Application of salazosulfapyridine in preparation of medicament for treating cholestatic liver disease

Abstract

The invention discloses an application of salazosulfapyridine in preparation of drugs for treating cholestatic liver diseases, which comprises the following steps: dissolving salazosulfapyridine in 0.5% sodium carboxymethyl cellulose to prepare a suspension, orally taking 150mg / kg of the suspension once a day, continuously taking the suspension for 21 days, and taking the suspension for later use. It is proved for the first time at the animal level that salazosulfapyridine can effectively improve subacute cholestatic liver injury constructed by intragastric ANIT-corn oil suspension, and a new medicine scheme with an improvement effect on cholestatic liver diseases is provided. The scheme has the advantages of safety, no toxicity, definite curative effect (obvious improvement effect), wide application prospect and the like. Pharmacological experiments are developed by using a cholestatic liver disease model, and results show that the salazosulfapyridine effectively relieves liver injury symptoms caused by cholestasis, specifically, the activity of ALT, AST, ALP, TBil and TBA in rat serum is reduced. Liver histomorphology HE and MASSON staining jointly show that salazosulfapyridine effectively improves liver inflammatory cell infiltration and the fibrosis degree of a manifold region.

Description

Technical Field

[0001] The present invention relates to the field of chemical medicine, and in particular to application of sulfasalazine in the preparation of a drug for treating cholestatic liver disease. Background Art

[0002] Cholestatic liver disease (CLD) is a dysfunction of bile formation and / or flow caused by genetic, immune, environmental or other causes. This biliary dysfunction in CLD can be confined to tiny hepatic ductules, intrahepatic bile ducts, segmental bile ducts or large intrahepatic and extrahepatic bile ducts. Excessive accumulation of toxic bile acids usually leads to progressive hepatobiliary damage. Most CLD patients eventually progress to cirrhosis and liver failure, requiring liver transplantation due to the lack of effective treatment. Primary sclerosing cholangitis (PSC) and primary biliary cholangitis (PBC) are the most prominent CLDs. Although ursodeoxycholic acid (UDCA) remains the baseline treatment for most cholestatic liver diseases, UDCA treatment puts approximately one-third of PBC patients and all PSC patients at risk of disease progression.

[0003] Sulfasalazine (SASP) is a prodrug that is currently used alone or in combination with other drugs in the clinic to treat a variety of diseases such as inflammatory bowel disease, rheumatoid arthritis and skin diseases. It has antibacterial, anti-inflammatory and immunosuppressive activities. SASP may produce anti-inflammatory effects by inhibiting the production of cytokines, prostaglandins and leukotrienes, reducing the adhesion and function of leukocytes, etc. The SASP mother and 5-ASA inhibit immune activity in many aspects by inhibiting the generation, activation and differentiation of lymphocytes. The SASP metabolite 5-ASA has a strong protective effect against oxidative damage and can also directly act as an antioxidant and free radical scavenger.

[0004] At present, there is no report on the use of sulfasalazine to treat cholestatic liver disease. Therefore, the present invention provides a use of sulfasalazine in the preparation of a drug for treating cholestatic liver disease. Summary of the invention

[0005] The technical problem to be solved by the present invention is to overcome the deficiencies in the prior art and provide an application of sulfasalazine in a drug for treating cholestatic liver disease.

[0006] The present invention first provides a new application of sulfasalazine, specifically, the application of sulfasalazine in treating liver diseases. The molecular formula of sulfasalazine is: C 18 H 14 N4O5S, molecular weight is 398.39, chemical structure is:

[0007]

[0008] Furthermore, the liver disease is cholestatic liver disease.

[0009] Furthermore, the sulfasalazine is combined with a pharmaceutical carrier to form a pharmaceutical preparation.

[0010] Furthermore, the pharmaceutical preparation includes: an oral solution.

[0011] Furthermore, the oral solution includes but is not limited to: oral solution, which is administered orally, and all dosage forms used are well known to ordinary technicians in the pharmaceutical field.

[0012] Furthermore, the administration route of the pharmaceutical preparation includes but is not limited to: oral administration.

[0013] Furthermore, the dosage of the pharmaceutical preparation is: 150 mg / kg.

[0014] Furthermore, the treatment plan is:

[0015] Sulfasalazine (Sigma, S0883) was dissolved in 0.5% sodium carboxymethylcellulose (CMC, Sigma, 419273) to prepare a suspension, which was orally administered once a day for 21 days.

[0016] The beneficial effects of the present invention are:

[0017] 1. The present invention has demonstrated for the first time at the animal level that sulfasalazine can effectively improve subacute cholestatic liver injury induced by intragastric administration of ANIT-corn oil suspension (80 mg / kg), providing a new drug regimen that has an improving effect on cholestatic liver disease. The regimen has the advantages of safety and non-toxicity, clear efficacy (obvious improvement effect), and broad application prospects.

[0018] 2. Pharmacological experiments were conducted using a cholestatic liver disease model. The results showed that sulfasalazine effectively alleviated the symptoms of liver damage caused by cholestasis, as shown by the decrease in the activities of ALT, AST, ALP, TBil, and TBA in rat serum, and the effect of reducing liver function biochemical indicators was better than that of the positive drug control. Liver tissue morphology HE and MASSON staining showed that sulfasalazine effectively improved the infiltration of liver inflammatory cells and the degree of portal fibrosis. BRIEF DESCRIPTION OF THE DRAWINGS

[0019] Figure 1 This is a graph showing the effects of sulfasalazine on biochemical indices of liver function in a rat model of cholestasis.

[0020] Figure 2 This is a diagram showing the effect of sulfasalazine on HE & MASSON in liver tissue of animal model rats. DETAILED DESCRIPTION

[0021] The present invention will be further described below in conjunction with specific embodiments. It should be understood that these embodiments are only used to illustrate the present invention and are not intended to limit the scope of the present invention. In addition, it should be understood that after reading the content recorded in the present invention, those skilled in the art can make various changes or modifications to the present invention, and these equivalent forms fall within the scope limited by the present invention equally.

[0022] The conventional reagents used in the following examples are all commercially available, and the biological experiments performed are all conventional biological experiments in the art and can be performed according to the instructions in the corresponding experimental manual or kit instructions.

[0023] Example 1

[0024] 1. Experimental materials: Clean-grade male SD rats, weighing about 100 g, were purchased from Vital River and raised under SPF conditions, with free access to food and water for 7 days to allow them to adapt to the environment.

[0025] 2. Animal modeling: After one week of adaptive feeding, the animals were gavaged with ANIT (α-naphthalene isothiocyanate)-corn oil suspension at a dose of 80 mg / kg to establish a subacute cholestasis model.

[0026] 3. Animal grouping: 40 rats were randomly divided into a normal control group (Contol), a model group (Model), an ursodeoxycholic acid group (UDCA, 100 mg / kg), and a sulfasalazine group (SASP, 150 mg / kg). The rats in the drug treatment groups were orally administered with the drug for 21 days.

[0027] 4. Administration method: ANIT-corn oil suspension was administered by gavage on the 1st, 7th, 14th and 21st days from the beginning of the experiment (the normal control group was given the same volume of corn oil), and the drug group was administered by gavage from the 2nd to the 22nd day from the beginning of the experiment (the normal control group and the model group were given the same volume of CMC).

[0028] 5. After the experiment, all experimental rats were killed and blood was taken from the heart for biochemical index detection. The rat chest skin was disinfected, and a needle was inserted at the strongest point of the apex beat to extract 5 mL of left ventricular arterial blood from the rat and serum was collected for serological testing (ALT, AST, ALP, TBili, ALB, TBA). Under sterile conditions, the skin was cut open to enter the abdominal cavity, the entire liver was quickly separated, and it was rinsed with saline. Part of the left lobe of the liver was cut out, the size was 5mm×5mm×2mm, fixed with paraformaldehyde, and routinely embedded in paraffin for sectioning for staining to observe the degree of liver tissue fibrosis.

[0029] 6. Experimental results:

[0030] Abnormalities of AST and ALT are commonly used indicators for evaluating liver function impairment. Figure 1The results showed that compared with the normal group, the serum AST and ALT levels of the model group rats were significantly increased, indicating liver cell damage. SASP and UDCA treatment can reduce the serum ALT and AST levels. Among them, SASP improves ALT and AST better than UDCA, indicating that SASP has an improving effect on the liver function of cholestatic rats and the effect is better than UDCA. Drug treatment can also reduce the abnormal increase of serum cholestasis indicators ALP, TBil, and TBA. The SASP group reduces ALP better than the UDCA group. Liver pathology results ( Figure 2 ) showed that the liver lobule structure was normal, the hepatocyte morphology was normal, no obvious bleeding, swelling, necrosis, and no obvious inflammatory cell infiltration were observed; in the model group, the liver lobule structure was destroyed, especially around the bile duct, with large areas of hepatocyte degeneration and necrosis, obvious collagen deposition around the portal area, accompanied by inflammatory cell infiltration and bile duct hyperplasia, and the hepatocyte degeneration area in the SASP group and UDCA group after intervention was reduced, necrosis decreased, and inflammatory cell infiltration alleviated.

[0031] Although the embodiments of the present invention have been shown and described above, it is understood that the above embodiments are exemplary and cannot be understood as limiting the present invention. A person skilled in the art may make some modifications and improvements based on the present invention. Therefore, modifications and improvements made without departing from the spirit of the present invention belong to the scope of protection claimed by the present invention.

Claims

1. Use of sulfasalazine in the preparation of a preparation for treating liver disease, wherein: The molecular formula of sulfasalazine is: 18 H 14 N4O5S, molecular weight is 398.39, chemical structure is:

2. The use according to claim 1, wherein: The liver disease is cholestatic liver disease.

3. The use according to claim 1, wherein: The sulfasalazine is combined with a pharmaceutical carrier to form a pharmaceutical preparation for treatment.

4. The use according to claim 3, wherein: The pharmaceutical preparation includes: oral solution.

5. The use according to claim 4, wherein: The oral solution includes: oral solution, which is administered orally.

6. The use according to claim 3, wherein: The administration route of the pharmaceutical preparation includes: oral administration.

7. The use according to claim 3, wherein: The dosage of the pharmaceutical preparation is: 150 mg / kg.

8. The use according to claim 3, wherein: The treatment regimen is: Sulfasalazine was dissolved in 0.5% sodium carboxymethylcellulose to prepare a suspension, which was orally administered once a day for 21 days.

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