CN110833556A

CN110833556A - Use of pyrazolopyrimidine derivatives for the treatment of hepatic fibrosis

Info

Publication number: CN110833556A
Application number: CN201810928311.2A
Authority: CN
Inventors: 李柏霖; 覃秋琳; 韦杰; 吴水华; 巫鑫; 陈丹
Original assignee: Guangxi Wuzhou Pharmaceutical Group Co Ltd
Current assignee: Guangxi Wuzhou Pharmaceutical Group Co Ltd
Priority date: 2018-08-15
Filing date: 2018-08-15
Publication date: 2020-02-25

Abstract

The invention relates to the technical field of medicines, in particular to application of pyrazolopyrimidine derivative in treatment of hepatic fibrosis. The liver fibrosis is related to liver tissue fibroplasia. The pyrazolopyrimidine derivative can obviously reduce the fibrosis degree of tissues, including reduction of the liver fibrosis level, reduction of tissue fibrosis proliferation and the like, and has a good treatment effect on hepatic fibrosis.

Description

Use of pyrazolopyrimidine derivatives for the treatment of hepatic fibrosis

Technical Field

The invention relates to the technical field of medicines, in particular to application of pyrazolopyrimidine derivative in treatment of hepatic fibrosis.

Background

Hepatic fibrosis is a reaction of liver to self-repair of various injuries, is a common pathological process for various chronic liver diseases to develop into cirrhosis, and is mainly characterized by unbalanced synthesis and degradation of extracellular matrix (ECM), resulting in massive deposition of EC in the perisinus space. Hepatic fibrosis is commonly seen in most chronic liver diseases with different etiological factors, if the etiological factors persist, hepatic fibrosis gradually worsens, causes reconstruction of hepatic lobules, blood vessels and the like, and pseudolobules and nodules are formed, liver cirrhosis is developed, and the health and the life of patients are seriously affected. Since various chronic liver diseases all cause hepatic fibrosis, and fibrosis is a necessary pathological process of serious fatal diseases such as liver cirrhosis, liver cancer and the like, the hepatic fibrosis resistance becomes a serious treatment of the chronic liver diseases.

The pyrazolopyrimidine derivative (formula I) is a small molecule inhibitor targeted on FLT3 kinase, is a novel compound independently developed by the applicant, and the patent of the compound is granted in China, the United states, Japan and other areas at present.

At present, only the pyrazolopyrimidine derivative is reported to treat acute myelogenous leukemia and psoriasis, and the pyrazolopyrimidine derivative has no application to the treatment of hepatic fibrosis.

Disclosure of Invention

In order to overcome the defects in the prior art, the invention provides the application of the pyrazolopyrimidine derivative in treating hepatic fibrosis.

The purpose of the invention can be realized by the following technical scheme:

application of pyrazolopyrimidine derivative shown as formula I or pharmaceutically acceptable salt and hydrate thereof in preparation of medicines for treating hepatic fibrosis

Preferably, the liver fibrosis is associated with fibroplasia of liver tissue.

Preferably, the pharmaceutically acceptable salts include hydrochloride, sulfate, mesylate, phosphate.

The invention also provides application of the pharmaceutical composition in preparing a medicament for treating hepatic fibrosis, which is characterized in that the pharmaceutical composition comprises pyrazolopyrimidine derivative shown as the formula I or pharmaceutically acceptable salt or hydrate thereof as an active ingredient and a pharmaceutically acceptable excipient.

Preferably, the pharmaceutical composition is an injection preparation, an oral preparation, an external preparation, a sustained release preparation, or a controlled release preparation.

Preferably, the oral preparation comprises tablets, granules and capsules.

Preferably, the pharmaceutical composition is a controlled release formulation, a sustained release formulation, an immediate release formulation.

Serum HA, LN, PCIII, IV-C four liver fibrosis terms are clinically ideal important serological indexes for diagnosing the disease development condition and the treatment effect of chronic liver disease patients and measuring the activity degree of inflammation and the fibrosis degree, the levels of the serum HA, LN, PCIII and IV-C gradually rise along with the inflammation and the liver fibrosis progression, and have positive correlation with the severity of the liver fibrosis.

The pyrazolopyrimidine derivative is originally an FLT3 inhibitor, and researches show that the pyrazolopyrimidine derivative can obviously reduce the fibrosis degree of tissues, including reduction of the level of four liver fibers, reduction of tissue fiber hyperplasia and the like, and has a good treatment effect on hepatic fibrosis.

Compared with the prior art, the method has the following advantages:

the pyrazolopyrimidine derivative provided by the invention is a novel medicament for treating hepatic fibrosis, not only expands the selectivity of the existing medicament for treating hepatic fibrosis and provides more medicament choices for the effective treatment of the hepatic fibrosis, but also further expands the application range of the pyrazolopyrimidine derivative as an FLT3 inhibitor.

Drawings

FIG. 1 is a histomorphological observation of rat liver tissue of a blank control group by HE staining.

FIG. 2 is a histomorphological observation of rat liver tissue by HE staining.

FIG. 3 is a histomorpheopathological observation of HE staining of rat liver tissue in the high dose group of pyrazolopyrimidine derivative.

FIG. 4 is histomorpheopathological observation of HE staining of liver tissue of rats in a low dose group of pyrazolopyrimidine derivative.

Detailed Description

The following examples are intended to illustrate the invention but are not intended to limit the scope of the invention.

Example 1 anti-rat immune liver fibrosis study

1. Experimental animals: male SD rats, clean grade, body mass (200. + -.20) g, purchased from Beijing Huafukang Biotech GmbH.

2. Experimental materials: pyrazolopyrimidine derivative was synthesized by the inventors. Pyrazolopyrimidine derivative was synthesized by the inventors. Mixing castor oil with ethanol 1: 1, passing through a sterile filter membrane of 0.22 mu m to obtain an ELE solution, weighing a certain mass of pyrazolopyrimidine derivative powder, dissolving the powder by using the ELE solution, adding a certain volume of sterilized water after the powder is completely dissolved, and uniformly mixing for later use. The volume ratio of the ELE solution to the sterile water is 1: 3. pig serum, Beijing Yanshengbo Biotech, Inc.; procollagen type III (PCIII), collagen type IV (CIV), Hyaluronic Acid (HA), Laminin (LN) radioimmunoassay kit, beijing north biotechnological institute.

3. Establishing a model: 60 rats were selected, 15 were left as blank control groups, and the remaining rats were operated as follows: each rat was given 0.5mL ip pig serum 2 times per week for the first 8 weeks, and 0.6mL ip0.6 times per week for the last 7 weeks, 2 times per week, for 15 weeks. Animals were sacrificed at random and pathological sections of liver tissue were HE stained to determine if the model replication was successful. All rats were given standard maintenance feed and drunk filtered tap water during the test period.

4. Animal grouping and administration: except for the blank control group, randomized into 3 groups: model group, pyrazolopyrimidine derivative high dose group (hereinafter referred to as high dose group), pyrazolopyrimidine derivative low dose group (hereinafter referred to as low dose group), and 15 of each group. After the molding is successful, 10mg/kg/d (1.0mg/mL liquid medicine) of the pyrazolopyrimidine derivative is administrated in a high-dose group by intragastric administration, and 5mg/kg/d (0.5mg/mL liquid medicine) of the pyrazolopyrimidine derivative is administrated in a low-dose group by intragastric administration; the model group and the blank control group are subjected to intragastric administration of 10mL/kg of mixed solution of an ELE solution and distilled water in a volume ratio of 1:3, 1 time per day and 4 weeks continuously.

5. Detecting the index

After a rat is anesthetized by 2% sodium pentobarbital ip, the abdominal cavity is immediately cut open, 6-9 mL of blood is quickly taken from the inferior vena cava, the rat is kept stand for 4 hours, and then the rat is centrifuged at 3000r/min for 10min, and the upper serum is taken. Detecting the HA, LN, PCIII and CIV levels of serum by a radioimmunoassay; taking part of liver right lobe tissue, placing in neutral formaldehyde solution for fixation, conventionally preparing liver tissue section, HE staining, and observing pathological change of liver tissue under a light microscope.

6. The statistical method comprises the following steps: the data were processed using SPSS17.0, the measured data were expressed as mean. + -. standard deviation, the comparisons between groups were analyzed using one-way anova, the comparisons between groups were examined using LSD, SNK, and P <0.05 was statistically significant for differences.

7. Results of the experiment

(1) Influence on serum hepatic fibrosis index of immune hepatic fibrosis rat

TABLE 1 Effect on immune hepatic fibrosis rat sera HA, LN, PCIII, CIV

Group of	HA/(ng·L^-1)	LN/(μg·L^-1)	PCIII/(μg·L^-1)	CIV/(μg·L^-1)
					Blank control group	40.28±3.18	53.68±2.37	42.16±10.28	10.43±1.53
Model set	71.59±4.22*	72.16±5.82*	75.64±14.31*	19.82±2.25*
					High dose group	46.94±3.51△	62.43±3.72△	45.93±7.65△	12.51±1.43△
Low dose group	62.08±3.74△	69.52±2.96	59.86±9.32△	17.36±1.64

P <0.05 compared to the blank control group and △ P <0.05 compared to the model group.

As shown in table 1, compared with the blank control group, the serum HA, LN, PCIII and CIV of the model group rats are significantly increased (P < 0.05); compared with the model group, the high-dose group HAs significantly reduced serum HA, LN, PCIII and CIV (P is less than 0.05); the low-dose group serum HA and PCIII are both reduced remarkably (P <0.05), and the effect on LN and CIV only shows a reduction trend. The results also show that the reduction degree of the pyrazolopyrimidine derivative on rat serum HA, LN, PCIII and CIV is in positive correlation with the dosage.

(2) Influence on pathological change of liver of immune liver fibrosis rat

As can be seen from FIGS. 1 and 2, the hepatic lobules of the blank control group have complete structure, the hepatic cells are arranged regularly, and no inflammatory cell infiltrates; the liver cells of the model group can be subjected to steatosis, balloon-like degeneration and spot necrosis, chronic inflammatory cell infiltration, liver cord arrangement disorder, fibrous connective tissue hyperplasia and liver lobule structure damage. As can be seen from fig. 3 and 4. The balloon-like degeneration, the steatosis, the inflammatory cell infiltration and the fibrous tissue hyperplasia of the liver cells of the high and low dose groups are lighter than those of the model group

Conclusion

The pyrazolopyrimidine derivative has a good treatment effect on hepatic fibrosis, and can obviously reduce the fibrosis degree of tissues, including reduction of the liver fibrosis level, reduction of tissue fibrosis proliferation and the like.

Although the invention has been described in detail hereinabove by way of general description, specific embodiments and experiments, it will be apparent to those skilled in the art that many modifications and improvements can be made thereto based on the invention. Accordingly, such modifications and improvements are intended to be within the scope of the invention as claimed.

Claims

1. Application of pyrazolopyrimidine derivative shown as formula I or pharmaceutically acceptable salt and hydrate thereof in preparation of medicines for treating hepatic fibrosis

2. Use according to claim 1, wherein the liver fibrosis is associated with fibroplasia of liver tissue.

3. Use according to claim 1, wherein the pharmaceutically acceptable salts comprise hydrochloride, sulfate, mesylate, phosphate.

4. Use of a pharmaceutical composition for preparing a medicament for treating liver fibrosis, wherein the pharmaceutical composition comprises the pyrazolopyrimidine derivative represented by formula i in claim 1, or a pharmaceutically acceptable salt or hydrate thereof, as an active ingredient, and a pharmaceutically acceptable excipient.

5. The use according to claim 4, wherein the pharmaceutical composition is an injection preparation, an oral preparation, an external preparation, a sustained-release preparation or a controlled-release preparation.

6. The use according to claim 5, wherein the oral formulation comprises tablets, granules, capsules.

7. The use according to claim 4, wherein the pharmaceutical composition is a controlled release formulation, a sustained release formulation, an immediate release formulation.