Curcumol derivative with therapeutic effects on acute lung injury

Abstract

This invention discloses a curcumin derivative with therapeutic effects on acute lung injury and its preparation method, comprising: dissolving curcumin in tetrahydrofuran, removing air from the reaction apparatus, and adding nitrogen to the reaction apparatus to fill it with nitrogen; then adding ethyl magnesium bromide and refluxing at a first preset temperature for 30 min; then adding succinic anhydride and refluxing at a second preset temperature for 4 h; stopping the reaction by adding water and adjusting the pH to 4 with hydrochloric acid; after extraction with ethyl acetate, purifying the upper layer using a preset method to obtain a yellow oily substance I-1; reacting curcumin succinate monoester I-1 with a 5% sodium bicarbonate solution, and evaporating the solvent to obtain curcumin succinate monoester sodium salt II. The raw material used in this invention has wide clinical applications and is safe and low in toxicity. The hydroxyl group of curcumin is combined with succinic anhydride through a Grignard reaction to form curcumin succinate monoester I-1, which is then reacted with sodium bicarbonate solution to obtain curcumin succinate monoester sodium salt II.

Description

Technical Field

[0001] This invention belongs to the field of turmeric alcohol derivative preparation technology, specifically relating to turmeric alcohol derivatives with therapeutic effects on acute lung injury and their preparation methods. Background Technology

[0002] Acute lung injury (ALI) is a common and complex clinical syndrome in critically ill patients, often progressing to acute respiratory distress syndrome (ARDS). It is characterized by rapid onset, frequently accompanied by non-cardiogenic pulmonary edema and persistent hypoxemia, and can lead to fatal respiratory failure. Due to the high prevalence and mortality of ALI / ARDS, identifying its causative factors and finding appropriate treatments is crucial. Currently, there are no specific drugs for treating acute lung injury; however, administering anti-inflammatory drugs to suppress inflammation is a potential treatment approach to improve ALI.

[0003] Curcumol is one of the main active ingredients in the traditional Chinese medicine Curcuma zedoaria. It exhibits good anti-inflammatory activity and can regulate multiple signaling pathways, including JAK1 / STAT3, PI3K / Akt / NF-κB, and TGF-β1 / Smads. Curcumol also shows potential in the treatment of lung diseases, influencing the development of asthma and promoting apoptosis. Its mechanism of action is mainly related to the inhibition of ERK / CREB pathway activation. Curcumol also has a certain therapeutic effect on chronic asthma, reducing lung inflammation in mice with chronic asthma and inhibiting abnormal activation of the Wnt / β-catenin pathway. Studies have also found that curcumol has good potential in treating acute lung injury; curcumol powder inhalation can effectively reduce the expression of inflammatory factors in ALI rats, with effects comparable to dexamethasone.

[0004] Although curcumin shows potential in treating acute lung injury, its extremely poor water solubility significantly inhibits its clinical application. Furthermore, the mechanism of action and molecular targets of curcumin in treating acute lung injury remain unclear and require further investigation. To address these issues, this paper proposes a curcumin derivative with therapeutic effects on acute lung injury and its preparation method. Summary of the Invention

[0005] The purpose of this invention is to provide a curcumin alcohol derivative with therapeutic effects on acute lung injury and a method for its preparation, so as to solve the problems mentioned in the background art.

[0006] To achieve the above objectives, the present invention provides the following technical solution: a curcuminol derivative with therapeutic effects on acute lung injury and a method for its preparation, wherein the curcuminol derivative has the following structure:

[0007]

[0008] Preferably, the synthetic route for the curcuminol derivative is as follows:

[0009]

[0010] A method for preparing the above-mentioned curcumin alcohol derivative includes the following steps:

[0011] S1: Dissolve curcumin in tetrahydrofuran, remove the air from the reaction apparatus, and add nitrogen to the reaction apparatus to fill it with nitrogen.

[0012] S2: Then add ethyl magnesium bromide and reflux for 30 min at the first preset temperature;

[0013] S3: Then add succinic anhydride and reflux at the second preset temperature for 4 hours;

[0014] S4: Add water to stop the reaction, and adjust the pH to 4 with hydrochloric acid;

[0015] S5: After extraction with ethyl acetate, the upper layer was purified using a pre-defined method to obtain a yellow oily substance I-1;

[0016] S6: Curcumol succinate monoester I-1 was reacted with 5% sodium bicarbonate solution, and the solvent was evaporated to obtain curcumol succinate monoester sodium salt II.

[0017] Preferably, in step S1, the amount of turmeric alcohol added is 0.4 mmol.

[0018] Preferably, in step S1, the amount of tetrahydrofuran added is 2.5 mL.

[0019] Preferably, in step S2, the amount of ethyl magnesium bromide added is 0.4 mmol.

[0020] Preferably, in step S2, the first preset temperature is 55°C.

[0021] Preferably, in step S2, the first preset temperature is 75°C.

[0022] Preferably, in step S3, the amount of succinic anhydride added is 1.2 mmol.

[0023] Preferably, in step S5, the yield of the yellow oily substance I-1 is 82%.

[0024] Preferably, in step S5, the preset method is silica gel column chromatography.

[0025] Compared with the prior art, the beneficial effects of the present invention are:

[0026] The raw materials used in this invention are widely used in clinical practice and are safe and low in toxicity. Through a Grignard reaction, the hydroxyl group of curcumin is combined with succinic anhydride to form curcumin succinate monoester I-1. This is then reacted with a 5% sodium bicarbonate solution to obtain curcumin succinate monoester sodium salt II. At a dose of 5 mg / kg, it can improve LPS-induced acute lung injury and reduce lung inflammation. Through proteomics, molecular docking, cell thermal transfer experiments, and protein imprinting experiments, it was determined that curcumin succinate monoester sodium salt II mainly regulates GSK3β expression to inhibit the activation of the mTOR signaling pathway, thereby exerting a therapeutic effect on acute lung injury. GSK3β is the direct target of curcumin succinate monoester sodium salt II. This invention demonstrates the potential of curcumin succinate monoester II as a lead compound in acute lung injury treatment strategies. GSK3β holds promise as a novel target for treating ALI, providing insights for the development of specific anti-ALI drugs. Attached Figure Description

[0027] Figure 1 This is a schematic diagram of the 1H NMR spectrum of the curcumin succinate monoester I-1 of the present invention.

[0028] Figure 2 This is a schematic diagram of the carbon NMR spectrum of the curcumin succinate monoester I-1 of the present invention.

[0029] Figure 3 This is a high-resolution mass spectrometry diagram of Curcumol Succinate Monoester I-1 of the present invention;

[0030] Figure 4 This is a schematic diagram of the experimental results of the proteomics analysis and protein imprinting experiments of this invention;

[0031] Figure 5 These are the experimental results of molecular docking analysis and cell thermal transfer experiments of this invention. Detailed Implementation

[0032] The technical solutions of the embodiments of the present invention will be clearly and completely described below with reference to the accompanying drawings. Obviously, the described embodiments are only some embodiments of the present invention, and not all embodiments. Based on the embodiments of the present invention, all other embodiments obtained by those skilled in the art without creative effort are within the scope of protection of the present invention.

[0033] Please see Figures 1-5 As shown, the present invention provides the following technical solution:

[0034] Example 1: Synthesis of Curcumol Succinate Monoester Sodium Salt II:

[0035] Curcumol derivatives, which have therapeutic effects on acute lung injury, are reacted via Grignard reaction to combine succinic anhydride with the hydroxyl group of curcumol, forming curcumol succinate monoester I-1. This is then reacted with 5% sodium bicarbonate solution to yield curcumol succinate monoester sodium salt II.

[0036] Furthermore, the synthetic route for the above-mentioned turmeric succinate monoester sodium salt is as follows:

[0037]

[0038] Furthermore, the preparation method of turmeric alcohol succinate monoester I-1 in this invention is as follows:

[0039] 0.4 mmol of curcumin was dissolved in 2.5 mL of tetrahydrofuran. The air in the reaction apparatus was evacuated, and the apparatus was filled with nitrogen. Then, 0.4 mmol of ethyl magnesium bromide was added, and the mixture was refluxed at 55 °C for 30 min. Next, 1.2 mmol of succinic anhydride was added, and the mixture was refluxed at 75 °C for 4 h. The reaction was stopped by adding water, and the pH was adjusted to 4 with hydrochloric acid. After extraction with ethyl acetate, the upper layer was purified by silica gel column chromatography to obtain a yellow oil I-1 in 82% yield. Curcumin succinate monoester I-1 was then dissolved in an equimolar amount of 5% sodium bicarbonate solution to obtain curcumin succinate monoester sodium salt II.

[0040] In addition, regarding the NMR and high-resolution mass spectrometry data of curcumin succinate monoester I-1 in this invention: 1H NMR (600MHz, CDCl3) δ4.89-4.85(m,2H,H-14),2.70-2.58(m,4H,H-2',H-3'),2.51(d,J=14.7Hz,1H, H-9a),2.34(td,J=11.9,11.2,6.4Hz,1H,H-9b),2.22-2.11(m,2H,H-1,H-6a),1.99-1.64(m,5H,H-2 a,H-3a,H-4,H-7,H-11),1.55-1.45(m,1H,H-2b),1.29-1.24(m,1H,H-3b),1.15(dd,J=12.3,6.5Hz, 1H,H-6b),1.00(d,J=6.6Hz,3H,H-15),0.94(d,J=6.4Hz,3H,H-12),0.86(d,J=6.5Hz,3H,H-13).13C NMR(150MHz, CDCl3)δ177.55(C-1'),169.36(C-16),144.43(C-10),113.1 8(C-14),109.40(C-8),89.73(C-5),54.55(C-1),51.61(C-7),39.42(C-4) ,36.82(C-9),33.49(C-6),30.86(C-3),30.26(C-3'),28.64(C-2'),28.34 (C-2,C-11),22.79(C-13),21.37(C-12),12.22(C-15).HR-MS(ESI):Calcd for C 19 H 27 O5(MH):335.1858; found:335.1855.

[0041] Furthermore, to verify the effect of turmeric succinate monoester sodium salt II on improving LPS-induced acute lung injury, the verification method is as follows:

[0042] Example 1

[0043] Thirty-six male C57BL / 6 mice, aged 6 weeks and acclimatized for one week, were randomly divided into six groups (n=6): a control group, a model group, three dose groups of turmeric succinate sodium monoester II (50, 25, and 5 mg / kg), and a dexamethasone group (DEX, 5 mg / kg). Mice were anesthetized by intraperitoneal injection of 1 μL / g tribromoethanol, followed by tracheal instillation of 50 μL of LPS at a concentration of 15 mg / kg. The control group received physiological saline. Two hours after LPS modeling, the high, medium, and low dose groups of turmeric succinate sodium monoester II and the dexamethasone group received the corresponding dose of the drug via tail vein injection, while the control and model groups received the same volume of physiological saline. Mice were sacrificed 24 hours after LPS administration, and bronchoalveolar lavage fluid was collected for later use.

[0044]

[0045] Table 1

[0046] The results are shown in Table 1. Curcumol succinate monoester sodium salt II can inhibit the expression of inflammatory factors in the bronchoalveolar lavage fluid of LPS-induced acute lung injury mice. At a dose of 5 mg / kg, it can significantly reduce the expression of TNF-α, IFN-γ and IL-17A, which is superior to the positive control drug dexamethasone (DEX).

[0047] Furthermore, to verify the mechanism of action and molecular target of turmeric succinate monoester sodium salt II in the treatment of acute lung injury through proteomics analysis and protein imprinting experiments, the verification method is as follows:

[0048] Example 2

[0049] 4D-label-free proteomics analysis was performed on mouse lung tissue from LPS-induced acute lung injury, including protein extraction, peptide digestion, data acquisition, and database retrieval. Liquid chromatography-tandem MS (LC-MS / MS) analysis was performed on a Nanoelute (Brook) coupled with a TimsTOFPro mass spectrometer. Qualitative and quantitative analysis of the mass spectrometry data was performed using MaxQuant 1.6.14 software. Proteins with significantly different expression levels (greater than 1.15 fold change and P-value < 0.05) were selected for screening.

[0050] Lung tissues from LPS-induced ALI mice in the Control, Model, and Curcumol succinate monoester sodium II groups were identified using proteomics, revealing 5383 differentially expressed proteins. Figure 4 In the A group, among which, turmeric succinate monoester sodium salt II showed 292 differentially expressed proteins compared to the model group. Figure 4(B in the text). GO functional analysis revealed that turmeric succinate monoester sodium salt II is mainly involved in metabolic processes, biological regulation of biological processes, and responses to stimuli. Figure 4 KEGG pathway enrichment analysis revealed that the mTOR signaling pathway may be a potential pathway for the anti-inflammatory activity of turmeric succinate monoester sodium salt II. Figure 4 (D in the text). In the mTOR pathway, compared with the model group, turmeric succinate monoester sodium salt II upregulated GSK3β protein expression, presumably exerting its therapeutic effect on acute lung injury by regulating GSK3β. Western blot results showed that turmeric succinate monoester sodium salt II can upregulate GSK3β expression and inhibit the expression of S6K protein downstream of the mTOR signaling pathway, indicating that turmeric succinate monoester sodium salt II can regulate GSK3β expression, thereby inhibiting the activation of the mTOR signaling pathway. Figure 4 (E in the middle).

[0051] Furthermore, to verify that molecular docking analysis and cell thermal transfer experiments indicate that GSK3β is the direct target of turmeric succinate monoester sodium salt II, the verification method is as follows:

[0052] Example 3

[0053] Molecular docking analysis: The potential interaction between the target compound and the protein GSK-3β was evaluated using Discovery Studio software (version v19.1.0.18287). Molecular docking calculations were performed using the CDOCKER module in the receptor-ligand interaction module to obtain the minimum binding energy and optimal binding site. The molecular docking results and 2D plots were also generated using Discovery Studio.

[0054] In the Cell Heat Transfer Assay (CETSA) assay, RAW264.7 cells were diluted to a density of 2 × 10⁶ cells, seeded in 6-well plates, and cultured for 24 hours. They were then treated with turmeric succinate monoester sodium salt II (50 μM) for 12 hours, while the control group received an equal volume of DMSO. Cells were heated for 3 minutes within a specified temperature range, followed by lysis, centrifugation, and Western blotting (WB) to assess GSK3β levels in these samples.

[0055] Depend on Figure 5 Molecular docking results indicate that turmeric succinate monoester sodium salt II exhibits a good binding affinity with GSK3β, with a binding free energy of -32.3184 kcal / mol. The terminal carbonyl group of turmeric succinate monoester sodium salt II also shows a strong hydrogen bond with LEU-88. Figure 5(A&B in the text). In the CETSA experiment, as the temperature increased, GSK3β in the DMSO group began to degrade at 55℃, while sodium turmeric succinate monoester II maintained the stability of GSK3β at high temperatures, effectively preventing its thermal degradation more effectively than the DMSO group. Figure 5 (C in the text). This result further validates the predictions of the molecular docking analysis, indicating that GSK3β is the direct target of turmeric succinate monoester sodium salt II for the treatment of LPS-induced ALI, providing a new approach for subsequent research on specific drugs for the treatment of ALI.

[0056] This invention selects curcumin, a widely used and safe, low-toxicity compound in clinical applications. Through a Grignard reaction, the hydroxyl group of curcumin is combined with succinic anhydride to obtain curcumin succinate monoester I-1. This is then reacted with 5% sodium bicarbonate solution to obtain curcumin succinate monoester sodium salt II. The synthetic method is universal, with a short reaction time and high yield. It can exert a therapeutic effect on acute lung injury at a dose of 5 mg / kg, superior to the positive control drug dexamethasone. Through proteomics, molecular docking, cell thermal transfer experiments, and protein imprinting experiments, it was determined that curcumin succinate monoester sodium salt II inhibits the activation of the mTOR signaling pathway to exert its therapeutic effect on acute lung injury, with GSK3β as its direct target. This invention introduces a curcumin derivative with therapeutic effects on acute lung injury and discovers GSK3β as a novel target for the treatment of acute lung injury, laying the foundation for further development of more valuable anti-inflammatory lead compounds and providing insights for the development of specific drugs for the treatment of acute lung injury.

[0057] Although embodiments of the invention have been shown and described, it will be understood by those skilled in the art that various changes, modifications, substitutions and alterations can be made to these embodiments without departing from the principles and spirit of the invention, the scope of which is defined by the appended claims and their equivalents.

Claims

1. Use of a derivative of elemaπol in the preparation of a medicament for acute lung injury, characterized in that: The turmeric alcohol derivative has the following structure: The dosage of the turmeric alcohol derivative is 2–50 mg / kg.

2. Use according to claim 1, characterized in that: The synthetic route for the curcuminol derivative is as follows:

3. Use according to claim 1, characterized in that: The turmeric alcohol derivative is prepared by the following method: S1: Dissolve curcumin in tetrahydrofuran, remove the air from the reaction apparatus, and add nitrogen to the reaction apparatus to fill it with nitrogen. S2: Then add ethyl magnesium bromide and reflux for 30 min at the first preset temperature; S3: Then add succinic anhydride and reflux at the second preset temperature for 4 hours; S4: Add water to stop the reaction, and adjust the pH to 4 with hydrochloric acid; S5: After extraction with ethyl acetate, the upper layer was purified using a pre-defined method to obtain a yellow oily substance I-1; S6: Curcumol succinate monoester I-1 was reacted with 5% sodium bicarbonate solution, and the solvent was evaporated to obtain curcumol succinate monoester sodium salt II.

4. Use according to claim 3, characterized in that: In step S1, the amount of turmeric alcohol added is 0.4 mmol.

5. Use according to claim 3, characterized in that: In step S1, the amount of tetrahydrofuran added is 2.5 mL.

6. Use according to claim 3, characterized in that: In S2, the amount of ethyl magnesium bromide added is 0.4 mmol.

7. Use according to claim 3, characterized in that: In step S2, the first preset temperature is 55°C.

8. Use according to claim 4, characterized in that: In step S2, the first preset temperature is 75°C.

9. Use according to claim 3, characterized in that: In S3, the amount of succinic anhydride added is 1.2 mmol.

10. Use according to claim 3, characterized in that: In step S5, the yield of the yellow oily substance I-1 is 82%.

11. Use according to claim 3, characterized in that: In step S5, the preset method is silica gel column chromatography.

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