Application of rilpivirine in preparation of medicine for preventing and treating trichinosis

Abstract

The invention discloses an application of rilpivirine (RPV) in preparation of a medicine for preventing and treating trichinosis, and particularly discloses an application of rilpivirine (RPV) in preparation of a medicine for preventing and treating trichinosis. The invention belongs to the technical field of biological medicine. According to the present invention, the trichinellosis mouse model is constructed by using the oral feeding method, such that the trichinellosis mouse model is constructed, after rilpivirine is orally taken, the number of larva sacs in skeletal muscle tissues of the trichina-infected mouse model, histopathology, ultrastructure change and the like are observed. Results show that rilpivirine (as shown in the formula I) has a remarkable effect of killing trichina larva cysts and can improve the degree of muscle tissue fibrosis caused by trichinosis. The invention provides a new technical means for treating trichinosis, and rilpivirine has a wide application prospect in the aspect of treating fibrosis-related diseases caused by parasitic infection. # imgabs0 #

Description

Technical Field

[0001] The present invention relates to a new use of rilpivirine (4-[[(1R)-6,7-dimethoxy-2-methyl-3,4-dihydro-1H-isoquinolin-1-yl]methyl]-2-[[(1R)-1-[(4-hydroxyphenyl)methyl]-6-methoxy-2-methyl-3,4-dihydro-1H-isoquinolin-7-yl]oxy]phenol), and particularly relates to the application of rilpivirine in the preparation of drugs for treating trichinellosis. The present invention belongs to the technical field of biomedicine. Background Art

[0002] Trichinella spiralis (T. spiralis) is simply called trichina. Its adult worms and larvae can parasitize in the small intestine and skeletal muscle cells of the human body at the same time, resulting in trichinellosis. Trichinella has extremely low host selectivity and can cause infections in many mammals and even avian animals. It is a common foodborne zoonotic parasitic disease, seriously endangering human health and livestock production. When humans accidentally ingest meat containing trichina larvae cysts, the larvae escape in the small intestine, causing intestinal inflammation and gastrointestinal symptoms such as nausea and vomiting. The newly born larvae of trichina enter the bloodstream and migrate to the host skeletal muscle cells to form larval cysts. During this process, systemic symptoms such as fever, edema, muscle pain, and limited mobility occur. In severe cases, important organs such as the heart and central nervous system may be involved, even endangering life.

[0003] Studies have confirmed that after the newly born larvae of trichina invade muscle cells, due to the mechanical stimulation of the larvae and the chemical stimulation of their metabolites, the muscle fibers are damaged, and a large number of inflammatory cells infiltrate around. The affected muscle cells are covered by a layer of collagen derived from the host, forming a collagen cyst. The synthesis of collagen starts on the 10th day after infection, and the collagen cyst continues to thicken until the 26th day after infection. During this process, the fibrous connective tissue directly connected around the muscle cells proliferates, and finally a fibrous layer is formed outside the collagen cyst. As the pathological damage of the muscle cells continues to develop, the patient will correspondingly experience systemic muscle pain, obvious tenderness and pain on palpation, and even a paralyzed state.

[0004] The currently used drugs for treating trichinellosis are mainly albendazole and mebendazole, but both of them are teratogenic, so they are contraindicated for pregnant women and children under 2 years old. Pyrantel can be recommended for pregnant women and children under 2 years old with trichinellosis, but its efficacy is currently unclear. Therefore, developing more effective, harmless and convenient drugs for treating trichinellosis remains a problem to be solved at present.

[0005] Rilpivirine (4-[[(1R)-6,7-dimethoxy-2-methyl-3,4-dihydro-1H-isoquinolin-1-yl]methyl]-2-[[(1R)-1-[(4-hydroxyphenyl)methyl]-6-methoxy-2-methyl-3,4-dihydro-1H-isoquinolin-7-yl]oxy]phenol) is a non-nucleoside HIV reverse transcriptase inhibitor and is widely used in the clinical treatment of acquired immunodeficiency syndrome. Research shows that rilpivirine can reduce the degree of liver fibrosis caused by chronic liver injury by selectively regulating STAT1-mediated apoptosis of hepatic stellate cells. There are also studies showing that rilpivirine has pharmacological properties such as antiplatelet and antitumor effects. However, there has been no research report on its use in trichinellosis so far.

[0006] Through studying the effect of rilpivirine on mice infected with Trichinella spiralis, the present invention found that rilpivirine can effectively reduce the number of larval cysts in the skeletal muscle tissue of mice infected with Trichinella spiralis, relieve the pathological and ultrastructural damage of skeletal muscle tissue, and improve the degree of muscle tissue fibrosis. Therefore, it can be used as a new type of drug for preventing and treating trichinellosis for the prevention and treatment of trichinellosis. Summary of the Invention

[0007] The object of the present invention is to provide the application of rilpivirine (4-[[(1R)-6,7-dimethoxy-2-methyl-3,4-dihydro-1H-isoquinolin-1-yl]methyl]-2-[[(1R)-1-[(4-hydroxyphenyl)methyl]-6-methoxy-2-methyl-3,4-dihydro-1H-isoquinolin-7-yl]oxy]phenol) in the preparation of drugs for preventing and treating trichinellosis.

[0008] To achieve the above object, the present invention adopts the following technical means:

[0009] In this invention, Trichinella cysts (100±5) per mouse were orally administered to infect mice, thereby establishing a Trichinella - infected mouse model. On the 10th day after infection, the mice were given oral rilpivirine (5 mg / kg / d) for 14 consecutive days. On the 28th day after infection, mouse samples were collected. The results showed that rilpivirine could significantly reduce the number of larval cysts in the skeletal muscle tissues of Trichinella - infected mice, relieve the pathological and ultrastructural damage of skeletal muscle tissues, and improve the degree of muscle tissue fibrosis. That is, through research, this invention found that rilpivirine (RPV) has the effect of treating trichinosis.

[0010] Therefore, based on the above - mentioned research, this invention proposes the application of rilpivirine in the preparation of drugs for preventing and treating trichinosis. The chemical structural formula of the rilpivirine is shown in Formula I:

[0011]

[0012] Preferably, the rilpivirine achieves the purpose of treating trichinosis by reducing the number of larval cysts in the skeletal muscle tissues of Trichinella - infected mice, relieving the pathological and ultrastructural damage of skeletal muscle tissues, and improving the degree of muscle tissue fibrosis.

[0013] Compared with the prior art, the beneficial effects of this invention are as follows:

[0014] This invention proposes a new drug for treating trichinosis - rilpivirine, which can be used for the prevention and treatment of trichinosis. Rilpivirine is a non - nucleoside HIV reverse transcriptase inhibitor approved by the FDA and is clinically used for the treatment of AIDS at an oral dose of 25 mg once a day. Therefore, at the recommended dose, this drug is safe and suitable for long - term use. In accordance with the principle of species - based body weight / body surface area conversion, rilpivirine can achieve the purpose of treating trichinosis by reducing the number of larval cysts in the skeletal muscle tissues of Trichinella - infected mice, relieving the pathological and ultrastructural damage of skeletal muscle tissues, and improving the degree of muscle tissue fibrosis. Therefore, the proposal of this invention provides a new technical means for the prevention and treatment of trichinosis and will also give rilpivirine broad application prospects in the treatment of fibrosis - related diseases caused by parasitic infections. BRIEF DESCRIPTION OF THE DRAWINGS

[0015] Figure 1 It shows the infection situation and counting results of larval cysts in the diaphragm tissues of Trichinella - infected mice;

[0016] Among them, A: Mouse diaphragm tissue (unstained, 4×), TS is the control group infected with Trichinella spiralis, and RPV is the group treated with rilpivirine after Trichinella spiralis infection; B: Larval cyst count in the diaphragm tissue of Trichinella spiralis-infected mice (*P<0.05); C: Histopathological observation of the diaphragm tissue of Trichinella spiralis-infected mice, indicating that rilpivirine can reduce the number of larval cysts in Trichinella spiralis-infected mice;

[0017] Figure 2 It is the observation result of the fibrosis degree of the diaphragm tissue of Trichinella spiralis-infected mice;

[0018] Among them, A: Masson staining of the diaphragm tissue of mice in the control group infected with Trichinella spiralis; B: Masson staining of the diaphragm tissue of mice in the rilpivirine treatment group infected with Trichinella spiralis; C: Percentage of collagen area in the larval cysts of Trichinella spiralis (**P<0.01), indicating that rilpivirine can significantly relieve the fibrosis degree of muscle tissue caused by Trichinella spiralis infection;

[0019] Figure 3 It is the observation result of the ultrastructure of the diaphragm tissue of Trichinella spiralis-infected mice;

[0020] Among them, A, C, and E are the ultrastructures of the larval cysts in the control group infected with Trichinella spiralis; B, D, and F are the ultrastructures of the larval cysts in the rilpivirine treatment group, indicating that rilpivirine can reduce the hyperplasia of fibrous tissue outside the larval cysts, thereby hindering the development of larvae in the cysts and destroying the morphological structure of the larvae. Detailed implementation manners

[0021] The present invention will be further described below in conjunction with specific examples, and the advantages and features of the present invention will become clearer as the description progresses. However, these examples are merely exemplary and do not constitute any limitation to the scope of the present invention. Those skilled in the art should understand that modifications or substitutions can be made to the details and forms of the technical solutions of the present invention without departing from the spirit and scope of the present invention, but such modifications and substitutions all fall within the protection scope of the present invention.

[0022] Example 1 Application of rilpivirine in the treatment of trichinosis

[0023] 1. Animals and experimental reagents:

[0024] Mice (strain: Kunming, purchased from Liaoning Changsheng Biotechnology Co., Ltd.).

[0025] 2. Establishment of an animal model of trichinosis:

[0026] Twenty-four healthy male Kunming mice (weighing 22 - 25 g) were used. Among them, 8 mice were used as the negative control group (NC group), and the remaining 16 mice were infected with Trichinella cysts by oral gavage. The number of cysts inoculated per mouse was (100 ± 5). An experimental animal model of Trichinella infection in mice was established. The mice infected with Trichinella were randomly divided into two groups according to the principle of body weight balance, namely the positive control group (TS group): saline was given on the 10th day after infection, and it was orally administered by gavage for 14 consecutive days; the rilpivirine treatment group (RPV group): rilpivirine (5 mg / kg / day) was given on the 10th day after infection, and it was orally administered by gavage for 14 consecutive days. On the 28th day after Trichinella infection, the diaphragmatic muscle tissues of mice in each group were aseptically collected for various index detections.

[0027] 3. Results

[0028] 3.1 Rilpivirine can reduce the number of larval cysts in Trichinella-infected mice

[0029] Twenty-eight days after mice were infected with Trichinella, the diaphragmatic muscle tissues of mice in each group were aseptically collected and placed under a microscope for counting of larval cysts. The results are as Figure 1 shown in A and 1B. The results showed that the number of cysts in the diaphragmatic muscle of mice in the RPV group was significantly reduced; the diaphragmatic muscle tissues were stained with HE, and the results are as Figure 1 shown in C. The results showed that in the TS group, the Trichinella larval cysts had a complete morphology, a large number of inflammatory cells infiltrated around, the worm body had a complete morphology, and a large amount of fibrosis of the cysts formed. While in the RPV group, the pathological damage degree of Trichinella cysts was significantly reduced, the worm body morphology was incomplete, the cyst wall was thinned, and the fibrosis area was reduced. It shows that rilpivirine can significantly reduce the number of larval cysts in Trichinella-infected mice.

[0030] 3.2 Rilpivirine can significantly relieve the degree of muscle tissue fibrosis caused by Trichinella infection

[0031] To further evaluate the effect of rilpivirine on muscle tissue fibrosis in Trichinella-infected mice, Masson staining was used to detect the content of collagen fibers outside the cysts in the diaphragmatic muscle of mice (muscle tissue and Trichinella were stained orange-red, while collagen fibers were stained blue). The results are as Figure 2 shown. In the diaphragmatic muscle of mice in the TS group, a large amount of hyperplasia of collagen fibers outside the cysts was found ( Figure 2 A); in the RPV group, the cyst wall of the larval cysts was significantly thinned, the staining became lighter, and the area of collagen fibers was significantly reduced ( Figure 2 B); calculating the percentage of the area of collagen fibers outside the cysts, the results are as Figure 2 shown in C. The results showed that the degree of muscle tissue fibrosis in the RPV group was significantly reduced (P = 0.0022).

[0032] 3.3 Rilpivirine can damage the ultrastructure of Trichinella larval cysts and hinder the development of the worm body

[0033] To further observe the changes in host muscle cells, Trichinella larvae cysts, and the worms, the diaphragm tissue was fixed in 2.5% glutaraldehyde to prepare electron microscopy specimens, and its ultrastructure was observed under a transmission electron microscope. The results are as follows Figure 3 shown. The results showed that the cysts in the TS group mice were formed completely and had thick cyst walls, and a large amount of collagen fibers were formed around the cysts ( Figure 3 A); the cysts in the RPV group were incompletely formed and had thin cyst walls, and there was a fusion phenomenon in the larval structures of the cysts ( Figure 3 B). At the same magnification, only partial structures of the worms could be observed in the TS group, while a complete worm could be seen in the RPV group, indicating that the worms in the RPV group were significantly smaller than those in the TS group.

[0034] The observation results of the larval structures in the cysts are as shown in Figure 3 C-E. The results showed that there were obvious gaps in the cysts of the TS group mice, and the epidermis and subcutaneous muscles of the worms were arranged neatly ( Figure 3 C), and the mitochondrial structures inside the worms were complete and in good shape ( Figure 3 indicated by the red arrow in E); the mitochondria inside the worms in the RPV group were swollen and accompanied by obvious vacuolization ( Figure 3 D).

Claims

1. Use of Rilpivirine (RPV) in the preparation of a drug for the prevention and treatment of trichinosis, wherein the chemical structure of the Rilpivirine (4-[4-[4-[(E)-2-Cyanovinyl]-2,6-dimethylphenylamino]pyrimidin-2-ylamino]benzonitrile) is as shown in Formula I:

2. The use according to claim 1, characterized in that: The rilpivirine achieves the purpose of treating trichinellosis by reducing the number of larval cysts in the skeletal muscle tissue of mice with trichinellosis, alleviating the damage of skeletal muscle tissue pathology and ultrastructure, and improving the degree of muscle tissue fibrosis.

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