Application of macastine B in preparation of medicine for resisting inflammation, promoting repair and resisting relapse and metastasis after liver cancer operation

Abstract

The invention belongs to the field of medicines, and discloses application of macastine B in preparation of anti-inflammatory, repair-promoting and relapse and metastasis-resisting medicines after liver cancer operation. The invention discloses an application of macastine B in preparation of a medicine for recovering a liver cancer patient after partial liver resection. The macaxantin B can promote liver function recovery, reduce the level of inflammatory factors of liver tissues and serum species, and reduce the tumor recurrence rate and the tumor visceral metastasis rate after tumor excision after an orthotopic liver cancer surgical operation, and compared with simultaneous administration of a liver protection drug and an anti-tumor drug, the macaxantin B has the advantages that the liver metabolism burden is reduced, and the curative effect is improved. And a novel drug treatment choice is provided for disease management of a liver cancer patient in a perioperative period. The invention discloses a medicine for promoting postoperative recovery of an orthotopic liver cancer surgical operation. The medicine comprises an effective therapeutic dose of macastine B and pharmaceutically acceptable auxiliary materials.

Description

Technical Field

[0001] This invention belongs to the pharmaceutical field and relates to the application of the lignan active ingredient manassatine B in the preparation of drugs that promote postoperative recovery of in situ liver cancer. Specifically, it relates to the application of manassatine B in the preparation of anti-inflammatory, repair-promoting, and anti-recurrence-metastasis drugs after liver cancer surgery, and particularly to the application of manassatine B in the preparation of drugs that reduce inflammatory response after liver resection of liver cancer and inhibit the growth and metastasis of residual liver cancer cells. Background Technology

[0002] Liver cancer is only the sixth most common primary cancer, with an incidence rate to mortality rate of nearly 1:1. [1] China accounts for more than 50% of the world's HCC (hepatic cell carcinoma) incidence rate, making it the second leading cause of cancer-related death after lung cancer. [2] Partial hepatectomy (PHx) is currently the only potentially curative treatment. [3] However, the incidence of postoperative metastasis within five years is 70%, with early recurrence (1-2 years postoperatively) being the most common. [4] These types of recurrent tumors typically exhibit poorer biological behavior, characterized by rapid growth and invasion. [5] Upregulation of immune evasion and enhanced treatment tolerance [6] The specific mechanisms mainly stem from the interaction between tumor cells and the liver: At the tumor cell level, after the primary tumor is removed, a subset of more proliferative and drug-resistant tumor stem cells escapes competitive inhibition and accelerates proliferation; simultaneously, residual tumor cells undergo a series of metastatic changes, such as activating EMT (epithelial-mesenchymal transition), degrading EMC (extracellular matrix ECM), and promoting microangiogenesis, thereby enhancing their invasive and metastatic capabilities. At the liver microenvironment level, niche remodeling is also a key influencing factor for recurrence and metastasis after HCC surgery, which can be summarized as follows: 1. The liver's strong regenerative capacity allows it to continuously release various growth factors during the wound repair process, non-specifically stimulating the proliferation of residual tumor cells; 2. The liver's inherent hepatogenic immune tolerance characteristics weaken local immune surveillance under pathological conditions, facilitating immune escape by tumor cells. [7] 3. The liver has a dual blood supply from the hepatic artery and portal vein, and the interstitial spaces within the hepatic sinusoidal endothelial cells provide a natural anatomical basis for the retention and escape of circulating tumor cells (CTCs), facilitating the colonization of metastatic lesions. [8] 4. Damage to liver tissue during the natural course of liver cancer and the perioperative period after liver resection, such as liver tissue necrosis and infection of adjacent liver tissue, are key drivers of the immune response. These factors can easily trigger excessive inflammation and postoperative complications, thereby promoting the survival and colonization of CTCs. [9] .

[0003] In summary, while PHx eradicates the primary tumor, it can exacerbate the malignancy of residual hepatocellular carcinoma cells and intensify the pro-inflammatory state and immunosuppressive tumor microenvironment in the liver. Against this backdrop, the core direction of postoperative treatment for liver cancer should focus on two aspects: first, directly eliminating microresidual lesions and eradicating activated tumor cells; second, reversing postoperative immunosuppression and alleviating chronic inflammation by regulating the liver microenvironment, promoting the restoration of normal liver physiological structure and function. Combining systemic clearance strategies with local microenvironment remodeling provides a crucial theoretical basis and clinical pathway for blocking postoperative metastasis cascades and improving long-term patient prognosis.

[0004] In recent years, the combination of adjuvant therapy and surgery has significantly improved the overall survival (OS) and recurrence-free survival (RFS) of patients with recurrent HCC. Guidelines for the diagnosis and treatment of primary liver cancer emphasize the role of targeted drugs and immunotherapy in the treatment and prevention of recurrent HCC. Clinical studies have found that postoperative targeted monotherapy as adjuvant therapy in surgical patients can significantly reduce recurrence and metastasis. [3] Sorafenib (Sora), a receptor tyrosine kinase inhibitor, exerts its anti-cancer effect by blocking angiogenesis and improving the tumor cell microenvironment. However, its direct inhibition of membrane surface receptor kinases leads to widespread drug resistance.

[10] And a high incidence of organ toxicity, specifically manifested as elevated liver enzymes, elevated levels of inflammatory factors, and dysregulation of antioxidant mechanisms.

[11] This can exacerbate postoperative liver damage and metabolic burden, limiting the effectiveness of treatment. Timely and appropriate use of medications with anti-inflammatory, choleretic, antioxidant, detoxifying, and hepatocyte membrane repair and protection effects can effectively help surgical patients recover liver function and reduce the probability of postoperative complications.

[12] However, adding hepatoprotective drugs to postoperative systemic antitumor therapy may pose potential risks of drug interactions or overdose. Currently, the emphasis is mainly on short-term, personalized medication under the guidance of a physician. Developing novel drugs with both anticancer and hepatoprotective functions to achieve safer and more effective comprehensive treatment of liver cancer is an urgent problem to be solved.

[0005] References: [1]Sung H, Ferlay J, Siegel RL, et al. Global cancer statistics2020: GLOBOCAN estimates of incidence and mortality worldwide for 36 cancers in 185 countries[J]. CA: A Cancer Journal for Clinicians, 2021, 71(3): 209-249. [2]Zheng R, Zhang S, Zeng H, et al. Cancer incidence and mortality in China, 2016[J]. Journal of the National Cancer Center, 2022, 2(1): 1-9. [3] National Health Commission of the People's Republic of China. Guidelines for the Diagnosis and Treatment of Primary Liver Cancer (2024 Edition) [J]. Journal of Clinical Hepatology, 2024, 40(5): 893-918. [4]Villanueva A. Hepatocellular carcinoma[J]. New England Journal ofMedicine, 2019, 380(15): 1450-1462. [5]Yang ZL, Gan YX, Xu JX, et al. Hyper-relapse disease is a special pattern of relapse of hepatocellular carcinoma after curativehepatectomy: A retrospective cohort study[J]. Journal of HepatocellularCarcinoma, 2025, 12: 1301-1314. [6]Sun Y, Wu L, Zhong Y, et al. Single-cell landscape of theecosystem in early-relapse hepatocellular carcinoma[J]. Cell, 2021, 184(2):404-421.e16. [7]Gautam J, Wu J, Lally J S V, et al. ACLY inhibition promotestumour immunity and suppresses liver cancer[J]. Nature, 2025, 645(8080): 507-517. [8]Sun Y F, Guo W, Xu Y, et al. Circulating tumor cells fromdifferent vascular sites exhibit spatial heterogeneity in epithelial andmesenchymal composition and distinct clinical significance in hepatocellularcarcinoma[J]. Clinical Cancer Research, 2018, 24(3): 547-559. [9]Chen K, Feng H, Zhang Y, et al. Tumor-derived CCL16 normalizestumor vasculature through macrophage ICAM-1 receptor and enhancesimmunotherapy efficacy in hepatocellular carcinoma[J]. Cancer Research, 2025,85(19): 3633-3650.

[10] Ladd A D, Duarte S, Sahin I, et al. Mechanisms of drug resistancein HCC[J]. Hepatology, 2024, 79(4): 926.

[11] AlAsmari A F, Ali N, AlAsmari F, et al. Elucidation of themolecular mechanisms underlying sorafenib-induced hepatotoxicity[J].Oxidative Medicine and Cellular Longevity, 2020, 2020(1): 7453406.

[12] Committee CP and TE, Association CSM E. Chinese expert consensus on the peri-operative management of hepatectomy for liver cancer(2021 edition)[J]. Chinese Journal of Oncology, 2021, 43(04): 414-430. Summary of the Invention

[0006] The inventors screened the in vitro anti-proliferation and anti-metastasis activity of manassatine B on HCCLM3 liver cancer cells. The results showed that manassatine B had the best anti-tumor proliferation and metastasis ability. The inventors further used LPS (lipopolysaccharide) to induce mouse mononuclear macrophage leukemia cells (RAW264.7) and human normal hepatocytes L02 to construct inflammation and liver injury models, respectively. ROS response, NO release, and liver enzyme levels were detected, revealing that manassatine B could alleviate inflammation and liver injury. Molecular biology techniques were used to explore the mechanism of action of manassatine B in anti-inflammatory, pro-repair, and anti-proliferation and metastasis of liver cancer cells. It was found that manassatine B downregulates the expression of GOLM1 (Golgi Membrane Protein 1), a key protein mediating chronic inflammatory responses and tumor metastasis, while simultaneously blocking the formation of the inflammatory microenvironment and the EMT (Epithelial-Mesenchymal Transition) process. Based on the above research results, the inventors further evaluated the intervention effect of manassatine B using animal models. An acute liver injury with inflammation model was established through liver resection accompanied by LPS induction, and a resection recurrence model was established by removing subcutaneous tumors in nude mice. In vivo experiments demonstrated that manassatine B can promote liver function recovery after orthotopic liver cancer surgery, reduce the levels of inflammatory factors in liver tissue and serum, and decrease the tumor recurrence rate and organ metastasis rate after tumor resection. This invention provides experimental support for the development of comprehensive liver cancer treatment drugs with both anti-inflammatory and anti-metastatic properties.

[0007] This invention is the first to discover that Manasartin B achieves a "dual function" by downregulating the expression of the key protein GOLM1, thereby inhibiting the proliferation and metastasis of residual tumor cells after surgery and improving acute liver injury after surgery. Compared with the simultaneous administration of hepatoprotective drugs and anti-tumor drugs, it reduces the metabolic burden on the liver and provides a novel drug treatment option for the perioperative management of liver cancer patients.

[0008] The purpose of this invention is to provide the application of manasatine B in the preparation of a recovery medicine for patients with liver cancer after partial hepatectomy.

[0009] Preferably, the application is the use of masatine B in the preparation of drugs that inhibit the proliferation and migration of liver cancer cells, reduce liver tissue inflammation, and promote recovery of liver cancer patients after partial hepatectomy.

[0010] Preferably, the application is the use of masatine B in the preparation of a medicament that promotes recovery after partial hepatectomy in patients with liver cancer by exerting the following effects: Downregulating the expression of proteins related to liver cancer proliferation and metastasis can reduce the proliferation and metastasis of residual liver cancer cells; downregulating the expression of inflammatory proteins and inhibiting inflammatory signaling pathways can reduce liver tissue inflammation.

[0011] Preferably, the application is the use of masatine B in the preparation of a drug that reduces the proliferation and metastasis of residual liver cancer cells by downregulating the expression of GOLM1, a protein related to liver cancer proliferation and metastasis, and inhibiting the overactivation of the EMT process.

[0012] Manasatine B reduces the expression of GOLM1 protein in residual tumor cells after tumor resection in liver cancer patients and inhibits the downregulation of EMT process by the Integrin signaling pathway, thereby reducing the proliferation and metastasis of residual liver cancer cells.

[0013] Preferably, the inflammation-related protein is the inflammation-related protein TRAF6 or GOLM1.

[0014] The application described is that of masatine B in the preparation of a drug that reduces liver tissue inflammation and promotes recovery after partial hepatectomy in patients with liver cancer by downregulating the expression of inflammatory-related proteins TRAF6 (tumor necrosis factor receptor-associated factor 6) and GOLM1 and inhibiting the overactivation of the NF-κB (nuclear factor kappa B) signaling pathway.

[0015] Manasatine B can reduce the secretion of liver injury markers in the serum of patients with hepatocellular carcinoma after partial hepatectomy. These liver injury markers are ALT (alanine aminotransferase), AST (aspartate aminotransferase), and LDH (lactate dehydrogenase).

[0016] Manasatine B can increase the level of repair cytokines in the liver of patients with hepatocellular carcinoma after partial hepatectomy.

[0017] The repair cytokine mentioned is IL-10.

[0018] Manasatine B can reduce the level of inflammatory factors in the liver after partial hepatectomy in patients with liver cancer.

[0019] Another object of the present invention is to provide the use of manasatine B in the preparation of a medicament for treating liver cancer or in the preparation of a medicament for inhibiting the proliferation and metastasis of residual liver cancer cells after tumor resection in liver cancer patients.

[0020] Preferably, the application is the use of masatine B in the preparation of drugs that reduce the proliferation and metastasis of residual liver cancer cells by downregulating the expression of proteins related to liver cancer proliferation and metastasis.

[0021] Preferably, the application is the use of masatine B in the preparation of a drug that reduces the proliferation and metastasis of residual liver cancer cells by downregulating the expression of GOLM1, a protein related to liver cancer proliferation and metastasis, and inhibiting the overactivation of the EMT process.

[0022] Manasatine B reduces the expression of GOLM1 protein in residual tumor cells after tumor resection in liver cancer patients and inhibits the downregulation of EMT process by the Integrin signaling pathway, thereby reducing the proliferation and metastasis of residual liver cancer cells.

[0023] Manasartin B reduces the level of inflammatory factors in the serum of liver cancer patients after tumor resection, increases the secretion of repair cytokines, and improves the body's inflammatory environment.

[0024] Manasartin B reduces the tumor recurrence rate and postoperative mortality rate in liver cancer patients after tumor resection, improves patient survival, and effectively improves prognosis.

[0025] Another object of the present invention is to provide the use of manasatine B in the preparation of a medicament for treating acute liver injury or liver failure.

[0026] Preferably, the application is the use of manasatine B in the preparation of a medicament for treating acute liver injury or liver failure caused by partial hepatectomy.

[0027] Another object of the present invention is to provide a medicament for promoting postoperative recovery after surgical treatment of hepatocellular carcinoma in situ, comprising an effective therapeutic dose of manasartine B and pharmaceutically acceptable excipients.

[0028] The drug can be administered via intravenous, intramuscular, or subcutaneous routes, or via oral, sublingual, or transdermal routes.

[0029] The drugs described are parenteral dosage forms, oral dosage forms, and transdermal dosage forms.

[0030] The parenteral drug delivery dosage forms include injectable solutions, lyophilized powder injections, and liposomes for injection.

[0031] The oral medications include tablets, granules, oral liquids, capsules, pills, and powders.

[0032] The transdermal drug delivery formulations include self-gelling agents, ointments, and transdermal patches. Attached Figure Description

[0033] Figure 1 The results were presented as mean ± SD. Compared with the control group, *P < 0.05, **P < 0.01, ***P < 0.001, ****P < 0.0001.

[0034] Figure 2 The effect of manasatine B on HCCLM3 proliferation was investigated; results are shown as mean ± SD; compared with the control group, *P < 0.05, **P < 0.01, ***P < 0.001, ****P < 0.0001.

[0035] Figure 3 The effect of manasatine B on HCCLM3 migration was investigated; the results are shown as mean ± SD; compared with the control group, *P < 0.05, **P < 0.01, ***P < 0.001, ****P < 0.0001.

[0036] Figure 4 The effect of manassatine B on the expression of HCCLM3 migration and invasion-related proteins was investigated. A: Changes in E-Cadherin and N-Cadherin expression; B: Quantitative analysis of E-Cadherin; C: Quantitative analysis of N-Cadherin; D: Changes in Integrin β1 and GOLM1 expression; E: Quantitative analysis of Integrin β1; F: Quantitative analysis of GOLM1. Results are presented as mean ± SD. Compared with the control group, *P < 0.05, **P < 0.01, ***P < 0.001, ****P < 0.0001.

[0037] Figure 5 The results show the fluorescence microscopy images (A, field of view: 10×) and quantitative analysis results of mean fluorescence intensity (B) of intracellular ROS generated by LPS-induced RAW264.7 cells after DCFH-DA incubation, representing the effect of manassatine B on ROS production. The results are shown as mean ± SD. Compared with the LPS group, *P < 0.05, **P < 0.01, ***P < 0.001, ****P < 0.0001.

[0038] Figure 6The results of flow cytometry fluorescence intensity (A) and flow cytometry fluorescence intensity quantitative analysis (B) are shown for the ROS production of RAW264.7 induced by LPS after treatment with the DCFH-DA probe, representing the amount of ROS generated by manasatine B. The results are shown as mean ± SD. Compared with the LPS group, *P < 0.05, **P < 0.01, ***P < 0.001, ****P < 0.0001.

[0039] Figure 7 The effect of manassatine B on LPS-induced NO release from RAW264.7 was investigated; results are shown as mean ± SD; compared with the LPS group, *P < 0.05, **P < 0.01, ***P < 0.001, ****P < 0.0001.

[0040] Figure 8 The effect and quantification of manassatine B on LPS-induced expression of inflammation-related proteins in RAW264.7 were investigated. Results are presented as mean ± SD. Compared with the Model group, *P < 0.05, **P < 0.01, ***P < 0.001, ****P < 0.0001; compared with the Control group, # P < 0.05, ### P < 0.001.

[0041] Figure 9 The effect of manassatine B on the expression of inflammation and damage-related proteins in LPS-induced L02 was investigated. Results are shown as mean ± SD. Compared with the Model group, *P < 0.05, **P < 0.01, ***P < 0.001, ****P < 0.0001.

[0042] Figure 10 The results of an in vivo pharmacodynamic study on liver injury in PHx+LPS C57 mice are shown. Among them, A: mouse body weight change curve; B: liver index 7 days after surgery; C: liver tissue 7 days after surgery.

[0043] Figure 11 Serum ALT levels in PHx+LPS C57 mice were measured on days 1, 3, 5, and 7 post-surgery; results are shown as mean ± SD; compared with the Model group, ***P < 0.001, ****P < 0.0001; compared with the Sham group, # P < 0.05, ## P < 0.01, #### P < 0.0001.

[0044] Figure 12Serum AST levels in PHx+LPS C57 mice were measured on days 1, 3, 5, and 7 post-surgery; results are shown as mean ± SD; compared with the Model group, *P < 0.05, **P < 0.01, ***P < 0.001, ****P < 0.0001; compared with the Sham group, #### P < 0.0001.

[0045] Figure 13 Serum LDH levels in PHx+LPS C57 mice were measured 7 days post-surgery; results are shown as mean ± SD; compared with the Model group, **P < 0.01, ****P < 0.0001; compared with the Sham group, #### P < 0.0001.

[0046] Figure 14 Postoperative histopathological results of PHx+LPS C57 mice.

[0047] Figure 15 The content of inflammatory factors in the supernatant of liver tissue homogenate from PHx+LPS C57 mice was measured; among them, (A) IL-1β; (B) TNF-α; (C) IL-10; the results are shown as mean ± SD; compared with the Model group, *P < 0.05, ***P < 0.001, ****P < 0.0001; compared with the Sham group, # P < 0.05, #### P < 0.0001, #### P < 0.0001.

[0048] Figure 16 The weight gain of mice after resection of BALBC-Nu subcutaneous hepatocellular carcinoma xenografts.

[0049] Figure 17 The results showed the tumor volume growth in mice after resection of BALBC-Nu subcutaneous hepatocellular carcinoma xenografts; the results are shown as mean ± SD; compared with the Model group, ****P < 0.0001.

[0050] Figure 18 These are representative images of subcutaneous tumor growth in mice after resection of BALBC-Nu subcutaneous hepatocellular carcinoma xenografts.

[0051] Figure 19 Representative tumor images (A, scale bar: 1 cm) and tumor volume (B) of anatomical tumor tissue after treatment in different groups; results are shown as mean ± SD; compared with the Model group, ****P < 0.0001.

[0052] Figure 20 The results of liver tissue pathology after treatment in different groups are shown.

[0053] Figure 21 The values ​​represent the serum inflammatory factor levels after treatment in different groups; where A: TNF-α, B: IL-1β, and C: IL-10; the results are shown as mean ± SD; compared with the Model group, *P < 0.05 and **P < 0.01. Detailed Implementation

[0054] All experimental data were processed using imageJ software. Statistical data were processed using GraphPad Prism 8.3.0 software. Experimental results are expressed as mean ± standard deviation (SD). One-way ANOVA was used to evaluate differences among groups. A p-value < 0.05 was considered statistically significant. Example 1

[0055] Investigation of the anti-proliferative and metastatic activity of manasatine B against liver cancer (I) The MTT assay was used to determine the cytotoxicity of manasatine B (MB) against human hepatocellular carcinoma cells HCCLM3. Preparation of MB-containing culture medium: Accurately weigh an appropriate amount of MB, add DMSO to dissolve it, and prepare an MB stock solution with a concentration of 50 mM; when administering the drug, dilute the MB stock solution with DMEM culture medium to obtain MB-containing culture medium with MB concentrations of 0.5, 5, 10, 25, 50, and 100 μM.

[0056] HCCLM3 cells in logarithmic growth phase were collected at a concentration of 8 × 10⁻⁶ cells / year. 3 Cells were seeded at a density of 10 cells / well in 96-well plates and cultured in DMEM medium containing 10% fetal bovine serum (FBS) for 24 h, after which the old medium was removed. For the control group, 200 μL of DMEM medium was added to each well, and for the MB groups, 200 μL of medium containing MB (0.5, 5, 10, 25, 50, 100 μM) was added to each well. Cells were incubated for 48 h, after which the medium was removed. 100 μL of 0.5 mg / mL MTT solution was added to each well, and the cells were incubated for 2 h. The absorbance was measured at 570 nm, and cell viability was calculated.

[0057] (II) Effect of MB on HCCM3 cell proliferation was determined using a plate colony assay. Preparation of complete culture medium containing MB: Accurately weigh an appropriate amount of MB, add DMSO to dissolve it, and prepare a 50mM MB stock solution; when administering the drug, dilute the MB stock solution with DMEM medium containing 10% fetal bovine serum (FBS) to obtain MB-containing complete culture medium with final MB concentrations of 10, 20, and 40 μM.

[0058] HCCLM3 cells were seeded at a density of 600 cells / well in 6-well plates, ideally with 2-4 single cells visible under a low-power microscope. After 72 hours of culture, the culture medium was removed. The Control group received 2 mL of DMEM medium containing 10% FBS, and the MB group received 2 mL of complete MB medium (final concentrations of 10, 20, and 40 μM, respectively). The medium was changed every 3 days, and cell status was observed. Cells were cultured for 14 days. Cells were washed once with PBS, fixed with 1 mL of 4% paraformaldehyde per well for 30 min, washed once with PBS, stained with 0.4% crystal violet for 20 min, air-dried at room temperature, and observed and photographed under a microscope for cell count. Data were processed using ImageJ to calculate the colony number (greater than 25 cells).

[0059] (III) Evaluation of the effect of MB on the motility of liver cancer cells using the Transwell migration assay Preparation of MB-containing culture medium: Accurately weigh an appropriate amount of MB, add DMSO to dissolve it, and prepare an MB stock solution with a concentration of 40 mM; when administering the drug, dilute the MB stock solution with DMEM to obtain MB-containing culture medium with MB concentrations of 100, 200 and 400 μM.

[0060] HCCLM3 cells in logarithmic growth phase were digested and resuspended into a cell suspension. 10 μL of the cell suspension was transferred to a cell counting chamber for cell counting. Based on the counting results, the suspension was diluted with DMEM medium to a concentration of 1 × 10⁻⁶ cells / mL. 6 Cell dilution buffer was prepared at 180 μL / mL. The MB group received 180 μL of cell dilution buffer, and the control group received 20 μL of complete culture medium containing MB. The lower chamber received 20 μL of DMEM medium instead of the complete MB medium. Cells were cultured for 48 h in DMEM medium containing 12% FBS. The chambers were first fixed with 4% paraformaldehyde for 30 min, then stained with 0.4% crystal violet for 30 min. The upper layer was gently wiped with a cotton swab to remove unmigrated cells, washed with water, and allowed to air dry appropriately. Cell migration was counted in three fields of view under a microscope.

[0061] (IV) The effect of MB on the expression of key regulatory proteins in the EMT process was determined by Western blotting. Preparation of MB-containing culture medium: Accurately weigh an appropriate amount of MB, add DMSO to dissolve it, and prepare an MB stock solution with a concentration of 40 mM; when administering the drug, dilute the MB stock solution with DMEM culture medium to obtain MB-containing culture medium with concentrations of 10, 20, and 40 μM.

[0062] HCCLM3 cells in logarithmic growth phase were fed at a rate of 3 × 10⁻⁶. 5Cells were seeded at a density of cells / well in 6-well plates and cultured for 24 h in DMEM medium containing 10% fetal bovine serum (FBS). The Control group received 2 mL of DMEM medium, and the MB groups received 2 mL of medium containing MB (final MB concentrations were 10, 20, and 40 μM, respectively). Cells were cultured for 48 h, washed twice with PBS, and 100 μL of RIPA cell lysis buffer containing protease and phosphatase inhibitors was added to each well. Cells were scraped off using a 1 mL pipette tip and collected into EP tubes, then lysed on ice for 30 min. After lysis, the cells were centrifuged at 12000 g for 10 min at 4 °C to remove the cell pellet, and the supernatant was collected to obtain the protein solution. Protein quantification was performed using a BCA (bicinchoninic acid) kit. The protein solution was then denatured by adding the corresponding volume of 5× loading buffer and boiling at 95 °C for 10 min to obtain the electrophoresis sample. Proteins were separated using 10% sodium dodecyl sulfate-polyacrylamide gel electrophoresis (SDS-PAGE). A suitable amount of polyvinylidene fluoride (PVDF) membrane was then transferred and blocked with 4% milk powder for 2 h. The membrane was washed three times for 10 min each time with Tris-HCl with Tween-20 (TBST). Primary antibody diluted with TBST was added, and the membrane was incubated on a shaker at room temperature for 2 h, then reacted overnight at 4°C, followed by washing with TBST. The PVDF membrane was then incubated with secondary antibody on a shaker at room temperature for 2 h, followed by washing with TBST. The membrane was developed using a Western blot imaging system, and the protein band grayscale values ​​were analyzed using ImageJ software. GAPDH and β-actin were used as internal reference proteins to detect the expression of E-cadherin, N-cadherin, Integrin β1 and GOLM1.

[0063] The results of the cytotoxicity assay and cell proliferation assay are as follows: Figure 1 , 2 As shown, MB has a significant inhibitory effect on the proliferation of HCCLM3 cells. MB at a low concentration of 10 μM showed significant anti-proliferative activity (P<0.001) and exhibited a certain dose-response relationship.

[0064] Transwell experimental results are as follows: Figure 3 As shown, MB dose-dependently inhibits HCCLM3 migration and invasion.

[0065] Western blotting results are as follows: Figure 4As shown, in HCCLM3 cells treated with different concentrations of MB (20 and 40 μM), E-Cadherin expression was higher than that in the control group, while N-Cadherin expression gradually decreased with increasing MB concentration, indicating that manassatine B can reverse the EMT process and inhibit HCCLM3 cell metastasis. After administration, the expression of key pivot proteins Integrin β1 and GOLM1 was downregulated, suggesting that the anti-metastasis effect of manassatine B may be related to the reduction of GOLM1 expression.

[0066] In summary, manasatine B exhibits good anticancer activity in vitro and effectively inhibits the proliferation and migration of HCCLM3 cells. Example 2

[0067] In vitro anti-inflammatory and liver function recovery activity of manasatine B was investigated. Preparation of MB-containing medium: Accurately weigh an appropriate amount of MB, add DMSO to dissolve it, and obtain an MB stock solution with a concentration of 20 mM. Dilute the MB stock solution with 5% FBS DMEM complete medium containing 1 μg / mL or 200 μg / mL LPS to obtain MB-containing medium with final MB concentrations of 2.5 μM, 5 μM, 10 μM, and 20 μM.

[0068] (I) Detection of intracellular reactive oxygen species (ROS) content: (1) Fluorescence microscopy imaging: When RAW264.7 cells grew to 80% confluence, RAW264.7 cells were imaged at 6.0 × 10⁻⁶ cells per cell. 5 Cells were seeded at a density of [number] cells / well in 6-well plates and cultured for 12 h. The culture medium was discarded. The drug-treated groups were treated with medium containing MB (final concentrations of 5 μM, 10 μM, and 20 μM, with LPS concentration of 1 μg / mL). The model group (LPS) was treated with DMEM medium containing 1 μg / mL LPS and 5% FBS, while the control group was treated with DMEM medium containing 5% FBS. After 12 h, the culture medium was removed, and the cells were washed twice with PBS. 1 mL of 10 μM DCFH-DA probe solution was added to each well, and the cells were incubated in the dark for 30 min. After incubation, the cells were washed three times with serum-free cell culture medium and observed and photographed under an inverted fluorescence microscope (Ex: 488 nm, Em: 525 nm). ImageJ was used for statistical analysis of the mean fluorescence intensity (Integrated Fluorescence Signal).

[0069] Experimental results show that labeling intracellular ROS with the fluorescent probe DCFH-DA and imaging analysis using fluorescence microscopy yields the following results: Figure 5 As shown, the intensity of green fluorescence in the model group cells was significantly enhanced compared to the control group, indicating that LPS successfully induced excessive ROS production. Compared with the model group, the dose-dependent fluorescence intensity decreased after MB intervention (P<0.01), suggesting that MB has an inhibitory effect on ROS production.

[0070] (2) Quantitative analysis by flow cytometry: When RAW264.7 cells reached 80% confluence, the RAW264.7 cells were cultured at 6.0 × 10⁻⁶ cells / cells. 5 Cells were seeded at a density of cells / well in 6-well plates and cultured for 12 h. The culture medium was discarded. The drug-treated groups were treated with medium containing MB (final concentrations of 5 μM, 10 μM, and 20 μM, with LPS concentration of 1 μg / mL). The model group (LPS) was treated with DMEM medium containing 1 μg / mL LPS and 5% FBS, while the control group was treated with DMEM medium containing 5% FBS. After 12 h of treatment, the culture medium was discarded, and the cells were washed twice with PBS. Cells were scraped from the cell pellet using a 1 mL pipette tip and collected in EP tubes. The tubes were centrifuged at 800 rpm for 4 min, and the supernatant was discarded. 1 mL of 10 μM DCFH-DA probe solution was added to each cell pellet, and the cells were incubated in the dark for 30 min, inverting every 5 min to ensure thorough contact between the probe and cells. After incubation, the cells were washed three times with PBS, centrifuged (800 rpm, 4 min), and resuspended in 1 mL PBS. Fluorescence intensity was measured using flow cytometry in the FITC channel.

[0071] ROS quantification was performed by flow cytometry, and the distribution of fluorescence intensity in each group of cells was as follows: Figure 6 As shown, the fluorescence signal in the model group shifted to the right compared to the blank group, while the signal in the drug-treated group gradually returned to near normal levels. Under different concentrations of MB treatment, the cell fluorescence intensity showed a dose-dependent decreasing trend (P < 0.001), indicating that MB has a significant inhibitory effect on LPS-induced ROS generation.

[0072] (II) Nitric oxide (NO) is an important mediator of inflammatory response; NO release was measured: When RAW264.7 cells reached 80% confluence, RAW264.7 cells were injected with 3.0 × 10⁻⁶ cells per cell line. 4Cells were seeded at a density of 1 cell / well in 96-well plates and cultured for 12 h. The culture medium was discarded. The drug treatment groups were treated with medium containing MB (final concentrations of 2.5 μM, 5 μM, 10 μM, and 20 μM, with LPS concentration of 1 μg / mL). A model group (LPS) and a blank group (Control) were also set up. The LPS group was added to DMEM medium containing 1 μg / mL LPS and 5% FBS, and the Control group was added to DMEM medium containing 5% FBS. After 24 h, the cell culture supernatant was collected. The NO content in the culture supernatant of each well was determined by the Griess method according to the NO kit instructions. The absorbance of each well was detected at 570 nm using an ELISA reader.

[0073] like Figure 7 As shown, RAW264.7 cells showed a significant increase in NO release after LPS induction, reaching twice that of the control group. However, the NO release was significantly reduced after MB intervention, indicating that MB has potential anti-inflammatory activity.

[0074] (III) Detection of expression of proteins regulating the inflammatory pathway in RAW264.7 cells: RAW264.7 cells in logarithmic growth phase were injected with 6.0 × 10⁻⁶ cells per cell line. 5 The cells were seeded at a density of [number] cells / well in 6-well plates. The drug treatment groups were treated with medium containing MB (final concentrations of 2.5 μM, 5 μM, 10 μM, and 20 μM, with LPS concentration of 1 μg / mL). The LPS group was treated with DMEM medium containing 5% FBS and 1 μg / mL LPS, while the control group was treated with complete DMEM medium containing 5% FBS. After 16 h, Western blotting was used to detect the expression of inflammation and key regulatory proteins, following the same procedure as in "Example 1 (IV): Determining the effect of MB on the expression of key regulatory proteins in the EMT process using Western blotting". GAPDH was used as an internal control protein to detect the expression of TRAF6 and p-NF-κB p65 proteins.

[0075] Detection of expression of L02 cell inflammation pathway-related proteins: L02 cells in logarithmic growth phase were inoculated at 3.0 × 10⁻⁶ cells per cell line. 4The cells were seeded at a density of cells / well in 6-well plates. The drug treatment groups were treated with medium containing MB (2.5 μM, 5 μM, 10 μM, 20 μM, LPS concentration 200 μg / mL), the LPS group was treated with DMEM medium containing 5% FBS and 200 μg / mL LPS, and the control group was treated with complete DMEM medium containing 5% FBS. After 48 h, Western blotting was used to detect the expression of inflammation and key regulatory proteins, following the same procedure as in "Example 1 (IV), Determining the Effect of MB on the Expression of Key Regulatory Proteins in the EMT Process using Western Blot". GAPDH was used as an internal control protein to detect the expression of GOLM1, Integrin β1, and p-NF-κB p65 proteins.

[0076] The results of the Western blot experiment are shown below. Figure 8 , Figure 9 This indicates that MB can inhibit LPS-induced overactivation of the NF-κB signaling pathway in Raw264.7 and L02 cells. Simultaneously, LPS-induced upregulation of GOLM1 and Integrin β1 expression in L02 cells suggests that GOLM1 is overexpressed in LPS-induced L02 cells, and MB can dose-dependently restore the expression of related proteins. In conclusion, MB exhibits good anti-inflammatory activity in vitro and has the potential to promote liver injury recovery. Example 3

[0077] In vivo activity of manasatine B in alleviating postoperative liver injury Animals: SPF-grade male C57BL / 6 mice (12 weeks old), Cavens (Changzhou) Biotechnology Co., Ltd., with laboratory animal production license number SCXK (Su) 2021-0013.

[0078] Establishment of an animal model of liver injury: C57BL / 6 mice were acclimatized for one week, then fasted overnight before being anesthetized by intraperitoneal injection of 4% chloral hydrate at a dose of 4 mL / kg. The mice were secured with medical tape to ensure their abdomens were facing upwards. The abdomen was disinfected by wiping with alcohol swabs. Using autoclaved surgical instruments, an incision of approximately 1 cm was made along the midline of the abdomen at the xiphoid process to expose the abdominal organs. The left lobe of the liver was ligated with medical sterile sutures. After the left lobe became ischemic and white, it was removed 2 mm from the ligation line. Hemostasis was achieved by applying pressure to the wound. The muscle layer and the outer skin layer were then sutured sequentially from the inside out, and the wound was disinfected with povidone-iodine. All procedures were performed on a sterile operating table in an SPF-grade animal facility, and all surgeries were performed between 8:00 AM and 12:00 PM. The sham-operated group (Sham) underwent only laparotomy without additional surgical procedures. After surgery, except for the sham-operated group, mice in the other groups were injected intraperitoneally with a low dose of 0.5 mg / kg LPS (prepared with physiological saline) to establish an acute liver injury with inflammation (PHx+LPS) model.

[0079] Experimental grouping: Mice were divided into 5 groups (n=8): model group (MOD), high-dose group (MB-H, MB dose 4 mg / kg), low-dose group (MB-L, MB dose 2 mg / kg), positive control group (magnesium isoglycyrrhizinate, MgIG, dose 10 mg / kg), and sham operation group.

[0080] Drug preparation: Accurately weigh an appropriate amount of the drug and dissolve it in a mixed solvent of DMSO, PEG300, Tween-80 and physiological saline in a volume ratio of 2:40:5:53 (MB-H: 1.2 mg / mL, MB-L: 0.6 mg / mL, MgIG: 3 mg / mL).

[0081] Administration regimen: 24 hours postoperatively, intraperitoneal injection was administered twice daily for a total of 7 days, with administration on days 1, 3, 5, and 7 postoperatively; the sham surgery group received intraperitoneal injection of normal saline.

[0082] Liver function test indicators: After modeling, blood was collected from the cheeks of mice at 12 h, 24 h, 72 h, 120 h, and 168 h. After standing at room temperature for 1 h, the mice were centrifuged at 4500 rpm for 15 min. The supernatant was the mouse serum. The mouse serum was aliquoted and tested immediately or stored at -80℃. The levels of aspartate aminotransferase (AST), alanine aminotransferase (ALT), and lactate dehydrogenase (LDH) in mouse serum were detected using the Amplex Red enzyme activity assay kit (Beyotime P2715S, P2711S, P0395S) to monitor the recovery of liver damage.

[0083] Liver index: After the last administration, all mice were euthanized by enucleation and cervical dislocation, and the livers were collected. The fat around the organs was removed, the livers were washed with physiological saline, the surface liquid was blotted dry with filter paper, and the livers were accurately weighed to calculate the proportion of liver to body mass (mg / g).

[0084] Histopathological observation and immunohistochemistry: Liver tissues from mice in each group were isolated to obtain samples for histopathological observation and immunohistochemical staining. The samples were fixed by immersing them in a 4% paraformaldehyde solution. Histopathological differences were observed under a light microscope using H&E staining.

[0085] Measurement of liver inflammatory factor levels: After the last administration, livers of mice in each group were collected. 100 mg of liver tissue was added to 500 mL of 1% α-toluenesulfonyl fluoride (PMSF) in PBS solution, and homogenized at 4℃ and 50 Hz for 60 s to obtain liver homogenate. The homogenate was centrifuged at 4℃ and 12000 rpm for 15 min, and the supernatant was collected as the liver tissue homogenate supernatant for subsequent detection. The protein concentration of the liver homogenate supernatant was quantified using a BCA kit, and the inflammatory factors TNF-α, IL-1β, and IL-10 were quantified according to the instructions using an ELISA kit to further assess the postoperative liver inflammation status of mice in each group.

[0086] Experimental results are as follows Figure 10 As shown. Weight changes in mice within 7 days post-surgery showed that the MOD group recovered slowly, while the drug-treated group showed significant weight gain on the first day post-surgery and returned to normal levels by the third day. Figure 10 A); Liver indices in each group indicated that 7 days post-surgery, the liver mass and volume in the MOD group had not yet returned to normal levels, which may be related to inflammation induced by low-dose LPS, while MB can effectively improve liver growth and recovery. Figure 10 B, C).

[0087] Serum LDH, AST, and ALT levels are important indicators of liver tissue damage. Serum ALT and AST levels in mice were monitored at different time points after surgery. The results are as follows: Figure 11 , 12 As shown, serum ALT and AST levels in both the drug-treated group and the MOD group peaked on postoperative day 1, with no significant difference, reflecting the acute liver injury in the acute liver injury with inflammation model. Serum ALT and AST levels in the drug-treated group were significantly lower than those in the MOD group on days 3, 5, and 7 (P<0.001). The liver injury recovery in the MB-H group was comparable to that in the MgIG group. Further studies showed that on postoperative day 7, MB (2 mg / kg, 4 mg / kg) effectively reduced the abnormal increase in serum LDH (P<0.01). Figure 13 ).

[0088] The results of the liver histological evaluation are shown in Figure 14 H&E results showed that the livers of mice in the MOD group were significantly damaged, characterized by widespread dilation of hepatic sinusoids, widening of cavities, and filling with erythrocytes, exhibiting a "congestion-like" appearance. Simultaneously, some hepatocytes showed increased cell volume, ballooning degeneration with cytoplasmic vacuolation, or fatty degeneration. Significant inflammatory cell infiltration was observed in the liver tissue. MB dose-dependently reduced the number of inflammatory cells and necrotic lobules in the livers of mice with acute liver injury and inflammation. Immunohistochemical staining of liver tissue sections from each group of mice was performed using the key protein GOLM1. Figure 14 The results showed that, compared with the Sham group, the MOD group mice exhibited abnormally elevated GOLM1 expression in liver tissue, accompanied by extensive loss of cellular structure. Overexpression of GOLM1 stimulated macrophage differentiation, leading to aggravated inflammatory responses. After treatment with different doses of MB and MgIG, GOLM1 expression levels rebounded, significantly decreasing compared to the MOD group, indicating that macrophage polarization was inhibited. After high-dose MB and MgIG treatment, GOLM1 expression levels were similar to those in the Sham group, and the liver tissue essentially returned to its normal physiological state.

[0089] Studies on the inflammatory state of the liver after treatment have shown that ( Figure 15 Compared to the Sham group, the MOD group showed a significant increase in TNF-α and IL-1β levels in the liver tissue homogenate supernatant. Following MB treatment, the levels of both TNF-α and IL-1β in the liver tissue homogenate supernatant were significantly reduced. In addition to reducing the levels of pro-inflammatory factors, MB administration also increased the levels of IL-10, a factor related to anti-inflammatory and cell repair functions, in the liver tissue homogenate supernatant.

[0090] In summary, MB can effectively improve liver damage and inhibit inflammation in mice. Example 4

[0091] In vivo activity of manasatine B in inhibiting postoperative recurrence and metastasis of liver cancer. Establishment of an animal model of recurrence after liver cancer surgery: A recurrence model of subcutaneous liver cancer xenograft resection was established using Balb / c nude mice (male, 6 weeks old, 20g). The specific steps were as follows: First, highly metastatic liver cancer cells HCCLM3 (1×10⁻⁶ mcg) were injected into the mice. 6 A tumor-bearing nude mouse model was established by subcutaneous injection of 0.1 mL PBS into mice; the tumor volume of the mice was monitored every other day, and the tumor volume was calculated (tumor volume calculation formula: length × width). 2 × 0.5). When the tumor volume reaches approximately 150 mm. 3At that time, the visible tumor tissue was surgically removed, the wound was sutured with 4-0 absorbable sutures, and disinfected with povidone-iodine. The nude mouse was then placed on a 37°C constant temperature heating pad to wait for it to wake up.

[0092] Animal grouping: After the operation, the mice were randomly divided into 5 groups (n=6): Model group, low-dose group (MB-L, 2 mg / kg), high-dose group (MB-H, 4 mg / kg), traditional Chinese medicine control group (95SACE, ethanol extract of Saussurea costatum, 25 mg / mL), and positive drug control group (Sora, 10 mg / kg).

[0093] Preparation of the ethanol extract of *Saururus chinensis*: 50g of *Saururus chinensis* herb (purchased from Hebei Jiaheng Lengbei Pharmaceutical Co., Ltd.) was placed in a round-bottom flask. An ethanol-water solution (95%) was added at a material-to-liquid ratio of 1:10g / mL. The mixture was soaked for 2 hours, then extracted twice by reflux, 2 hours each time. The extracts were combined, and most of the solvent was removed by rotary evaporation until no alcohol odor remained, yielding a concentrated solution. The concentrated solution was frozen overnight at -20°C, then lyophilized to obtain a lyophilized powder, which was stored at -20°C.

[0094] Drug preparation: Accurately weigh an appropriate amount of the drug and prepare a suspension using a mixed solvent of DMSO, PEG300, Tween-80 and physiological saline in a volume ratio of 2:40:5:53 (MB-H: 1.2 mg / mL; MB-L: 0.6 mg / mL; Sorafenib, Sora: 3 mg / mL; Saururus chinensis ethanol extract: 7.5 mg / mL).

[0095] Administration regimen: 24 h after surgery, mice in the MB-L, MB-H and Sora groups were administered the drug via intraperitoneal injection twice a day for a total of 30 days; mice in the Model group were administered the drug via intraperitoneal injection of physiological saline twice a day for a total of 30 days; mice in the 95SACE group were administered the drug via gavage once a day for a total of 30 days.

[0096] Local tumor recurrence and mortality in mice were monitored during drug administration. Local recurrence was defined as the presence of a visible subcutaneous nodule (approximately 20 mm in volume) in the mouse skin. 3 Mice were recorded for body weight and tumor volume every three days. After the last administration, all mice were euthanized by enucleation, cervical dislocation, and whole blood, serum, organs, and tumor tissue were collected.

[0097] Histopathological observation and immunohistochemistry: Heart, liver, spleen, lung, kidney, and tumor tissue samples from mice in each group were fixed in 4% paraformaldehyde solution, decalcified with 10% EDTA, routinely dehydrated, embedded in paraffin, and sectioned. Histopathological differences were observed under a light microscope using H&E staining to assess organ metastasis of the tumor. Serum inflammatory factors TNF-α, IL-1β, and IL-10 were quantified using an ELISA kit according to the manufacturer's instructions to assess the inflammatory status of the mice.

[0098] Further in vivo evaluation of the anti-tumor recurrence and metastasis effects of MB: 30-day weight change curve ( Figure 16 The results showed that there was no significant difference in body weight among the groups of mice during postoperative treatment, indicating that MB-induced systemic toxicity was low. Monitoring changes in recurrent tumor volume over 30 days (…) Figure 17 The growth rate of recurrent tumor volume in the Model group mice was significantly higher than that in all treatment groups. On day 30, the recurrent tumor volume in the MB-L group, MB-H group, 95SACE group, and Sora group mice was significantly lower than that in the Model group. This is consistent with... Figure 18 , 19 The results were highly consistent. Histopathological observation and hematoxylin-eosin (H&E) staining were also observed. Figure 20 This further confirmed the anti-metastatic effect of MB. In the Model group mice, there was extensive necrosis of liver tissue and extensive liver tumor tissue was observed. MB could reduce the area of ​​abnormal liver tissue in a dose-dependent manner. Among them, high-dose MB and 95SACE showed similar effects to the positive control drug (Sora), indicating that the drug effectively inhibited liver metastasis of tumors.

[0099] Figure 21 The study showed that the systemic inflammatory status of mice in each group was significantly increased in the serum of the Model group. After MB treatment, the levels of TNF-α and IL-1β in the liver of mice decreased, and the levels in the MB-H group were significantly lower than those in the Model group, indicating that MB treatment can alleviate postoperative systemic inflammation. In addition to reducing the levels of pro-inflammatory factors, high-dose MB administration significantly increased the level of IL-10, a factor related to anti-inflammation and cell repair, in the serum of mice, indicating that high-dose MB treatment is beneficial for the body to recover to a normal physiological state after surgery.

[0100] In summary, MB can effectively improve postoperative recurrence and metastasis of liver cancer, inhibit the chronic inflammatory environment around the tumor, and promote recovery after partial hepatectomy.

Claims

1. Use of manassantin B in the preparation of a drug for promoting recovery after partial hepatectomy in a liver cancer patient.

2. Use according to claim 1, characterized in that: The use is the use of manassantin B in the preparation of a drug for inhibiting proliferation and migration of liver cancer cells, reducing inflammation of liver tissue, and promoting recovery after partial hepatectomy in a liver cancer patient.

3. Use according to claim 1 or 2, characterized in that: The use is the use of manassantin B in the preparation of a drug for promoting recovery after partial hepatectomy in a liver cancer patient by exerting the following effects: down-regulating expression of liver cancer proliferation and metastasis-related proteins to reduce proliferation and metastasis of residual liver cancer cells; down-regulating expression of inflammatory-related proteins and inhibiting inflammatory signaling pathways to reduce inflammation of liver tissue.

4. Use according to claim 3, characterized in that: The use is the use of manassantin B in the preparation of a drug for promoting recovery after partial hepatectomy in a liver cancer patient by exerting the following effects: down-regulating expression of liver cancer proliferation and metastasis-related protein GOLM1 and inhibiting excessive activation of EMT process to reduce proliferation and metastasis of residual liver cancer cells; down-regulating expression of inflammatory-related proteins TRAF6 and GOLM1 and inhibiting excessive activation of NF-κB signaling pathways to reduce inflammation of liver tissue.

5. Use of manassantin B in the preparation of a drug for treating liver cancer or a drug for inhibiting proliferation and metastasis of residual liver cancer cells after tumor resection in a liver cancer patient.

6. Use according to claim 5, characterized in that: The use is the use of manassantin B in the preparation of a drug for reducing proliferation and metastasis of residual liver cancer cells by down-regulating expression of liver cancer proliferation and metastasis-related proteins.

7. Use of manassantin B in the preparation of a drug for treating acute liver injury or liver failure.

8. Use according to claim 7, characterized in that: The use is the use of manassantin B in the preparation of a drug for treating acute liver injury or liver failure caused by partial hepatectomy.

9. A medicament for facilitating post-surgical recovery from hepatocarcinoma in situ, characterized by: The drug comprises an effective therapeutic dose of manassantin B and pharmaceutically acceptable excipients.

10. The medicament according to claim 9, characterized in that: The drug is in a parenteral administration form, an oral administration form, or a transdermal administration preparation. The drug is in a parenteral administration form, an oral administration form, or a transdermal administration preparation.

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