CN116942656B - Application of naringenin derivative in preparation of anti-pancreatitis medicine - Google Patents

Abstract

The invention belongs to the technical field of medicines, and particularly relates to application of naringenin derivatives in preparation of anti-pancreatitis medicines. In order to develop a novel medicine with obvious severe acute pancreatitis resistance, the invention provides application of naringenin derivative 7-hydroxy-2- (4-hydroxyphenyl) -5- (3-methylbut-2-en-1-oxyl) benzopyran-4-one or salt thereof in preparing medicines for preventing and/or treating pancreatitis. The compound or the salt thereof can obviously reduce the level of serum amylase and serum pancrelipase caused by severe acute pancreatitis, lighten the edema, the inflammation and the necrosis of pancreatic tissues when the severe acute pancreatitis occurs, provide a new medicine source for treating the pancreatitis and provide a new choice for developing and applying anti-pancreatitis medicines.

Description

Use of naringenin derivatives in preparing anti-pancreatitis drugs

Technical Field

The invention belongs to the field of medical technology, and particularly relates to use of naringenin derivatives in preparing anti-pancreatitis drugs.

Background Art

Severe acute pancreatitis has an acute onset and develops rapidly, seriously endangering the patient's physical and mental health. It is a difficult and difficult disease in clinical treatment. According to statistics, severe acute pancreatitis is often accompanied by multiple complications such as pancreatic abscess, pancreatic pseudocyst, shock, organ failure, etc. Severe cases may be secondary to abdominal, respiratory, urinary tract infections, and the spread of infection causes sepsis, which imposes a huge burden on patients, their families, and society. In recent years, the incidence of severe acute pancreatitis has increased year by year worldwide. Because the pathogenesis of its cell damage has not been fully elucidated and there is no specific drug to prevent or treat it, conducting research on the mechanism of severe acute pancreatitis and developing specific drugs for severe acute pancreatitis are major scientific issues that need to be solved globally. In summary, it is crucial to discover new therapeutic drugs and to deeply explore the pathogenesis and treatment of severe acute pancreatitis.

Naringenin, with a molecular formula of C ₁₅ H ₁₂ O ₅ , is a flavonoid compound with multiple biological functions such as anti-tumor, anti-aging, and regulation of fat metabolism. Existing evidence shows that naringenin can reduce myocardial damage in mice with severe acute pancreatitis. However, due to its limited therapeutic effect, its clinical development potential is greatly limited. Therefore, it is necessary to research and develop naringenin derivatives with significant anti-severe acute pancreatitis effects and obvious pancreatic targeting.

Summary of the invention

The object of the present invention is to provide a use of a naringenin derivative (a compound shown in Formula I, defined as compound 1, named YZJ122) or a salt thereof in the preparation of a drug for preventing and/or treating pancreatitis, and in particular, to find that compound YZJ122 has a good therapeutic and preventive effect on severe acute pancreatitis.

The present invention provides the use of a compound represented by formula I or a salt thereof in the preparation of a drug for preventing and/or treating pancreatitis:

Furthermore, the pancreatitis includes at least one of acute pancreatitis or chronic pancreatitis; the acute pancreatitis includes at least one of mild acute pancreatitis, moderate acute pancreatitis or severe acute pancreatitis; the chronic pancreatitis includes at least one of alcoholic, biliary, recurrent, hereditary, autoimmune or idiopathic chronic pancreatitis.

Preferably, the pancreatitis is severe acute pancreatitis.

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

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

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.

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

Furthermore, the auxiliary material is at least one of glyceryl behenate, hypromellose or magnesium stearate.

Furthermore, the preparation is an oral preparation, a nasal mucosal preparation, an oral mucosal preparation or an injection preparation.

In the preferred technical scheme of the present invention, the effective dose of the compound represented by Formula I or its salt can be changed according to the mode of administration, the age and weight of the patient, the severity of the disease and other relevant factors. The recommended dose for oral administration is 100-1000 mg/time, 1-3 times a day; the recommended dose for injection is 15-45 mg/time, once a day; the recommended dose for spray inhalation administration is 500-1000 mg/time, 1-3 times a day.

Beneficial effects: The present invention provides the use of a naringenin derivative YZJ122 or a salt thereof in the preparation of a drug for preventing and/or treating pancreatitis. Animal experiments show that, compared with (S)-naringenin, compound YZJ122 can significantly reduce the levels of serum amylase and serum pancreatic lipase caused by severe acute pancreatitis, and reduce the edema, inflammation and necrosis of pancreatic tissue during severe acute pancreatitis, which indicates that the compound YZJ122 of the present invention can be used to develop drugs that reduce the levels of serum amylase and serum pancreatic lipase and reduce pancreatic tissue damage during the onset of pancreatitis. The application of the present invention can provide a new source of drugs for the treatment of pancreatitis, and has potential significant economic and social benefits. The preparation prepared by compound YZJ122 has the prospect of being used as a drug for the treatment of pancreatitis, and is expected to become an innovative drug for the treatment of pancreatitis with high efficiency and low toxicity, and has broad prospects for industrialization.

BRIEF DESCRIPTION OF THE DRAWINGS

FIG1 is the ¹ H NMR spectrum of compound 1 of Example 1;

FIG2 is the ¹ H NMR spectrum of compound 3 of Example 1;

FIG3 is the ¹ H NMR spectrum of compound 4 of Example 1;

FIG4 is the ¹ H NMR spectrum of compound 5 of Example 1;

FIG5 is a graph showing the effects of (S)-naringenin and compounds 1, 3, 4, and 5 on the degree of pancreatic edema in severe acute pancreatitis under the NAT model in Experimental Example 2;

FIG6 is a graph showing the effects of (S)-naringenin and compounds 1, 3, 4, and 5 on pancreatic inflammation caused by severe acute pancreatitis in Experimental Example 3 under the NAT model;

FIG7 is a graph showing the effects of (S)-naringenin and compounds 1, 3, 4, and 5 on tissue necrosis caused by severe acute pancreatitis in Experimental Example 4 under the NAT model;

FIG8 is a graph showing the effects of (S)-naringenin and compounds 1, 3, 4, and 5 on serum amylase levels caused by acute pancreatitis in the NAT model in Experimental Example 5;

FIG. 9 is a graph showing the effects of (S)-naringenin and compounds 1, 3, 4, and 5 on serum pancreatic lipase levels caused by acute pancreatitis under the NAT model in Experimental Example 6.

(ns: P>0.05, *: P<0.05, **: P<0.01, ***: P<0.001, ****: P<0.0001, n=4)

DETAILED DESCRIPTION

The applicant designed four derivatives based on the structure of (S)-naringenin, namely compounds 1, 3, 4, and 5, as shown below. The applicant found that although compound 1 and compounds 3, 4, and 5 have similar structures, compound 1 exhibits significant anti-severe acute pancreatitis activity and has more efficient anti-severe acute pancreatitis activity in vitro and in vivo compared with (S)-naringenin (PCM). Animal experiments have confirmed that the therapeutic effect of compound 1 at the lowest dose of 25 mg/kg for mice is significantly better than the therapeutic effects of (S)-naringenin, compounds 3, 4, and 5 at 200 mg/kg.

Definition of terms:

The compounds and derivatives provided by the present invention can be named according to the IUPAC (International Union of Pure and Applied Chemistry) or CAS (Chemical Abstracts Service, Columbus, OH) nomenclature system.

The term "pharmaceutically acceptable" means that a carrier, vehicle, diluent, excipient, and/or formed salt is generally chemically or physically compatible with the other ingredients that constitute a pharmaceutical dosage form and physiologically compatible with the receptor.

The term "pharmaceutically acceptable salt" refers to acidic and/or basic salts formed by the compounds of the present invention with 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 (e.g., equivalent). These salts may form a precipitate in the solution and be collected by filtering, or be recovered after solvent evaporation, or be obtained by freeze drying after reaction in an aqueous medium. The salt described 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.

The administration route of the compound of the present invention is not particularly limited, and representative administration routes include, but are not limited to, oral, parenteral (intravenous, intramuscular or subcutaneous), and topical administration.

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: (a) fillers or solubilizers, for example, starches, lactose, sucrose, glucose, mannitol, and silicic acid; (b) binders, for example, hydroxypropyl methylcellulose, hydroxymethylcellulose, alginates, gelatin, polyvinyl pyrrolidone, sucrose, and acacia; (c) humectants, for example, glycerol; (d) disintegrants, for example, agar, calcium carbonate, potato or tapioca starch, alginic acid, certain complex silicates, and sodium carbonate; (e) solubilizers, for example, paraffin; (f) absorption accelerators, for example, quaternary ammonium compounds; (g) wetting agents, for example, cetyl alcohol, glyceryl monostearate and glyceryl behenate; (h) adsorbents, for example, kaolin; and (i) lubricants, for example, talc, calcium stearate, magnesium stearate, solid polyethylene glycols, sodium lauryl sulfate, or mixtures thereof. In the case of capsules, tablets and pills, the dosage forms may also comprise buffering agents.

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.

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.

Besides such inert diluents, the composition may 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 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.

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.

Dosage forms for topical administration of the compounds of the invention include ointments, powders, patches, sprays and inhalants. The active ingredient is mixed under sterile conditions with a physiologically acceptable carrier and any preservatives, buffers, or propellants that may be required.

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

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.

The scheme of the present invention will be explained below in conjunction with the embodiments. It will be appreciated by those skilled in the art that the following embodiments are only used to illustrate the present invention and should not be considered as limiting the scope of the present invention. Where specific techniques or conditions are not indicated in the embodiments, the techniques or conditions described in the literature in this area or the product specifications are used. The reagents or instruments used are not indicated by the manufacturer and are all conventional products that can be obtained commercially.

Example 1 Preparation of Compound YZJ122

Compound YZJ122 is a derivative obtained by optimizing the structure of (S)-naringenin. The precursor compound 2 of this compound has a known synthetic route (RSC Adv., 2021, 11, 28934–28939).

Compound 1: 7-hydroxy-2-(4-hydroxyphenyl)-5-(3-methylbut-2-en-1-oxy)benzopyran-4-one, molecular weight: 340.

Compound 3: 4-(7-acetoxy-5-hydroxy-8-(3-methylbut-2-en-1-yl)-4-oxacyclohexan-2-yl)phenyl acetate, molecular weight: 424.

Compound 4: 5,7-dihydroxy-2-(4-hydroxyphenyl)-8-(3-methylbut-2-en-1-yl)benzopyran-4-one, molecular weight: 340.

Compound 5: 4-(7-acetoxy-5-methoxy-8-(3-methylbut-2-en-1-yl)-4-oxo-2-yl)phenyl acetate, molecular weight: 438.

Synthesis of compound 1: Weigh compound 2 (30 mg) and dissolve it in methanol (1.5 mL), add solid NaOH (6 mg), and react at room temperature. After the reaction is completed by TLC monitoring, add 1M HCl to adjust the pH to neutral, extract with ethyl acetate three times, combine the organic phases, dry over anhydrous sodium sulfate, filter, concentrate under reduced pressure, and purify on a silica gel column to obtain compound 4 (23 mg, 93%).

Synthesis of compound 3: Compound 2 (300 mg) was weighed and dissolved in 1,2-dichloroethane (2 mL), Eu(fod) ₃ (80 mg) was added, and the mixture was reacted at 80° C. After the reaction was completed by TLC monitoring, the reaction mixture was cooled to room temperature and purified by silica gel column to obtain compound 3 (183 mg, 61%).

Synthesis of compound 4: Weigh compound 3 (50 mg) and dissolve it in methanol (1.5 mL), add solid NaOH (9 mg), and react at room temperature. After TLC monitoring, add 1M HCl to adjust the pH to neutral, extract with ethyl acetate three times, combine the organic phases, dry over anhydrous sodium sulfate, filter, concentrate under reduced pressure, and purify on a silica gel column to obtain compound 4 (38 mg, 95%).

Synthesis of compound 5: Weigh compound 3 (100 mg) and dissolve it in ether (2.5 mL), add silver oxide (218 mg) and iodomethane (88 uL), and react at room temperature. TLC monitors the reaction completion, filter the reaction solution, concentrate the filtrate, and purify it on a silica gel column to obtain compound 5 (65 mg, 62%).

Compound 1: ¹ H NMR (400MHz, DMSO) δ7.28 (d, J = 11.6 Hz, 2H), 6.78 (d, J = 8.5 Hz, 2H), 5.98 (s, 1H), 5.86 (s, 1H) ,5.39(t,J＝7.0Hz,1H),5.28(dd,J＝12.6,3.0Hz,1H),4.47(d,J＝6.5Hz,2H),2.92(dd,J＝16.3,12.6Hz, 1H), 2.46 (dd, J＝16.4, 3.1Hz, 1H), 1.74 (s, 3H), 1.69 (s, 3H).

Compound 3: ¹ H NMR (400MHz, DMSO) δ11.80 (s, 1H), 7.65–7.49 (m, 2H), 7.21 (d, J = 8.6Hz, 2H), 6.36 (s, 1H), 5.72 ( dd,J＝13.1,2.9Hz,1H),5.00(t,J＝7.1Hz,1H),3.40(dd,J＝17.1,13.1Hz,1H),3.09(d,J＝6.6Hz,2H), 2.93(dd,J＝17.2,3.1Hz,1H),2.29(s,6H),1.59(s,3H),1.53(s,3H).

Compound 4: ¹ H NMR (400MHz, DMSO) δ12.11 (s, 1H), 10.74 (s, 1H), 9.55 (s, 1H), 7.31 (d, J = 8.6Hz, 2H), 6.80 (d, J＝8.6Hz,2H),5.97(s,1H),5.42(dd,J＝12.4,3.1Hz,1H),5.09(t,J＝6.5Hz,1H),3.20(dd,J＝17.1,12.4 Hz, 1H), 3.09 (d, J = 7.3Hz, 2H), 2.72 (dd, J = 17.1, 3.1Hz, 1H), 1.57 (d, J = 19.1Hz, 6H).

Compound 5: ¹ H NMR (400MHz, CDCl ₃ ) δ7.45 (d, J = 8.8 Hz, 2H), 7.13 (d, J = 8.6 Hz, 2H), 6.29 (s, 1H), 5.43 (d, J ＝16.1Hz,1H),5.14–5.01(m,1H),3.87(s,3H),3.19(d,J＝7.1Hz,2H),3.07–2.94(m,1H),2.85(d,J＝ 16.4Hz,1H),2.32(s,6H),1.65(s,3H),1.58(s,3H).

The ¹ H NMR spectra of compounds 1, 3, 4 and 5 are shown in Figures 1-4, respectively.

Experimental Example 1 Establishment of Acute Pancreatitis Model

Eight-week-old female Balb/c mice were randomly divided into a blank group, a model group (NaT), a (S)-naringenin (PCM) treatment group (NaT+PCM), and different concentrations of naringenin derivatives treatment groups (YZJ122 25mg/kg+NaT, YZJ122 50mg/kg+NaT, YZJ122 75mg/kg+NaT, YZJ122 200mg/kg+NaT, compound 3 200mg/kg+NaT, compound 4 200mg/kg+NaT, compound 5 200mg/kg+NaT). The results of histopathological examination showed that the (S)-naringenin (PCM) treatment group (NaT+PCM) and different concentrations of naringenin derivatives treatment groups did not show visible toxic side effects.

The mice in the NaT group, PCM group, and naringenin derivative treatment group were injected with 3.0% sodium taurocholate by retropancreaticobiliary injection. The blank group was injected with an equal volume of 0.90% saline. The naringenin derivative treatment group was intraperitoneally injected with therapeutic drugs 1h, 3h, and 6h after modeling, and samples were collected 24h later. The pancreas of the experimental group mice was fixed, dehydrated, stained, dewaxed, transparent, and sealed by pathological staining to observe the tissue integrity, pancreatic edema, inflammatory cell infiltration, and necrosis of the pancreatic tissue. The iodine/starch colorimetric method was used to determine the level of serum amylase in mice. The biotin double antibody sandwich enzyme-linked immunosorbent assay (ELISA) was used to determine the level of serum pancreatic lipase in mice.

Experimental Example 2: Experiment on the effect of YZJ122 on reducing pancreatic edema in severe acute pancreatitis

Pancreatic edema is one of the most intuitive changes in the pancreas when severe acute pancreatitis occurs. The degree of edema can reflect the severity of acute pancreatitis. Its cause is mainly the recruitment and infiltration of a large number of immune cells in the pancreas when acute pancreatitis occurs. Among these large numbers of infiltrating immune cells, the activation of neutrophils plays an important role in the pathological process of acute pancreatitis. The degree of pancreatic edema is usually measured by the ratio of the mass difference between wet weight and dry weight to wet weight.

As shown in the results of Figure 5, the degree of pancreatic tissue edema was evaluated by comparing the weight of fresh pancreatic samples (wet weight) and the weight of dried pancreatic samples (dry weight), and the edema scores of the blank group, model group (NaT), drug-treated groups (YZJ, 3, 4, 5) and (S)-naringenin (PCM) group were compared. The results showed that compared with the model group (NaT), drug-treated groups (3, 4, 5) and (S)-naringenin (PCM), the compound YZJ122 of the present invention can significantly improve the pancreatic edema caused by severe acute pancreatitis, and the effect is significantly better than (S)-naringenin and derivatives 3, 4, 5.

The degree of pancreatic edema in the NaT group (2.30) was significantly higher than that in the blank group (0.58) (P < 0.0001). Compared with the NaT group, after treatment with (S)-naringenin derivative YZJ122, the levels of pancreatic edema in 25 mg/kg+NaT, 50 mg/kg+NaT, 75 mg/kg+NaT, and 200 mg/kg+NaT groups decreased to 1.61 (P < 0.0001), 0.99 (P < 0.0001), 0.63 (P < 0.0001), and 1.11 (P < 0.0001), respectively, compared with the NaT group. However, after treatment with (S)-naringenin (PCM), compound 3, compound 4, and compound 5, the levels of pancreatic edema only decreased to 1.89 (P < 0.001), 2.05 (P > 0.05), 2.11 (P > 0.05), and 1.95 (P < 0.01). It can be concluded that among derivatives 1, 3, 4, and 5, only compound YZJ122 significantly improved the edema of pancreatic tissue in severe acute pancreatitis after treatment, improved severe acute pancreatitis, and the therapeutic effect of compound YZJ122 at the lowest dose of 25 mg/kg was significantly better than the therapeutic effect of (S)-naringenin (PCM) 200 mg/kg (P < 0.05).

Experimental Example 3: Experiment on the effect of YZJ122 on reducing the severity of pancreatic inflammation in severe acute pancreatitis

By comparing the inflammatory scores of the blank group, the model group (NaT), the drug-treated groups (YZJ, 3, 4, 5) and the (S)-naringenin (PCM) group, the results showed that compared with the model group (NaT), the drug-treated groups (3, 4, 5) and (S)-naringenin (PCM), the compound YZJ122 of the present invention can significantly improve the inflammation caused by severe acute pancreatitis, and the effect is significantly better than (S)-naringenin and derivatives 3, 4, 5.

As shown in Figure 6, the pancreatic inflammatory score of the NaT group (1.83) was significantly higher than that of the blank group (0.43) (P < 0.0001). Compared with the NaT group, after treatment with the (S)-naringenin derivative YZJ122, the pancreatic inflammatory score of the 25 mg/kg+NaT, 50 mg/kg+NaT, 75 mg/kg+NaT, and 200 mg/kg+NaT groups decreased to 1.32 (P < 0.0001), 0.84 (P < 0.0001), 0.40 (P < 0.0001), and 0.94 (P < 0.0001), respectively, compared with the NaT group, while after treatment with (S)-naringenin (PCM), compound 3, compound 4, and compound 5, the scores only decreased to 1.65 (P < 0.01), 1.71 (P > 0.05), 1.74 (P > 0.05), and 1.58 (P < 0.001). It can be concluded that among derivatives 1, 3, 4, and 5, only compound YZJ122 significantly improved the inflammatory degree of pancreatic tissue in severe acute pancreatitis after treatment, improved severe acute pancreatitis, and the therapeutic effect of YZJ122 at the lowest dose of 25 mg/kg was significantly better than the therapeutic effect of (S)-naringenin (PCM) 200 mg/kg (P < 0.0001).

Experimental Example 4: Experiment on the effect of YZJ122 on reducing pancreatic tissue necrosis in severe acute pancreatitis

By comparing the tissue necrosis scores of the blank group, model group (NaT), drug-treated groups (YZJ, 3, 4, 5) and (S)-naringenin (PCM) group, the results showed that compared with the model group (NaT), drug-treated groups (3, 4, 5) and (S)-naringenin (PCM), the compound YZJ122 of the present invention can significantly improve tissue necrosis caused by severe acute pancreatitis, and the effect is significantly better than (S)-naringenin and derivatives 3, 4, 5.

As shown in Figure 7, the pancreatic tissue necrosis score of the NaT group (2.53) was significantly increased compared with the blank group (0.10) (P < 0.0001). After treatment with the (S)-naringenin derivative YZJ122, the necrosis score of the 25 mg/kg+NaT, 50 mg/kg+NaT, 75 mg/kg+NaT, and 200 mg/kg+NaT groups decreased to 1.64 (P < 0.0001), 0.76 (P < 0.0001), 0.12 (P < 0.0001), and 0.65 (P < 0.0001) respectively compared with the NaT group, while that of (S)-naringenin (PCM), compound 3, compound 4, and compound 5 only decreased to 2.06 (P < 0.0001), 2.25 (P < 0.05), 2.45 (P > 0.05), and 2.08 (P < 0.0001) after treatment. It can be concluded that compound YZJ122 treatment significantly improved the inflammatory degree of pancreatic tissue in severe acute pancreatitis, improved severe acute pancreatitis, and the therapeutic effect of the lowest dose of YZJ122, 25 mg/kg, was significantly better than the therapeutic effect of (S)-naringenin (PCM) 200 mg/kg (P < 0.0001).

Experimental Example 5: Experiment on the effect of YZJ122 on reducing serum amylase levels in severe acute pancreatitis

Existing data show that the serum amylase content of more than 90% of patients with acute pancreatitis will increase. Therefore, the present invention uses a serum amylase kit to measure the amylase level in mouse serum. By comparing the serum amylase levels of the blank group, model group (NaT), drug-treated groups (YZJ, 3, 4, 5) and (S)-naringenin (PCM) group, the results show that compared with the model group (NaT), drug-treated groups (3, 4, 5) and (S)-naringenin (PCM), YZJ122 can significantly reduce the increase in serum amylase caused by severe acute pancreatitis, and the effect is significantly better than (S)-naringenin and derivatives 3, 4, 5.

Amylase can hydrolyze starch to produce glucose, maltose and dextrin. When the substrate concentration is known and in excess, iodine solution is added to the unhydrolyzed starch to produce a blue complex. The amount of hydrolyzed starch can be calculated based on the depth of the blue color, thereby calculating the activity. As shown in Figure 8, the serum amylase activity of the NaT group (17446U/L) was significantly higher than that of the blank group (4066U/L) (P < 0.0001). After YZJ122 treatment, the serum amylase activity of the 25mg/kg+NaT, 50mg/kg+NaT, 75mg/kg+NaT, and 200mg/kg+NaT groups decreased to 10124U/L (P < 0.0001), 6972U/L, respectively, compared with the NaT group. The serum amylase activity of mice with severe acute pancreatitis was significantly downregulated after treatment with compound YZJ122, thereby improving severe acute pancreatitis, and the therapeutic effect of the lowest dose of YZJ122, 25 mg/kg, was significantly better than that of (S)-naringenin (PCM) 200 mg/kg (P < 0.001).

Experimental Example 6: Experiment on reducing serum pancreatic lipase activity in patients with severe acute pancreatitis with YZJ122

In addition to severe acute pancreatitis, serum amylase can also be elevated in other diseases, such as gastric perforation, pancreatic tumors, etc., so the diagnosis of severe acute pancreatitis with serum amylase alone has limitations. Serum pancreatic lipase activity increases after the onset of severe acute pancreatitis and can also be used to measure acute pancreatitis. It is usually more specific and lasts longer, so serum pancreatic lipase is measured to verify the accuracy of the conclusion.

The level of serum pancreatic lipase in mice was determined using a lipase kit. The serum pancreatic lipase levels in the blank group, model group (NaT), drug-treated groups (YZJ, 3, 4, 5) and (S)-naringenin (PCM) were compared. The results showed that compared with the model group (NaT), drug-treated groups (3, 4, 5) and (S)-naringenin (PCM), YZJ122 could significantly reduce the increase in serum pancreatic lipase caused by severe acute pancreatitis, and the effect was significantly better than (S)-naringenin and its derivatives 3, 4, 5.

The level of mouse pancreatic lipase was determined by biotin double antibody sandwich enzyme-linked immunosorbent assay (ELISA). After adding lipase to the enzyme-labeled wells pre-coated with mouse pancreatic lipase monoclonal antibodies, biotin-labeled antibodies were added, and then combined with avidin HRP to form immune complexes. After incubation and washing, unbound enzymes were removed, and then substrates A and B were added to produce blue, which was finally converted to yellow under the action of acid. The depth of color is positively correlated with the concentration of mouse pancreatic lipase in the sample.

As shown in Figure 9, the serum pancreatic lipase activity in the NaT group (639 U/L) was significantly higher than that in the blank group (43 U/L) (P < 0.0001). After YZJ122 treatment, the 25 mg/kg+NaT, 50 mg/kg+NaT, 75 mg/kg+NaT, and 200 mg/kg+NaT groups decreased to 281 U/L (P < 0.0001), 142 U/L (P < 0.0001), and 142 U/L (P < 0.0001) respectively. L (P < 0.0001), 52U/L (P < 0.0001) and 138U/L (P < 0.0001), while (S)-naringenin (PCM), compound 3, compound 4 and compound 5 only decreased to 475U/L (P < 0.0001), 513U/L (P < 0.001), 586U/L (P> 0.05) and 489U/L (P < 0.0001). It can be concluded that the serum pancreatic lipase activity of mice with severe acute pancreatitis was significantly downregulated after YZJ122 treatment, thereby improving severe acute pancreatitis, and the therapeutic effect of the lowest dose of YZJ122, 25mg/kg, was significantly better than the therapeutic effect of (S)-naringenin (PCM) 200mg/kg (P < 0.0001).

Claims (7)

1. Use of the compound represented by formula I or its salt in the preparation of a drug for preventing and/or treating pancreatitis: 2. The use according to claim 1, characterized in that: the pancreatitis includes at least one of acute pancreatitis or chronic pancreatitis; the acute pancreatitis includes at least one of mild acute pancreatitis, moderate acute pancreatitis or severe acute pancreatitis; the chronic pancreatitis includes at least one of alcoholic, biliary, recurrent, hereditary, autoimmune or idiopathic chronic pancreatitis. 3. The use according to claim 2, characterized in that: the pancreatitis is severe acute pancreatitis. 4. 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. 5. The use according to claim 4, characterized in that the preparation is a sustained-release agent or a controlled-release agent. 6. The use according to claim 4, characterized in that the auxiliary material is at least one of glyceryl behenate, hypromellose or magnesium stearate. 7. 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.