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

Abstract

The invention provides application of isorhamnetin in preparation of a medicine 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 of the embodiment show that compared with a model group, the isorhamnetin can significantly reduce the levels of serum amylase and serum pancreatic lipase caused by severe acute pancreatitis, alleviate edema, inflammation, apoptosis and necrosis of pancreatic tissues when acute pancreatitis occurs, and reduce the incidence of acute pancreatitis. Therefore, the compound isorhamnetin can be used for developing drugs for reducing the levels of serum amylase and serum pancreatic lipase and relieving pancreatic tissue damage in the pancreatitis pathogenesis process.

Description

Technical Field

[0001] The invention belongs to the technical field of medicines, and particularly relates to use of isorhamnetin in preparing a medicament for preventing and treating acute pancreatitis. Background Art

[0002] Acute pancreatitis is the most common gastrointestinal disease requiring hospitalization. It is associated with sudden inflammation of the pancreas and is characterized by premature production of intracellular trypsinogen and premature activation of inflammatory cells, which cause necrosis of acinar cells and local or systemic inflammation and organ failure. The etiology of acute pancreatitis is related to pancreatic duct obstruction caused by gallstones, alcoholism, obesity, diabetes, and drug-induced. The incidence of acute pancreatitis is increasing year by year. About one-fifth of patients will develop moderate or severe pancreatitis, accompanied by necrosis of peripancreatic tissue and multiple organ failure, with a mortality rate of up to 20%. In the past few decades, the treatment strategy for acute pancreatitis has made great progress, especially symptomatic treatment within 72 hours, such as moderately aggressive fluid resuscitation, anti-inflammatory treatment, anti-infective treatment, and surgical treatment, but there is still a lack of targeted drugs for causal treatment, and the clinical efficacy is limited.

[0003] Isorhamnetin is a flavonoid compound with a chemical structure shown in Formula I and a molecular formula of C 16 H 12 O7. Isorhamnetin is widely found in the flowers, fruits and leaves of many plants such as Ginkgo, Seabuckthorn, Bupleurum, Pollen Typhae and Lotus. Pharmacological studies in recent years have found that isorhamnetin has multiple cardiovascular and cerebrovascular protective effects, including endothelial protection, anti-atherosclerosis, anti-myocardial ischemia, protection of myocardial cells, inhibition of myocardial fibrosis, blood pressure reduction, anti-thrombosis and platelet aggregation, and anti-oxidation. It also has multiple effects such as inhibiting adipocyte differentiation, hypoxia resistance, anti-tumor, hypoglycemic, anti-inflammatory and antiviral. However, there are no reports on the use of isorhamnetin in the preparation of drugs for the prevention and treatment of acute pancreatitis.

[0004] Summary of the invention

[0005] The purpose of the present invention is to provide the use of isorhamnetin in preparing a medicament for preventing and treating acute pancreatitis.

[0006] Furthermore, 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.

[0007] Furthermore, the drug reduces serum amylase activity and serum pancreatic lipase activity.

[0008] Furthermore, the drug reduces pancreatic edema, inflammation and necrosis when severe acute pancreatitis occurs.

[0009] Furthermore, the drug is a preparation prepared by using the compound represented by Formula I or its salt as an active ingredient and adding pharmaceutically acceptable excipients or auxiliary ingredients.

[0010] Furthermore, the preparation is a sustained-release preparation or a controlled-release preparation.

[0011] Furthermore, there is no particular limitation on the administration of the compounds of the present invention, and representative administration methods include but are not limited to: the preparation is administered orally, by nasal mucosa, oral mucosa or injection. In the preferred technical scheme of the present invention, the effective dose of the compound or its salt shown in Formula I can be changed according to the administration method, the patient's age and weight, the severity of the disease and other related factors.

[0012] The present invention provides the use of isorhamnetin or its salt in the preparation of a drug for preventing and / or treating acute pancreatitis. Animal experiments show that compared with a model group, isorhamnetin can significantly reduce the levels of serum amylase and serum pancreatic lipase caused by severe acute pancreatitis, and reduce edema, inflammation, apoptosis and necrosis of pancreatic tissue during acute pancreatitis, which indicates that the compound isorhamnetin of the present invention can be used to develop drugs for reducing the levels of serum amylase and serum pancreatic lipase and reducing pancreatic tissue damage during the onset of pancreatitis.

[0013] The term "pharmaceutically acceptable" in the specification refers to a carrier, vehicle, diluent, excipient, and / or salt formed that is generally chemically or physically compatible with other ingredients constituting a pharmaceutical dosage form and physiologically compatible with the receptor.

[0014] The term "pharmaceutically acceptable salt" in the specification refers to acidic and / or basic salts formed by the compounds of the present invention and inorganic and / or organic acids and bases, and also includes zwitterionic salts (inner salts), and also includes quaternary ammonium salts, such as alkylammonium salts. These salts can be directly obtained in the final separation and purification of the compound. It can also be obtained by mixing the above-mentioned compound with a certain amount of acid or base appropriately (for example, equivalent). These salts may form a precipitate in the solution and be collected by filtering, or be recovered after the solvent is evaporated, or be obtained by freeze drying after reaction in an aqueous medium.

[0015] The salt in the present invention can be a hydrochloride, sulfate, citrate, benzenesulfonate, hydrobromide, hydrofluoride, phosphate, acetate, propionate, succinate, oxalate, malate, succinate, fumarate, maleate, tartrate or trifluoroacetate of the compound.

[0016] Solid dosage forms for oral administration include capsules, tablets, pills, powders and granules. In these solid dosage forms, the active compound is mixed with at least one conventional inert excipient or carrier, such as sodium citrate or dicalcium phosphate, or with the following ingredients:

[0017] (a) fillers or solubilizers, for example, starch, lactose, sucrose, glucose, mannitol and silicic acid;

[0018] (b) binders, for example, hypromellose, hydroxymethylcellulose, alginate, gelatin, polyvinylpyrrolidone, sucrose and gum arabic;

[0019] (c) humectants, for example, glycerin;

[0020] (d) disintegrating agents, for example, agar, calcium carbonate, potato or tapioca starch, alginic acid, certain complex silicates, and sodium carbonate;

[0021] (e) a buffering solvent, such as paraffin;

[0022] (f) absorption accelerators, for example, quaternary ammonium compounds;

[0023] (g) wetting agents, for example cetyl alcohol, glyceryl monostearate and glyceryl behenate;

[0024] (h) adsorbents, for example, kaolin;

[0025] (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 form may also contain a buffering agent.

[0026] Solid dosage forms such as tablets, pills, capsules, pills and granules are prepared using coatings and shell materials, such as enteric coatings and other materials known in the art. They may contain opacifiers, and the release of the active compound or compounds in such compositions may be delayed in a certain part of the digestive tract. Examples of embedding components that may be used are polymeric substances and waxes. If necessary, the active compound may also be in microencapsulated form with one or more of the above-mentioned excipients.

[0027] Liquid dosage forms for oral administration include pharmaceutically acceptable emulsions, solutions, suspensions, syrups or tinctures. In addition to the active compound, the liquid dosage form may contain an inert diluent conventionally used in the art, such as water or other solvents, solubilizers and emulsifiers, for example, ethanol, isopropanol, ethyl carbonate, ethyl acetate, propylene glycol, 1,3-butylene glycol, dimethylformamide and oils, in particular cottonseed oil, peanut oil, corn germ oil, olive oil, castor oil and sesame oil or mixtures of these substances.

[0028] Besides such inert diluents, the composition may also include adjuvants such as wetting agents, emulsifying and suspending agents, sweetening, flavoring, and perfuming agents.

[0029] Suspensions, in addition to the active compounds, may contain suspending agents such as, for example, ethoxylated isostearyl alcohols, polyoxyethylene sorbitol and sorbitan esters, microcrystalline cellulose, aluminum methanol and agar, or mixtures of these substances, and the like.

[0030] Compositions for parenteral injection may include 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 non-aqueous carriers, diluents, solvents or excipients include water, ethanol, polyols and suitable mixtures thereof.

[0031] The pharmaceutically acceptable excipients described in the present invention refer to substances contained in the dosage form in addition to the active ingredients.

[0032] The pharmaceutically acceptable auxiliary ingredients of the present invention have certain physiological activities, but the addition of the ingredients will not change the dominant position of the above-mentioned pharmaceutical composition in the treatment of diseases, but only play auxiliary effects, which are merely the utilization of the known activity of the ingredients and are conventional auxiliary treatment methods in the medical field. If the above-mentioned auxiliary ingredients are used in combination with the pharmaceutical composition of the present invention, they should still fall within the scope of protection of the present invention. BRIEF DESCRIPTION OF THE DRAWINGS

[0033] Figure 1 is the pancreatic index of mice (ns p>0.05, *p<0.05, **p<0.01, ***p<0.001);

[0034] Figure 2 This is a picture of HE staining of mouse pancreatic tissue (scale bar 100 μm);

[0035] Figure 3 The results of TUNEL staining of mouse pancreatic tissue (***p<0.001, scale bar 100μm);

[0036] Figure 4 is the serum amylase content of mice (***p<0.001);

[0037] Figure 5 is the serum lipase content of mice (***p<0.001). DETAILED DESCRIPTION

[0038] The present invention is described in detail below by way of examples. However, the examples provided herein are only for illustrative purposes and are not intended to limit the present invention.

[0039] Twenty-four C57BL / 6 mice were selected and adaptively fed for 1 week, and then randomly divided into four groups: blank control group (Control group), acute pancreatitis model group (AP group), low-dose isorhamnetin group (L group), and high-dose isorhamnetin group (H group), with 6 mice in each group.

[0040] On days 1-3, the Control group and AP group were gavaged with 0.5% sodium carboxymethylcellulose solution, and the L group and H group were gavaged with 10 mg / kg or 20 mg / kg isorhamnetin, respectively, once a day.

[0041] On the 4th day, the AP, L and H groups were intraperitoneally injected with 50 μg / kg caerulein saline solution, once every 1 hour (h), for a total of 7 times. At 1, 3, and 5 hours after the injection of caerulein, the AP group was gavaged with 0.5% sodium carboxymethylcellulose, the L and H groups were gavaged with 10 mg / kg or 20 mg / kg isorhamnetin, and the Control group was given the vehicle. 6 hours after the last injection of caerulein, the mice were weighed and anesthetized with tribromoethanol, blood was taken from the eyeballs and left at room temperature for 30 minutes, and the supernatant was centrifuged to detect the levels of serum amylase and lipase; the pancreatic tissue was weighed, fixed with 4% paraformaldehyde, embedded in paraffin, and sections were stained with hematoxylin-eosin (HE) or TUNEL for subsequent pathological analysis.

[0042] Example 1

[0043] Experiment on the effect of isorhamnetin on the pancreatic edema during acute pancreatitis:

[0044] The pancreatic index was calculated by calculating the ratio of pancreatic weight to body weight (PW / BW). A larger pancreatic index indicated a greater degree of pancreatic edema. Figure 1 As shown, according to Figure 1 The results showed that compared with the Control group, the pancreatic index of mice in the AP group was significantly increased, while the pancreatic index of the high-dose isorhamnetin group was significantly decreased, indicating that the prevention and treatment of acute pancreatitis by isorhamnetin is related to the inhibition of pancreatic edema.

[0045] Example 2

[0046] Experiment on the inhibition of pancreatic tissue inflammation and necrosis by isorhamnetin during acute pancreatitis:

[0047] Mouse pancreatic tissue was fixed with 4% paraformaldehyde, dehydrated with gradient ethanol (30%, 50%, 70%, 80%, 100%), permeabilized with xylene, embedded in paraffin, sectioned, stained with HE, and randomly collected images under an optical microscope, such as Figure 2 As shown. Figure 2It can be seen that the pancreatic histology of the Control group was intact, and obvious pathological damage was observed in the AP group, such as cytoplasmic shrinkage, destruction of acinar cell structure, vacuolation, inflammation, necrosis, and acinar edema. Compared with the AP group, the pathological damage of pancreatic tissue in the isorhamnetin-treated group was significantly improved.

[0048] The pancreatic tissue was pathologically analyzed according to the Rongione scoring method. The scoring criteria are as follows:

[0049] Necrosis: 20% to 50% extensive pancreatic parenchymal necrosis, 3 points;

[0050] 5% to 20% focal necrosis, 2 points;

[0051] >50% periductal necrosis, scored 1 point;

[0052] If there are no above characteristics, 0 points will be given;

[0053] Inflammatory cells appeared in >50% of the lobules in the pancreatic parenchyma, scored 3 points;

[0054] <50% leaflets, 2 points;

[0055] Inflammatory cells appearing around the duct or in the ductal area were scored as 1 point;

[0056] The absence of the above characteristics was scored as 0 points. The results are shown in Table 1.

[0057] Table 1 Scores of pancreatic tissue necrosis and inflammation in mice

[0058]

[0059] Note: ns p>0.05, *p<0.05, **p<0.01, ***p<0.001, n=6.

[0060] Referring to the results in Table 1, it can be seen that compared with the Control group, the area of ​​necrosis and inflammation in the pancreatic tissue of the AP group was significantly increased, while the area of ​​necrosis and inflammation in the pancreatic tissue of the isorhamnetin group showed a decreasing trend, and there was a statistically significant difference between the high-dose isorhamnetin group and the AP group, indicating that the effect of isorhamnetin in preventing and treating pancreatitis is related to the inhibition of pancreatic necrosis and inflammation.

[0061] Example 3

[0062] Experiment on the inhibition of pancreatic tissue apoptosis by isorhamnetin during acute pancreatitis:

[0063] The pancreatic tissues of mice in each group were fixed with 4% paraformaldehyde, dehydrated with gradient ethanol (30%, 50%, 70%, 80%, 100%), permeabilized with xylene, embedded in paraffin, sliced, and stained according to the instructions of the TUNEL kit. The images were collected and analyzed using a fluorescence microscope. Figure 3 As shown, from Figure 3 It can be seen that compared with the Control group, the apoptosis rate of cells in the pancreatic tissue of the AP group was significantly increased; compared with the AP group, the apoptosis rate of cells in the pancreatic tissue of the isorhamnetin group was significantly decreased, indicating that the effect of isorhamnetin in preventing and treating pancreatitis is related to the inhibition of pancreatic cell apoptosis.

[0064] Example 4

[0065] Experiment of isorhamnetin reducing serum amylase in acute pancreatitis:

[0066] The MNCHIP animal-specific fully automatic biochemical analyzer Celercare V5 was used to detect the serum amylase content of mice in each group. Figure 4 As shown, from Figure 4 It can be seen that compared with the Control group, the serum amylase content in the AP group was significantly increased; compared with the AP group, the serum amylase content in the isorhamnetin-treated group was significantly decreased, indicating that the effect of isorhamnetin in preventing and treating acute pancreatitis is related to the inhibition of the release of pancreatic amylase.

[0067] Example 5

[0068] Experiment on the effect of isorhamnetin on reducing serum lipase in acute pancreatitis:

[0069] The MNCHIP animal-specific fully automatic biochemical analyzer Celercare V5 was used to detect the serum lipase content of mice in each group. Figure 5 As shown, from Figure 5 It can be seen that compared with the Control group, the serum lipase content in the AP group was significantly increased; compared with the AP group, the serum lipase content in the isorhamnetin-treated group was significantly decreased, indicating that the effect of isorhamnetin in preventing and treating acute pancreatitis is related to the inhibition of the release of pancreatic lipase.

Claims

1. Use of isorhamnetin in the preparation of a medicament for preventing or treating acute pancreatitis, characterized in that: Isorhamnetin is a flavonoid compound, and its chemical structure 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 medicine is a preparation prepared by taking the compound shown in 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 preparation or a controlled-release preparation.

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.

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