CN119405651A - Application of isorhamnetin in preparation of medicines for preventing and treating acute pancreatitis - Google Patents

Abstract

The invention provides application of isorhamnetin in preparing medicines for preventing or treating acute pancreatitis. The acute pancreatitis comprises at least one of mild acute pancreatitis, moderate acute pancreatitis or severe acute pancreatitis. Animal experiments in examples show that compared with a model group, the isorhamnetin Li Suke can obviously reduce the levels of serum amylase and serum pancreatic lipase caused by severe acute pancreatitis, and reduce edema, inflammation, apoptosis and necrosis of pancreatic tissues when acute pancreatitis occurs, so that the isorhamnetin compound can be used for developing medicines for reducing the levels of serum amylase and serum pancreatic lipase and reducing pancreatic tissue injury in the incidence process of pancreatitis.

Description

Application of isorhamnetin in preparation of medicines for preventing and treating acute pancreatitis

Technical Field

The invention belongs to the technical field of medicines, and particularly relates to application of isorhamnetin in preparation of a medicine for preventing and treating acute pancreatitis.

Background

Acute pancreatitis (Acute pancreatitis) is the most common gastrointestinal disorder requiring hospitalization, associated with sudden inflammation of the pancreas, characterized by premature production of intracellular trypsinogens and premature activation of inflammatory cells, leading to necrosis of acinar cells and local or systemic inflammation and organ failure. The etiology of acute pancreatitis is related to pancreatic duct obstruction, alcoholism, obesity, diabetes, and drug induction caused by gallstones. The incidence of acute pancreatitis has increased year by year, with about one fifth of patients developing moderate or severe pancreatitis, with necrosis of surrounding tissues of the pancreas and multiple organ failure, with mortality rates as high as 20%. In the last decades, the treatment strategies of acute pancreatitis have been greatly developed, and especially symptomatic treatment modes such as moderate aggressive fluid resuscitation, anti-inflammatory treatment, anti-infection treatment, surgical treatment and the like within 72 hours, but the targeted drugs for treatment are still lacking, and the clinical effect is limited.

Isorhamnetin (isorhamnetin) is a flavonoid compound, and has a chemical structural formula shown in formula I and a molecular formula of C ₁₆H₁₂O₇. Isorhamnetin is widely present in flowers, fruits and leaves of many plants such as ginkgo, sea buckthorn, bupleurum, pollen typhae, lotus and the like. In recent years, pharmacodynamic researches show that the isorhamnetin has the effects of protecting endothelium of mice Li Suyou, resisting atherosclerosis, resisting myocardial ischemia, protecting myocardial cells, inhibiting myocardial fibrosis, reducing blood pressure, resisting thrombus, platelet aggregation, resisting oxidization and the like, has the effects of inhibiting adipocyte differentiation, resisting hypoxia, resisting tumors, reducing blood glucose, resisting inflammation, resisting viruses and the like, but has no report on the preparation of medicines for preventing and treating acute pancreatitis.

Disclosure of Invention

The invention aims to provide application of isorhamnetin in preparing medicines for preventing and treating acute pancreatitis.

Further, the acute pancreatitis includes at least one of mild acute pancreatitis, moderate acute pancreatitis, or severe acute pancreatitis. Preferably, the pancreatitis is severe acute pancreatitis.

Further, the medicament reduces serum amylase activity and serum pancreatic lipase activity.

Further, the medicament reduces pancreatic edema, inflammation and necrosis when severe acute pancreatitis occurs.

Further, the medicine is a preparation prepared by taking a compound shown in a formula I or salt thereof as an active ingredient and adding pharmaceutically acceptable auxiliary materials or auxiliary ingredients.

Further, the preparation is a slow release agent or a controlled release agent.

Further, the mode of administration of the compounds of the present invention is not particularly limited, and representative modes of administration include, but are not limited to, oral, nasal, oral, or injectable administration of the formulation. In a preferred embodiment of the present invention, the effective dosage of the compound of formula I or a salt thereof may vary depending on the mode of administration, the age and weight of the patient, the severity of the condition, and other related factors.

The invention provides application of isorhamnetin or a salt thereof in preparing medicines for preventing and/or treating acute pancreatitis. Animal experiments show that compared with a model group, the isorhamnetin Li Suke can obviously reduce the level of serum amylase and serum pancreatic lipase caused by severe acute pancreatitis, and reduce edema, inflammation, apoptosis and necrosis of pancreatic tissues when the acute pancreatitis occurs, so that the isorhamnetin can be used for developing medicines for reducing the level of serum amylase and serum pancreatic lipase and reducing pancreatic tissue injury in the incidence process of pancreatitis.

The term "pharmaceutically acceptable" in the specification means that the carrier, cargo, diluent, adjuvant, and/or salt formed is generally chemically or physically compatible with the other ingredients comprising the pharmaceutical dosage form, and physiologically compatible with the recipient.

The term "pharmaceutically acceptable salts" in the specification refers to acid and/or base salts of the compounds of the invention with inorganic and/or organic acids and bases, and also includes zwitterionic salts (inner salts) and also includes quaternary ammonium salts, for example alkylammonium salts. These salts may be obtained directly in the final isolation and purification of the compounds. Or by mixing the above-mentioned compound with a certain amount of an acid or a base as appropriate (for example, equivalent). These salts may be obtained by precipitation in solution and collected by filtration, or recovered after evaporation of the solvent, or by lyophilization after reaction in an aqueous medium.

The salts of the present invention may be the hydrochloride, sulfate, citrate, benzenesulfonate, hydrobromide, hydrofluoric, phosphate, acetate, propionate, succinate, oxalate, malate, succinate, fumarate, maleate, tartrate or trifluoroacetate salts of the compounds.

Solid dosage forms for oral administration include capsules, tablets, pills, powders and granules. In these solid dosage forms, the active compound is admixed with at least one conventional inert excipient or carrier, such as sodium citrate or dicalcium phosphate, or with the following ingredients:

(a) Fillers or solubilisers, for example starch, lactose, sucrose, glucose, mannitol and silicic acid;

(b) Binders, for example, hypromellose, hydroxymethyl cellulose, alginate, gelatin, polyvinylpyrrolidone, sucrose and acacia;

(c) Humectants, for example, glycerin;

(d) Disintegrants, for example, agar-agar, calcium carbonate, potato or tapioca starch, alginic acid, certain complex silicates, and sodium carbonate;

(e) Slow solvents such as paraffin;

(f) Absorption accelerators, for example quaternary amine compounds;

(g) Wetting agents, such as cetyl alcohol, glyceryl monostearate and glyceryl behenate;

(h) Adsorbents, such as kaolin;

(i) Lubricants, for example, talc, calcium stearate, magnesium stearate, solid polyethylene glycol, sodium lauryl sulfate, or mixtures thereof. In capsules, tablets and pills, the dosage forms may also comprise buffering agents.

Solid dosage forms such as tablets, dragees, capsules, pills and granules are prepared with coatings and shells, such as enteric coatings and other materials well known in the art. They may contain opacifying agents and the release of the active compound or compounds in such compositions may be released in a delayed manner in a certain part of the digestive tract. Examples of embedding components that can be used are polymeric substances and waxes. The active compound may also be in the form of microcapsules with one or more of the above excipients, if desired.

Liquid dosage forms for oral administration include pharmaceutically acceptable emulsions, solutions, suspensions, syrups or tinctures. In addition to the active compound, the liquid dosage forms may contain inert diluents commonly used in the art such as, for example, water or other solvents, solubilizing agents and emulsifiers such as ethyl alcohol, isopropyl alcohol, ethyl carbonate, ethyl acetate, propylene glycol, 1, 3-butylene glycol, dimethylformamide and oils, in particular, cottonseed, groundnut, corn germ, olive, castor and sesame oils or mixtures of these substances and the like.

In addition to these inert diluents, the compositions can also include adjuvants such as wetting agents, emulsifying and suspending agents, sweetening, flavoring, and perfuming agents.

Suspensions, in addition to the active compounds, may contain suspending agents as, for example, ethoxylated isostearyl alcohols, polyoxyethylene sorbitol and sorbitan esters, microcrystalline cellulose, aluminum methoxide and agar-agar or mixtures of these substances, and the like.

Compositions for parenteral injection may comprise physiologically acceptable sterile aqueous or anhydrous solutions, dispersions, suspensions or emulsions, and sterile powders for reconstitution into sterile injectable solutions or dispersions. Suitable aqueous and nonaqueous carriers, diluents, solvents or excipients include water, ethanol, polyols and suitable mixtures thereof.

The pharmaceutically acceptable auxiliary materials of the invention refer to substances contained in the dosage form except for active ingredients.

The pharmaceutically acceptable auxiliary component has certain physiological activity, but the addition of the component does not change the predominance of the pharmaceutical composition in the disease treatment process, but only plays auxiliary effects, and the auxiliary effects are only the utilization of the known activity of the component and are auxiliary treatment modes commonly used in the medical field. If the auxiliary components are used together with the pharmaceutical composition of the present invention, the auxiliary components still fall within the scope of the present invention.

Drawings

Fig. 1 shows the pancreatic index of mice (ns p >0.05, < p <0.01, < p < 0.001);

FIG. 2 is a photograph of a mouse pancreatic tissue HE staining (scale 100 μm);

Fig. 3 shows the TUNEL staining results of mouse pancreatic tissue (×p <0.001, scale 100 μm);

Fig. 4 is serum amylase content of mice (×p < 0.001);

fig. 5 shows the serum lipase content of mice (×p < 0.001).

Detailed Description

The present invention is described in detail below by way of examples. However, the examples provided herein are for illustrative purposes only and are not intended to limit the present invention.

24C 57BL/6 mice were selected and fed adaptively for 1 week, and were randomly divided into four groups, namely a blank Control group (Control group), an acute pancreatitis model group (AP group), a mice Li Sudi dose group (L group) and a mice Li Sugao dose group (H group), and 6 mice each.

On days 1-3, the Control group and the AP group were perfused with 0.5% sodium carboxymethyl cellulose solution, and the L group and the H group were perfused with 10mg/kg or 20mg/kg isorhamnetin, respectively, 1 time per day.

On day 4, 50 μg/kg of ranpirin physiological saline solution was intraperitoneally injected into the AP, L and H groups, respectively, 1 time every 1 hour (H) at intervals of 7 times, and the vehicle was administered in the 1,3,5H, AP, 10mg/kg or 20mg/kg isorhamnetin, control groups, respectively, of ranpirin, and in the 1,3,5H, AP, lavage, 0.5% sodium carboxymethyl cellulose, L and H groups, respectively. After 6h of final injection of rana chensinensis, mice were weighed and anesthetized with tribromoethanol, the eyeballs were allowed to stand at room temperature for 30min, the supernatants were centrifuged to detect serum amylase and lipase levels, pancreatic tissues were weighed, 4% paraformaldehyde fixed, paraffin embedded, and sections were stained with hematoxylin-eosin (HE) or TUNEL for subsequent pathology analysis.

Example 1

Experiment of the extent of pancreatic edema in the event of isorhamnetin inhibition of acute pancreatitis:

The ratio of the pancreatic weight to the body weight of the mice (P.W/B.W) was calculated to give a pancreatic index, which indicates a large degree of pancreatic edema. The pancreas index of each group is shown in fig. 1, and the pancreas index of the mice in the AP group is obviously up-regulated compared with that in the Control group, while the pancreas index of the mice in the Li Sugao dose group is obviously down-regulated according to the result of fig. 1, which indicates that the isorhamnetin can prevent and treat acute pancreatitis and inhibit pancreatic edema.

Example 2

Experiment of isorhamnetin in inhibiting inflammation and necrosis of pancreatic tissue when acute pancreatitis occurs:

Mouse pancreatic tissue, 4% paraformaldehyde fixed, gradient ethanol (30%, 50%, 70%, 80%, 100%) dehydrated, xylene permeabilized, paraffin embedded, sectioned, HE stained, light microscopy randomly acquired images as shown in figure 2. From fig. 2, it can be seen that the Control group pancreas is histologically intact, and that the AP group observes significant pathological lesions such as cytoplasmic shrinkage, destruction of acinar cell structure, cavitation, inflammation, necrosis and acinar edema. Compared with the AP group, the isorhamnetin treatment group can observe that the pathological damage of pancreatic tissues is obviously improved.

Pancreatic tissue was subjected to pathological analysis with reference to Rongione scoring method, scoring criteria:

necrosis is 20% -50% of extensive pancreas parenchymal necrosis, counting 3 minutes;

5% -20% of focal necrosis, counting for 2 minutes;

>50% pericatheter necrosis, 1 minute;

Without the above features, count 0 points;

Inflammatory cells appear in >50% of the leaflets of the pancreas, counting 3 points;

<50% leaflets, score 2;

inflammatory cells appear around or in the area of the catheter, 1 minute;

The results are shown in Table 1 without the above characterizations 0 points.

TABLE 1 mouse pancreatic tissue necrosis and inflammation score

Note that ns p >0.05, p <0.01, p <0.001, n=6.

Referring to the results in Table 1, the areas of necrosis and inflammation were significantly up-regulated in the pancreatic tissue of the AP group compared to the Control group, while the areas of necrosis and inflammation were reduced in the pancreatic tissue of the isomouse Li Suzu, and the dose group of isomouse Li Sugao was statistically different from the AP group, indicating that the effect of isorhamnetin on preventing pancreatitis was related to the inhibition of necrosis and inflammation of the pancreas.

Example 3

Experiment of isorhamnetin to inhibit apoptosis of pancreatic tissue when acute pancreatitis occurs:

Taking pancreatic tissues of each group of mice, fixing 4% paraformaldehyde, dehydrating gradient ethanol (30%, 50%, 70%, 80% and 100%) and permeabilizing xylene, embedding paraffin, slicing, staining according to TUNEL kit instructions, collecting pictures by a fluorescence microscope and analyzing, as shown in figure 3, wherein the apoptosis rate of the pancreatic tissues of the AP group is obviously up-regulated compared with the Control group, and the apoptosis rate of the cells of the pancreatic tissues of the mice Li Suzu compared with the AP group is obviously down-regulated, so that the effect of isorhamnetin in preventing pancreatitis is related to the inhibition of pancreatic apoptosis.

Example 4

Experiment of isorhamnetin to reduce serum amylase in acute pancreatitis:

The serum amylase content of each group of mice is detected by adopting a MNCHIP animal-dedicated full-automatic biochemical analyzer CELERCARE V, as shown in fig. 4, the serum amylase content of the AP group is obviously up-regulated compared with the Control group, and the serum amylase content of the isorhamnetin treatment group is obviously down-regulated compared with the AP group, so that the effect of isorhamnetin on preventing and treating acute pancreatitis is related to the inhibition of the release of pancreatic amylase.

Example 5

Experiment of isorhamnetin to reduce serum lipase in acute pancreatitis:

The serum lipase content of each group of mice is detected by adopting a MNCHIP animal-dedicated full-automatic biochemical analyzer CELERCARE V, as shown in fig. 5, the serum lipase content of the AP group is obviously up-regulated compared with the Control group, and the serum lipase content of the isorhamnetin treatment group is obviously down-regulated compared with the AP group, so that the effect of isorhamnetin on preventing and treating acute pancreatitis is related to the release of pancreatic lipase inhibition.

Claims (5)

1. Use of isorhamnetin in preparing a drug for preventing or treating acute pancreatitis, characterized in that isorhamnetin is a flavonoid compound, and its chemical structural formula is shown in the following formula I: . 2. The use according to claim 1, characterized in that: the acute pancreatitis includes at least one of mild acute pancreatitis, moderate acute pancreatitis or severe acute pancreatitis. 3. The use according to claim 1, characterized in that the drug is a preparation prepared by using the compound represented by formula I or its salt as the active ingredient and adding pharmaceutically acceptable excipients. 4. The use according to claim 3, characterized in that the preparation is a sustained-release agent or a controlled-release agent. 5. The use according to claim 4, characterized in that the preparation is an oral preparation, a nasal mucosal preparation, an oral mucosal preparation or an injection preparation.