New application of meclozine and analogues thereof

Abstract

The invention relates to the technical field of novel application of compounds, in particular to novel application of meclozine and analogues thereof. The invention relates to an application of at least one of meclozine and analogues thereof or pharmaceutically acceptable salts or esters thereof in preparation of drugs for preventing and / or treating pancreatitis. The meclozine and the analogue thereof can obviously improve pancreatic tissue necrosis, inflammatory response and oxidative stress of pancreatitis, inhibit alveolar cell death and improve lung injury caused by pancreatitis, so that the meclozine and the analogue thereof can be used as a medicine for treating pancreatitis.

Description

Technical Field

[0001] This invention relates to the field of new applications of compounds, and more specifically, to new applications of meclozine and its analogues. Background Technology

[0002] Pancreatitis is an inflammation of the pancreas caused by various pathogenic factors, and is classified into acute pancreatitis and chronic pancreatitis according to its course. With improved living standards and changes in dietary structure, the incidence of pancreatitis is increasing year by year, and it has become the most common reason for hospitalization of gastrointestinal patients. Pancreatitis is caused by abnormal activation of trypsinogen within the pancreas, leading to the digestion and hydrolysis of pancreatic tissues, causing an inflammatory reaction in the pancreas or affecting surrounding organs. Acute pancreatitis is a common acute abdominal condition, often presenting with sudden onset of persistent and severe acute upper abdominal pain, accompanied by nausea, vomiting, abdominal distension, jaundice, fever, and altered mental status. In severe cases, pancreatic tissue damage and necrosis may occur, with abnormally elevated serum amylase and lipase levels. Approximately 80% of acute pancreatitis cases have mild symptoms, affecting only the pancreas, and resolve spontaneously within a few days without organ dysfunction or local or systemic complications; this is defined as mild acute pancreatitis, and the mortality rate for these patients is low. However, approximately 20% of patients with acute pancreatitis may develop severe acute pancreatitis, characterized by pancreatic or peripancreatic tissue necrosis or organ failure. This condition progresses rapidly, has a poor prognosis, and a mortality rate as high as 30%. Furthermore, severe acute pancreatitis can be complicated by lung injury of varying severity, ranging from mild hypoxemia to acute respiratory distress syndrome (ARDS). Lung injury is one of the most common complications of severe acute pancreatitis and a leading cause of death in pancreatitis patients.

[0003] Acute pancreatitis has multiple contributing factors, with gallstones obstructing the common bile duct (38%) and excessive alcohol intake (36%) being the main causes. Due to the unique physiological structure of the pancreatic duct and bile duct openings, bile duct stones obstructing the pancreaticobiliary opening cause bile reflux, which, combined with internal pancreatic digestion, leads to pancreatic damage and inflammation of varying degrees. The pathogenesis of alcohol-induced pancreatitis is complex, involving damage and dysfunction of acinar and ductal cells in the exocrine pancreas. Furthermore, hypertriglyceridemia is also a predisposing factor, ranking as the third leading cause of pancreatitis. More than 15% of patients with severe hypertriglyceridemia will develop acute pancreatitis, exhibiting more severe clinical manifestations and often experiencing multiple organ failure. Notably, recent studies have shown an increased risk of developing pancreatitis associated with diabetes. Pancreatitis is also associated with factors such as medications, infections, genetic diseases, surgical trauma, and gender.

[0004] Because the etiology of pancreatitis is complex and the specific pathogenesis is not yet fully understood, there are currently no clearly defined targeted drugs or effective intervention methods for its treatment. Currently, the clinical treatment strategies and measures for acute pancreatitis are mainly symptomatic, including: first, fasting to reduce gastrointestinal stress and providing nutritional support through intravenous fluids; second, using anti-inflammatory drugs to reduce the inflammatory response caused by acute pancreatitis; and third, using trypsin inhibitors, such as ulinastatin, to inhibit trypsin activity and reduce pancreatic damage. However, there are currently no effective specific drugs for acute pancreatitis, especially severe acute pancreatitis and its complications such as lung damage. Treatment options are limited, clinical efficacy is poor, leading to slow patient recovery, long hospital stays, heavy financial burdens, and even death. Therefore, effective treatment drugs for acute pancreatitis are urgently needed.

[0005] In view of this, the present invention is proposed. Summary of the Invention

[0006] The purpose of this invention is to provide new applications for meclopramide and its analogues. The meclopramide and its analogues provided in the embodiments of this invention can significantly improve pancreatic tissue necrosis, inflammatory response, oxidative stress, inhibit acinar cell death, and improve lung injury caused by pancreatitis in pancreatitis, and can therefore be used as drugs for the treatment of pancreatitis.

[0007] This invention is implemented as follows: In a first aspect, the present invention provides the use of at least one of meclopramide and its analogues or pharmaceutically acceptable salts or esters thereof in the preparation of medicaments for the prevention and / or treatment of pancreatitis.

[0008] In an optional embodiment, the meclozine and its analogues are selected from chiral isomers or mixtures of chiral isomers of compounds shown in the following structural formulas, or pharmaceutically acceptable salts thereof: In this context, R1 represents hydrogen or halogen; R2 represents any one of hydrogen, C1-C3 unsubstituted alkyl, C1-C3 unsubstituted carboxyl, and C1-C3 unsubstituted amide; and R3 represents C1-C3 unsubstituted alkylene or C3-C5 alkenyl.

[0009] In an optional embodiment, R1 represents any one of hydrogen, chlorine, and bromine; R2 represents any one of hydrogen, methyl, ethyl, acetate, COOH, formamide, and acetamide; and R3 represents methylene or 2-propenyl.

[0010] In an optional embodiment, the meclozine and its analogues are selected from at least one of the compounds shown in the following structural formulas: , , , , ,

[0011] , and ; Preferred or .

[0012] In an optional implementation, the pancreatitis includes acute pancreatitis.

[0013] In an optional implementation, the pancreatitis includes severe acute pancreatitis.

[0014] In an optional embodiment, the drug is a drug for improving lung damage caused by pancreatitis.

[0015] In an optional embodiment, the drug is a drug for relieving upper abdominal pain caused by pancreatitis.

[0016] In an optional embodiment, the drug may further include pharmaceutically acceptable excipients.

[0017] In an optional embodiment, the pharmaceutically acceptable salt includes at least one of hydrochloride, hydrobromide, sulfate, phosphate, acetate, citrate, lactate, ascorbate, maleate, tartrate, malate, gluconate, sulfonate, citrate, benzoate, benzenesulfonate, carbonate, methanesulfonate, stearate, nitrate, valerate, and succinate.

[0018] The present invention has the following beneficial effects: the meclozine and its analogues provided in the embodiments of the present invention can significantly improve pancreatic tissue necrosis, inflammatory response, oxidative stress, inhibit acinar cell death, and improve lung injury caused by pancreatitis in pancreatitis, and can therefore be used as drugs for the treatment of pancreatitis, especially with good therapeutic effects on acute pancreatitis. Attached Figure Description

[0019] To more clearly illustrate the technical solutions of the embodiments of the present invention, the accompanying drawings used in the embodiments will be briefly introduced below. It should be understood that the following drawings only show some embodiments of the present invention and should not be regarded as a limitation on the scope. For those skilled in the art, other related drawings can be obtained based on these drawings without creative effort.

[0020] Figure 1 The figure shows the effect of meclozar and its analogues on AR42J amylase in rat-derived pancreatic acinar cells induced by taeniacin; compared with the control group: *P <0.05, **P<0.01, ***P <0.001, ****P <0.0001; compared with the model group: #P <0.05, ##P <0.01, ###P <0.001, ####P <0.0001; Figure 2 Figure 1 shows the effects of meclopramide and compound i on amylase and lipase in rat-derived pancreatic acinar cells AR42J induced by taeniacin. Specifically, A shows the effect of meclopramide on taeniacin-induced AR42J cell amylase, B shows the effect of meclopramide on taeniacin-induced AR42J cell lipase, C shows the effect of meclopramide on taeniacin-induced AR42J cell viability, D shows the effect of compound i on taeniacin-induced AR42J cell amylase, E shows the effect of compound i on taeniacin-induced AR42J cell lipase, and F shows the effect of compound i on taeniacin-induced AR42J cell viability. Compared with the Control group: *P <0.05, **P <0.01, ***P <0.001, ****P <0.0001; compared with the model group: #P <0.05, ##P <0.01, ###P <0.001, ####P <0.0001; Figure 3 Figure A shows the effect of meclopramide on L-arginine-induced cell death in rat-derived pancreatic acinar cells AR42J; Figure B shows the effect of meclopramide on L-arginine-induced cell viability; Figure C shows the effect of meclopramide on L-arginine-induced PI staining; Compared with the Control group: *P <0.05, **P <0.01, ***P <0.001, ****P <0.0001; compared with the model group: #P <0.05, ##P <0.01, ###P <0.001, ####P <0.0001; Figure 4Figure A shows the effect of meclopramide on taurocholate-induced cell death in rat-derived AR42J pancreatic acinar cells; Figure B shows the effect of meclopramide on taurocholate-induced cell viability; Figure C shows the effect of meclopramide on taurocholate-induced PI staining; Compared with the Control group: *P <0.05, **P <0.01, ***P <0.001, ****P <0.0001; compared with the model group: #P <0.05, ##P <0.01, ###P <0.001, ####P <0.0001; Figure 5 Figure A shows the effect of meclozine and compound i on pancreatic damage induced by scutellarin combined with lipopolysaccharide in C57BL / 6 mice with severe acute pancreatitis; where A is the effect of meclozine on pancreatic damage induced by scutellarin combined with lipopolysaccharide in C57BL / 6 mice with severe acute pancreatitis, and B is the effect of compound i on pancreatic damage induced by scutellarin combined with lipopolysaccharide in C57BL / 6 mice with severe acute pancreatitis. Figure 6 Figure A shows the effect of meclozine and compound i on lung injury induced by scutellarin combined with lipopolysaccharide in C57BL / 6 mice with severe acute pancreatitis; where A is the effect of meclozine on lung injury induced by scutellarin combined with lipopolysaccharide in C57BL / 6 mice with severe acute pancreatitis, and B is the effect of compound i on lung injury induced by scutellarin combined with lipopolysaccharide in C57BL / 6 mice with severe acute pancreatitis. Figure 7 The figures show the effects of meclopramide and compound i on serum and pancreatic tissue amylase and lipase levels in C57BL / 6 mice with severe acute pancreatitis induced by taeniacin combined with lipopolysaccharide. Figures A and C show the effects of meclopramide on serum and pancreatic tissue amylase levels; figures B and D show the effects of meclopramide on serum and pancreatic tissue lipase levels; figure E shows the effects of compound i on serum amylase levels; and figure F shows the effects of compound i on serum lipase levels. Compared to the control group: *P <0.05, **P <0.01, ***P <0.001, ****P <0.0001; compared with the model group: #P <0.05, ##P <0.01, ###P <0.001, ####P <0.0001; Figure 8 Figure 1 shows the effects of meclopramide and compound i on inflammatory factors in C57BL / 6 mice induced by lipopolysaccharide combined with omega-3 fatty acids (MPO). Figure 2 shows the effects of meclopramide on serum IL-6, B on serum MPO, C on pancreatic MPO, D on serum IL-6, E on serum IL-1β, and F on serum MPO. Compared with the control group: *P <0.05, **P <0.01, ***P <0.001, ****P <0.0001; compared with the model group: #P <0.05, ##P <0.01, ###P <0.001, #### P <0.0001; Figure 9 Figure 1 shows the effect of meclopramide on oxidative stress induced by lipopolysaccharide combined with taeniacin in C57BL / 6 mice with severe acute pancreatitis; Figure 2 shows the effect of meclopramide on MDA (malondialdehyde) in mouse serum, and Figure 3 shows the effect of meclopramide on MDA (malondialdehyde) in mouse pancreatic tissue; compared with the control group: *P <0.05, **P <0.01, ***P <0.001, ****P <0.0001; compared with the model group: #P <0.05, ##P <0.01, ###P <0.001, #### P <0.0001; Figure 10 The figure shows the effect of meclopramide on apoptosis induced by crotonin combined with lipopolysaccharide in pancreatic tissue cells of C57BL / 6 mice with severe acute pancreatitis; where A is the effect of meclopramide on the expression level of Caspase-7 protein in mouse pancreatic tissue, and GAPDH is the internal control protein; B is the quantitative result of the expression level of Caspase-7 protein in each group with different cleavage forms; compared with the control group: *P <0.05, **P <0.01, ***P <0.001, ****P <0.0001; compared with the model group: #P <0.05, ##P<0.01, ###P <0.001, ####P <0.0001; Figure 11 Figure showing the effect of meclozine on pancreatic damage in L-arginine-induced severe acute pancreatitis in C57BL / 6 mice; Figure 12 The figures show the effects of meclopramide on serum and pancreatic tissue amylase and lipase levels in L-arginine-induced severe acute pancreatitis in C57BL / 6 mice. Figures A and C show the effects of meclopramide on serum and pancreatic tissue amylase levels, while figures B and D show the effects of meclopramide on serum and pancreatic tissue lipase levels. Compared to the control group: *P <0.05, **P <0.01, ***P <0.001, ****P <0.0001; compared with the model group: #P <0.05, ##P <0.01, ###P <0.001, ####P <0.0001; Figure 13 Figure 1 shows the effect of meclopramide on the levels of inflammatory factors in L-arginine-induced severe acute pancreatitis in C57BL / 6 mice; where A represents the effect of meclopramide on TNF-α in mouse serum, B represents the effect of meclopramide on IL-1β in mouse serum, and C represents the effect of meclopramide on IL-6 in mouse serum; compared with the control group: *P <0.05, **P <0.01, ***P <0.001, ****P <0.0001; compared with the model group: #P <0.05, ##P <0.01, ###P <0.001, ####P <0.0001; Figure 14 Figure 1 shows the effect of meclopramide on oxidative stress in L-arginine-induced severe acute pancreatitis in C57BL / 6 mice. Figure 2 shows the effect of meclopramide on serum catalase in mice; Figure 3 shows the effect of meclopramide on serum MDA in mice; Figure 4 shows the effect of meclopramide on MDA in mouse pancreatic tissue; Figure 5 shows the effect of meclopramide on serum hydrogen peroxide; Figure 6 shows the effect of meclopramide on hydrogen peroxide in pancreatic tissue. Compared with the control group: *P <0.05, **P <0.01, ***P<0.001, ****P <0.0001; compared with the model group: #P <0.05, ##P <0.01, ###P <0.001, ####P <0.0001. Detailed Implementation

[0021] To make the objectives, technical solutions, and advantages of the embodiments of the present invention clearer, the technical solutions in the embodiments of the present invention will be clearly and completely described below. Where specific conditions are not specified in the embodiments, conventional conditions or conditions recommended by the manufacturer shall apply. Reagents or instruments whose manufacturers are not specified are all conventional products that can be purchased commercially.

[0022] Currently, severe acute pancreatitis is a dangerous condition with a high mortality rate, but there is a lack of specific drugs in clinical practice. Treatment measures are still limited to nutritional support and the use of anti-inflammatory drugs, with limited efficacy, long treatment cycles, and reliance on the patient's own recovery. New treatment drugs are urgently needed.

[0023] Meclozine (Meclizine), also known as chlorobenzamine, is an antihistamine used to treat motion sickness, dizziness, and vertigo. Its chemical structure is shown in Formula 1. Meclozine is an H1 histamine receptor antagonist and is used to treat motion sickness, dizziness, and vertigo. Meclozine has a good safety profile with mild side effects, including drowsiness, dry mouth, constipation, and fatigue.

[0024] Formula 1 The inventors discovered through research that at least one of meclopramide and its analogues, or pharmaceutically acceptable salts or esters thereof, has a good therapeutic effect on pancreatitis, especially acute pancreatitis, such as severe acute pancreatitis, and related symptoms. These related symptoms include lung injury, pancreatic injury, and upper abdominal pain induced by severe acute pancreatitis.

[0025] Furthermore, meclozidine and its analogues are selected from chiral isomers or mixtures of chiral isomers of compounds shown in the following structural formulas, or pharmaceutically acceptable salts thereof: In this context, R1 represents hydrogen or halogen; R2 represents any one of hydrogen, C1-C3 unsubstituted alkyl, C1-C3 unsubstituted carboxyl, and C1-C3 unsubstituted amide; and R3 represents C1-C3 unsubstituted alkylene or C3-C5 alkenyl.

[0026] For example, R1 represents any one of hydrogen, bromine, chlorine, and fluorine. R2 represents any one of hydrogen, methyl, ethyl, propyl, acetate, COOH, propionate, formamide, and acetamide; R3 represents methylene or 2-propenyl.

[0027] Furthermore, meclozine and its analogues are selected from at least one of the compounds shown in the following structural formulas: a, b、 b、 d、 e f g、 h and i; Preferred h or i.

[0028] It should be noted that (1) the compound number corresponds to the structural formula of the compound shown.

[0029] (2) Compounds b and c represent chiral compounds respectively, while compound a represents a mixture of compounds b and c. Compounds e, f, g, and h are all mixtures of chiral compounds.

[0030] (3) All compounds a to i were purchased from commercially available products. For example, compound ag was purchased from WuXi AppTec, compound h was purchased from Aladdin, and compound i was purchased from Macklin.

[0031] Furthermore, meclozine and its analogues or pharmaceutically acceptable salts thereof include at least one of the following: hydrochloride, hydrobromide, sulfate, phosphate, acetate, citrate, lactate, ascorbate, maleate, tartrate, malate, gluconate, sulfonate, citrate, benzoate, benzenesulfonate, carbonate, methanesulfonate, stearate, nitrate, valerate, and succinate.

[0032] The drug also includes pharmaceutically acceptable excipients. These include, but are not limited to, fillers, binders, disintegrants, lubricants, coating materials, preservatives, flavoring agents, and colorants. For example, fillers include starch, sucrose, lactose, microcrystalline cellulose, and pregelatinized starch. Binders include starch paste, sodium carboxymethyl cellulose (CMC-Na), hydroxypropyl methylcellulose (HPMC), and povidone (PVP). Disintegrants include dry starch, sodium carboxymethyl starch (CMS-Na), and crospovidone (PVPP). Lubricants include magnesium stearate, micronized silica gel, and talc. Coating materials include gastric-soluble (e.g., HPMC, PVP), enteric-soluble (e.g., CAP, acrylic resin), and water-insoluble (e.g., ethyl cellulose). Preservatives include sodium benzoate; flavoring agents include sucrose and stevia; colorants include iron oxide; solubilizers include surfactants; and co-solvents include potassium iodide.

[0033] The features and performance of the present invention will be further described in detail below with reference to embodiments.

[0034] I. Cellular Level Experiments Experimental Example 1 Determining the effect of meclozin and its analogues on amylase in AR42J cells induced by taeniacin AR42J cells in logarithmic growth phase were seeded into 96-well plates at 500,000 cells per ml and grown overnight at 37°C. After treatment with different concentrations of meclozidine or meclozidine-like compounds (AI) for 1 hour in medium containing 1% FBS, cytosine (30 nM) was added for a total treatment of 24 hours. Cell viability was assessed using the MTT assay to detect the toxicity of the compounds to the cells, and the amylase content in the cell culture supernatant was determined using an amylase assay kit.

[0035] See results Figure 1 The results for meclopramide and its analogues ai correspond one-to-one. Figure 1 AI in the middle. According to Figure 1 It is known that meclozine and its analogues significantly inhibit the secretion of amylase induced by taeniacin without significant cytotoxicity. Among them, meclozine has the most significant inhibitory effect on amylase.

[0036] Experiment Example 2 Determining the effects of meclozazine and compound i on amylase and lipase secreted by AR42J induced by taeniacin. AR42J cells in logarithmic growth phase were seeded into 96-well plates at 500,000 cells per ml and grown overnight at 37°C. After treatment with meclozazine (2.5 μM, 5 μM, 10 μM, 15 μM) and compound i (10 nM, 100 nM, 500 nM, 5 μM) for 1 hour in medium containing 1% FBS, cytosine (30 nM) was added for a further 24 hours of co-treatment. Cell viability was assessed using the MTT assay to detect the toxicity of the compounds to the cells. The levels of amylase and lipase in the cell culture supernatant were determined using amylase and lipase assay kits.

[0037] See results Figure 2 ,according to Figure 2 It can be seen that meclozine and compound i significantly inhibited the secretion of amylase in rat pancreatic acinar cells AR42J cells induced by taeniacin. Figure 2 China A and Figure 2 (D), and can simultaneously inhibit the secretion of lipase in AR42J cells induced by taurine (D). Figure 2 China B and Figure 2 (Middle E). Furthermore, MTT results showed that meclozine and compound i had no significant toxicity to AR42J cells ( Figure 2 C and Figure 2 (Middle F).

[0038] Experimental Example 3 Determining the effect of meclozamide on L-arginine-induced AR42J cell death AR42J cells in logarithmic growth phase were seeded into 96-well plates at 500,000 cells per ml and grown overnight at 37°C. After treatment with 5 μM meclomethasone in F12K medium containing 1% FBS for 1 hour, L-arginine (25 mM) was added for a total treatment of 24 hours. Cell viability was determined by the MTT assay; cell necrosis was detected by PI staining; the level of lactate dehydrogenase (LDH) in the cell culture supernatant was determined using an LDH assay kit; and mitochondrial membrane potential was detected using a TMRM probe.

[0039] See results Figure 3 ,like Figure 3 As shown, AR42J cells exhibited a significant decrease in cell viability and extensive cell death 24 hours after L-arginine induction, while meclozine significantly reversed cell death and improved cell viability. Figure 3 (A). L-arginine induces pancreatic acinar cell necrosis; PI staining detects a large number of PI-positive cells; meclopramide significantly reduces the number of PI-positive cells and decreases cell necrosis ( Figure 3(B) Following L-arginine induction, LDH release from cells significantly increased, indicating cell membrane damage and cellular injury. Meclozazine itself had no significant effect on LDH release, but it significantly reduced L-arginine-induced LDH release and significantly alleviated L-arginine-induced cell damage. Figure 3 (C) Mitochondrial membrane potential (TMRM) probe staining showed that L-arginine induction led to a significant decrease in mitochondrial membrane potential (significantly reduced red fluorescence); meclopramide itself had no significant effect on TMRM fluorescence, but it could significantly inhibit the L-arginine-induced decrease in red fluorescence, suggesting that meclopramide can restore mitochondrial membrane potential and significantly alleviate L-arginine-induced mitochondrial damage. Figure 3 (D).

[0040] Experiment Example 4 Determining the effect of meclozazine on sodium taurocholate-induced AR42J cell death AR42J cells in logarithmic growth phase were seeded into 96-well plates at a density of 500,000 cells per milliliter and grown overnight at 37°C. After treatment with 5 μM meclomethasone in F12K medium containing 1% FBS for 1 hour, sodium taurocholate (200 μM) was added for a total treatment of 48 hours. Cell viability was determined by the MTT assay; cell necrosis was detected by PI staining; the level of lactate dehydrogenase in the cell culture supernatant was determined using an LDH assay kit; and mitochondrial membrane potential was detected using a TMRM probe.

[0041] See results Figure 4 ,like Figure 4 As shown, AR42J cells showed decreased cell viability after 48 hours of treatment with sodium taurocholate, while meclozine was able to reverse cell death and increase cell viability. Figure 4 (A). Sodium taurocholate induced cell necrosis and detected PI-positive cells. Meclopramide significantly reduced the number of PI-positive cells and decreased cell necrosis (…). Figure 4 (B) Treatment with sodium taurocholate significantly increased cellular LDH release. Meclozine itself had no significant effect on LDH release, but it significantly reduced sodium taurocholate-induced LDH release and alleviated sodium taurocholate-induced cell damage. Figure 4 (C). Sodium taurocholate significantly induced a decrease in mitochondrial membrane potential (reduced red fluorescence of TMRM), while meclozine significantly inhibited the sodium taurocholate-induced decrease in mitochondrial membrane potential (increased red fluorescence of TMRM) and alleviated sodium taurocholate-induced mitochondrial damage. Figure 4 (D).

[0042] II. Animal-level experiments Experimental Example 5 Determining the effects of meclozine and compound i on improving severe acute pancreatitis induced by crotonin combined with lipopolysaccharide in mice. 1. Animal grouping and administration After one week of acclimatization, male C57 / BL6 mice (18-20 g) were randomly divided into five groups of six mice each: a blank control group, a model group induced by linalool combined with lipopolysaccharide (linalool 50 μg / kg, lipopolysaccharide 15 mg / kg), a positive control group of ulinastatin (40000 U / kg), a low-dose group of meclozine (5 mg / kg), a high-dose group of meclozine (12.5 mg / kg), a low-dose group of compound i (5 mg / kg), and a high-dose group of compound i (25 mg / kg).

[0043] Mice were fasted for 12 hours before the experiment, but had free access to water. After weighing, mice were randomly divided into groups. The model group received intraperitoneal injections of 50 μg / kg of lindane, once per hour for a total of seven times. Immediately after the last injection of lindane, mice were given an intraperitoneal injection of lipopolysaccharide (15 mg / kg). After modeling, the control and model groups received intraperitoneal injections of physiological saline at a dose volume of 10 mL / kg; the positive control group received intraperitoneal injections of ulinastatin (40000 U / kg); the meclopramide groups received high (12.5 mg / kg) and low (5 mg / kg) doses via gavage, and the compound i groups received high (25 mg / kg) and low (5 mg / kg) doses via gavage. Animals were provided with ample food and water after administration. A second dose was administered 12 hours after modeling, and samples were collected 48 hours after modeling. 2. Animal euthanasia, sampling, and sample processing Forty-eight hours after establishing the pancreatitis model in mice, blood was collected by enucleation and allowed to clot naturally at room temperature. Pancreatic and lung tissues were quickly harvested, rinsed in physiological saline, and the surface adipose tissue and mucosa were carefully removed. The tail portion of the pancreatic tissue was fixed in 4% paraformaldehyde for histopathological examination; the left lung was also fixed in 4% paraformaldehyde for histopathological examination. The remaining pancreatic tissue and right lung were labeled, flash-frozen in liquid nitrogen, and then stored at -80 °C for long-term preservation for other relevant indicator detection.

[0044] 3. Histopathological examination Pancreatic and lung tissues were fixed in 4% paraformaldehyde for 72 hours, placed in embedding cassettes, and numbered. After dehydration with a gradient of alcohols, clearing with xylene, embedding in liquid paraffin, and cutting into 6 μm thick sections, the tissues were dewaxed with xylene, dissolved in a gradient of alcohols of different concentrations, stained with hematoxylin and eosin, mounted with neutral resin, and observed, photographed, and analyzed under a light microscope.

[0045] 4. Oxidative stress level detection Whole blood was collected from mice, and after clotting at room temperature for 30 minutes, it was centrifuged (3500 rpm, 15 min, 4℃) to obtain the supernatant serum. Pancreatic tissue from mice was collected and homogenized with pre-chilled physiological saline at a weight-to-volume ratio of 1:9. The homogenate was collected and stored on ice for later use. MDA levels in mouse serum and pancreatic tissue were detected using an MDA detection kit according to the manufacturer's instructions.

[0046] 5. Inflammatory factor detection According to the kit manufacturer's instructions, the levels of IL-6 and MPO in mouse serum and pancreatic tissue were detected using the inflammatory factor enzyme-linked immunosorbent assay (ELISA) kit and the Nanjing Jiancheng MPO detection kit.

[0047] 6. Serum and pancreatic tissue amylase and lipase detection Whole blood was collected from mice and centrifuged at room temperature for 30 minutes to obtain the supernatant serum (3500 rpm, 15 min, 4℃). Pancreatic tissue from mice was homogenized with pre-cooled physiological saline at a weight-to-volume ratio of 1:9 and centrifuged (4000 rpm, 10 min, 4℃) to obtain the tissue homogenate. Amylase and lipase levels in serum and pancreatic tissue were measured using an amylase and lipase assay kit, following all manufacturer's instructions.

[0048] 7. Western blot analysis of the expression levels of Caspase 7 and GAPDH proteins in mouse pancreatic tissue. Mouse pancreatic tissue was homogenized by adding an appropriate amount of RIPA lysis buffer containing phosphatase inhibitors and PMSF. The homogenate was centrifuged at 12000 rpm for 15 min at 4°C, and the supernatant was collected as the total protein extract. Protein concentration was determined using a BCA protein quantification kit according to the manufacturer's instructions. Proteins were separated by SDS-PAGE gel electrophoresis and transferred to a PVDF membrane. The membrane was blocked with 5% skim milk for 2 h at room temperature and washed three times with TBST for 5 minutes each time. Primary antibody was incubated overnight at 4°C according to experimental requirements and the target protein molecular weight. The next day, the membrane was incubated with HRP-goat anti-rabbit secondary antibody (1:5000) for 2 h at room temperature and washed three times with TBST for 5 minutes each time. In a dark room, luminescent solutions A and B were mixed 1:1 and applied to the PVDF membrane. The target protein bands were visualized using a gel imaging system, and the target protein intensity was quantified using analysis software.

[0049] The results are as follows: (1) To determine the protective effect of meclozidine and compound i on the pancreas of mice with severe acute pancreatitis, H&E staining was performed on pancreatic tissue from mice with a severe acute pancreatitis model induced by the combined use of crotonin and LPS. See the results below. Figure 5The efficacy results of meclopramide are as follows Figure 5 As shown in Figure A. The pancreatic tissue of the control group (Control) mice was intact, with normal acinar structure, homogeneous texture, and almost no inflammatory cells, indicating a normal physiological state. In contrast, the pancreas of mice in the severe acute pancreatitis group showed extensive inflammatory cell infiltration, varying degrees of edema and necrosis, and increased interacicular spacing, suggesting severe damage and necrosis of the pancreatic tissue. The pancreatic pathological changes in the positive control group (40,000 U / kg) mice showed improvement, with reduced cell necrosis and inflammatory cell infiltration. Oral administration of low-dose (5 mg / kg) and high-dose (12.5 mg / kg) meclozide significantly reduced pancreatic tissue necrosis, significantly decreased interacicular spacing, reduced inflammatory cell infiltration, and significantly improved tissue integrity. In particular, the high-dose group was almost indistinguishable from the Control group, and its effect was significantly superior to that of the positive control group (ulinastatin). The efficacy results of compound i are as follows: Figure 5 As shown in Figure B, the pancreatic tissue of mice in the severe acute pancreatitis group showed severe damage and necrosis, with extensive inflammatory cell infiltration, varying degrees of edema and necrosis, and increased interacicular spacing. After gavage administration of compound i, the necrosis of the pancreatic tissue in mice was improved, the acinar structure remained intact, the interacicular spacing decreased, and the efficacy was superior to that of the positive control drug ulinastatin. These experimental results demonstrate that meclopramide and compound i can significantly improve the pathological damage of pancreatic tissue in mice with severe acute pancreatitis, and their efficacy is superior to that of the positive control drug ulinastatin.

[0050] (2) After the onset of severe acute pancreatitis, distal organs can be affected, causing lung damage. To determine the effects of meclozidine and compound i on lung damage in mice with severe acute pancreatitis, H&E staining was performed on mouse lung tissue. See the results below. Figure 6 The efficacy results of meclopramide are as follows Figure 6 As shown in Figure A, the lung tissue of the control group mice was intact, with normal alveolar structure, occasional bronchial congestion, and very few inflammatory cells, indicating a normal physiological state. The lungs of the model group mice showed pulmonary hemorrhage, alveolar wall thickening, alveolar cavity atrophy, and extensive inflammatory cell infiltration, suggesting severe lung damage. The lung damage in the positive control group (40,000 U / kg) mice improved, but alveolar congestion and alveolar cavity atrophy remained, although the alveolar wall structure was slightly improved compared to the model group. Oral administration of low-dose (5 mg / kg) and high-dose (12.5 mg / kg) meclozide significantly improved lung damage in mice, with occasional alveolar congestion, significantly reduced inflammatory cell infiltration, and restoration of normal alveolar structure. The high-dose group showed particularly significant improvement in lung tissue structure, demonstrating better efficacy than the positive control group (ulinastatin). The efficacy results of compound i are as follows: Figure 6As shown in Figure B, the alveolar structure of mice in the control group remained intact and in a normal state, while the model group mice showed increased pulmonary hemorrhage, alveolar wall thickening, alveolar cavity collapse, and severe lung damage. Gavage administration of low-dose (5 mg / kg) and high-dose (25 mg / kg) of compound i significantly improved lung damage in mice, with occasional alveolar congestion, significantly reduced inflammatory cell infiltration, and restoration of normal alveolar structure. The high-dose group showed particularly significant improvement in lung tissue structure, demonstrating a superior effect compared to the positive control drug ulinastatin. These experimental results indicate that meclozazine and compound i can significantly improve lung damage in mice with severe acute pancreatitis, and their effects are superior to those of the positive control drug ulinastatin.

[0051] (3) To evaluate the effects of meclozidine and compound i on severe acute pancreatitis, the levels of amylase and lipase in mouse serum and pancreatic tissue were measured. See results below. Figure 7 For the efficacy results of meclopramide, please refer to [link / reference]. Figure 7 Chinese AD. (e.g., [likely referring to a specific language or technology]) Figure 7 China A and Figure 7 As shown in Figure C, the serum and pancreatic tissue amylase levels in mice with severe acute pancreatitis induced by taeniacin combined with LPS were significantly higher than those in the control group, with the mean increase being approximately three times that of the control group. Serum amylase levels in the meclozamide treatment group and the positive control drug ulinastatin group were significantly lower than those in the model group, and the efficacy of high-dose meclozamide was comparable to that of the positive control drug ulinastatin. Serum and pancreatic tissue lipase assay results are shown below. Figure 7 As shown in Figures B and D, the serum and pancreatic tissue lipase levels in the model group were significantly higher than those in the control group. After treatment with meclopramide and the positive control drug ulinastatin, serum lipase levels decreased significantly, and the efficacy of meclopramide and the positive control drug ulinastatin was comparable. See the efficacy results for compound i for details. Figure 7 The results of serum amylase level measurements in E and F are as follows: Figure 7 As shown in Figure E, the amylase level in the model group mice was significantly higher than that in the control group. After treatment with compound i via gavage, the serum amylase level was significantly lower than that in the model group. The high- and low-dose compound i treatment groups were superior to the positive control drug ulinastatin group. Serum lipase measurement results are shown below. Figure 7 As shown in Figure F, after treatment with compound i and the positive control drug ulinastatin, serum lipase levels were significantly reduced, and the high-dose compound i group was significantly better than the positive control drug ulinastatin.

[0052] (4) To determine the effects of meclopramide and compound i on the inflammatory response in mice with severe acute pancreatitis, the serum levels of inflammatory factors IL-6, IL-1β, and MPO were measured. See the results for meclopramide efficacy. Figure 8 AC, such as Figure 8 China A and Figure 8As shown in Figure B, serum IL-6 and MPO levels were significantly elevated in mice in the severe acute pancreatitis model group, while IL-6 and MPO levels were significantly decreased in the positive control group (ulinastatin) and the meclozamide treatment group. Figure 8 As shown in Figure C, MPO levels in the pancreatic tissue of mice with severe acute pancreatitis were significantly elevated. Compared with the model group, treatment with the positive control drugs ulinastatin and meclozazine significantly reduced the MPO level in the pancreatic tissue of mice. See [link to compound i] for efficacy results. Figure 8 DF, such as Figure 8 D, Figure 8 China E and Figure 8 As shown in Figure F, serum levels of IL-6, IL-1β, and MPO were significantly elevated in mice in the severe acute pancreatitis model group. Compared with the model group, the positive control drug ulinastatin and compound i significantly reduced serum levels of IL-6, IL-1β, and MPO in mice. These experimental results demonstrate that meclozidine and compound i can significantly alleviate the inflammatory response in mice with severe acute pancreatitis.

[0053] (5) Pancreatitis is often accompanied by varying degrees of oxidative stress, which exacerbates the progression of the disease. To investigate the effect of meclopramide on oxidative stress during the pathogenesis of severe acute pancreatitis, the lipid peroxidation marker MDA in mouse serum and pancreatic tissue was measured. See the results below. Figure 9 ,like Figure 9 China A and Figure 9 As shown in Figure B, malondialdehyde (MDA) was significantly elevated in the model group, indicating that severe acute pancreatitis is accompanied by significant oxidative stress. The positive control group showed a decrease compared to the model group, while the meclopramide group showed a significant decrease compared to the model group, with better efficacy than the positive control group. These experimental results demonstrate that meclopramide can significantly alleviate oxidative stress in mice with severe acute pancreatitis. (6) Apoptosis of pancreatic tissue cells occurs after severe acute pancreatitis. To investigate the effect of meclozamide on apoptosis induced by severe acute pancreatitis, the expression level of the apoptosis marker Caspase-7 in pancreatic tissue cells was measured. See the results below. Figure 10 ,like Figure 10 China A and Figure 10 As shown in Figure B, compared with the control group, the cleaved form of Caspase-7 in the pancreatic tissue of the model group mice was significantly increased; compared with the model group, the cleaved form of Caspase-7 in both the positive control group (ulinastatin) and the meclopramide group was significantly decreased. These results indicate that meclopramide has a significant inhibitory effect on pancreatic tissue apoptosis in severe acute pancreatitis.

[0054] Experimental Example 6 The therapeutic effect of meclozidine on L-arginine-induced severe acute pancreatitis in mice 1. Animal grouping and administration After being acclimatized to their environment for one week, C57 / BL6 male mice (18-20 g) were randomly divided into five groups of six mice each: control group, L-arginine model group (L-arginine 3.5 g / kg), positive control drug ulinastatin group (40000 U / kg), low-dose meclozine group (1 mg / kg), and high-dose meclozine group (2.5 mg / kg). Mice were fasted for 12 hours before the experiment, but had free access to water. After weighing, mice were randomly assigned to groups. The model group received L-arginine intraperitoneally at a dose of 3.5 g / kg, injected once per hour for two hours. The control group received the same frequency and dose of saline intraperitoneally. After modeling, the control and model groups received saline intraperitoneally at a dose of 10 mL / kg; the positive control group received ulinastatin intraperitoneally; and the meclopramide group received meclopramide intraperitoneally. Animals were provided with ample food and water after administration. Samples were collected 24 hours after modeling for subsequent experiments.

[0055] 2. Animal euthanasia, sampling, and sample processing Twenty-four hours after establishing the pancreatitis model in mice, blood was collected by enucleation and allowed to clot naturally at room temperature. Pancreatic tissue was quickly harvested, washed in physiological saline, and the surface adipose tissue and mucosa were carefully removed. The tail portion of the pancreatic tissue was fixed in 4% paraformaldehyde for histopathological examination. The remaining pancreatic tissue was labeled, flash-frozen in liquid nitrogen, and then stored at -80°C for long-term preservation and to detect other relevant indicators. 3. Histopathological examination Pancreatic tissue was fixed in 4% paraformaldehyde for 72 hours, then placed in embedding cassettes and numbered. After dehydration with a gradient of alcohols, clearing with xylene, embedding in liquid paraffin, and cutting into 6 μm thick sections, the tissue was dewaxed with xylene, dissolved in a gradient of alcohols of different concentrations, stained with hematoxylin and eosin, mounted with neutral resin, and observed, photographed, and analyzed under a light microscope.

[0056] 4. Oxidative stress level detection Whole blood was collected from mice, and after clotting at room temperature for 30 minutes, it was centrifuged (3500 rpm, 15 min, 4℃) to obtain the supernatant serum. Pancreatic tissue from mice was collected and homogenized with pre-chilled physiological saline at a weight-to-volume ratio of 1:9. The homogenate was collected and stored on ice. Following the manufacturer's instructions, the levels of MDA, ROS, and catalase in mouse serum and pancreatic tissue were measured using an MDA detection kit, Amplex Red fluorescent dye (hydrogen peroxide detection reagent), and a catalase detection kit.

[0057] 5. Inflammatory factor detection According to the kit manufacturer's instructions, the levels of pro-inflammatory factors IL-6, IL-1β, TNF-α and MPO were measured using the enzyme-linked immunosorbent assay (ELISA) kit for inflammatory factors and the Nanjing Jiancheng MPO detection kit.

[0058] 6. Serum and pancreatic tissue amylase and lipase detection Whole blood was collected from mice and allowed to clot at room temperature for 30 minutes before centrifugation (3500 rpm, 15 min, 4℃) to obtain the supernatant serum. Pancreatic tissue from mice was collected and homogenized with pre-cooled physiological saline at a weight-to-volume ratio of 1:9, then centrifuged (4000 rpm, 10 min, 4℃) to obtain the tissue homogenate. Amylase and lipase levels in serum and pancreatic tissue were measured using an amylase and lipase assay kit, with all steps performed according to the manufacturer's instructions.

[0059] The results are as follows: (1) To determine the protective effect of meclozamide on the pancreas of mice with severe acute pancreatitis, H&E staining was performed on pancreatic tissue from L-arginine-induced severe acute pancreatitis model mice. See the results below. Figure 11 ,like Figure 11 As shown in Figure A, the pancreatic tissue of the control group mice was structurally intact, with normal acinar structure, homogeneous texture, and very little inflammatory cell infiltration. In contrast, the pancreatic tissue of the model group mice showed extensive inflammatory cell infiltration, varying degrees of edema and necrosis, and increased interacinar spacing, indicating severe pancreatic tissue damage and necrosis. The pancreatic necrosis and inflammatory cell infiltration in the positive control group (40,000 U / kg) improved. Intraperitoneal injection of low-dose (1 mg / kg) and high-dose (2.5 mg / kg) meclozide resulted in reduced pancreatic tissue necrosis, decreased interacinar spacing, and relatively intact tissue structure in mice, with the high-dose group showing significantly better results than the positive control group. These experimental results demonstrate that meclozide can significantly improve pancreatic tissue damage in mice with severe acute pancreatitis, and its efficacy is superior to that of the positive control group (ulinastatin).

[0060] (2) To investigate the protective effect of meclozamide against L-arginine-induced severe acute pancreatitis in mice, the levels of amylase and lipase in mouse serum and pancreatic tissue were measured. See results below. Figure 12 ,like Figure 12 China A and Figure 12 As shown in Figure C, the serum and pancreatic tissue amylase levels in the model group mice were significantly higher than those in the control group. Compared with the model group, treatment with meclopramide significantly reduced serum and pancreatic tissue amylase levels in mice, and the high-dose meclopramide group was more effective than the positive control drug ulinastatin. Figure 12 China B and Figure 12As shown in Figure D, serum lipase levels were significantly elevated in the model group mice compared to the control group. Serum lipase levels were significantly reduced after treatment with meclozidine and the positive control drug ulinastatin, compared to the model group. These experimental results demonstrate that meclozidine can significantly inhibit the increase of amylase and lipase in the severe acute pancreatitis model, and its inhibitory effect on amylase is superior to that of the positive control drug ulinastatin.

[0061] (3) To determine the effect of meclozamide on the inflammatory response in mice with L-arginine-induced severe acute pancreatitis, the serum levels of inflammatory factors IL-6, IL-1β, and TNF-α were measured. See results below. Figure 13 , Figure 13 The results from the AC study showed that, compared with the control group, the serum levels of IL-6, IL-1β, and TNF-α in the model group mice were significantly increased. Compared with the model group, the positive control drugs ulinastatin and meclozamine significantly reduced the levels of IL-6, IL-1β, and TNF-α. These experimental results indicate that meclozamine can significantly improve the inflammatory response in mice with severe acute pancreatitis.

[0062] (4) Oxidative stress is often present after the onset of pancreatitis, which is an important factor contributing to the aggravation of the condition. To investigate the effect of meclopramide on oxidative stress induced by L-arginine-induced severe acute pancreatitis, the levels of lipid peroxidation markers MDA, catalase, and hydrogen peroxide in mouse serum and pancreatic tissue were measured. See results below. Figure 14 ,like Figure 14 As shown in Figure A, compared to the control group, the catalase level in the model group mice was significantly reduced, while the high-dose meclopramide group significantly restored catalase levels. Figure 14 China B and Figure 14 As shown in Figure C, compared with the control group, the MDA levels in serum and pancreatic tissue of the model group were significantly increased. Compared with the model group, the MDA level in the high-dose meclozamide group was significantly reduced, and its efficacy was significantly better than that of the positive control drug ulinastatin. The levels of hydrogen peroxide in serum and pancreatic tissue can directly reflect the level of oxidative stress. Figure 14 China D and Figure 14 The results showed that, compared with the control group, the serum and pancreatic tissue levels of the model group mice were significantly increased, indicating severe oxidative stress. Compared with the model group, the hydrogen peroxide levels in the positive control group (ulinastatin) and the meclopramide treatment group were significantly decreased, and the antioxidant effect of meclopramide was superior to that of ulinastatin. These experimental results indicate that meclopramide can significantly improve the antioxidant capacity and reduce oxidative stress in mice with severe acute pancreatitis.

[0063] In the embodiments of this invention, CER represents taurine; LPS represents lipopolysaccharide; L-Arg represents L-arginine; MDA represents malondialdehyde; MPO represents myeloid peroxidase; and TLCS represents sodium taurocholate.

[0064] In summary, meclozine significantly improved pancreatic damage in mice with severe acute pancreatitis, reduced serum amylase and lipase levels, improved pancreatic pathological damage, reduced inflammatory response and oxidative stress, and inhibited pancreatic acinar cell death, further demonstrating that meclozine and its derivatives can treat pancreatitis.

[0065] The above description is merely a preferred embodiment of the present invention and is not intended to limit the invention. Various modifications and variations can be made to the present invention by those skilled in the art. Any modifications, equivalent substitutions, improvements, etc., made within the spirit and principles of the present invention should be included within the scope of protection of the present invention.

Claims

1. The use of at least one of meclozidine and its analogues or pharmaceutically acceptable salts or esters thereof in the preparation of medicaments for the prevention and / or treatment of pancreatitis.

2. The application according to claim 1, characterized in that, The meclopramide and its analogues are selected from chiral isomers or mixtures of chiral isomers of the compounds shown in the following structural formulas, or pharmaceutically acceptable salts thereof: In this context, R1 represents hydrogen or halogen; R2 represents any one of hydrogen, C1-C3 unsubstituted alkyl, C1-C3 unsubstituted carboxyl, and C1-C3 unsubstituted amide; and R3 represents C1-C3 unsubstituted alkylene or C3-C5 alkenyl.

3. The application according to claim 2, characterized in that, R1 represents any one of hydrogen, chlorine, and bromine; R2 represents any one of hydrogen, methyl, ethyl, acetate, COOH, formamide, and acetamide; R3 represents methylene or 2-propenyl.

4. The application according to any one of claims 1-3, characterized in that, The meclozine and its analogues are selected from at least one of the compounds shown in the following structural formulas: 、 、 、 、 、 , and ; Preferred or .

5. The application according to claim 1, characterized in that, The pancreatitis mentioned includes acute pancreatitis.

6. The application according to claim 5, characterized in that, The pancreatitis mentioned includes severe acute pancreatitis.

7. The application according to claim 1, characterized in that, The drug is used to improve lung damage caused by pancreatitis.

8. The application according to claim 1, characterized in that, The medication is for relieving upper abdominal pain caused by pancreatitis.

9. The application according to claim 1, characterized in that, The drug also includes pharmaceutically acceptable excipients.

10. The application according to claim 1, characterized in that, The pharmaceutically acceptable salts include at least one of the following: hydrochloride, hydrobromide, sulfate, phosphate, acetate, citrate, lactate, ascorbate, maleate, tartrate, malate, gluconate, sulfonate, citrate, benzoate, benzenesulfonate, carbonate, methanesulfonate, stearate, nitrate, valerate, and succinate.

Images