Cordycepin-glycyrrhizic acid derivatives and their application in the preparation of products for preventing and treating lung injury / pulmonary fibrosis

Abstract

The present invention relates to the field of pharmaceutical technologies, and specifically discloses cordycepin-glycyrrhizic acid derivatives, a preparation method thereof, and an application thereof in the preparation of products for preventing and treating lung injury / pulmonary fibrosis. The cordycepin-glycyrrhizic acid derivatives are obtained by connecting the A-terminal site of the glycyrrhizic acid monomer structure shown by formula Ia with any one or more sites of the Bi to Biii terminals of the cordycepin monomer structure shown by formula Ib. The cordycepin-glycyrrhizic acid derivatives provided by the present invention have the characteristics of good safety, high bioavailability, and multiple mechanisms. They can not only overcome the structural defects of cordycepin itself, but also strengthen the treatment of lung injury / pulmonary fibrosis through a synergistic effect, and can be effectively applied to the prevention and treatment of clinical major diseases such as lung injury / pulmonary fibrosis.

Description

Technical Field

[0001] The present invention relates to the field of pharmaceutical technology, and specifically relates to cordycepin-glycyrrhizic acid derivatives, a preparation method thereof, and an application thereof in the preparation of products for preventing and treating lung injury / pulmonary fibrosis. Background Art

[0002] Cordyceps is a traditional Chinese medicine. Cordycepin, as an active ingredient of Cordyceps, is a precious natural product with various biological activities such as anti-tumor, anti-viral, immunomodulatory, and anti-inflammatory effects. It has been found that cordycepin can induce apoptosis of tumor cells; inhibit apoptosis and renal fibrosis of cells in diabetic nephropathy mice by promoting autophagy, and reduce the histopathological damage of diabetic complications; have a protective effect on various inflammatory diseases such as hepatitis by regulating signaling pathways such as NF-κB and Nrf2 / HO-1. In addition, cordycepin can enhance the transcriptional activity of Nrf2, induce the expression of antioxidant enzymes and phase II detoxifying enzyme HO-1, and reverse LPS-induced lung injury. The above results indicate that cordycepin can effectively reduce inflammation and improve tissue fibrosis, and has good preventive and improvement effects on pneumonia, lung injury, tissue fibrosis, etc. However, there are many problems in the research on the drug properties and industrialization of cordycepin, which seriously restrict its in-depth development and scientific application. First of all, natural cordyceps is expensive and the extraction cost is high, up to more than $200 per 0.1 gram. Most domestic research on cordycepin stays at the level of artificial cultivation of Cordyceps fruiting bodies and mycelia, with low technical content. Cordyceps products with high cordycepin content are scarce, and mainly rely on scale to promote efficiency, unable to compete with foreign similar products. Therefore, empowering the extraction of active components in traditional Chinese medicine with biomanufacturing technology can greatly increase the content and extraction efficiency of natural active components, and then have significant market competitiveness and generate huge economic benefits. On the other hand, as an analogue of adenosine, cordycepin has a short metabolic cycle in vivo, is easily hydrolyzed by adenosine deaminase, and is rapidly converted into biologically inactive 3'-deoxyinosine. Its in vivo efficacy is not as expected. Therefore, modifying or derivatizing cordycepin to improve the drug activity of cordycepin and enhance or prolong the drug effect is an important direction in the research on the drug properties of cordycepin.

[0003] To stabilize the structure of cordycepin, reactions such as acylation, phosphorylation, and esterification are usually carried out to add side chains, groups, or ions with anti-deamination effects to construct cordycepin derivatives with reasonable structures. The N-acyl cordycepin derivatives synthesized by Wei et al. can protect its amino group and improve its biological activity; Shimada et al. reported that substituting the hydrogen at the 4'-position with fluorine can delay its metabolism and reduce its cytotoxicity. Currently, the types of cordycepin derivatives obtained are limited, and although adding certain side chain groups to cordycepin can delay deamination, it still cannot completely block the deamination by ADA. Therefore, it is still necessary to improve the types and positions of the side chains and groups of cordycepin derivatives. Therefore, the present invention provides a cordycepin-glycyrrhizic acid derivative, a preparation method thereof, and an application thereof in the preparation of products for preventing and treating lung injury / lung fibrosis. Summary of the Invention

[0004] Object of the Invention: The technical problem to be solved by the present invention is to provide a cordycepin-glycyrrhizic acid derivative in view of the deficiencies of the prior art.

[0005] Another technical problem to be solved by the present invention is to provide a preparation method of the cordycepin-glycyrrhizic acid derivative.

[0006] Another technical problem to be solved by the present invention is to provide a composition containing the above-mentioned cordycepin-glycyrrhizic acid derivative.

[0007] A further technical problem to be solved by the present invention is to provide an application of the above-mentioned cordycepin-glycyrrhizic acid derivative.

[0008] Idea of the Invention: Starting from the pathogenesis of lung injury / lung fibrosis, through the early design of target prediction and molecular docking technology, the present invention uses cordycepin, a characteristic bio-manufactured product, as the parent nucleus and glycyrrhizic acid as the auxiliary molecule to synthesize the innovative compound cordycepin-glycyrrhizic acid derivative. And through techniques such as molecular pharmacology, pharmacokinetics, and cell biology, its efficacy, mechanism of action, and in vivo process are comprehensively evaluated to provide a theoretical basis and technical support for the development of candidate new drugs.

[0009] To solve the above first technical problem, the present invention discloses a cordycepin-glycyrrhizic acid derivative, or its stereoisomer, or its pharmaceutically acceptable salt.

[0010] In some embodiments, the cordycepin-glycyrrhizic acid derivative is obtained by connecting the A-terminal site of the glycyrrhizic acid monomer structure shown by formula Ia with any one or more sites of the Bi to Biii terminals of the cordycepin monomer structure shown by formula Ib.

[0011]

[0012] In some embodiments, the cordycepin-glycyrrhizic acid derivative is selected from any one of I-1 to I-7.

[0013] In some embodiments, the cordycepin-glycyrrhizic acid derivative is selected from I-2, I-6 or I-7.

[0014] In some embodiments, the cordycepin-glycyrrhizic acid derivative is selected from I-2 or I-6.

[0015] In some embodiments, the cordycepin-glycyrrhizic acid derivative is selected from I-2.

[0016]

[0017]

[0018] To solve the above second technical problem, the present invention discloses a method for preparing the above cordycepin-glycyrrhizic acid derivative.

[0019] In some embodiments, the method for preparing the cordycepin-glycyrrhizic acid derivative comprises the following steps: (1) Synthesis of the active molecule glycyrrhetinyl chloride (the first reaction): Glycyrrhizic acid is subjected to acyl chlorination modification with oxalyl chloride, and the reaction is carried out until glycyrrhizic acid is completely dissolved. After the reaction is completed, distillation under reduced pressure and recrystallization are carried out to obtain glycyrrhetinyl chloride shown in Formula II; (2) Synthesis of the series of cordycepin-glycyrrhizic acid derivatives (the second reaction): The obtained glycyrrhetinyl chloride is subjected to a one-pot reaction with the compound shown in Formula III to prepare a mixture of the above cordycepin-glycyrrhizic acid derivatives I-2, I-6 and I-7.

[0020]

[0021] Wherein, R 1 and R 2 are each independently selected from a hydroxyl group or a protecting group, R 3 is selected from an amino group or a protecting group; and R 1 , R 2 are not simultaneously selected from protecting groups, and there is also no case where only R 2 is a protecting group; the protecting group includes but is not limited to -OTBS, -TBDPSO, -OAc, -NHCbz.

[0022] In the first reaction,

[0023] In some embodiments, the molar ratio of oxalyl chloride to glycyrrhizic acid is 1-5:1; in some embodiments, the molar ratio of oxalyl chloride to glycyrrhizic acid is 1-2:1; in some embodiments, the molar ratio of oxalyl chloride to glycyrrhizic acid is 2:1.

[0024] In some embodiments, the reaction further includes a catalyst; in some embodiments, the catalyst is N,N-dimethylformamide; in some embodiments, the amount of the catalyst used is 10-200 μL, in some embodiments, the amount of the catalyst used is 10-100 μL, in some embodiments, the amount of the catalyst used is 40-60 μL, in some embodiments, the amount of the catalyst used is 50 μL.

[0025] In some embodiments, the reaction is carried out in an organic solvent or a solvent-free environment; in some embodiments, the organic solvent is dichloromethane; in some embodiments, the molar volume ratio of glycyrrhizic acid to the organic solvent is 1 mmol:5-100 mL; in some embodiments, the molar volume ratio of glycyrrhizic acid to the organic solvent is 1 mmol:5-35 mL; in some embodiments, the molar volume ratio of glycyrrhizic acid to the organic solvent is 1 mmol:20 mL.

[0026] In some embodiments, the temperature of the reaction is 20-100 °C; in some embodiments, the temperature of the reaction is 40-70 °C; in some embodiments, the temperature of the reaction is the reflux temperature.

[0027] In the second reaction,

[0028] In some embodiments, when R in the compound shown in Formula III 1 is selected from hydroxyl, R 2 is selected from hydroxyl, R 3 is selected from amino, the prepared cordycepin-glycyrrhizic acid derivative is a mixture of Compounds I-2, I-6 and I-7; the specific preparation method is that cordycepin (the compound shown in Formula III, R 1 is selected from hydroxyl, R 2 is selected from hydroxyl, R 3 is selected from amino) reacts directly with glycyrrhl chloride to obtain a mixture of Compounds I-2, I-6 and I-7.

[0029] In some embodiments, when R in the compound shown in Formula III 1 , R 2 or R 3 any one or two groups are selected from protecting groups, the prepared cordycepin-glycyrrhizic acid derivatives are Compounds I-1, I-3, I-4 and I-5 respectively; the specific preparation method is as follows: using cordycepin as a raw material, the compound shown in Formula III (R 1 , R 2 or R 3Any one or two groups are selected from protecting groups). After the compound shown in Formula III reacts with glycyrrhoyl chloride, the protecting group is removed to obtain the cordycepin-glycyrrhizic acid derivative. Taking cordycepin-glycyrrhizic acid derivative I-5 as an example, the specific reaction route is shown as follows.

[0030]

[0031] In some embodiments, the molar ratio of the glycyrrhoyl chloride to the compound shown in Formula III is 1-5:1; in some embodiments, the molar ratio of the glycyrrhoyl chloride to the compound shown in Formula III is 1-3:1; in some embodiments, the molar ratio of the glycyrrhoyl chloride to the compound shown in Formula III is 2:1.

[0032] In some embodiments, the solvent for the reaction is an organic solvent; in some embodiments, the organic solvent is anhydrous pyridine; in some embodiments, the molar volume ratio of the compound shown in Formula III to the organic solvent is 1 mmol:10-200 mL; in some embodiments, the molar volume ratio of the compound shown in Formula III to the organic solvent is 1 mmol:10-50 mL; in some embodiments, the molar volume ratio of the compound shown in Formula III to the organic solvent is 1 mmol:20 mL.

[0033] In some embodiments, the reaction further includes triethylamine; in some embodiments, the molar ratio of the compound shown in Formula III to triethylamine is 1:4-8; in some embodiments, the molar ratio of the compound shown in Formula III to triethylamine is 1:6.

[0034] In some embodiments, the temperature of the reaction is 60-100 °C; in some embodiments, the temperature of the reaction is 70-90 °C; in some embodiments, the temperature of the reaction is 75-85 °C; in some embodiments, the temperature of the reaction is 80 °C.

[0035] In some embodiments, after the second reaction, the product is identified by modern analytical methods including MS, HPLC-MS, NMR, and FTIR; in some embodiments, if the obtained product is a mixture, according to the difference in the polarity of the molecular structure, a continuous chromatographic separation method is used to separate and purify the mixture, and the high-efficiency separation and purification of the glycyrrhizic acid-cordycepin series derivatives are achieved through the adsorption effect of different types of resins on the cordycepin derivatives; in some embodiments, if the obtained product is a mixture, after the second reaction, separation and purification are carried out by a rapid liquid chromatography preparative instrument with n-hexane and ethyl acetate as the mobile phase to obtain compounds I-2, I-6, and I-7 respectively.

[0036] To solve the above-mentioned third technical problem, the present invention discloses a pharmaceutical composition comprising (i) the cordycepin-glycyrrhizic acid derivative in the above-mentioned first technical problem, or its stereoisomer, or its pharmaceutically acceptable salt, and (ii) a pharmaceutically acceptable carrier, excipient or adjuvant.

[0037] In some embodiments, the dosage form of the pharmaceutical composition is a capsule, granule, tablet, injection, chewable tablet, patch, etc.

[0038] To solve the above-mentioned fourth technical problem, the present invention discloses the application of the above-mentioned cordycepin-glycyrrhizic acid derivative, or its stereoisomer, or its pharmaceutically acceptable salt, or the above-mentioned pharmaceutical composition in the preparation of products for treating and / or preventing lung injury or pulmonary fibrosis, and in the preparation of products for treating and / or preventing pneumonia.

[0039] In some embodiments, the products include but are not limited to health products, drugs, etc.

[0040] Beneficial effects: Compared with the prior art, the present invention has the following advantages:

[0041] The cordycepin-glycyrrhizic acid derivative provided by the present invention has the characteristics of good safety, high bioavailability and multiple mechanisms. It can not only overcome the structural defects of cordycepin itself, but also strengthen the treatment of lung injury / pulmonary fibrosis through synergistic effects, and can be effectively applied to the prevention and treatment of major clinical diseases such as lung injury / pulmonary fibrosis. BRIEF DESCRIPTION OF THE DRAWINGS

[0042] The following further describes the present invention in detail with reference to the drawings and specific embodiments, and the above and / or other advantages of the present invention will become clearer.

[0043] Figure 1 It is the synthetic route diagram of the cordycepin-glycyrrhizic acid derivative in the present invention.

[0044] Figure 2 It is the effect of the cordycepin-glycyrrhizic acid derivative on the lung index of LPS-induced lung injury mice.

[0045] Figure 3 It is the effect of the cordycepin-glycyrrhizic acid derivative on the pathological changes of the lungs of LPS-induced lung injury mice.

[0046] Figure 4 It is the effect of the cordycepin-glycyrrhizic acid derivative on the lung index of bleomycin-induced pulmonary fibrosis mice.

[0047] Figure 5 It is the effect of the cordycepin-glycyrrhizic acid derivative on TGF-β1 in the lungs of bleomycin-induced pulmonary fibrosis mice. DETAILED DESCRIPTION OF THE EMBODIMENTS

[0048] In the following examples, the experimental methods are conventional methods unless otherwise specified; the reagents and materials can be obtained from commercial sources unless otherwise specified.

[0049] Example 1: Preparation of Cordycepin-Glycyrrhizic Acid Derivatives

[0050] (1) Synthesis of the active molecule Glycyrrhetinyl Chloride:

[0051] Weigh glycyrrhizic acid (10 mmol, 8.09 g) into a reaction flask, add 200 mL of dichloromethane as the solvent, add oxalyl chloride (20 mmol, 2.54 g), and additionally add 50 μL of N,N-dimethylformamide. The reaction is carried out under reflux at 50 °C until glycyrrhizic acid is completely converted. After the reaction is completed, the reaction solution is subjected to vacuum distillation, recrystallized with n-hexane, and filtered to obtain the active molecule Glycyrrhetinyl Chloride. The NMR and mass spectrometry detections are as follows:

[0052] 1 H NMR(400MHz,CDCl 3 )5.77(S,1H),4.47-5.60(m,4H),3.60-4.11(m,5H),3.03(m,1H),1.12-2.10(m,31H),0.94(s,1H),0.91(s,3H),0.89(s,6H); 13 C NMR(100MHz,CDCl 3 )δ203.2,174.7,173.2,171.2,128.3,114.5,109.8,92.5,86.9,80.7,77.9,76.8,74.8,73.8,72.7,66.4,59.5,54.6,50.6,47.5,42.7,41.1,38.8,38.6,36.9,35.9,35.0,34.9,34.0,32.0,31.7,27.4,26.7,26.6,25.2,23.7,23.1,20.9,19.3,14.2.MSI-MS:811.36[M+H] + .

[0053] (2) Synthesis of Cordycepin-Glycyrrhizic Acid Derivative Series:

[0054] The derivative of cordycepin-glycyrrhizic acid was prepared by a one-pot method. Cordycepin (10 mmol, 2.54 g) and glycyrrhetinyl chloride (20 mmol, 16.5 g) were dissolved in 200 mL of anhydrous pyridine. The reaction was carried out under the protection of nitrogen or argon. In addition, triethylamine (60 mmol, 7.6 mL) was added for the one-pot reaction. The reaction was carried out at 80 °C. The reaction dynamics were monitored by a high-performance liquid chromatograph. When the cordycepin was completely converted, a reaction solution containing a mixture of glycyrrhizic acid-coupled cordycepin compounds I-2, I-6, and I-7 was obtained. The reaction solution was filtered, extracted with water and ethyl acetate, the organic phase was collected, and concentrated under reduced pressure to obtain a mixture.

[0055] Separation and purification were carried out by a rapid liquid chromatography preparative instrument using n-hexane and ethyl acetate as the mobile phase, and the effluent of the corresponding components was collected and concentrated to obtain the corresponding glycyrrhizic acid-cordycepin coupling compound. The rapid liquid chromatography preparative instrument is Sepabean of Santai Technology Changzhou Co., Ltd. M machine - standard model, ELSD detector, gradient elution with ethyl acetate and n-hexane (1:100 to 2:1).

[0056] Compound I-2: 1 H NMR (400 MHz, CD 3 OD) δ8.56 (s, 1H), 8.35 (s, 1H), 6.16 (d, 1H), 3.84 - 5.73 (m, 12H), 4.01 (m, 1H), 3.51 - 3.75 (m, 3H), 3.04 (m, 1H), 1.02 - 2.10 (m, 32H), 0.94 (s, 1H), 0.89 - 0.91 (m, 9H); 13 C NMR (100 MHz, CD 3 OD) δ203.1, 173.2, 171.2, 170.8, 152.4, 151.8, 149.8, 140.3, 128.3, 123.5, 112.3, 109.8, 99.2, 92.5, 83.6, 82.4, 80.7, 77.9, 76.8, 74.8, 73.8, 73.0, 72.7, 61.1, 59.5, 54.6, 47.5, 42.7, 41.7, 38.8, 38.6, 37.4, 36.9, 35.9, 35.6, 35.3, 34.5, 34.1, 31.7, 27.4, 26.7, 26.6, 25.2, 23.7, 23.6, 20.9, 19.3, 14.2. MSI-MS: 1043.2[M+H] + 。

[0057] Compound I-6: 1 H NMR (400 MHz, CD3 OD) δ 8.56 (s, 1H), 8.35 (s, 1H), 6.16 (d, 1H), 3.83 - 5.79 (m, 23H), 4.02 (m, 1H), 3.51 - 3.77 (m, 3H), 3.04 (m, 2H), 1.02 - 2.10 (m, 62H), 0.94 (m, 2H), 0.89 - 0.92 (m, 18H).

[0058] Compound I-7: 1 H NMR (400 MHz, CD 3 OD) δ 8.56 (s, 1H), 8.35 (s, 1H), 6.16 (d, 1H), 3.83 - 5.77 (m, 34H), 4.0 (m, 1H), 3.51 - 3.75 (m, 3H), 3.05 (m, 3H), 1.02 - 2.10 (m, 92H), 0.94 (m, 3H), 0.88 - 0.91 (m, 27H).

[0059] Application Example 1: Study on the ameliorating effect of cordycepin - glycyrrhizic acid derivatives on LPS - induced lung injury in mice

[0060] 1. Experimental materials and methods

[0061] (1) Experimental animals and main reagents

[0062] SPF - level C57BL / 6 male mice;

[0063] LPS, cordycepin, cordycepin - glycyrrhizic acid derivatives I - 2, I - 6 and I - 7 prepared in Example 1, positive drug (dexamethasone).

[0064] (2) LPS - induced acute lung injury model in mice

[0065] Male C57BL / 6 mice were adaptively fed for 1 week before the experiment to adapt to the environment. They were randomly divided into the following groups, with 6 mice in each group, namely the normal group, the model group, the cordycepin group (100 mg / kg), the cordycepin-glycyrrhizic acid derivative group (50 mg / kg and 100 mg / kg I-2, I-6 and I-7), and the positive drug dexamethasone group (1 mg / kg). The model group was given solvent control by gavage, the cordycepin group was given 100 mg / kg cordycepin by gavage, the cordycepin-glycyrrhizic acid derivative group was given 50 mg / kg and 100 mg / kg cordycepin-glycyrrhizic acid derivatives I-2, I-6 and I-7 by gavage respectively, and the positive drug dexamethasone group was given 1 mg / kg dexamethasone by intraperitoneal injection. After 3 hours of administration, LPS (15 mg / kg i.p.) was given to establish the model. Cordycepin (100 mg / kg), cordycepin-glycyrrhizic acid derivative (50 mg / kg and 100 mg / kg), and positive drug dexamethasone (1 mg / kg) were continuously given at 12, 36, and 60 hours after modeling. The mice were sacrificed 72 hours after modeling.

[0066] 2. Detection indexes

[0067] (1) Organs and organ indexes

[0068] After the mice in each group were sacrificed, the lungs were taken out, precisely weighed, and the organ indexes were calculated.

[0069] Lung index (mg / g) = lung weight (mg) / body weight (g)

[0070] (2) H&E staining pathological analysis

[0071] 3. Experimental results

[0072] (1) Effects of cordycepin-glycyrrhizic acid derivative on the lung index of LPS-induced lung injury mice

[0073] As Figure 2 shown, compared with the normal control group, the lung index of LPS-induced lung injury mice increased significantly (p < 0.01); compared with the model group, the lung indexes of the cordycepin-glycyrrhizic acid derivative group, the cordycepin group, and the dexamethasone (positive drug) group of mice decreased significantly and approached the normal control group; among them, the decrease in the cordycepin-glycyrrhizic acid derivative I-2 group was the most obvious and was closest to the normal control group.

[0074] (2) Effects of cordycepin-glycyrrhizic acid derivative on the pathological changes of the lungs of LPS-induced lung injury mice

[0075] As Figure 3As shown, obvious septal thickening, inflammatory cell infiltration, interstitial and alveolar edema were observed in the lung tissues of the mice in the model group. Cordycepin-glycyrrhic acid derivatives group (I-2) and dexamethasone could alleviate the pathological changes in mice with LPS-induced lung injury, reduce the alveolar septal thickness, decrease the degree of inflammatory cell infiltration, relieve interstitial edema, and significantly improve the symptoms of lung injury.

[0076] 4. Experimental conclusions

[0077] The cordycepin-glycyrrhic acid derivatives group could significantly reduce the lung index of mice with LPS-induced lung injury, making it approach normal, indicating that the cordycepin-glycyrrhic acid derivatives group could alleviate lung edema in mice with LPS-induced lung injury; the cordycepin-glycyrrhic acid derivatives group could also alleviate the pathological changes in mice with LPS-induced lung injury, reduce the alveolar septal thickness, decrease the degree of inflammatory cell infiltration, relieve interstitial edema, and significantly improve the pathological changes of lung injury in mice.

[0078] Application Example 2: Study on the improvement effect of cordycepin-glycyrrhic acid derivatives on bleomycin-induced pulmonary fibrosis in mice

[0079] 1. Experimental materials and methods

[0080] (1) Experimental animals and main reagents

[0081] SPF-grade C57BL / 6 male mice;

[0082] Bleomycin, cordycepin-glycyrrhic acid derivatives I-2, I-6 and I-7 prepared in Example 1, positive drug (pirfenidone).

[0083] (2) Bleomycin-induced mouse pulmonary fibrosis model

[0084] C57BL / 6 male mice were adaptively fed for 1 week before the experiment to adapt to the environment. They were randomly divided into the following groups, with 6 mice in each group, namely the normal group, the model group, the cordycepin-glycyrrhic acid derivatives group (50 mg / kg and 100 mg / kg I-2, I-6 and I-7), and the positive drug pirfenidone (400 mg / kg). The model group was given solvent control by gavage, the cordycepin-glycyrrhic acid derivatives group was given cordycepin derivatives 50 mg / kg and 100 mg / kg cordycepin-glycyrrhic acid I-2, I-6 and I-7 by gavage respectively, and the positive drug pirfenidone 400 mg / kg. After 7 days of administration, the mice were modeled. On the 1st, 4th, 8th, 11th, 15th, 18th, 22nd, and 25th days of modeling, bleomycin (50 mg / kg i.p.) was given, and on the 40th and 60th days of modeling, the mice were sacrificed. During the modeling period, the drugs were continued to be administered once a day.

[0085] 2. Detection indicators

[0086] (1) Lung index

[0087] After sacrificing each group of mice, the lungs were removed, precisely weighed, and the organ index was calculated.

[0088] Lung index (mg / g) = lung weight (mg) / body weight (g).

[0089] (2) Detection of TGF-β1 in lung tissue

[0090] Alveoli were collected and detected using an Elisa kit.

[0091] 3. Experimental results

[0092] (1) Effects of cordycepin-glycyrrhizic acid derivatives on the lung index of bleomycin-induced pulmonary fibrosis mice

[0093] As Figure 4 shown, compared with the normal control group, the lung index of the lungs of bleomycin-induced pulmonary fibrosis mice increased significantly (p < 0.01); compared with the model group, the lung index of the mice in the cordycepin-glycyrrhizic acid derivative group decreased significantly, approaching that of the normal control group (p < 0.01), especially the decrease in the cordycepin-glycyrrhizic acid derivative I-2 group was the most obvious and was closest to the normal control group; in addition, compared with the model group, the lung index of the mice in the pirfenidone (positive drug) group also decreased significantly and approached the normal control group (p < 0.01).

[0094] (2) Effects of cordycepin-glycyrrhizic acid derivatives on TGF-β1 in bleomycin-induced pulmonary fibrosis mice

[0095] As Figure 5 shown, compared with the normal control group, TGF-β1 in bleomycin-induced pulmonary fibrosis mice increased significantly (p < 0.01); compared with the model group, the lung index of the mice in the cordycepin-glycyrrhizic acid derivative group decreased significantly, especially the decrease in the cordycepin-glycyrrhizic acid derivative I-2 group was the most obvious and was closest to the normal control group; in addition, compared with the model group, the mice in the pirfenidone (positive drug) group also decreased significantly (p < 0.01).

[0096] The present invention provides the idea and method of cordycepin-glycyrrhizic acid derivatives and their application in the preparation of products for preventing and treating lung injury / pulmonary fibrosis. There are many methods and ways to specifically implement this technical solution. The above is only the preferred embodiment of the present invention. It should be noted that for those of ordinary skill in the art, without departing from the principle of the present invention, several improvements and refinements can be made, and these improvements and refinements should also be regarded as the protection scope of the present invention. Each component not clearly defined in this embodiment can be realized by existing technologies.

Claims

1. Cordycepin-glycyrrhizic acid derivative, or a pharmaceutically acceptable salt thereof, Characterized in that, The cordycepin-glycyrrhizic acid derivative is obtained by connecting the A-terminal site of the glycyrrhizic acid monomer structure shown by formula Ia with any one or more sites of the Bi to Biii terminals of the cordycepin monomer structure shown by formula Ib; 2. The cordycepin-glycyrrhizic acid derivative according to claim 1, or a pharmaceutically acceptable salt thereof, Characterized in that, The cordycepin-glycyrrhizic acid derivative is selected from any one of I-1 to I-7; 3. Preparation method of the cordycepin-glycyrrhizic acid derivative according to claim 1 or 2, Characterized in that, Glycyrrhizic acid reacts with oxalyl chloride in a first reaction to obtain glycyrrhizoyl chloride shown by formula II; the obtained glycyrrhizoyl chloride reacts with the compound shown by formula III in a second reaction to prepare the cordycepin-glycyrrhizic acid derivative; Wherein, R 1 and R 2 are each independently selected from a hydroxyl group or a protecting group, R 3 is selected from an amino group or a protecting group; and R 1 , R 2 are not simultaneously selected from protecting groups, and there is also no case where only R 2 is a protecting group; when R 1 or R 2 is selected from a protecting group, they are each independently selected from -OTBS, -OAc; when R 3 is selected from a protecting group, R 3 is -NHCbz.

4. The preparation method according to claim 3, Characterized in that, In the first reaction, the molar ratio of the oxalyl chloride to the glycyrrhizic acid is 1 to 5:

1.

5. The preparation method according to claim 3, Characterized in that, In the first reaction, the molar ratio of the oxalyl chloride to the glycyrrhizic acid is 1 to 2:

1.

6. The preparation method according to claim 3, Characterized in that, In the first reaction, the reaction also includes a catalyst, and the catalyst is N,N-dimethylformamide.

7. The preparation method according to claim 6, Characterized in that, The dosage of the catalyst is 10 to 200 μL.

8. The preparation method according to claim 6, Characterized in that, The dosage of the catalyst is 10 to 100 μL.

9. The preparation method according to claim 6, Characterized in that, The dosage of the catalyst is 50 μL.

10. The preparation method according to claim 3, Characterized in that, In the first reaction, the reaction is carried out in an organic solvent or a solvent-free environment.

11. The preparation method according to claim 10, Characterized in that, The organic solvent is dichloromethane.

12. The preparation method according to claim 10, Characterized in that, The molar volume ratio of the glycyrrhizic acid to the organic solvent is 1 mmol: 5 to 100 mL.

13. The preparation method according to claim 3, Characterized in that, In the first reaction, the reaction temperature is 20 to 100 °C.

14. The preparation method according to claim 3, Characterized in that, In the second reaction, the molar ratio of the glycyrrhizoyl chloride to the compound shown by formula III is 1 to 5:

1.

15. The preparation method according to claim 3, Characterized in that, In the second reaction, the molar ratio of the glycyrrhizoyl chloride to the compound shown by formula III is 1 to 3:

1.

16. The preparation method according to claim 3, Characterized in that, In the second reaction, the solvent for the reaction is an organic solvent, and the molar volume ratio of the compound shown by formula III to the organic solvent is 1 mmol: 10 to 200 mL.

17. The preparation method according to claim 16, Characterized in that, The molar volume ratio of the compound shown by formula III to the organic solvent is 1 mmol: 10 to 30 mL.

18. The preparation method according to claim 16, wherein, the organic solvent is anhydrous pyridine.

19. The preparation method according to claim 3, wherein, in the second reaction, the reaction temperature is 60-100 °C.

20. The preparation method according to claim 3, wherein, in the second reaction, the reaction temperature is 75-85 °C.

21. A pharmaceutical composition, wherein, it comprises (i) the cordycepin-glycyrrhizic acid derivative according to claim 1 or 2, or a pharmaceutically acceptable salt thereof, and (ii) a pharmaceutically acceptable carrier.

22. Use of the cordycepin-glycyrrhizic acid derivative according to claim 1 or 2, or a pharmaceutically acceptable salt thereof, or the pharmaceutical composition according to claim 21 in the preparation of a product for treating and / or preventing lung injury or pulmonary fibrosis.

23. Use of the cordycepin-glycyrrhizic acid derivative according to claim 1 or 2, or a pharmaceutically acceptable salt thereof, or the pharmaceutical composition according to claim 21 in the preparation of a product for treating and / or preventing pneumonia.

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