1.96(m,12H),1.69(s,3H),1.57(s,3H),1.38–1.19(m,2H). 13 C NMR (100MHz, CDCl3): δ=177.5,166.7,147.8,146.8,144.7,131.9,129.8,127.1,122.9,116.9,114.7,109. 8,88.6,82.5,58.2,56.9,55.9,51.4,45.9,44.9,36.7,35.3,30.4,27.3,23.9,18.5.HRMS(ESI):m / z[M+H] + Calcd for ChemicalFormula:C 27 H 36 NO6:470.2537; found:470.2538.

[0073] As attached Fig.28 As shown, the preparation of compound 5:

[0074] At room temperature, citric acid (1.0 mmol) was added to a solution of compound 2 (1.0 mmol) in ethyl acetate (10 mL). The reaction mixture was then stirred at room temperature for 30 minutes, the white solid in the mixture was filtered off, and washed with ethyl acetate to obtain compound 5. Yield: 77%. mp: 96-98°C. 1H NMR (400MHz, DMSO-d6): δ = 9.58 (s, 1H), 7.57 (d, J = 15.8Hz, 1H), 7.28 (d, J = 1.8Hz, 1H), 7. 08(dd,J=8.2,1.8Hz,1H),6.78(d,J=8.1Hz,1H),6.31(d,J=15.8Hz,1H),4.0–3.95(m,1H) ,3.81(s,3H),3.07(d,J=9.8Hz,1H),2.83–2.76(m,3H),2.65–2.49(m,4H),2.48–2.28(m ,8H),2.27–2.00(m,5H),1.95–1.87(m,1H),1.68(s,3H),1.49(s,3H),1.36–1.29(m,1H). 13 C NMR (100MHz, DMSO-d6): δ=177.5,176.9,170.8,166.4,149.7,148.4,145.4,132.1,130.1,126.2,123.5,116.4,115.9,111. 6,88.7,82.2,71.9,57.4,56.3,56.1,50.9,45.4,44.4,43.6,36.8,35.2,30.3,26.8,24.3,21.2,14.6.HRMS(ESI):m / z[M+H] + Calcd for Chemical Formula:C 27 H 36 NO 13 :470.2537; found:470.2539.

[0075] Embodiment 3:

[0076] Attached Fig.29 The synthetic route of compound 6

[0077] As attached Fig.29 As shown, the preparation of compound 3:

[0078] Compound 11 (5 mmol), ferulic acid (7.5 mmol), DIAD (5 mmol) and PPh3 (7.5 mmol) were added to anhydrous THF (50 mL) at room temperature and nitrogen. The reaction solution was stirred at room temperature for 4 h. The reaction was quenched with saturated ammonium chloride solution and extracted with ethyl acetate (3×30 mL). The combined organic layer was washed with saturated brine, dried over anhydrous Na2SO4, filtered and concentrated. The crude product was purified on a silica gel column (PE:EA=2:1) to obtain compound 3 as a yellow solid. Yield: 55%;mp: 96-98℃.1 H NMR (400MHz, CDCl3) δ = 7.60 (d, J = 15.9Hz, 1H), 7.05 (dd, J = 8.2, 1.8Hz, 1H), 6.99 (d, J = 1.8Hz, 1H), 6.91 ( d,J=8.2Hz,1H),6.24(dd,J=9.7,6.2Hz,2H),6.03(s,1H),5.73(t,J=8.1Hz,1H),5.55(d,J=3.2Hz,1H), 4.76(d,J=12.5Hz,1H),4.59(d,J=12.5Hz,1H),3.92(s,3H),3.87(t,J=9.3Hz,1H),3.01(m,J=9.1,4.7H z,1H),2.90(d,J=9.4Hz,1H),2.47-2.16(m,6H),1.72–1.65(m,1H),1.55(s,3H),1.12(t,J=12.6Hz,1H).

[0079] As attached Fig.29 As shown, the preparation of compound 12:

[0080] Compound 3 (1 mmol), K2CO3 (15 mmol) and Me2NH·HCl (8 mmol) were added to DCM (10 mL) dried at room temperature and stirred at this temperature for 5 h. The solid in the mixture was filtered off and concentrated under reduced pressure. The residue was dissolved in CH2Cl2 and then rinsed with water. The organic layer was dried over Na2SO4 and concentrated under reduced pressure. The crude product was purified by column chromatography on silica gel using ethyl acetate-petroleum ether as eluent to obtain the desired product 12 as a yellow oil. Yield: 96%. 1 H NMR (400MHz, CDCl3): δ = 7.62 (d, J = 15.9Hz, 1H), 7.08 (dd, J = 8.2, 1.8Hz, 1H), 7.03–7.02 (m, 1H), 6.92 (d ,J=8.2Hz,1H),6.30(d,J=15.9Hz,1H),5.66(t,J=8.0Hz,1H),4.87(d,J=12.8Hz,1H),4.66(d,J=12.9H z,1H),3.92(s,3H),3.90–3.84(m,2H),2.81(d,J=9.4Hz,1H),2.78–2.74(m,1H),2.64(dd,J=12.9,5.7 Hz,1H),2.51–2.28(m,6H),2.24(s,6H),2.18–2.12(m,2H),1.59-1.54(m,4H),1.10(t,J=12.7Hz,1H). 13C NMR (100MHz, CDCl3) δ = 176.9, 166.8, 148.1, 146.8, 145.2, 135.9, 128.5, 126.8, 123.1, 114.9, 114.7, 109. 3,81.2,66.2,63.8,59.8,58.2,55.9,45.7,44.4,42.9,36.9,26.9,24.5,23.7,17.9.HRMS(ESI):m / z[M+H] + Calcd for Chemical Formula:C 27 H 36 NO7:486.2486; found:486.2485.

[0081] As attached Fig.29 As shown, the preparation of compound 6:

[0082] To a solution of compound 12 (1 mmol) in ethyl acetate (10 mL) was added citric acid (1 mmol) at room temperature. The reaction mixture was then stirred at 70°C for 8 hours. The white solid in the mixture was filtered off and washed with ethyl acetate to obtain compound 6. Yield: 65%. 1 H NMR (400MHz, D2O): δ=7.49 (d, J=15.9Hz, 1H), 7.08 (d, J=1.7Hz, 1H), 7.01 (dd, J=8.2, 1.6Hz, 1H), 6.82 (d, J=8.2Hz, 1H) ,6.27(d,J=16.0Hz,1H),5.56(t,J=8.0Hz,1H),4.59(d,J=13.1Hz,1H),4.43(d,J=13.0Hz,1H),4.21(t,J=9.6Hz,1H), 3.78(s,3H),3.45–3.39(m,1H),3.28(dd,J=13.2,3.5Hz,1H),3.16–3.06(m,1H),2.92–2.84(m,7H),2.79–2.63(m,4H) ,2.38–2.12(m,4H),2.09–1.95(m,4H),1.76–1.70(m,1H),1.49(s,3H),1.16(t,J=7.2Hz,1H),0.93(t,J=12.2Hz,1H). 13C NMR (100MHz, D2O): δ=177.8,177.4,174.0,168.7,148.2,147.7,146.2,134.4,129.5,126.5,123.2,115.6,114.2,111.2,82. 2,73.5,66.6,63.4,62.7,61.7,55.8,55.5,43.4,42.4,41.3,35.7,24.9,24.0,22.9,20.5,16.6,13.2.HRMS(ESI):m / z[M+H] + Calcd for Chemical Formula:C 27 H 36 NO7:486.2486; found:486.2481.

[0083] Embodiment 4:

[0084] Attached Fig.30 The synthetic route of compound 4

[0085] As attached Fig.30 As shown, the preparation of compound 13:

[0086] Compound 11 (3 mmol), K2CO3 (45 mmol) and Me2NH·HCl (24 mmol) were added to DCM (25 mL) dried at room temperature, stirred at this temperature for 5 h, the solid in the mixture was filtered off, and concentrated under reduced pressure. The residue was dissolved in CH2Cl2 and then washed with water. The organic layer was dried over Na2SO4 and concentrated under reduced pressure. The crude product was purified by column chromatography on silica gel with ethyl acetate-petroleum ether as eluent to obtain the desired product 13 as a yellow oil. Yield: 95%; 1 H NMR (400MHz, CDCl3): δ = 5.57 (t, J = 8.0Hz, 1H), 4.09 (dd, J = 31.7, 13.1Hz, 2H), 3.84 (t, J = 9.0Hz, 1H), 2.81-2.78 (m, 1H), 2.74–2.68 (m, 1 H),2.65–2.55(m,1H),2.49–2.23(m,6H),2.23-2.20(m,7H),2.14-2.07(m,2H),1.62–1.54(m,1H),1.51(s,3H),1.06(t,J=12.8Hz,1H). 13C NMR (100MHz, CDCl3): δ=176.9,140.8,127.0,81.4,65.8,63.9,59.9,57.5,45.6,44.0,42.0,36.9,27.3,25.6,23.6,17.8.

[0087] As attached Fig.30 As shown, the preparation of compound 14:

[0088] At room temperature, DPPA was added to a solution of compound 13 (1 mmol) and DBU (2 mmol) in THF (10 mL). The reaction mixture was stirred at room temperature for 8 h. The organic layer was removed under vacuum to obtain a yellow oil. The yellow oil was then dissolved in EtOAc (10 mL) / H2O (10 mL) at room temperature. The reaction was extracted with ethyl acetate (3×30 mL). The combined organic layers were washed with saturated brine, dried over Na2SO4, and concentrated. The crude product was purified on a silica gel column (PE:EA=1:1) to obtain compound 14 as a colorless oil. Yield: 65%. 1 H NMR (400MHz, CDCl3): δ = 5.60 (t, J = 8.1Hz, 1H), 3.95 (d, J = 13.2Hz, 1H), 3.87-3.83 (m, 2H), 2.76-2.56 (m, 2H), 2. 61-2.56(m,1H),2.47–2.35(m,3H),2.34–2.08(m,11H),1.64–1.56(m,1H),1.54(s,3H),1.07(t,J=12.6Hz,1H). 13 C NMR (100MHz, CDCl3): δ = 176.8, 135.7, 129.6, 81.0, 63.9, 59.8, 58.1, 55.4, 45.6, 44.1, 42.6, 36.9, 26.6, 24.4, 23.7, 17.8. HRMS (ESI): m / z [M+H] + Calcd for Chemical Formula:C 17 H 27 N4O3:335.2083; found:335.2085.

[0089] As attached Fig.30 As shown, the preparation of compound 15:

[0090] PPh3 (0.44 mmol) and H2O (45 μL) were added to a solution of compound 14 (0.4 mmol) in THF (3 mL). The reaction mixture was stirred at room temperature overnight. The organic layer was removed under vacuum to obtain a yellow oil. The crude product was purified on a silica gel column (DCM: CH2Cl2 = 6:1) to obtain compound 15 as a colorless oil. Yield: 36%. 1 H NMR (400MHz, CDCl3): δ = 5.50 (t, J = 7.9Hz, 1H), 3.82 (d, J = 9.5Hz, 1H), 3.45 (s, 1H), 3.33 (d, J = 6.3Hz, 1H), 2.80–2.71 (m, 2H), 2.57 (dd, J = 1 2.9,6.5Hz,1H),2.49(s,2H),2.44–2.34(m,3H),2.28-2.25(m,3H),2.22(s,6H),2.18–2.08(m,3H),1.53(s,3H),1.06(t,J=12.3Hz,1H). 13 C NMR (100MHz, CDCl3): δ = 176.9, 141.2, 124.8, 81.2, 63.7, 60.0, 58.4, 45.7, 45.4, 44.3, 43.2, 37.1, 26.8, 24.6, 23.7, 17.9. HRMS (ESI): m / z [M+H] + Calcd for Chemical Formula:C 17 H 29 N2O3:309.2173; found:309.2174.

[0091] As attached Fig.30 As shown, the preparation of compound 16:

[0092] Acryl ketone 17 (1 mmol) was added to a solution of ferulic acid (1.1 mmol) in DCE (5 mL) at room temperature. The reaction mixture was stirred at 80°C until compound 17 was completely consumed. The reaction mixture was purified by silica gel column chromatography (PE:EA=5:1) to obtain compound 16 as a brown oil. Yield: 75%. 1H NMR (400MHz, CDCl3): δ=7.95–7.92(m,2H),7.74(d,J=15.9Hz,1H),7.57–7.50(m,1H),7.47–7.43(m,2H),7.13–7.10(m,1H) ),7.06(d,J=1.6Hz,1H),6.95–6.90(m,2H),6.36(d,J=15.9Hz,1H),6.17–6.09(m,1H),3.92(s,3H),2.48(d,J=0.8Hz,3H); 13 C NMR (100MHz, CDCl3): δ=190.4,164.5,164.2,148.6,147.3,146.9,138.7,132.7,128. 5,128.1,126.4,123.5,114.9,113.9,113.3,109.6,55.9,19.1.HRMS(ESI):m / z[M+H] + Calcd for Chemical Formula:C 20 H 18 NaO5:361.1046; found:361.1045.

[0093] As attached Fig.30 As shown, the preparation of compound 4:

[0094] Compound 16 was added to a solution of compound 15 (2 mmol) in DMF (10 mL) at room temperature. The reaction mixture was stirred at room temperature until compound 16 was completely consumed. The reaction was quenched with brine and extracted with ethyl acetate (3×15 mL). The combined organic layers were washed with saturated brine, dried over anhydrous Na2SO4, filtered and concentrated. The crude product was purified on a silica gel column (DCM:CH2Cl2=15:1) to give compound 4 as a yellow solid. mp:132~134°C. Yield: 50%. 1H NMR (400MHz, DMSO-d6): δ = 9.40 (s, 1H), 8.00 (t, J = 5.8Hz, 1H), 7.34 (d, J = 15.7Hz, 1H), 7.13 (d, J = 1.8 Hz,1H),6.99(dd,J=8.2,1.8Hz,1H),6.78(d,J=8.1Hz,1H),6.49(d,J=15.7Hz,1H),5.41-5.29(m,1H ),4.01(t,J=9.5Hz,1H),3.90-3.84(m,1H),3.80(s,3H),2.72(d,J=9.5Hz,1H),2.64–2.55(m,3H),2 .47–2.19(m,5H),2.17(s,6H),2.08–2.00(m,3H),1.64–1.54(m,1H),1.47(s,3H),0.94–0.84(m,1H). 13 C NMR (100MHz, DMSO-d6): δ=177.8,165.7,148.7,148.3,139.6,138.3,126.9,124.1,121.9,119.3,116.1,111 .2,81.1,63.7,60.2,58.7,55.9,45.9,43.9,42.8,42.3,37.4,26.2,24.9,23.6,17.9.HRMS(ESI):m / z[M+H] + Calcdfor Chemical Formula:C 27 H 37 N2O6:485.2646; found:485.2641.

[0095] (I) The present application discusses the anti-inflammatory activity of the compounds on RAW264.7 cells as follows:

[0096] 1. Experimental Principle

[0097] Inflammatory factors include TNF-α, IL-1β and IL-6, which participate in and accelerate the inflammatory response during the progression of inflammation. In order to determine the efficacy of these compounds in inhibiting the inflammatory process, the present application tested the pro-inflammatory cytokines released by RAW264.7 cells after LPS stimulation. The present application first preliminarily evaluated the inhibitory effects of all compounds on LPS-induced RAW264.7 inflammatory response-related factors.

[0098] 2. Experimental Methods

[0099] (1) Cell culture and treatment

[0100] Mouse RAW264.7 macrophages were maintained at 37°C and 5% CO2 atmosphere in Dulbecco's modified Eagle's medium (DMEM) supplemented with 10% fetal bovine serum (FBS) and 1% streptomycin-penicillin.

[0101] Isolation of total RNA and quantitative real-time polymerase chain reaction (qRT-PCR)

[0102] Total RNA was isolated using TRIzol reagent (Takara, JPN) and then transcribed into cDNA using the Transcripton cDNA Synthesis Kit (TransGen Biotech, China) according to the manufacturer's instructions. Real-time quantitative PCR was performed using Green qPCR SuperMix (TransGen Biotech, China).

[0103] (2) Enzyme-linked immunosorbent assay (ELISA)

[0104] Tumor necrosis factor α (TNF-α) and interleukin 1β (IL-1β) in the cell culture supernatant were detected using commercially available ELISA kits (Biolegend, US). The plate was read on a Bio-Rad microplate reader at a wavelength of 450 nm.

[0105] 3. Experimental results

[0106] Preliminary results showed that compounds 1 and 3 significantly reduced the levels of pro-inflammatory cytokines in macrophages compared with LPS stimulation (see Fig.31 ). It is worth noting that although FA-MCL hybrid molecule 1 had a significant inhibitory effect on proinflammatory cytokines, its corresponding Michael addition product 2 barely reduced the expression of cytokines (p>0.99). Similarly, FA-PTL amide hybrid molecule 4 and prodrug 5 showed no anti-inflammatory activity. Therefore, compound 3 was used for subsequent studies.

[0107] Salt 6 is a water-soluble prodrug of compound 12. Compound 6 has the potential to effectively reduce the expression of proinflammatory cytokines IL-1β and IL-6 (see Fig.31 Next, the present application detected the protein levels of TNF-α and IL-6 by ELISA method, and found that compound 6 can also inhibit the release of pro-inflammatory cytokines in vitro (see Appendix Fig.32 ). Therefore, compound 6 with better comprehensive performance was selected for further study.

[0108] (II) Cytotoxicity test

[0109] Experimental methods:

[0110] (1) Cell viability assay

[0111] The toxicity of these compounds was determined by CCK-8 method. RAW264.7 cells were seeded in 96-well plates at a density of 2×105 / mL (others were 5×104 / mL) and a volume of 100 μL / well. After incubation at 37°C for 12 h, the compounds were incubated for 48 h with concentrations of 1.25, 2.5, 5, 10, 20, 40 and 80 μM, respectively, and 10 μL of CCK-8 solution was added to each well, and incubation was continued at 37°C for 2 h. The absorbance at a wavelength of 450 nm was measured using a microplate reader (Bio-Rad Laboratories, Inc., Hercules, CA, USA).

[0112] (2) Experimental conclusion

[0113] The non-cytotoxic doses of these derivatives on RAW264.7 cells were determined using CCK-8 cell proliferation and cytotoxicity assays (CCK-8). As shown in Table 1 (see attached figure of the specification), compound 6 is safe at a concentration of 2 μM. Compound 6 was further tested as a novel anti-inflammatory drug for the treatment of acute lung injury in an animal model of acute lung injury.

[0114] (III) Compound 6 can increase the body weight of BLM-induced mice

[0115] To further confirm the anti-inflammatory effect of compound 6, we established an acute lung injury model in C57 / 6j mice using bleomycin. A partial increase in body weight as a function of time was observed in the control group, and pulmonary administration of bleomycin (BLM, 2.5 mg / kg) resulted in a significant decrease in body weight. In contrast, daily intravenous (ip) administration of compound 6 (10 mg / kg) to treat mice with BLM-induced lung injury significantly prevented weight loss and alleviated BLM-induced acute lung injury ( Fig.33 ). However, treatment with compound 6 did not reverse the survival rate of BLM mice. Note that during the acute exudative phase (i.e., days 3-7), the effects of dexamethasone and compound 6 were not observed as we expected. To further investigate the protective effect of compound 6 against BLM-induced acute lung injury, we investigated the pathological changes in lung tissue (Supplementary Fig.34 ). The results of lung H&E staining showed that there was a large amount of inflammatory cell infiltration in the lung tissue of mice in the BLM group, accompanied by edema and destruction of alveolar structure. Compound 6 can reduce inflammatory cell infiltration and edema in the lung tissue of mice and improve alveolar structure. The above results show that compound 6 can significantly inhibit the infiltration of lung tissue cells and has a good protective effect on BLM-induced acute lung injury in mice.

[0116] IC values ​​of anti-inflammatory activities of several compounds 50 The values ​​are shown in Table 1 and Table 2:

[0117]

[0118] Table 1

[0119]

[0120] Table 2

[0121] This application can be administered in the form of injection, tablet or capsule:

[0122] Preparation of injection: After the compound prepared in the above example is dissolved in a small amount of DMSO, injection water is added according to the routine method, fine filtration is performed, and the injection is prepared by filling and sterilization.

[0123] Tablet preparation: The compound prepared in the above examples of the present application and the excipient are added to the excipient in a weight ratio of 5:1, granulated and tableted to obtain tablets.

[0124] Capsule preparation: The compound prepared in the above examples of the present application and the excipient are added to the excipient at a weight ratio of 5:1 to prepare capsules. Sesquiterpene lactones are a very important class of natural products with a variety of pharmacological properties.

[0125] The present invention uses natural products involving sesquiterpene lactones, including parthenolide, Michelia lactone, iso-inulinolide, iso-inulinolide epoxidation products, dehydrocostus lactone and the like, wherein parthenolide (PTL) is a natural sesquiterpene lactone compound extracted from Feverfew (Tanacetum parthenium L.). Parthenolide can selectively kill cancer stem cells by targeting NF-kB, redox thiol, HDAC or DNMT. And PTL is easily converted into MCL in the presence of p-toluenesulfonic acid, and the yield is excellent. Compared with PTL, MCL has a more stable structure, better in vivo pharmacokinetic characteristics, and also has good anti-inflammatory and anti-tumor properties. It can effectively inhibit the growth of leukemia cells and the occurrence of tumors. And through the preparation of the present application, the pharmaceutical composition can be given three modes of administration, namely, injection, tablets and capsules, which effectively overcomes the disadvantages of poor water solubility and oral bioavailability of traditional semiterpene lactone derivatives, and is more conducive to its clinical application.

[0126] The above description is only a preferred embodiment of the present invention and does not limit the present invention in any form. Any simple modification, equivalent change and modification made to the above embodiment according to the technical essence of the present invention without departing from the technical solution of the present invention shall fall within the scope of the technical solution of the present invention.

Claims

1. A ferulic acid-sesquiterpene lactone hybrid molecule, characterized in that: Its general structural formula is the structure shown in Formula I: Where: R1 is hydrogen or deuterium, R2 is R3 represents an amine compound.

2. A ferulic acid-sesquiterpene lactone hybrid molecule according to claim 1, characterized in that: When R3 is an amine compound, it is any one of alkylamine, arylamine, dialkylamine, diarylamine, cyclic amine or heterocyclic amine.

3. A ferulic acid-sesquiterpene lactone hybrid molecule according to claim 2, characterized in that: The R3 represents any one functional group or a combination of multiple functional groups of alkylthiol, arylthiol, alkoxy or aryloxy.

4. The ferulic acid-sesquiterpene lactone hybrid molecule according to claim 1, characterized in that: The pharmaceutically acceptable salt is a salt formed by a compound of formula I and any of the following inorganic or organic acids: hydrofluoric acid, hydrochloric acid, hydrobromic acid, hydroiodic acid, sulfuric acid, phosphoric acid, nitric acid, phosphorous acid, sulfurous acid, carbonic acid, boric acid, phosphomolybdic acid, selenious acid, methanesulfonic acid, substituted methanesulfonic acid, phenylsulfonic acid, substituted phenylsulfonic acid, fumaric acid, citric acid, maleic acid, tartaric acid, oxalic acid, D-malic acid, L-malic acid, DL-malic acid, L-lactic acid, D-lactic acid, DL-lactic acid, formic acid, substituted formic acid, acetic acid, propionic acid, butyric acid, valeric acid, oleic acid, lauric acid, p-toluenesulfonic acid, 1-naphthalenesulfonic acid, 2-naphthalenesulfonic acid, phthalic acid, malonic acid, succinic acid, glycolic acid, mercaptoic acid, glycine, sarcosine, sulfonic acid, nicotinic acid, methylpyridine acid, isonicotinic acid, dichloroacetic acid, benzoic acid or substituted benzoic acid.

5. A pharmaceutical composition, characterized in that: Contains the ferulic acid-sesquiterpene lactone hybrid molecule according to claim 1 and a pharmaceutically acceptable pharmaceutical carrier, excipient or other drug for treating acute lung injury.

6. The pharmaceutical composition according to claim 5, characterized in that: The pharmaceutical carrier or excipient is one or more solid, semisolid, liquid preparations and pharmaceutical product adjuvants. The pharmaceutical composition is administered in two forms: injection or oral administration. The injection is through intravenous injection or intramuscular injection, and the oral dosage form is tablets or capsules.

7. The use of the pharmaceutical composition according to claim 5, characterized in that: Used in drugs for the treatment of acute lung injury, especially bleomycin-induced lung injury.

8. A method for preparing a ferulic acid-sesquiterpene lactone hybrid molecule, characterized in that: The synthetic route is as follows: S1. Ferulic acid and TBDMSCl were added to a round-bottom flask, followed by CH2Cl2 and 30 mmol DIPEA for reaction at room temperature for 4 h, wherein the molar ratio of ferulic acid, TBDMSCl and DIPEA was 1:2.5:

3. The crude product after concentration was dissolved in 100 mL of ethyl acetate, washed, dried and distilled under reduced pressure to obtain a yellow oily initial product. The crude product and K2CO3 were then added to a mixture of THF and H2O at room temperature for sufficient reaction for 1 h, and then extracted, washed and dried to obtain compound 7. S2. At 0°C and nitrogen, SOCl2 and DMF were added to the toluene solution of compound 7 for sufficient reaction. After the reaction mixture was stirred at 80°C for 1 hour, the excess SOCl2 was removed under vacuum to obtain the acid chloride 8 for standby use. Subsequently, DIPEA was added to the anhydrous CH2Cl2 solution of Michelia lactone at 0°C and nitrogen for reaction, wherein the millimolar ratio of Michelia lactone to DIPEA was 1:

2. Then, the anhydrous CH2Cl2 solution of acid chloride 8 was added dropwise at this temperature and stirred at room temperature for 6 hours. The reaction solution was quenched with a saturated NH4Cl aqueous solution. After three extractions, washing, drying, filtering and concentration, a crude oil was obtained. The crude product was purified to obtain a colorless oily compound 9 for standby use. S3, respectively select compound 3, compound 9 and compound 11, then mix any of the above compounds with K2CO3 and Me2NH·HCl respectively, add them to dry DCM, filter the solid after sufficient reaction, concentrate the filtered solution under reduced pressure, dissolve it in CH2Cl2, wash, dry, and concentrate under reduced pressure again, then elute and purify to obtain the corresponding compound 10, compound 12 or compound 13 for standby use; S4. At 0°C, TBAF was added to a THF solution of compound 9 or compound 10, and the reaction mixture was stirred at room temperature for 40 min. The reaction was quenched with a saturated ammonium chloride solution, and extracted with ethyl acetate three times. The three organic layers were combined, washed, dried, and concentrated to obtain a crude product, and the crude product was purified to obtain the corresponding compound 1 or compound 2; S5, adding citric acid to the ethyl acetate solution of compound 2 or compound 12, and then stirring, filtering and washing the reaction mixture to obtain the corresponding compound 5 or compound 6; S6. At room temperature, DPPA was added to a THF solution of compound 13 and DBU, wherein the millimolar ratio of compound 13 to DBU was 1:

2. After sufficient reaction, the organic layer was removed under vacuum to obtain a yellow oily substance. The yellow oily substance was then dissolved in an EtOAc / H2O solution at room temperature, wherein the volume ratio of EtOAc to H2O was 1:

1. The compound 14 was then purified on a silica gel column after extraction, washing, and drying. S7. Add PPh3 / H2O solution to the THF solution of compound 14, stir the reaction mixture at room temperature overnight, remove the organic layer under vacuum to obtain a yellow oily crude product, and purify the crude product on a silica gel column to obtain compound 15; S8. Add compound 17 to the ferulic acid and DCE solution, stir the reaction mixture at 80°C until compound 17 is completely consumed, purify the reaction mixture by silica gel column chromatography to obtain compound 16; then add compound 16 to the DMF solution of compound 15 at room temperature, stir the reaction mixture at room temperature until compound 16 is completely consumed, quench the reaction with brine, extract with ethyl acetate three times, wash, dry, filter, concentrate and purify on a silica gel column to obtain compound 4.

9. The method for preparing a ferulic acid-sesquiterpene lactone hybrid molecule according to claim 8, characterized in that: The washing in step S1 is performed by mixing 1M HCl and brine in a volume ratio of 1:1, the washing in S2 and S3 is performed by brine, the drying in the above steps is performed by anhydrous Na2SO4, and the purification is performed by column chromatography on a silica gel column.

10. The method for preparing a ferulic acid-sesquiterpene lactone hybrid molecule according to claim 8, characterized in that: The compound 17 is olefinic, and the compound 11 is a PTL allylic oxidation product.

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