Glycyrrhetinic acid urea derivative as well as preparation method and application thereof

Abstract

The invention belongs to the field of preparation of medical compounds, and relates to a glycyrrhetinic acid urea derivative as well as a preparation method and application thereof. The invention discloses a glycyrrhetinic acid urea derivative. The structure of the glycyrrhetinic acid urea derivative is shown as a general formula (I). The invention further discloses a preparation method of the glycyrrhetinic acid urea derivative and application of the glycyrrhetinic acid urea derivative as an anti-inflammatory drug, and the glycyrrhetinic acid urea derivative can be specifically applied to drugs or pharmaceutical compositions for treating and / or preventing acute kidney injury, acute lung injury, chronic obstructive pulmonary disease, hypertoxemia and other related diseases. # imgabs0 #

Description

Technical Field

[0001] The invention belongs to the field of preparation of pharmaceutical compounds, and in particular relates to a glycyrrhetinic acid urea derivative and a preparation method and use thereof. Background Art

[0002] Inflammation is a complex physiological and pathological response of the body to tissue damage or invasion of pathogenic factors, and is the basic defense mechanism of the immune system. Moderate inflammatory response is an important process for host defense against harmful stimuli and healing recovery, but acute inflammation may lead to tissue damage, sepsis, cancer (Immunobiology, 2009, 214 (9-10): 761-777) and shock (Heiss E, Herhaus C, et al., J Biolchem, 2001, 276, 32008-32015) and other lesions. Under the action of inflammatory factors, monocytes will migrate to extravascular tissues and differentiate into macrophages, which will produce tumor necrosis factor (TNF-α), interleukins (TL-1β, TL-6), prostaglandin E2 (PGE2) and nitric oxide (NO) after being activated. The large-scale expression of these factors will have a profound impact on endothelial cells and epithelial cells, and perform host defense, tissue reconstruction or repair in the mesenchymal cells of the local microenvironment (Yona S, Jung S. et al., Curr Opin Hematol 2010, 17, 53-59).

[0003] Nonsteroidal anti-inflammatory drugs (NSAIDs) have long played an important role in analgesia and anti-inflammation, but long-term use of NSAIDs can lead to gastrointestinal adverse events.

[0004] Licorice was first recorded in Shennong's Herbal Classic. It is also known as sweet grass root, Ural licorice, honey grass, etc. It is often used to treat inflammation, gastric ulcers, liver disease, adrenal insufficiency and tumors (Monograph. Glycyrrhiza glabra [J]. Altern. Med. Rev, 2005, 10: 230-237.). Glycyrrhizic acid is the main biologically active ingredient in licorice. Glycyrrhetinic acid is obtained by hydrolyzing two molecules of glucuronic acid. A large number of studies (Baltina, L.A. Stolyarova, O.V. Baltina, L.A., et al., Pharma. Chem. J, 2010, 44: 299–302. Shetty, A.V. Thirugnanam, S. Dakshinamoorthy, G., et al., Int. J. Oncol, 2011, 39: 635–640. Xie, C. Li, X. Wu, J., et al., Inflammation, 2015, 38: 1639–1648.) have shown that glycyrrhetinic acid can inhibit the activation of key metabolic enzymes of arachidonic acid (AA), including phospholipid A2 (PLA2), cyclooxygenase (COXs) and 5-lipoxygenase (LOX), and exert anti-inflammatory and anti-cancer effects. However, glycyrrhetinic acid (GA) is insoluble in water, making it difficult to absorb when made into preparations, and has low bioavailability.

[0005] A series of novel glycyrrhizic acid urea compounds were disclosed in Chinese scientific literature (Design, synthesis and therapeutic effects of novel glycyrrhizic acid urea compounds on acute kidney injury, master's thesis, Anhui Medical University, 2021). It uses glycyrrhizic acid as the core structure and introduces urea units into the structure of GA. Through splicing design and synthesis, a series of glycyrrhizic acid urea derivatives were synthesized, and the pharmacological effects of novel glycyrrhizic acid urea compounds were explored. It was found that introducing urea units into the structure of GA can improve the efficacy and utilization.

[0006] The present invention, through structural modification, enolizes the 2-position of glycyrrhetinic acid, and then transforms the 20-position carboxyl group into a substituted urea structure, which can enhance the various pharmacological activities of glycyrrhetinic acid, increase its water solubility, and improve its bioavailability, thus providing a new approach for the research and development of glycyrrhetinic acid derivatives. Summary of the invention

[0007] In order to enhance the pharmacological activity of glycyrrhetinic acid compounds, further increase their water solubility and improve their bioavailability, the present invention provides a glycyrrhetinic acid urea derivative and a preparation method and use thereof. The present invention provides a preparation method of the glycyrrhetinic acid urea derivative and also discloses the use of the glycyrrhetinic acid urea derivative as an anti-inflammatory drug, which can be specifically applied to drugs or pharmaceutical compositions for treating and / or preventing related diseases such as acute kidney injury, acute lung injury, chronic obstructive pulmonary disease, and septicemia.

[0008] The present invention achieves the above technical effects through the following technical solutions:

[0009] The present invention aims to provide a glycyrrhetinic acid urea derivative and a preparation method and use thereof;

[0010] The present invention relates to a compound of general formula (I) or a stereoisomer, tautomer, derivative, prodrug or pharmaceutically acceptable salt thereof:

[0011]

[0012] In formula (I), R is a mono-substituted or di-substituted amino group;

[0013] The monosubstituent of the amino group is selected from substituted or unsubstituted C1-C6 alkyl, C3-C6 cycloalkyl, substituted or unsubstituted C6-C8 aryl;

[0014] The disubstituted amino group is The n is selected from 1-2; the Z is selected from C, O or N; the position of Z is any reasonable position on the ring; and the R1 is selected from H, C1-C3 alkyl.

[0015] Furthermore, the substituent of the C1-C6 alkyl group is selected from hydroxyl, carboxyl, Z is O or N; the position of Z is any reasonable position on the ring;

[0016] The substituent of the C6-C8 aryl group is selected from halogen and C1-C3 alkyl;

[0017] Said In the above, when n is 1, Z is C and R1 is H; when n is 2, Z is selected from C, N, O, and Z is in the para position to the N on the ring, and R1 is selected from H and methyl.

[0018] Furthermore, the Wherein Z is O, and Z is in adjacent positions to another substituent on the ring; Wherein Z is N, and Z is in the ortho, para or meta position relative to another substituent on the ring.

[0019] Specifically, R is selected from n-propylamino, cyclopropylamino, cyclopentylamino, cyclohexylamino, hydroxyethylamino, carboxymethylamino, 4-fluorophenylamino, 2-ethylphenylamino, tetrahydropyrrolyl, piperidinyl, morpholinyl, N-methylpiperazinyl, 2-pyridylmethylamino, 3-pyridylmethylamino, 4-pyridylmethylamino, and 2-furanylmethylamino.

[0020] The present invention also provides a method for preparing the above-mentioned compound or its stereoisomers, tautomers, derivatives, prodrugs or pharmaceutically acceptable salts, comprising the following steps:

[0021] Using glycyrrhetinic acid as a raw material, according to the literature (Chemische Berichte, 1957, vol. 90, p. 601, 604) and (European Journal of Medicinal Chemistry, 2009, 44 (10): 4112-4120.), 2-hydroxyene-3-glycyrrhetinic acid ketone (II) was prepared by reaction.

[0022] Step (1) 2-Hydroxy-3-glycyrrhetinic acid ketone (II) is acylated with acetic anhydride to generate 2-acetoxy-3-glycyrrhetinic acid ketone (III)

[0023]

[0024] The solvent used in the reaction is an aprotic solvent, and the reaction temperature is -10 to 50°C.

[0025] Step (2) 2-acetoxyene-3-glycyrrhetinic acid ketone (III) reacts with thionyl chloride to generate 2-acetoxyene glycyrrhetinic acid chloride-3-ketone (IV), which is then reacted with sodium azide to generate 2-acetoxyene glycyrrhetinic acid azide-3-ketone (V), which is then heated and rearranged to obtain isocyanate (VI)

[0026]

[0027] The reaction solvent for converting the carboxyl group into the acyl chloride is an aprotic solvent, and the reaction temperature is 30 to 80°C; the solvent used for converting the acyl chloride into the azido ester is a mixed solvent of THF or acetone and water, and the ratio of the two solvents is 1:5 to 5:1 (volume ratio), and the reaction temperature is -5 to 30°C; the solvent for the azido ester rearrangement reaction is an aprotic solvent, and the reaction temperature is 30 to 100°C.

[0028] Step (3) isocyanate (VI) reacts with substituted amine to obtain 2-acetoxyene-glycyrrhetinic acid urea (VII)

[0029]

[0030] i: R1 and R2 are not H at the same time. When one of them is H, the other is n-propyl, cyclopropyl, cyclopentyl, cyclohexyl, hydroxyethyl, carboxymethyl, 4-fluorophenyl, 2-ethylphenyl, 2-pyridylmethyl, 3-pyridylmethyl, 4-pyridylmethyl, or 2-furylmethyl;

[0031] ii: When neither R1 nor R2 is H, represents tetrahydropyrrole, morpholine, piperidine, N-methylpiperazine;

[0032] The solvent used in the reaction is an aprotic solvent, and the reaction temperature is 0-60°C.

[0033] Step (4) 2-acetoxyene-glycyrrhetinic acid urea (VII) is hydrolyzed under alkaline conditions to obtain glycyrrhetinic acid urea derivative (I)

[0034]

[0035] The reagent used in the hydrolysis reaction is an inorganic base, the solvent is an alcohol solvent, and the reaction temperature is 0-60°C.

[0036] A further object of the present invention is to provide the use of the above-mentioned compound or its stereoisomers, tautomers, derivatives, prodrugs or pharmaceutically acceptable salts in the preparation of anti-inflammatory drugs and anti-inflammatory drug compositions;

[0037] The pharmaceutical composition refers to a composition containing the above-mentioned compound or its stereoisomers, tautomers, derivatives, prodrugs or pharmaceutically acceptable salts, and a pharmaceutically acceptable carrier.

[0038] Another object of the present invention is to provide the use of the above-mentioned compound or its stereoisomers, tautomers, derivatives, prodrugs or pharmaceutically acceptable salts in the preparation of drugs or pharmaceutical compositions for treating and / or preventing acute kidney injury, acute lung injury, chronic obstructive pulmonary disease, septicemia and other related diseases.

[0039] The technical effect of the present invention is that: glycyrrhetinic acid is used as a raw material, C-3 hydroxyl group is oxidized to carbonyl group, C-2 is enolized and esterified, C-20β carboxyl group is converted into isocyanate, and then addition reaction is carried out with substituted amine, and then alkaline hydrolysis is used to obtain glycyrrhetinic acid urea derivatives; the present invention further improves the solubility and bioavailability of glycyrrhetinic acid compounds after modifying the glycyrrhetinic acid compounds. Pharmacological efficacy experiments show that glycyrrhetinic acid urea derivatives (I) have no obvious cytotoxicity, can effectively bind to STING (stimulator of interferon genes) protein, block STING / NF-κB signal transduction related to inflammation, reduce the secretion of inflammatory factors and macrophage polarization; and have a significant protective effect on acute tubular necrosis in mice caused by cisplatin. BRIEF DESCRIPTION OF THE DRAWINGS

[0040] Figure 1 The synthetic route of glycyrrhetinic acid urea derivatives

[0041] Figure 2 Cytotoxicity evaluation of glycyrrhetinic acid urea derivatives. MTT assay was used to detect the cytotoxicity of drugs on mouse macrophage RAW.264.7 cells. Cis, cisplatin; NC, normal control group.

[0042] Figure 3The HK-2 cell model was established with cisplatin, and then glycyrrhetinic acid urea derivatives were added. The MTT method was used to detect the protective effect of the drugs on the cells. Cis, cisplatin; NC, normal control group.

[0043] Figure 4 The cell thermal shift assay was used to investigate compound I 15 Binding ability to STING. (A) Cellular thermal shift assay-melting curve. (B) Dose-response relationship of cellular thermal shift assay.

[0044] Figure 5 Glycyrrhetinic acid urea derivatives I 15 Effects of LPS on STING / NF-κB signal transduction induced by LPS in RAW264.7 cells. 15 After pretreatment with LPS (2.5, 5, 10 μM) for 1 hour, LPS (0.5 μg / mL) was incubated for 24 hours, and protein expression in RAW264.7 cells was detected by Western blotting. (A) Western blot analysis of STING, PP65 / P65, and iNOS. (B, C, D) Quantitative data of proteins.

[0045] Figure 6 Glycyrrhetinic acid urea derivatives I 15 Protective effect of glycyrrhetinic acid urea derivative I on cisplatin-induced AKI in C57BL / 6 mice 15 After pretreatment with (30, 60 mg / kg), mice were intraperitoneally injected with cisplatin (20 mg / kg). (A, B) renal function markers (BUN and Scr), (C, D, E) pro-inflammatory factors (IL-1β, TNF-α and IL-6) were detected. Cis, cisplatin; NC, normal control group.

[0046] Figure 7 Glycyrrhetinic acid urea derivatives I 15 Effects on cisplatin (20 mg / kg)-induced pathological changes in kidney and liver tissues. Tissues were examined histologically using H&E. Cis, cisplatin; NC, normal control group; H&E, hematoxylin and eosin. DETAILED DESCRIPTION

[0047] The beneficial effects of the present invention are now further described by the following examples. It should be understood that these examples are only for illustrative purposes and do not limit the scope of the present invention. At the same time, obvious changes and modifications made by ordinary technicians in the field according to the present invention are also included in the scope of the present invention.

[0048] The glycyrrhizic acid urea derivatives involved in the specific embodiments of the present invention are shown in Table 1, wherein R represents different substituents possessed by the compound:

[0049] Table 1: Glycyrrhetinic acid urea derivatives Ⅰ1~Ⅰ 16

[0050]

[0051]

[0052]

[0053] Example 1 to Example 16

[0054] Glycyrrhetinic acid derivatives I1~I 16 Preparation (synthetic route as Figure 1 (shown)

[0055] The compound 2-hydroxyene-3-glycyrrhetinic acid copper (II) was prepared according to the method reported in the literature.

[0056] Weigh compound II (482 mg, 1.0 mmol) and dissolve it in ethyl acetate (200 mL). Add 4-dimethylaminopyridine (DMAP, 366 mg, 3.0 mmol) and drop acetic anhydride (0.38 mL, 4.0 mmol) in 30 minutes. Stir at room temperature for 2 hours, wash with dilute hydrochloric acid (0.1 mol / L, 50 mL) and water (50 mL). Collect the organic phase, dry over anhydrous sodium sulfate, concentrate under reduced pressure, and separate and purify by silica gel column chromatography with gradient elution to obtain compound III (white powder, 466 mg, yield 89%).

[0057] Weigh compound III (524 mg, 1.0 mmol) and dissolve it in dichloromethane (60 mL). Add thionyl chloride (0.29 mL, 4.0 mmol) and heat under reflux for 2 hours. Concentrate the reaction solution under reduced pressure, add ether to make pulp, filter, wash and dry to obtain 2-acetylhydroxy-3-keto glycyrrhetinic acid chloride, dissolve it in tetrahydrofuran (100 mL), dissolve sodium azide (200 mg, 3.1 mmol) in water (30 mL), add it dropwise at 0-5 ° C, and complete the addition in 30 minutes. Stir for 1 hour, extract with ethyl acetate twice, wash with water twice, wash with saturated brine, dry over anhydrous sodium sulfate, filter to remove anhydrous sodium sulfate, heat the filtrate to 65 ° C, stir for 6 hours, concentrate the reaction solution under reduced pressure, add acetonitrile to recrystallize to obtain compound VI (white powder, 349 mg, yield 67%).

[0058] Weigh compound VI (1 mmol), dissolve it in ethyl acetate (20 mL), take the corresponding amine (1.5 mmol), stir at room temperature, react for 3-5 hours, monitor the reaction by TLC thin layer chromatography, after the reaction of compound VII is complete, wash it twice with distilled water and once with saturated brine, collect the organic phase, dry it with anhydrous sodium sulfate for 1 hour, and concentrate it to dryness under reduced pressure to obtain a crude compound VII.

[0059] The crude compound VII obtained in the previous step was dissolved in ethanol (20 mL), and sodium hydroxide powder (50 mg, 1.25 mmol) was added. The mixture was stirred at room temperature overnight. After monitoring by TLC thin-layer chromatography, the reaction of compound VIII was complete. 20 mL of ethyl acetate and 30 mL of distilled water were added to separate the mixture, and the mixture was washed twice with distilled water and once with saturated brine. The organic phase was collected, dried over anhydrous sodium sulfate for 1 hour, concentrated under reduced pressure, and separated and purified by silica gel column chromatography (petroleum ether: ethyl acetate, 2:1→1:3) to obtain the corresponding compound I1-I 16 .

[0060] The above-mentioned glycyrrhetinic acid urea derivative I1-I 16 The spectrum analysis of is shown below:

[0061] Example 1

[0062] Compound I1: white powder, yield 86%; 1 H NMR (500MHz, CDCl3) δ7.08 (s, 1H, OH), 6.26 (d, J = 19.2Hz, 1H, CH = C), 5.68 (s, 1H, NH) ,5.27(s,1H,C=CH),5.15(s,1H,NH),3.07–3.02(m,3H,NCH2),2.67(s,1H),2.22(s, 3H),2.03(s,3H),1.80(s,1H),1.56(s,2H),1.43(s,4H),1.31(s,5H),1.28(s,5H), 1.19(s,4H),1.10(s,4H),1.13(s,4H),1.00–0.99(m,1H),0.86(s,2H),0.81(s,3H); 13 C NMR(126MHz, CDCl3)δ200.56(C=O),199.66(C=O),171.73(C=CH),158.67(NC=O),143.49 (C-OH),131.21(CH=CO),127.71(CH=C),56.96,53.15,52.27,46.73,45.69,44.24,43.7 6,43.65(NCH2),41.93,37.85,35.88,32.23,31.94,31.34,29.73,28.86,28.53,27.03, 26.44(2C),26.21,23.49,23.39,21.81,20.97,19.07,18.02,11.54(CH3,propylamine).

[0063] Example 2

[0064] Compound I2: white powder, yield 89%; 1 H NMR (400MHz, CDCl3) δ7.07(s,1H,OH),6.00(s,1H,OH),5.79(s,1H,C=CH),5.75(s,1H,CH=C),5.43(s,1H,NH),5. 30(s,1H,NH),3.70(s,2H,NCH2),3.31(s,2H,CH2OH),2.72(d,J=6.6Hz,1H),2.27(d,J=10.5Hz,1H),2.05(d,J=1 8.8Hz,3H),1.82(d,J=13.7Hz,2H),1.69(d,J=13.7Hz,2H),1.58(s,2H),1.49(s,1H),1.45(s,3H),1.36(d,J=6. 6Hz,4H),1.32(s,4H),1.25(s,1H),1.23(s,3H),1.16(s,3H),1.13(s,3H),1.02(d,J=12.1Hz,1H),0.84(s,4H); 13 C NMR (101MHz, CDCl3) δ200.51(C=O), 200.28(C=O), 172.34(C=CH), 159.54(NC=O ),143.56(C-OH),130.77(CH=CO),127.72(CH=C),63.35(CH2OH),57.12,53.57 ,53.31,52.56,46.97,45.88,44.25,43.90,43.15,43.04,37.87,35.93,32.35 ,32.07,28.80,28.66,27.11,26.57,26.30,23.45,21.88,21.08,19.15,18.08.

[0065] Example 3

[0066] Compound I3: white powder, yield 81%; 11H NMR (500 MHz, CDCl3) δ 7.10 (s, 1H, OH), 5.94 (s, 1H, C=CH), 5.66 (s, 1H, CH=C), 5.48 (d, J=19.7 Hz, 1H, NH), 4.87 (s, 1H, NH), 2.69 (s, 1H), 2.40 (d, J=3.1 Hz, 2H), 2.12 (s, 1H), 2.07 (s.1H), 2.03 (s, 1H, NCH), 1.85 (s, 2H), 1.72 (s, 1H), 1.57 (s, 2H), 1.45 (s, 2H), 1.35 (s, 5H), 1.26 (s, 1H), 1.21 (s, 4H), 1.16 (s, 5H), 1.19 (s, 4H), 1.05 (s, 1H), 0.87 (s, 6H), 0.75–0.73 (m, 2H, cyclopropyl), 0.57 (s, 2H, cyclopropyl); 13 13C NMR (126 MHz, CDCl3) δ 200.52 (C=O), 198.97 (C=O), 175.67 (C=CH), 158.96 (NC=O), 143.48 (C-OH), 131.02 (CH=CO), 128.08 (CH=C), 56.99, 53.26, 52.70, 47.40, 45.70, 44.17, 43.76, 42.66, 37.83, 35.96, 32.30, 32.00, 31.91, 28.77, 28.60, 27.09, 26.46, 26.25, 23.47, 22.57, 21.83, 20.99, 19.10, 18.06, 7.66 (2C, cyclopropyl).

[0067] Example 4

[0068] Compound I4: white powder, yield 90%; 1H NMR (400MHz, CDCl3) δ7.12 (s, 1H, OH), 6.41 (s, 1H, NCH), 5.69 (d, J = 4.0 Hz, 2H, NH), 5.30 (s, 1H, C = CH), 4.87 (s, 1H, CH = C), 4.17 (s, 1H), 3.83 (d, J = 7.0 Hz, 1H), 3.43 (t, J = 6.3 Hz, 1H), 2.70 (d, J = 6.9 Hz, 1H), 2.18 (s, 1H), 2.07 (s, 2H), 1.95 (s, 1H), 1.85(s,1H),1.75(d,J=4.0Hz,1H),1.69(s,1H),1.58(s,2H),1.47(s,3H),1.41(s,4H),1.34(d,J=1.8Hz,3H),1.34–1.3 2(m,3H),1.25(s,2H),1.23(s,3H),1.17(s,5H),1.14(s,2H),1.12(s,2H),1.03(d,J=13.3Hz,1H),0.87(d,J=3.3Hz,6H); 13 CNMR(101MHz, CDCl3)δ200.90(C=O),200.55(C=O),170.03(C=CH),157.75(NC=O),143.50(C-OH),1 36.59(CH=CO),128.07(CH=C),57.04,54.33,53.34,52.70(NCH),47.18,45.75,45.64,44.41,44.21 ,43.81,37.89,37.53,35.94,33.90(cyclopentyl),33.83(cyclopentyl),32.08,28.79,27.11,26 .52,24.31,23.93(cyclopentyl),23.48(cyclopentyl),22.14,21.87,21.65,21.03,19.16,18.11.

[0069] Example 5

[0070] Compound I5: white powder, yield 79%; 1H NMR (500MHz, CDCl3) δ7.12(s,1H,OH),5.96(s,1H,C=CH),5.70(s,1H,CH=C),5.30(s,1H,NH),4.84(s,1H,NH),4.46(s,1H,NCH),3. 42(d,J=11.0Hz,1H),2.70(s,1H),2.18(t,J=5.4Hz,1H),2.13(d,J=13.5Hz,1H),2.07(s,2H),2.04(dd,J=7.8,5.8Hz,1H),1.96(s ,1H),1.92(s,2H),1.86–1.81(m,1H),1.72–1.69(m,5H),1.59(d,J=7.5Hz,2H),1.46(s,4H),1.42(dd,J=8.4,2.3Hz,1H),1.35(s, 3H),1.33(s,4H),1.29(d,J=3.6Hz,1H),1.26–1.24(m,1H),1.22(s,3H),1.16(s,4H),1.13(s,4H),1.04–1.00(m,1H),0.86(s,4H); 13 C NMR(126MHz, CDCl3)δ200.57(C=O),199.29(C=O),170.92(C=CH),157.41(NC=O),143.48(C-OH),131.0 5(CH=CO),127.96(CH=C),57.02,53.29,52.53,49.32(NCH),47.01,45.73,44.21,43.79,43.40,37.87, 35.91,34.06(cyclohexyl),33.97(cyclohexyl),33.32,32.32,32.05,28.82,28.65,27.09,26.50,26. 29(cyclohexyl),25.74(cyclohexyl),25.62(cyclohexyl),25.08,23.50,21.86,21.00,19.13,18.08.

[0071] Example 6

[0072] Compound I6: white powder, yield 89%; 1H NMR (500MHz, CDCl3) δ7.12(s,1H,OH),5.89(s,1H,C=CH),5.69(s,1H,CH=C),5.30(s,1H,NH),3.80(s,1H, NCH2),3.31–3.30(m,2H,NCH2),2.71(s,1H,NCH2),2.13(s,2H),2.07(d,J=4.6Hz,1H),2.04(s,1H),1.92( d,J=2.6Hz,2H),1.85(s,1H),1.73(t,J=13.4Hz,4H),1.59(s,2H),1.47(s,3H),1.37–1.35(m,7H),1.33(s ,1H),1.29–1.28(m,2H),1.24(d,J=9.0Hz,7H),1.17(d,J=3.3Hz,3H),1.14(s,3H),0.88(d,J=2.4Hz,3H); 13 C NMR(126MHz, CDCl3)δ200.56(C=O),198.92(C=O),170.29(C=CH),156.12(NC=O),14 3.49(C-OH),131.03(CH=CO),128.14(CH=C),57.01,53.33,52.79,47.42,45.74,45. 71(2C,NCH2),44.20,43.81,43.17,37.87,36.03,32.36,32.11,31.98,29.01,28.83 ,27.12,26.53,26.34,25.76(2C,pyrrolidine),23.51,21.87,21.02,19.13,18.12.

[0073] Example 7

[0074] Compound I7: white powder, yield 82%; 1H NMR (500MHz, CDCl3) δ7.12(s,1H,OH),5.69(s,2H,C=CH,CH=C),5.30(s,1H,NH),4.06(s,2H,NCH2) ,3.29(d,J=5.5Hz,2H,NCH2),2.71(s,1H),2.08(d,J=6.4Hz,4H),1.87–1.85(m,2H),1.72(s,2H), 1.59(s,3H),1.55(s,3H),1.47(s,3H),1.44(s,1H),1.43(s,1H),1.39–1.32(m,6H),1.28(d,J=3. 1Hz,1H),1.24(s,1H),1.23(s,4H),1.17(s,4H),1.14(s,3H),1.04(d,J=13.9Hz,1H),0.87(s,4H); 13 C NMR (126MHz, CDCl3) δ200.56 (C=O), 198.93 (C=O), 170.25 (C=CH), 156.90 (NC=O), 143. 49(C-OH),131.03(CH=CO),128.14(CH=C),57.00,53.33,52.73,47.53,45.74,45.44( 2C,NCH2),44.19,43.81,42.93,37.87,36.08,32.36,32.11,29.83,28.87,28.81,27. 12,26.53,26.34,25.83(2C,piperidine),24.62,23.48,21.86,21.02,19.13,18.12.

[0075] Example 8

[0076] Compound I8: white powder, yield 85%; 1H NMR (400MHz, CDCl3) δ7.10(s,1H,OH),5.93(s,1H,NH),5.67(s,1H,C=CH),5.29(s,1H,CH= C),4.15(s,1H,NCH2),3.69(s,4H,morpholine),3.30(s,3H,morpholine),2.70(s,1H),2. 06(d,J=9.1Hz,3H),1.85(d,J=13.1Hz,2H),1.72(s,1H),1.57(s,3H),1.45(s,4H),1.35–1 .34(m,5H),1.26(s,1H),1.21(s,6H),1.15(s,4H),1.12(s,3H),1.07(s,1H),0.86(s,4H); 13 C NMR(101MHz, CDCl3)δ200.52(C=O),199.02(C=O),170.25(C=CH),157.00(NC=O),143.45 (C-OH),130.98(CH=CO),128.10(CH=C),66.63(2C,OCH2,morpholine),56.98,53.26,52. 97,47.37,45.71,44.35(2C,NCH2,morpholine),44.17,43.77,42.67,37.83,35.99,32. 29,32.06,32.01,28.85,28.66,27.09,26.46,26.28,23.47,21.84,21.00,19.09,18.06.

[0077] Example 9

[0078] Compound I9: white powder, yield 84%; 1H NMR (500MHz, CDCl3) δ7.10(s,1H,NH),5.67(s,1H,C=CH),4.08(s,1H,CH=C),3.39–3.30(m,4H,NCH2 ,methylpiperazine),2.69(s,1H),2.37(t,J=5.0Hz,4H),2.30(s,3H,NCH3),2.11–1.99(m,4H),1. 90–1.81(m,2H),1.71(t,J=13.8Hz,2H),1.58(d,J=4.6Hz,3H),1.45(s,3H),1.40(s,3H),1.34(d,J =6.7Hz,6H),1.23–1.18(m,5H),1.15(d,J=4.1Hz,3H),1.12(s,3H),1.06–1.00(m,1H),0.86(s,3H); 13 CNMR(126MHz, CDCl3)δ200.51(C=O),198.88(C=O),170.14(C=CH),156.75(NC=O),143.50(C-OH ),131.00(CH=CO),128.11(CH=C),56.98,54.78(2C,NCH2,methylpiperazine),53.27,52.87,4 7.43,46.19,45.70,44.18,44.04,43.77(NCH3,methylpiperazine),42.72,37.82,36.01,32.3 1,32.07,28.83,28.70,27.10,27.00,26.47,26.29,25.46,23.46,21.83,20.99,19.09,18.08.

[0079] Example 10

[0080] Compound I 10 : White powder, yield 83%; 1H NMR(500MHz,CDCl3)δ8.49(s,1H,NCH,pyridine),7.71–7.65(m,1H,pyridine),7.32(d,J=7.8Hz,1H,pyridine),7.21– 7.17(m,1H,pyridine),7.09(d,J=2.3Hz,1H,OH),5.86(d,J=40.0Hz,1H),5.64(s,1H),5.34–5.03(m,2H),4.48–4.36(m, 2H,NCH2),2.68(s,1H),2.20(dd,J=13.2,3.7Hz,1H),2.08–2.02(m,2H),1.82(s,1H),1.70(s,1H),1.57(s,3H),1.47–1. 45(m,3H),1.33(s,6H),1.27–1.21(m,8H),1.13(d,J=10.1Hz,6H),1.00(d,J=12.6Hz,1H),0.84(dd,J=22.5,4.1Hz,5H); 13 CNMR(126MHz, CDCl3)δ199.45(C=O),197.95(C=O),169.73(C=CH),157.43,156.86(NC =O),147.61,142.44(C-OH),136.23,129.95(CH=CO),126.77(CH=C),121.39(2C),55.8 6,52.15,51.48,45.73,44.77,44.56(NCH2),43.10,42.62,41.86,36.70,34.64,31.19 ,30.87,28.68,27.63,27.43,26.01,25.37,25.16,22.35,20.72,19.87,17.99,16.96.

[0081] Embodiment 11

[0082] Compound I 11 : White powder, yield 86%; 1H NMR(500MHz,CDCl3)δ8.55(s,1H,NCH,pyridine),8.49–8.45(m,1H,NCH,pyridine),7.71(d,J=7.9Hz,1H,pyridine),7.29(dd,J=7.8, 4.9Hz,1H,pyridine),7.06(s,1H,OH),5.54(d,J=17.8Hz,2H),5.30(s,1H,NH),4.78(s,1H,NH),4.38(m,2H,NCH2),2.69(s,1H),2.12(d ,J=11.5Hz,1H),2.05–2.01(m,1H),1.84–1.79(m,1H),1.71(d,J=13.3Hz,2H),1.58(s,3H),1.46(d,J=9.8Hz,4H),1.42–1.38(m,1H),1. 35(d,J=9.6Hz,6H),1.31(s,1H),1.28–1.25(m,4H),1.22(s,3H),1.14(d,J=4.7Hz,5H),1.01(d,J=13.4Hz,1H),0.85(d,J=29.6Hz,4H); 13 C NMR (126MHz, CDCl3) δ200.53(C=O), 199.48(C=O), 171.17(C=CH), 157.56(NC=O), 148.64(NCH, p yridine),148.37(NCH,pyridine),143.55(C-OH),137.99,136.08,130.77(CH=CO),127.75(CH =C),123.99,57.05,53.29,52.73,47.16,45.76(NCH2),44.23,43.85,41.65,37.84,35.86,32. 34,32.08,31.62,29.84,28.84,28.64,27.12,26.51,26.24,23.45,21.88,21.08,19.16,18.08.

[0083] Example 12

[0084] Compound I 12 : White powder, yield 85%; 1H NMR(400MHz, CDCl3) δ8.52(d,J=5.6Hz,2H,NCH,pyridine),7.29(d,J=5.3Hz,2H,pyridine),7.05(s,1H ,OH),6.02(s,1H),5.47(s,1H),5.30(s,1H,NH),5.08(s,1H,NH),4.38(s,2H,NCH2),2.67(s,1H),2.18– 2.01(m,3H),1.85–1.76(m,1H),1.68(d,J=13.3Hz,2H),1.59–1.57(m,2H),1.44(s,4H),1.33(d,J=10.6 Hz, 6H), 1.28 (s, 1H), 1.23 (d, J = 10.1Hz, 7H), 1.13 (d, J = 6.2Hz, 6H), 1.01 (d, J = 11.6Hz, 1H), 0.81 (s, 4H).

[0085] Embodiment 13

[0086] Compound I 13 : White powder, yield 89%; 1 H NMR(400MHz, CDCl3)δ7.69(s,1H),7.15(s,1H,NH),6.97–6.94(m,3H,benzene),5.70(s,2H),5.30(s,2H,NH),2 .70(s,1H),2.38(d,J=14.1Hz,1H),2.27(d,J=13.7Hz,1H),2.20–2.15(m,1H),2.09(s,3H),2.05–2.01(m,1H),1 .86–1.81(m,1H),1.72(d,J=12.2Hz,2H),1.60(s,2H),1.47(s,3H),1.36(d,J=5.5Hz,2H),1.32(d,J=7.6Hz,4H) ,1.28(s,1H),1.23(s,3H),1.18(s,1H),1.15(s,5H),1.05(d,J=7.3Hz,1H),0.89(d,J=3.9Hz,1H),0.80(s,4H).

[0087] Embodiment 14

[0088] Compound I 14 : White powder, yield 90%; 1H NMR (400MHz, CDCl3) δ7.19(s,2H,benzene),7.15(s,1H,OH),6.96(t,J=5.2Hz,1H,benzene),6.78–6.67(m,1H,benze ne),6.28(s,1H),6.18(s,1H),5.60(d,J=45.8Hz,1H,NH),5.30(s,1H,NH),2.72–2.51(m,8H),2.06(d,J=11.6Hz,2H) ,1.93(d,J=15.4Hz,1H),1.85(d,J=10.5Hz,1H),1.74(d,J=10.2Hz,2H),1.61(s,3H),1.45(s,6H),1.39(d,J=4.0Hz, 2H),1.36(s,2H),1.27–1.23(m,4H),1.20–1.16(m,4H),1.0252(s,1H),0.86(d,J=27.1Hz,6H),0.77(d,J=4.2Hz,1H).

[0089] Embodiment 15

[0090] Compound I 15 : White powder, yield 89%; 1 H NMR (400MHz, CDCl3) δ7.32(s,1H,OCH,furan),7.09(s,1H,OH),6.29–6.25(m,1H,furan),6.19–6.16(m,1H,furan),5.6 8(s,1H,NH),5.34(d,J=39.8Hz,1H,NH),4.93(d,J=25.6Hz,1H),4.31(dd,J=13.8,3.7Hz,2H,NCH2),3.47(s,1H),2.68(s ,1H),2.17(d,J=5.4Hz,1H),2.06–2.01(m,1H),1.82(d,J=4.4Hz,1H),1.68(d,J=13.2Hz,2H),1.57(s,2H),1.45(s,6H) ,1.32(d,J=6.4Hz,10H),1.25–1.24(m,1H),1.22(s,3H),1.15(s,3H),1.12(s,3H),1.01(d,J=12.1Hz,1H),0.82(s,3H); 13C NMR (101MHz, CDCl3) δ200.58(C=O), 199.55(C=O), 171.24(C=CH), 157.66(NC=O), 152.87(CO, furan),143.49(C-OH),141.99(OCH,furan),131.02(CH=CO),127.80(CH=C),110.50(furan), 106.77(furan),57.02,53.27,52.63,46.88,45.74,44.22,43.79,43.47,37.86,37.51,35.84 ,32.31,32.01,31.41,28.79,28.55,27.08,26.48,26.23,23.45,21.86,20.99,19.14,18.06.

[0091] Example 16

[0092] Compound I 16 : White powder, yield 86%; 1 H NMR (500MHz, CDCl3) δ7.08(s,1H,OH),5.98(s,2H,NH),5.49(s,1H,C=CH),4.10(d,J=17 .5Hz,1H,CH=C),3.74–3.64(m,2H,NCH2),2.75(s,1H),2.33–2.28(m,1H),2.14–2.04(m ,2H),1.86–1.70(m,4H),1.60(d,J=7.9Hz,4H),1.45(s,5H),1.40(s,3H),1.36(s,3H), 1.21(d,J=14.1Hz,6H),1.16(s,3H),1.13(s,3H),1.00(d,J=13.4Hz,1H),0.81(s,3H); 13 C NMR(126MHz, CDCl3)δ201.58(C=O),200.48(C=O),172.97(C=CH),160.73(COOH) ,159.25(NC=O),143.55(C-OH),130.79(CH=CO),127.73(CH=C),57.03,53.25,52 .67,47.20,45.99,44.24,43.86,42.52(NCH2),37.88,35.50,32.40,32.15,28.8 4,28.70,27.15,26.75,26.75,26.15,25.46,23.23,21.88,21.24,19.27,18.07.

[0093] Example 17 Glycyrrhetinic acid urea derivative I1-I 16 Cytotoxicity evaluation:

[0094] Cell viability and cisplatin resistance were determined by MTT assay. RAW264.7 cells or HK2 cells in good logarithmic phase growth were taken, the original culture medium was discarded, washed three times with PBS, and fresh culture medium was added. The cells were blown evenly with a 1 ml pipette, counted on a cell counting plate, and then seeded in a 96-well plate (1×10 4 / well) and placed in a cell culture incubator for 24 hours before removal. The original culture medium was discarded, and freshly prepared culture medium containing different concentration gradient compounds was added to the well plate seeded with RAW264.7 cells and cultured for another 24 hours. After HK2 cells were treated with the freshly prepared title compound (100 μM) for 12 hours, cisplatin (20 μM) was added to each well and incubated for 24 hours.

[0095] Add 20 μL MTT solution (5 mg / mL) in dark and incubate for another 4 h. Discard the liquid in the wells and add 150 μL dimethyl sulfoxide (DMSO) to dissolve the formazan. After 15 min, measure the OD values ​​of all wells at a wavelength of 490 nm.

[0096] The results are as follows Figure 2 As shown, most compounds did not affect cell viability, indicating that the new glycyrrhizic acid urea compounds had little toxicity to mouse macrophages and would not affect the subsequent anti-inflammatory activity determination experiments.

[0097] Example 18 Glycyrrhetinic acid urea derivative I1-I 16 Evaluation of anti-inflammatory activity

[0098] NO is an important pro-inflammatory mediator. Excessive production of NO in the body may lead to inflammation-related diseases. Existing literature reports that NO inhibitors can be used to treat inflammation-related diseases. In order to detect the glycyrrhetinic acid urea derivatives I1-I 16 The anti-inflammatory effect of RAW264.7 cells was detected by Griess reagent. 5RAW264.7 cells were inoculated into 10 cells / well and cultured for 20 hours, and then the cells were pretreated with a compound at a concentration of 20uM, and then stimulated with LPS (1ug / ml) after 2 hours of preparation in serum-free medium. After 24 hours of LPS stimulation, the nitrite level was detected using Griess reagent (Beyotime, China) according to the instructions to determine the amount of NO produced. The results showed that compared with the normal group, the NO level of the cells treated with LPS increased significantly. Compared with the cells treated with LPS, the compounds prepared by the present invention can reduce the NO content in the cell supernatant, among which compounds (I1, I3, I5, I6, I8, I9) 11 , I 15 ) significantly reduced the NO content in the cell supernatant, which shows that the glycyrrhetinic acid urea compounds prepared by the present invention have good anti-inflammatory activity, among which compounds (I1, I3, I5, I6, I8, I 11 , I 15 ) has a more significant anti-inflammatory activity.

[0099] Table 2 Glycyrrhetinic acid urea derivatives I1-I 16 Inhibitory effect on the production of inflammatory factor NO

[0100] compound 20μM (inhibition rate%) compound 20μM (inhibition rate%) <![CDATA[I1]]> 59.17±3.19 <![CDATA[I9]]> 30.31±4.85 <![CDATA[I2]]> 22.48±1.10 <![CDATA[I 10 ]]> 16.20±3.15 <![CDATA[I3]]> 66.93±2.03 <![CDATA[I 11 ]]> 51.57±2.24 <![CDATA[I4]]> 39.53±5.02 <![CDATA[I 12 ]]> 30.77±1.85 <![CDATA[I5]]> 81.65±4.67 <![CDATA[I 13 ]]> 45.30±0.70 <![CDATA[I6]]> 95.09±0.97 <![CDATA[I 14 ]]> 27.39±2.56 <![CDATA[I7]]> 41.03±9.13 <![CDATA[I 15 ]]> 102.94±2.5 <![CDATA[I8]]> 55.30±1.32 <![CDATA[I 16 ]]> 22.79±2.45

[0101] Note: Glycyrrhetinic acid urea derivatives I1-I 16 The inhibitory effect of compounds I1-I on NO production in LPS-stimulated RAW 264.7 cells at a concentration of 20 μM was compared with that of compounds I1-I 16 The NO inhibition rate of compound I was found 15 The NO inhibition rate was the highest.

[0102] Example 19 Glycyrrhetinic acid urea derivative I1-I 16 Protective effect on renal tubular epithelial cells

[0103] In patients with acute kidney injury (AKI) induced by cisplatin, renal tubular epithelial cells are severely damaged in the early stage, mainly manifested by apoptosis, autophagy, cell cycle protein dysregulation, activation of mitogen-activated protein kinase (MAPK) signaling pathway, direct toxicity to renal tubular epithelial cells, DNA damage, etc. Damage to the renal tubules ultimately leads to structural and functional disorders of the kidneys, and in severe cases can lead to renal failure or even death of the patient. Therefore. Compounds with protective effects on renal tubular epithelial cells can improve cisplatin-induced AKI to a certain extent.

[0104] The MTT method was used to determine the protective effect of the compounds on cisplatin-induced HK2 renal tubular epithelial cell injury. HK2 cells were treated with the same concentration (50 μM) of glycyrrhetinic acid urea compounds 12 h in advance and then treated with cisplatin for 24 h. Figure 3 As shown, compared with the normal group, the cisplatin group significantly reduced cell activity. 10 ,I 14 ,I 15 and I 16 ) after treatment, the cell viability was significantly improved compared with the cisplatin group, indicating that glycyrrhizic acid urea compounds can play a certain protective role on renal tubular epithelial cells damaged by cisplatin.

[0105] Example 20 Glycyrrhetinic acid urea derivative I 15 Binding to STING (stimulator of interferon genes) enhances thermal stability

[0106] The cellular thermal shift assay (CETSA) method can provide key target contact information for a range of different drug targets. We use the CETSA method to first treat cells with drugs or controls, lyse the cells and remove cell debris and aggregates by centrifugation, and finally use denaturing gel electrophoresis and specific antibodies for the target protein to detect the remaining thermally stable target protein by western blot. This method can evaluate the thermal stability of proteins with or without ligands. The specific principle is that the binding of drugs to target proteins enhances the thermal stability of target proteins, and unbound proteins are more likely to denature and precipitate. Figure 4 As shown, compound I 15 The processed STING protein has better thermal stability. 15 The temperature-induced aggregation curve of the group was generally higher than that of the DMSO group, and was most obvious at 55℃, 60℃ and 65℃. 15 The amount of TLR4 protein in the treated cells indicated that 15 It binds to the STING protein in a dose-dependent manner to reduce protein degradation. 15 It can act as a ligand to interact with STING and improve its thermal stability.

[0107] Example 21 Glycyrrhetinic acid urea derivative I 15 Inhibition of LPS-induced inflammation in RAW264.7 cells by blocking the STING / NF-κB pathway

[0108] Acute kidney injury (AKI) is closely related to both intrarenal and systemic inflammation, involving a variety of different molecular mechanisms including the STING / NF-κB pathway. Abnormal activation of the STING / NF-κB pathway is closely related to inflammation, so preventing this activation is of great significance for the treatment of AKI.

[0109] RAW264.7 cells in good condition were inoculated into 6-well plates and treated with different concentration gradients of glycyrrhetinic acid urea derivative I 15After 1 hour of treatment, the cells were exposed to LPS (0.5 μg / mL) or cisplatin (10 μM) for 24 hours. The broken kidney tissue and cells were lysed with pre-cooled RIPA lysis buffer (containing PMSF and phosphatase inhibitors) for 30 minutes, and then centrifuged at 12,000 rpm for 20 minutes in a low-temperature centrifuge (4°C). The supernatant was added to the protein buffer and mixed well, and then boiled for 10 minutes (100°C) to obtain the protein sample, which was used directly or stored in a -20°C refrigerator.

[0110] The prepared protein samples were electrophoresed at 80V low voltage for 30min in SDS-PAGE polyacrylamide gel, then adjusted to 120V for 50min, and then transferred to PVDF membrane at appropriate current intensity (200mA, 60min). After blocking with rapid blocking solution for 10-15 minutes, the membrane was incubated with primary antibodies against β-actin (1:1 1500, Affinity, USA), KIM-1 (1:1 1000, imounway, USA), STING (1:1 1000, Proteintech, USA), NFκB-p65 (1:1 1000, Proteintech, USA), p-NFκB-p65 (1:1000, Abcam, USA), iNOS (1:1 1000, Proteintech, USA) and at 4°C overnight. Finally, the membranes were incubated with enzyme-labeled secondary antibody (1:1 5000, Affinity, USA) at room temperature for 1 h and exposed to light using enhanced chemiluminescence detection reagent.

[0111] Figure 5 Western blot showed that compared with the control group, the LPS group significantly increased the expression of STING and iNOS and NF-κB phosphorylation, activating the STING / NF-κB pathway. 15 The results showed that glycyrrhizic acid urea derivative I 15 It can play an anti-inflammatory role by blocking STING / NF-κB signaling.

[0112] Example 22 Glycyrrhetinic acid urea derivative I 15 Significantly reduced the levels of various inflammatory factors in cisplatin-induced AKI mice.

[0113] Thirty-two 8-week-old male C57BL / 6 mice (20-25 g) were purchased from Hangzhou Ziyuan Laboratory Animal Technology Co., Ltd. Before the experiment, the mice were adaptively fed at appropriate temperature and humidity for 7 days. The mice were randomly divided into 4 groups (8 mice in each group): control group, model group (cisplatin 20 mg / kg), low-dose I 15Group (5 mg / kg) + cisplatin, high dose I 15 Group I (10 mg / kg) + cisplatin. 15 The mice in group A were intragastrically administered with compound I for 7 consecutive days. 15 (30mg / kg), and the other groups were given an equal amount of normal saline. One hour after the end of the oral administration on the third day, except for the normal group mice which were intraperitoneally injected with cisplatin solution (20mg / kg), the normal group mice were intraperitoneally injected with the same dose of normal saline. The animals were killed after anesthesia on the seventh day, and blood, kidney and liver tissues were collected. The blood sample was placed on ice for 30 minutes, centrifuged at 1200rmp for 5 minutes, and the upper serum was taken and stored at -80℃. When used, it can be thawed at room temperature. Half of the kidney was stored at -80℃, and the other part was stored in 4% paraformaldehyde solution to fix the tissue, which was convenient for subsequent sectioning and observation of tissue morphology.

[0114] Take out the ELISA kit from the -20℃ refrigerator in advance and place it at room temperature for more than 30 minutes to warm it up. Centrifuge the standard and dilute it with the sample diluent according to the concentration gradient given in the instructions to obtain eight different concentrations of standard solutions. Add the collected mouse serum and standard solution to a 96-well plate containing a solid phase carrier, incubate in a 37℃ constant temperature shaker for 2 hours, discard the liquid in the well, shake dry, directly add 100μL of biotin antibody working solution, and incubate at 37℃ for 2 hours. Discard the liquid in the well, wash the plate twice and shake it off, add 100μL of horseradish peroxidase working solution to all wells, and incubate in a 37℃ constant temperature shaker for 2 hours. Discard the liquid in the well, wash the plate 5 times and shake it off. Add 90μL of substrate solution to each well and incubate in a 37℃ constant temperature shaker in the dark for 30 minutes. After taking out the 96-well plate, add the stop solution to each well to terminate the reaction. At a wavelength of 450nm, the absorbance in the well was measured with an enzyme-labeled instrument. The standard curve was calculated, and the protein concentrations of IL-1β, IL-6, and TNF-α were obtained by substituting the absorbance into the standard curve.

[0115] Add 20 μL of double distilled water, 10 mmol / L BUN standard solution and collected mouse serum to the buffered enzyme solution. Mix the solution and incubate at 37°C for 10 minutes. Add 1 mL of phenol colorimetric agent and 1 mL of alkaline sodium hypochlorite. Mix thoroughly and incubate at 37°C for 10 minutes. Add 200 μL of solution to each well of a 96-well plate, detect the absorbance of each well at a wavelength of 640 nm and calculate the urea nitrogen content. All operations followed the manufacturer's protocol, and Scr and BUN levels were measured using creatinine and urea assay kits.

[0116] In cisplatin-induced AKI, the secretion of various inflammatory cytokines is increased, such as IL-1β, IL-6, IL-18, monocyte chemoattractant protein-1, expressed and secreted by activated normal T cells, macrophage inflammatory protein-2, intercellular cell adhesion molecule-1, and transforming growth factor-β.

[0117] like Figure 6 Glycyrrhetinic acid urea derivative I 15 After treatment, the level of inflammatory factors was significantly reduced. 15 Can significantly reduce inflammation.

[0118] Example 23 Glycyrrhetinic acid urea derivative I 15 Protective effect on cisplatin-induced acute kidney injury

[0119] One of the important characteristics of AKI is renal tubular damage or even necrosis. 15 The therapeutic and preventive effects of cisplatin-induced AKI in mice were evaluated.

[0120] According to Example 22, the kidney tissues soaked in 4% paraformaldehyde solution for 24 hours were sliced ​​and stained with H&E, and then the histological morphology was observed using a slide scanner. Figure 7 As shown in the figure, the renal tubular epithelial cells in the normal group were normal in morphology, without necrosis or inflammatory cell infiltration. In the kidney sections of the cisplatin group, it was observed that the renal tubular epithelial cells were swollen and necrotized, with obvious interstitial spaces and inflammatory cell infiltration, and the pathological damage of the renal tissue was significant. 15 After administration of high dose I 15 The renal tubular epithelial cells in the group were normal in morphology, without obvious fragmentation or necrosis, and the glomerular morphology was normal, indicating that the glycyrrhetinic acid urea derivative I 15 It has a significant protective effect on cisplatin-induced acute kidney injury.

[0121] Example 24 Glycyrrhetinic acid derivative I1-I 16 Application examples

[0122] The following formulations were prepared in a conventional manner:

[0123] Name of raw materials and auxiliary materials Dosage Active ingredients 10～100g Compressible starch 50～200g Magnesium Stearate 1.5g Polyvinylpyrrolidone 15～30g Hydroxypropyl methylcellulose 15～30g 75% ethanol solution 80~100mL

[0124] The glycyrrhizic acid urea derivatives prepared in Examples 1 to 16 are passed through a 120-mesh sieve, and the compressible starch is passed through a 100-mesh sieve. The prescribed amount of compressible starch is weighed and mixed with the raw materials in equal increments; polyvinyl pyrrolidone and hydroxypropyl methylcellulose are dissolved in a 75% ethanol solution, and the mixed powder of the above active ingredients and compressible starch is taken, and the above solution is used as a wetting agent to make a soft material, granulated with a 30-mesh sieve, dried at 60°C for 2 to 3 hours, and sieved with a 26-mesh sieve. Take the above-prepared granules, add the prescribed amount of magnesium stearate, mix well, determine the intermediate content, and compress into tablets or capsules to obtain the finished preparation.

[0125] Glycyrrhizic acid urea derivatives can also be made into sustained-release tablets, dripping pills, granules, injections, etc. The clinically recommended dose is 30-100 mg / day, orally, twice a day.

[0126] The above embodiments are only for illustrating the technical concept and features of the present invention, and their purpose is to enable people familiar with the technology in this field to understand the content of the present invention and implement it, and they cannot be used to limit the protection scope of the present invention. Any equivalent changes or modifications made according to the spirit of the present invention should be within the protection scope of the present invention.

Claims

1. A compound or a stereoisomer, tautomer, derivative, prodrug or pharmaceutically acceptable salt thereof, characterized in that: It has the following general formula: In formula (I), R is a mono-substituted or di-substituted amino group; The monosubstituent of the amino group is selected from substituted or unsubstituted C1-C6 alkyl, C3-C6 cycloalkyl, substituted or unsubstituted C6-C8 aryl; The disubstituted amino group is The n is selected from 1-2; the Z is selected from C, O or N; the position of Z is any reasonable position on the ring; and the R1 is selected from H, C1-C3 alkyl.

2. The compound according to claim 1 or its stereoisomer, tautomer, derivative, prodrug or pharmaceutically acceptable salt, characterized in that: The substituents of the C1-C6 alkyl group are selected from hydroxyl, carboxyl, The Z is selected from O or N; the position of the Z is any reasonable position on the ring; the substituent of the C6-C8 aryl is selected from halogen and C1-C3 alkyl.

3. The compound according to claim 1 or its stereoisomer, tautomer, derivative, prodrug or pharmaceutically acceptable salt, characterized in that Said In the above, when n is 1, Z is C and R1 is H; when n is 2, Z is selected from C, N, O, and Z is in the para position to the N on the ring, and R1 is selected from H and methyl.

4. The compound according to claim 2 or its stereoisomer, tautomer, derivative, prodrug or pharmaceutically acceptable salt, characterized in that: Said Wherein Z is O, and Z is in adjacent positions to another substituent on the ring; Wherein Z is N, and Z is in the ortho, para or meta position relative to another substituent on the ring.

5. The compound according to claim 1 to 4 or its stereoisomer, tautomer, derivative, prodrug or pharmaceutically acceptable salt, characterized in that: R is selected from n-propylamino, cyclopropylamino, cyclopentylamino, cyclohexylamino, hydroxyethylamino, carboxymethylamino, 4-fluorophenylamino, 2-ethylphenylamino, tetrahydropyrrolyl, piperidinyl, morpholinyl, N-methylpiperazinyl, 2-pyridylmethylamino, 3-pyridylmethylamino, 4-pyridylmethylamino, and 2-furanylmethylamino.

6. A method for preparing the compound of claims 1 to 5 or its stereoisomers, tautomers, derivatives, prodrugs or pharmaceutically acceptable salts, characterized in that: The preparation method comprises the following steps: (1) 2-Hydroxy-3-glycyrrhetinic acid ketone (II) is acylated with acetic anhydride to generate 2-acetoxy-3-glycyrrhetinic acid ketone (III) (2) 2-acetoxy-3-glycyrrhetinic acid ketone (III) reacts with thionyl chloride to generate 2-acetoxy-3-glycyrrhetinic acid chloride-3-ketone (IV), which then reacts with sodium azide to generate 2-acetoxy-3-glycyrrhetinic acid azide-3-ketone (V), which is then heated and rearranged to obtain isocyanate (VI) (3) Isocyanate (VI) reacts with substituted amine to obtain 2-acetoxyethylene-glycyrrhetinic acid urea (VII) i: R1 and R2 are not H at the same time. When one of them is H, the other is n-propyl, cyclopropyl, cyclopentyl, cyclohexyl, hydroxyethyl, carboxymethyl, 4-fluorophenyl, 2-ethylphenyl, 2-pyridylmethyl, 3-pyridylmethyl, 4-pyridylmethyl, or 2-furylmethyl; ii: When neither R1 nor R2 is H, represents tetrahydropyrrole, morpholine, piperidine, N-methylpiperazine; (4) 2-Acetoxyene-glycyrrhetinic acid urea (VII) is hydrolyzed under alkaline conditions to obtain glycyrrhetinic acid urea derivative (I) 7. The preparation method according to claim 6, characterized in that The solvent used in step (1) is an aprotic solvent, the acylating agent is acetic anhydride, and the reaction temperature is -10 to 50°C; the solvent for the reaction of 2-acetoxygenyl glycyrrhetinyl chloride-3-one (IV) with sodium azide in step (2) is a mixed solution of tetrahydrofuran or acetone and water, and the ratio of the two solvents is 1:5 to 5:1 (volume ratio), and the reaction temperature is -5 to 30°C; the temperature of the rearrangement reaction of 2-acetoxygenyl glycyrrhetinic acid azide-3-one (V) is 30 to 100°C; the reagent used for the hydrolysis of 2-acetoxygenyl-glycyrrhetinic acid urea (VII) in step (4) is an inorganic base, the solvent is an alcohol solvent, and the reaction temperature is 0 to 60°C.

8. A pharmaceutical composition, characterized in that The pharmaceutical composition contains the compound according to any one of claims 1 to 5 or its stereoisomer, tautomer, derivative, prodrug or pharmaceutically acceptable salt, and a pharmaceutically acceptable carrier.

9. Use of the compound according to any one of claims 1 to 5 or its stereoisomers, tautomers, derivatives, prodrugs or pharmaceutically acceptable salts in the preparation of anti-inflammatory drugs or anti-inflammatory pharmaceutical compositions.

10. Use of the compound according to any one of claims 1 to 5 or its stereoisomers, tautomers, derivatives, prodrugs or pharmaceutically acceptable salts in the preparation of drugs or pharmaceutical compositions for treating and / or preventing acute kidney injury, acute lung injury, chronic obstructive pulmonary disease, septicemia and other related diseases.

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