Use of dipeptide derivative

Abstract

Provided in the present invention is the use of a dipeptide derivative or a pharmaceutically acceptable salt thereof in the preparation of a drug for reducing the CK-18M30 level in serum during liver injuries. The dipeptide derivative for injection of the present invention can reduce the CK-18M30 level, ALT level, AST level, liver stiffness, etc., and can be used for treating different types of liver injuries.

Description

Use of a dipeptide derivative

[0001] Cross-reference to Related Applications

[0002] The present application claims priority to the prior application with the patent application number 202411058617.9, filed on August 02, 2024, to the State Intellectual Property Office of China, and the title of the prior application is "Use of a dipeptide derivative". The prior application is incorporated by reference in its entirety into the present application. TECHNICAL FIELD

[0003] The present application relates to the technical field of peptide-containing biological medicine formulations, and in particular, to the use of a dipeptide derivative. BACKGROUND

[0004] The liver is an important organ for material synthesis, biological transformation and detoxification in the human body, and is usually the first target organ where toxic side effects occur and the main target organ. Liver injury has become the most common liver disease, seriously affecting developed countries in Europe and the United States and developing countries such as China.

[0005] According to different causes, liver injury includes the following types: drug-induced liver injury, alcoholic liver injury, viral liver injury, liver injury caused by non-alcoholic fatty liver disease, autoimmune liver injury, ischemic liver injury, etc.

[0006] The current treatment of liver injury has the following problems: (1) limited treatment options: some liver injuries lack specific treatment drugs, such as NAFLD, autoimmune liver disease, and drug-induced liver injury. Especially drug-induced liver injury (DILI) caused by various prescription or non-prescription drugs, health products, natural drugs, biological agents, and herbal and dietary supplements (HDS). Unlike fatty liver disease with clear histological features, liver biopsy of DILI cases can show a variety of histological results, such as inflammation, fibrosis, vascular damage, cholestasis, necrosis, nodular regeneration, and duct destruction, so it is difficult to establish specific treatment for DILI.

[0007] (2) Poor patient compliance: especially in treatment regimens that require long-term lifestyle intervention or alcohol abstinence.

[0008] (3) Drug resistance problem: the problem of drug resistance in antiviral therapy increases the difficulty of treatment.

[0009] (4) Treatment side effects: long-term use of immunosuppressants and antiviral drugs can cause serious side effects.

[0010] CK-18M30 is a fragment of cytokeratin 18 (CK-18) produced by caspase cleavage during hepatocyte apoptosis. It is a specific marker of hepatocyte apoptosis and is particularly suitable for assessing the therapeutic effect of non-alcoholic fatty liver disease (NAFLD) and non-alcoholic steatohepatitis (NASH), drug-induced liver injury and viral hepatitis. It is also an early biomarker of liver injury and is particularly suitable for assessing the therapeutic effect of early non-alcoholic fatty liver disease (NAFLD) and non-alcoholic steatohepatitis (NASH), early drug-induced liver injury and early viral hepatitis. Therefore, the development of drugs that reduce the level of CK-18M30 is of great significance for the treatment of various types of liver injury and the treatment of early liver injury.

[0011] However, there are currently few relevant drugs for clinical use. Although there are many drugs for treating liver injury, they aim to improve liver function as a whole, reduce inflammation and fibrosis, and do not necessarily reduce the level of CK-18M30, and are mainly for the treatment of a certain type of liver injury that causes liver injury. For example, antiviral drugs such as entecavir, tenofovir, sofosbuvir are used to treat viral hepatitis; metabolic modulators such as pioglitazone are used to treat non-alcoholic fatty liver disease (NAFLD) and non-alcoholic steatohepatitis (NASH). Some studies have shown that certain drugs can improve liver function but do not necessarily reduce the level of CK-18M30. For example, a study (Biomedicines, special issue: New Challenges in the Study of Liver Diseases: From Molecular Pathogenesis to Therapeutic Approaches) found that free Omega-3 fatty acids and fenofibrate can increase the levels of biomarkers of liver cell damage AST and CK-18M30, rather than reduce them. Therefore, there is an urgent need for a clinical drug for treating liver injury that can reduce the level of CK-18M30.

[0012] CN102603865A discloses that F573 has obvious inhibitory effect on rat fulminant liver failure caused by acute liver injury of rats. CN102600452A further discloses that F573 changes related indexes in liver failure subjects. However, whether F573 can be used for treating liver injury, how the treatment mechanism is, whether F573 can be used as a clinical drug for reducing CK-18M30 level to specifically treat liver injury and treat early liver injury, and whether F573 is effective for different types of liver injury (including drug-induced liver injury, alcohol-induced liver injury, virus-induced liver injury, non-alcoholic fatty liver disease-induced liver injury, autoimmune liver disease-induced liver injury, ischemic liver injury, etc.) are not disclosed. SUMMARY

[0013] In view of the deficiencies in the prior art, the application provides an application of a dipeptide derivative or a pharmaceutically acceptable salt thereof, which can be used as a drug for reducing CK-18M30 level to treat different types of liver injury and has the advantages of good effect, good compliance, and high safety.

[0014] In one aspect of the application, an application of a dipeptide derivative or a pharmaceutically acceptable salt thereof in preparing a drug for reducing CK-18M30 level in serum during liver injury is provided.

[0015] The chemical structure of the dipeptide derivative is shown in Formula I:

[0016] In one embodiment of the application, the application further includes an application in preparing at least one drug for the following purposes:

[0017] (1) reducing alanine aminotransferase and / or aspartate aminotransferase in serum during liver injury;

[0018] (2) inhibiting hepatocyte apoptosis during liver injury; preferably, caspase-mediated hepatocyte apoptosis;

[0019] (3) reducing liver stiffness during liver injury.

[0020] In one embodiment of the application, the liver injury includes drug-induced liver injury (DILI) or other types of liver injury reaching the same degree of liver injury.

[0021] In one embodiment of the application, the other types of liver injury include alcohol-induced liver injury, virus-induced liver injury, non-alcoholic liver injury, autoimmune liver disease-induced liver injury, genetic and metabolic liver disease-induced liver injury, liver vascular disease-induced liver injury, liver tumor-induced liver injury, and cirrhosis-induced liver injury.

[0022] In one embodiment of the present application, the alcoholic liver injury includes, but is not limited to, alcoholic fatty liver, alcoholic hepatitis, and alcoholic cirrhosis-induced liver injury. The autoimmune liver disease includes, but is not limited to, autoimmune hepatitis, primary biliary cholangitis, and primary sclerosing cholangitis. The genetic and metabolic liver disease includes, but is not limited to, Wilson's disease, hemochromatosis, and alpha 1-antitrypsin deficiency. The liver vascular disease includes, but is not limited to, Budd-Chiari syndrome, and hepatic veno-occlusive disease. The liver tumor includes, but is not limited to, benign tumors such as liver hemangioma, and malignant tumors such as hepatocellular carcinoma.

[0023] In one embodiment of the present application, the liver injury is early-stage liver injury. In one embodiment of the present application, the liver injury does not include liver failure.

[0024] In one embodiment of the present application, the drug is administered at a dose of 0.125-2.5 mg / kg, preferably 0.5-2 mg / kg. In one embodiment of the present application, the drug is administered at a dose of 0.125 mg / kg, 0.25 mg / kg, 0.5 mg / kg, 0.6 mg / kg, 0.7 mg / kg, 0.8 mg / kg, 0.9 mg / kg, 1.0 mg / kg, 1.1 mg / kg, 1.2 mg / kg, 1.3 mg / kg, 1.4 mg / kg, 1.5 mg / kg, 1.6 mg / kg, 1.7 mg / kg, 1.8 mg / kg, 1.9 mg / kg, 2.0 mg / kg, 2.1 mg / kg, 2.2 mg / kg, 2.3 mg / kg, 2.4 mg / kg, 2.5 mg / kg. In one preferred embodiment of the present application, the drug is administered at a dose of 2.0 mg / kg.

[0025] In one embodiment of the present application, the dosage form of the drug includes tablets, capsules, injections, or pills.

[0026] In one embodiment of the present application, the administration of the drug includes intravenous injection, intramuscular injection, subcutaneous injection, intraperitoneal injection, or intrahepatic injection.

[0027] In one embodiment of the present application, the composition comprises 0.01-99% of the compound of Formula I or a pharmaceutically acceptable salt thereof, based on the total weight.

[0028] In one embodiment of the present application, the composition further comprises other therapeutic agents.

[0029] In one embodiment of the present application, the other therapeutic agents include antiviral drugs, immunosuppressants, antidotes, anti-inflammatory drugs, cholagogues, metabolic modulators, antioxidants, anti-fibrotic drugs, hepatoprotective drugs, anti-tumor drugs, supportive therapy drugs, vaccines.

[0030] In one embodiment of the present application, the antiviral drugs include nucleos(t)ide analogs (e.g. entecavir, tenofovir) and interferons (e.g. interferon alpha, pegylated interferon) for viral hepatitis. The immunosuppressive agents include corticosteroids (e.g. prednisone), calcineurin inhibitors (e.g. tacrolimus, cyclosporine A) for autoimmune liver diseases. The detoxifying agents include N-acetylcysteine (NAC) for drug- or toxin-induced liver injury. The anti-inflammatory drugs include non-steroidal anti-inflammatory drugs (NSAIDs) and corticosteroids for controlling liver inflammation. The choleretic agents include ursodeoxycholic acid (UDCA) for biliary diseases. The metabolic modulators include insulin sensitizers (e.g. metformin), fibric acid drugs. The antioxidants include vitamin E, S-adenosylmethionine (SAMe) for protecting hepatocytes from oxidative stress. The anti-fibrotic agents include colchicine, testosterone propionate for slowing down or reversing the progression of liver fibrosis. The hepatoprotective agents include fat emulsions, magnesium sulfate, heparin. The anti-tumor agents include chemotherapeutic drugs, targeted therapy drugs (e.g. sorafenib) and immunotherapeutic drugs for liver tumors. The supportive care agents include branched-chain amino acids, lactulose, rifampicin. The vaccines include hepatitis B vaccine.

[0031] In one embodiment of the present application, the other therapeutic agent is diammonium glycyrrhizinate.

[0032] In one embodiment of the present application, the dipeptide derivative or a pharmaceutically acceptable salt thereof and the other therapeutic agent are co-administered. The co-administration includes continuous administration in any order or at any time interval, so that the two or more therapeutic agents exert their biological activities at the same time. Preferably, the co-administration produces a synergistic therapeutic effect.

[0033] In one embodiment of the present application, the composition further comprises a pharmaceutically acceptable excipient.

[0034] In an embodiment of the present application, the adjuvants include buffers selected from, for example, acetate, Tris, phosphate, citrate and other organic acids; antioxidants include ascorbic acid and methionine; preservatives selected from, for example, octadecyldimethylbenzyl ammonium chloride, hexamethonium chloride, benzalkonium chloride, benzotonic chloride, phenol, butylparaben, alkyl parabens such as methyl or propyl parabens, catechol, resorcinol, cyclohexanol; proteins selected from, for example, serum albumin, gelatin or immunoglobulins; hydrophilic polymers selected from, for example, polyvinylpyrrolidone; amino acids such as glycine, glutamine, asparagine, histidine, arginine or lysine; chelating agents selected from, for example, EDTA; sugars selected from, for example, glucose, mannose, dextrins, sucrose, mannitol, trehalose or sorbitol; surfactants selected from, for example, polysorbates; metal complexes selected from, for example, zinc-protein complexes; non-ionic surfactants selected from, for example, Tween or polyethylene glycol (PEG); liposomes, albumin microspheres, polyesters, micelles, sustained-release matrices and the like.

[0035] In a second aspect of the present application, there is provided a use of a composition in the preparation of a medicament for treating liver injury, wherein the composition comprises a dipeptide derivative or a pharmaceutically acceptable salt thereof, and diammonium glycyrrhizinate.

[0036] In an embodiment of the present application, the chemical structure of the dipeptide derivative is shown in Formula I:

[0037] In a third aspect of the present application, there is provided a use of a substance for detecting CK-18M30 marker in the preparation of a product; the function of the product is to evaluate the therapeutic effect of the above-mentioned dipeptide derivative and pharmaceutically acceptable salt thereof on a liver injury subject.

[0038] In an embodiment of the present application, the product is a detection kit.

[0039] In an embodiment of the present application, the product further comprises a substance for detecting an ALT marker and a substance for detecting an AST marker.

[0040] In an embodiment of the present application, the sample to be detected is serum.

[0041] Compared with the prior art, the present application has the following beneficial effects:

[0042] (1) The effectiveness of the test drug of the present application on early liver injury and different types of liver injury: the dipeptide derivative injection (2.0 mg / kg dose) shows significant therapeutic effect in improving the liver function (ALT level, AST level and liver stiffness) of liver injury subjects caused by various causes.

[0043] (2) The specificity of the test drug of the present application: the test drug of the present application can reduce the CK-18M30 level of the liver damage object caused by NAFLD, NASH, viral hepatitis and drug and other causes.

[0044] (3) Compliance: the test object has good compliance to the dipeptide derivative reagent for injection, and the patient administration duration is 7-14 days, which can be used for clinical treatment.

[0045] (4) Safety of the test drug of the present application: the safety of the dipeptide derivative reagent for injection in each dose group is good, almost no serious adverse events, and is not related to the test drug, which can be used for clinical treatment. BRIEF DESCRIPTION OF DRAWINGS

[0046] Figure 1 is a diagram of the mean value of ALT baseline and the mean value of the change from baseline after administration;

[0047] Figure 2 is a diagram of the mean value of AST baseline and the mean value of the change from baseline after administration;

[0048] Figure 3 is a diagram of the mean value of TBil baseline and the mean value of the change from baseline after administration;

[0049] Figure 4 is a diagram of the mean value of ALP baseline and the mean value of the change from baseline after administration;

[0050] Figure 5 is a diagram of the mean value of AFP baseline and the mean value of the change from baseline after administration;

[0051] Figure 6 is a diagram of the mean value of INR baseline and the mean value of the change from baseline after administration;

[0052] Figure 7 is a diagram of the mean value of PTA baseline and the mean value of the change from baseline after administration;

[0053] Figure 8 is a diagram of the mean value of CK-18M30 baseline and the mean value of the change from baseline after administration;

[0054] Figure 9 is a diagram of the mean value of Caspase 3 baseline and the mean value of the change from baseline after administration;

[0055] Figure 10 is a diagram of the mean value of Caspase 1 baseline and the mean value of the change from baseline after administration;

[0056] Figure 11 is a diagram of the mean value of HGF baseline and the mean value of the change from baseline after administration. DETAILED DESCRIPTION

[0057] The technical solutions of the present application will be further described in detail below in combination with specific examples. It should be understood that the following examples are only illustratively described and explained the present application, and should not be interpreted as limiting the scope of protection of the present application. Any technology realized based on the above description of the present application is covered within the scope intended to be protected by the present application.

[0058] The raw materials and reagents used in the following examples are commercially available or can be prepared by known methods, unless otherwise specified. The detection methods used in the present application are well known in the art. The detection method of alanine aminotransferase (ALT): enzyme method (such as enzyme-linked immunosorbent assay ELISA or colorimetric method), detection reagent includes commercially available ALT kit, common brands are Roche, Abbott, Beckman Coulter, etc. The detection method of aspartate aminotransferase (AST): enzyme detection method (such as colorimetric method or fluorescence method), detection reagent includes also using commercially available AST kit, common brands are Roche, Abbott, Beckman Coulter, etc. The detection method of total bilirubin (TBIL): chemical or enzyme detection method (such as dichlorobenzene diazonium salt method), detection reagent includes TBIL kit, common brands are Beckman Coulter, Roche, Abbott, etc. The detection method of alkaline phosphatase (ALP): enzyme method (such as colorimetric method), detection reagent includes ALP kit, common brands are Roche, Abbott, Beckman Coulter, etc. The detection method of alpha-fetoprotein (AFP): enzyme-linked immunosorbent assay (ELISA), detection reagent includes AFP kit, common brands are Roche, Abbott, Beckman Coulter, etc. The detection method of international normalized ratio (INR): prothrombin time (PT) detection, INR is calculated, detection reagent includes INR / PT detection kit, common brands are Stago, Siemens, Sysmex, etc. The detection method of prothrombin activity (PTA): prothrombin time (PT) detection, detection reagent includes PT detection kit, common brands are Stago, Siemens, Sysmex, etc. The detection method of CK-18M30 (cytokeratin 18 fragment): enzyme-linked immunosorbent assay (ELISA), detection reagent includes CK-18M30 ELISA kit, common brands are PEVIVA, Abbexa, etc. The detection method of Caspase 1: ELISA, Western blot or colorimetric method, detection reagent includes Caspase 1 ELISA kit or colorimetric kit, common brands are Abeam, Thermo Fisher Scientific, etc. The detection method of Caspase 3: ELISA, Western blot or colorimetric method, detection reagent includes Caspase 3 ELISA kit or colorimetric kit, common brands are Abeam, Thermo Fisher Scientific, etc.The detection method of hepatocyte growth factor (HGF) is enzyme-linked immunosorbent assay (ELISA), and the detection reagent includes HGF ELISA kit, and common brands include R&D Systems, Thermo Fisher Scientific, etc.

[0059] In the present application, the change of each visit each examination compared with baseline = each visit each examination result - baseline each examination result.

[0060] Example clinical trial

[0061] 1. Screening of subjects

[0062] (1) Screening criteria

[0063] Age ≥ 18 and ≤ 60 years old, gender unrestricted;

[0064] Diagnosis of liver injury: patients with clinical diagnosis of hepatocellular injury type or mixed type of liver injury or CHB (chronic hepatitis B) patients with hepatitis B virus infection for more than 6 months.

[0065] Serum ALT of subjects: 2-10 times the upper limit of normal (ULN), TBil <5 times ULN;

[0066] DILI patients: duration of abnormal liver biochemical indicators [ALT, AST, ALP, γ-glutamyl transpeptidase (GGT), TBil, albumin, prothrombin time] is not more than 90 days;

[0067] Subjects (including partners) are willing to take effective contraceptive measures from screening to 6 months after the last administration of trial drug;

[0068] Sign the informed consent form and be able to comply with the requirements of the program; if the subject cannot sign the informed consent form, the legal guardian or witness shall sign according to the requirements of the regulations.

[0069] (2) Exclusion criteria

[0070] ① According to the judgment of the researcher, the subject is a patient with cholestatic liver injury;

[0071] ② Diagnosed as cirrhosis in the past or liver stiffness measurement (LSM) ≥ 12.4 kPa at the time of screening;

[0072] ③ Patients with severe or life-threatening heart, lung, brain, kidney, gastrointestinal and systemic diseases, and patients with malignant tumors;

[0073] ④ Have the following laboratory test values or abnormal test values:

[0074] a. Routine blood test: platelet (PLT) <75 x 109 / L, hemoglobin (HGB) <90 g / L;

[0075] b. Prothrombin activity <40%, prothrombin time (PT) prolongation >5 s;

[0076] c. Left ventricular ejection fraction (LVEF) <50%;

[0077] 5. Allergic or intolerant to the test drug, or allergic constitution;

[0078] 6. Subjects who are unable to express their own complaints, such as mental illness and severe neurosis;

[0079] 7. Poor compliance and non-cooperators;

[0080] 8. Pregnant women, lactating women or women of childbearing age who are ready to get pregnant;

[0081] 9. Participants in other clinical trials within 3 months;

[0082] 10. Use of liver-protecting drugs other than ursodeoxycholic acid or adenosylmethionine within 3 days before randomization;

[0083] Any condition deemed unsuitable for inclusion by the investigator.

[0084] 2. Randomization process

[0085] This trial adopts a randomized, double-blind, placebo-controlled design. The specific randomization process is as follows:

[0086] (1) Randomization scheme design:

[0087] 16 patients with liver damage (DILI patients and other types of patients with the same degree of liver damage) were enrolled. The subjects received 0.5, 1.0, 2.0 mg / kg of the test drug or placebo treatment according to the ratio of 1:1:1:1, once a day, for 7 or 14 consecutive days. Then 9 patients with CHB were enrolled for open-label treatment with the test drug, and the dosage was determined based on the results of efficacy and safety tests in the first 16 patients. The subjects also need to take basic treatment drug diammonium glycyrrhizinate enteric-coated capsules. After drug withdrawal, the subjects are followed up for 28 days for safety. During the study, the subjects need to visit the planned visit points, receive clinical laboratory tests, vital signs, physical examinations, 12-lead electrocardiogram examinations, abdominal B-ultrasound, cardiac color Doppler ultrasound, biomarker detection, MELD score, AARC score, survival status assessment, while monitoring AE, and recording concomitant / combined medication of the subjects.

[0088] (2) Randomization: Randomization was performed by a central randomization system. Each subject was randomly assigned to the corresponding treatment group at the time of enrollment.

[0089] (3) Blinding design: This study adopted a double-blind design, i.e., neither the researchers nor the subjects knew the assigned group information (except for the 9 CHB patients enrolled in the subsequent phase, who will receive open-label treatment of the trial drug). Since sterile water for injection was used as the control drug and the trial drug needed to be dissolved and prepared, the researchers responsible for the preparation of the trial drug remained open during the treatment period, and the remaining researchers and subjects remained blinded. The blinding process of the study should be documented in writing.

[0090] 3. Study process

[0091] (1) Baseline assessment: All enrolled subjects underwent baseline assessment before starting treatment, including medical history collection, physical examination, laboratory tests (such as blood routine, blood biochemistry, coagulation function, etc.), imaging examination (such as abdominal B-ultrasound, liver stiffness determination, etc.).

[0092] (2) Treatment regimen: 16 subjects were randomly assigned to receive trial drug or placebo treatment. 0.5 mg / kg, 1.0 mg / kg, 2.0 mg / kg of trial drug or equal volume of placebo, the subsequent 9 cases of CHB patients were determined according to the efficacy and safety test results of the first 16 patients enrolled in the comprehensive consideration, the drug volume was 2 mL, which was administered by intramuscular injection, once a day, for 1-2 weeks of continuous administration. Basic treatment: received diammonium glycyrrhizinate enteric-coated capsules, the drug dosage was 150 mg, 3 times a day. The first 3 subjects were given 14 drug treatments, and the subsequent subjects were given 7 drug treatments, with good subject compliance.

[0093] (3) Follow-up and monitoring: Subjects were followed up and monitored regularly during and after treatment. The main follow-up time points were treatment days 1 (D1), 2 (D2), 3 (D3), 7 (D7), 14 (D14), and 28 days after treatment. Follow-up content (V2.0) included clinical symptom evaluation, laboratory tests, and imaging examinations, etc.

[0094] (4) Safety monitoring: During the entire study, all adverse events (AEs) and serious adverse events (SAEs) of all subjects were recorded and reported in a timely manner. Researchers need to assess the safety of subjects regularly and adjust or terminate treatment if necessary.

[0095] 4. Efficacy definition

[0096] (1) Biochemical markers: including alanine aminotransferase (ALT), aspartate aminotransferase (AST), total bilirubin (TBIL), alkaline phosphatase (ALP), alpha-fetoprotein (AFP) and other liver function related indicators.

[0097] (2) Coagulation function markers: international normalized ratio (INR), prothrombin activity (PTA).

[0098] (3) Other biomarkers: including CK-18M30 (cytokeratin 18 fragment), Caspase 1, Caspase 3, HGF (hepatocyte growth factor) and others.

[0099] (4) Changes in liver stiffness: changes in liver stiffness were evaluated by transient elastography and two-dimensional shear wave elastography.

[0100] (5) Adverse events (AEs) and serious adverse events (SAEs).

[0101] 5. Statistical methods

[0102] (1) General principles

[0103] All statistical analyses will be programmed and calculated using SAS 9.4 or higher statistical analysis software.

[0104] Continuous variables will be statistically described using the number of cases, mean, median, standard deviation, minimum and maximum; categorical and ordinal variables will be statistically described using the frequency and percentage of each category or level.

[0105] (2) Statistical analysis data set

[0106] Full analysis data set (FAS): subjects who were randomized and used at least one trial drug. FAS is used for efficacy analysis.

[0107] Safety data set (SS): includes all subjects who were randomized and used at least one trial drug. Among them, those who used the trial drug incorrectly are evaluated for safety according to the actual treatment group received by the subjects.

[0108] The determination of all analysis sets will be determined at the data review meeting.

[0109] (3) Safety analysis

[0110] Adverse events were coded using the International Medical Terminology (MedDRA).

[0111] The number of cases, number of times and percentage of AEs, AEs related to the test drug, SAEs, SAEs related to the test drug, SUSARs, AEs leading to drug discontinuation, AEs leading to withdrawal, and AEs leading to death were summarized and listed according to the severity. The AEs related to the test drug that occurred only in the test group, in both the test group and the control group, and only in the control group were listed.

[0112] All AEs that occurred during the study were coded according to MedDRA, and the AEs, AEs related to the test drug, SAEs, SAEs related to the test drug, SUSARs, AEs leading to drug discontinuation, AEs leading to withdrawal, and AEs leading to death were summarized according to the system organ class (SOC) and preferred term (PT), with each subject counted at most once.

[0113] The adverse events were summarized according to the different severity of SOC and PT, and the number of cases, number of times and incidence rate were calculated. Each subject was counted at most once in the same severity level of the same term (SOC or PT).

[0114] The mean, median, standard deviation, minimum and maximum of the laboratory examination results were summarized by group, and the amount of change relative to the baseline was described, and the paired t-test or non-parametric test was used for pre- and post-dose comparison. The clinical significance judgment cross table of pre- and post-dose laboratory examinations was summarized, and the results of subjects who were normal before the test but abnormal after treatment were listed.

[0115] The detection results of vital signs and ECG indicators were described by group. For continuous variables, the mean, median, standard deviation, minimum and maximum were also described, and the amount of change relative to the baseline was described. The clinical significance judgment cross table of pre- and post-dose ECG indicators and vital signs was summarized, and the results of subjects who were normal before the test but abnormal after treatment were listed.

[0116] The clinical significance judgment cross table of pre- and post-dose physical examination was described by group, and the results of subjects who were normal before the test but abnormal after treatment were listed.

[0117] (4) Efficacy and biomarker analysis

[0118] Descriptive statistical analysis was used with FAS. For measurement data, the mean, standard deviation, median, maximum and minimum were listed, and for count data and ordinal data, the frequency (proportion) was listed.

[0119] 6. Efficacy results:

[0120] (1) ALT level changes: as shown in Table 1, Figure 1.

[0121] Table 1 ALT data indicators

[0122] From the results, after 28 days of drug withdrawal, the ALT levels of the subjects in each group were decreased to different degrees compared with the baseline. The specific changes were as follows: 0.5 mg / kg group: ALT level decreased to 126.77 U / L, decreased by 34.94% compared with the baseline. 1.0 mg / kg group: ALT level decreased to 105.10 U / L, decreased by 18.77% compared with the baseline. 2.0 mg / kg group: ALT level decreased to 49.52 U / L, decreased by 72.23% compared with the baseline. Placebo group: ALT level decreased to 111.90 U / L, decreased by 31.24% compared with the baseline. Statistical analysis showed that the decrease in ALT level in the 2.0 mg / kg group was significantly higher than that in the other groups (P<0.05).

[0123] (2) AST level changes: as shown in Table 2, Figure 2.

[0124] Table 2 AST data indicators

[0125] From the results, after 28 days of drug withdrawal, the AST levels of the subjects in each group were decreased to different degrees compared with the baseline. The specific changes were as follows: 0.5 mg / kg group: AST level decreased to 79.52 U / L, decreased by 34.57% compared with the baseline. 1.0 mg / kg group: AST level decreased to 52.37 U / L, decreased by 16.38% compared with the baseline. 2.0 mg / kg group: AST level decreased to 33.67 U / L, decreased by 68.18% compared with the baseline. Placebo group: AST level increased to 165.35 U / L, increased by 31.35% compared with the baseline. Statistical analysis showed that the decrease in AST level in the 2.0 mg / kg group was significantly higher than that in the other groups (P<0.05).

[0126] (3) Changes in other biochemical indicators: as shown in Tables 3-5, Figures 3-5.

[0127] Table 3 TBil data indicators

[0128] Table 4 ALP data indicators

[0129] Table 5 AFP data indicators

[0130] From the results, it can be seen that the total bilirubin (TBIL) and alkaline phosphatase (ALP) levels of all treatment groups had no significant change compared with the baseline. The downward trend of AFP level did not reach the statistical significant level (e.g. p>0.05), indicating that the change might be caused by random fluctuation only.

[0131] (4) Coagulation function indicators: As shown in Table 6, Table 7, Figure 6, and Figure 7.

[0132] Table 6 INR data indicators

[0133] Table 7 PTA data indicators

[0134] From the results, it can be seen that INR and PTA in each group generally fluctuated within the normal value or the range of abnormal data without clinical significance.

[0135] (5) Other biomarkers: As shown in Table 8-Table 11, and Figure 8-Figure 11.

[0136] Table 8 CK-18M30 data indicators

[0137] Table 9 Caspase 3 data indicators

[0138] Table 10 Caspase 1 data indicators

[0139] Table 11 HGF data indicators

[0140] From Table 8, it can be seen that on D2, the CK-18M30 levels of all dose groups and the placebo group decreased, but the percentage of decrease in the 2.0 mg / kg group was the highest (74%). On D3, the percentage of decrease in the CK-18M30 level of the 2.0 mg / kg group was still the highest (67%), while the percentage of decrease in the placebo group was the lowest (22%). On D7, the percentage of decrease in the CK-18M30 level of the 2.0 mg / kg group was still the highest, but the decrease in all groups was reduced. After 28 days of drug withdrawal, the CK-18M30 levels of the 0.5 mg / kg and 1.0 mg / kg groups increased, while the CK-18M30 level of the 2.0 mg / kg group continued to decrease (69%), and the CK-18M30 level of the placebo group also decreased slightly (14%). It can be seen that the CK-18M30 level decreased the most in the 2.0 mg / kg group of the test drug (P<0.05).

[0141] From Table 9 to Table 11, the change trend of Caspase 1 and Caspase 3 in each dose group of the test drug was not obvious compared with the baseline. There was no obvious regularity in the change of HGF in each dose group of the test drug compared with the baseline.

[0142] (6) Change of liver stiffness: as shown in Table 12.

[0143] Table 12 Liver stiffness change data

[0144] From Table 12, the Mean (SD) value 28 days after drug withdrawal showed the change of liver stiffness after the end of treatment. The average value of the 2.0 mg / kg group decreased to 6.40, the average change amount was the largest (-1.48), the change percentage was the highest (19%), and the decrease amplitude was the largest (P < 0.05), indicating that it had a significant reducing effect on liver stiffness. The liver stiffness of the 1.0 mg / kg group (-0.90, 11%) and the 0.5 mg / kg group (-0.33, 5%) also decreased, but not as significant as the 2.0 mg / kg group. The average value of the liver stiffness of the placebo group increased slightly (8.33), indicating no treatment effect. The liver stiffness showed a dose-dependent decrease.

[0145] (7) Safety results

[0146] During the entire study, the adverse events (AE) and serious adverse events (SAE) of all subjects were recorded in detail. The following are the adverse event statistics of each group: as shown in Table 13 to Table 15.

[0147] Table 13 TEAE data indicators

[0148] Table 14 TRAE data indicators

[0149] Table 15 SAE data indicators

[0150] From the results, the occurrence of adverse events (TEAE, TRAE) was not related to the dose, indicating that most adverse events may not be directly related to treatment, or a clear causal relationship could not be established at the time of evaluation. There was only 1 case of serious adverse event (SAE) in the placebo group, which was not related to the test drug. This indicates that the test drug is safe and not related to the dose.

[0151] Finally, it is to be explained that the above embodiments are only used to illustrate the technical solutions of the present application but not to limit the present application. Although the present application has been described in detail with reference to the preferred embodiments, those skilled in the art should understand that the technical solutions of the present application can be modified or equivalently replaced without departing from the purpose and scope of the present application, and all of them should be covered in the scope of the claims of the present application.

Claims

1. Use of a dipeptide derivative or a pharmaceutically acceptable salt thereof in the manufacture of a medicament for reducing serum CK-18 M30 level in liver injury; wherein The chemical structure of the dipeptide derivative is shown in Formula I:

2. Use according to claim 1, wherein The use also includes use in the manufacture of a medicament for at least one of the following purposes: (1) reducing serum alanine aminotransferase and / or aspartate aminotransferase in liver injury; (2) inhibiting hepatocyte apoptosis in liver injury; preferably caspase-mediated hepatocyte apoptosis; (3) reducing liver stiffness in liver injury.

3. Use according to claim 1 or 2, characterized in that, The liver injury includes drug-induced liver injury (DILI) or other types of liver injury reaching the same degree of liver injury; Preferably, the other types of liver injury include damage caused by alcoholic liver disease, liver injury caused by viral hepatitis, liver injury caused by non-alcoholic fatty liver, liver injury caused by autoimmune liver disease, liver injury caused by genetic and metabolic liver disease, liver injury caused by liver vascular disease, liver injury caused by liver tumor, liver injury caused by cirrhosis, liver injury caused by bile duct stones; more preferably, the other types of liver injury include damage caused by alcoholic liver disease, liver injury caused by viral hepatitis; Preferably, the liver injury does not include liver failure; Preferably, the liver injury is early liver injury.

4. Use according to claim 1 or 2, wherein the compound is ###0002### The dose of the medicament is 0.125-2.5 mg / kg, preferably 0.5-2 mg / kg, more preferably, the dose of the medicament is 2.0 mg / kg.

5. Use according to claim 1 or 2, wherein the compound is ###0002### The dosage form of the medicament includes tablets, capsules, injections, or pills.

6. Use according to claim 1 or 2, wherein the compound is ###0002### The administration of the medicament includes intravenous injection, intramuscular injection, subcutaneous injection, intraperitoneal injection, or intrahepatic injection.

7. Use according to claim 1 or 2, wherein the compound is ###00002### The medicament is a composition, Preferably, the composition includes 0.01-99% of the compound of formula I or a pharmaceutically acceptable salt thereof based on the total weight.

8. Use according to claim 7, wherein the compound is ###0002### The composition also includes other therapeutic agents; Preferably, the other therapeutic agents include antiviral drugs, immunosuppressants, detoxifying agents, anti-inflammatory drugs, cholagogues, metabolic modulators, antioxidants, anti-fibrotic drugs, hepatoprotective drugs, anti-tumor drugs, supportive therapy drugs, vaccines; Preferably, the other therapeutic agent is diammonium glycyrrhizinate; Preferably, the dipeptide derivative or a pharmaceutically acceptable salt thereof and the other therapeutic agent are administered in combination; Preferably, the combination administration includes continuous administration in any order or at any time interval, so that two or more therapeutic agents exert their biological activities at the same time; Preferably, the combination administration produces a synergistic therapeutic effect; Preferably, the composition also includes pharmaceutically acceptable excipients.

9. Use of a composition in the manufacture of a medicament for treating liver injury, the composition including a dipeptide derivative or a pharmaceutically acceptable salt thereof, diammonium glycyrrhizinate; wherein The chemical structure of the dipeptide derivative is shown in Formula I:

10. Use of a substance for detecting CK-18 M30 marker in the manufacture of a product; the function of the product is to evaluate the therapeutic effect of a dipeptide derivative and a pharmaceutically acceptable salt thereof on a liver injury subject; Preferably, the product is a detection kit, Preferably, the product also includes a substance for detecting an ALT marker and a substance for detecting an AST marker; Preferably, the sample for detection is serum.

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