CN113398134A

CN113398134A - Application of insulin-like growth factor 1 receptor inhibitor BMS536924 in preparation of medicine for treating cystic echinococcosis

Info

Publication number: CN113398134A
Application number: CN202110770074.3A
Authority: CN
Inventors: 吕国栋; 林仁勇; 李锦田; 刘辉; 毕晓娟; 杨宁
Original assignee: First Affiliated Hospital of Xinjiang Medical University
Current assignee: First Affiliated Hospital of Xinjiang Medical University
Priority date: 2021-07-08
Filing date: 2021-07-08
Publication date: 2021-09-17

Abstract

The invention relates to the technical field of echinococcosis medicaments, in particular to application of an insulin-like growth factor 1 receptor inhibitor BMS536924 in preparing a medicament for treating cystic echinococcosis; the invention discloses the application of BMS536924 in preparing a medicament for treating cystic echinococcosis for the first time; the in vivo and in vitro experimental data show that BMS536924 is a high-efficiency anti-echinococcosis drug molecule; the traditional Chinese medicine composition has obvious treatment effect on cystic echinococcosis, the effect is superior to albendazole sulfoxide serving as an albendazole metabolite, and the combined drug effect of the BMS536924 and the ABZSO drugs is higher than that of the BMS536924 and the ABZSO drugs which are independently administered; meanwhile, the dosage of the combined albendazole of BMS536924 is reduced by 1/4-1/2 compared with the dosage of the albendazole sulfoxide alone, so that the dosage and the drug resistance problem are greatly reduced, and a good treatment effect is achieved.

Description

Application of insulin-like growth factor 1 receptor inhibitor BMS536924 in preparation of medicine for treating cystic echinococcosis

Technical Field

The invention relates to the technical field of echinococcosis drugs, and relates to application of an insulin-like growth factor 1 receptor inhibitor BMS536924 in preparation of a drug for treating cystic echinococcosis.

Background

Cystic Echinococcosis (CE) is caused by Echinococcus granulosus (Echinococcus granulosus)Echinococcus granulosus, Eg) The serious zoonosis parasitic disease caused by parasitism on human bodies belongs to the world neglected disease, the disease is reported to be an important public health problem again by the world health organization in 2010, and the prevention and the development of a novel treatment approach are required to be strengthened. The initial infection of cystic echinococcosis in the intermediate host is achieved by orally taking infectious eggs, which pass through the intestinal wall after incubating the larvae in the stomach of the intermediate host, and mainly parasitize the liver organs. Due to the difficulties in controlling the infection source, the lack of efficient means for preventing and controlling the hydatid disease and the like, the data of the circulation in the last 5 years shows that seven provinces (Sinkiang, Qinghai, Ningxia, Gansu, Tibet, Sichuan and inner Mongolia) in the western China are still high circulation areas or extra-high circulation areas, the prevalence rate of the hydatid disease of the crowd is 0.05-0.63%, and the prevalence rate of part of the areas is as high as 8.93-12.38%, so that the life safety of people and livestock husbandry is seriously harmed, and huge economic loss is caused.

At present, the method for treating echinococcosis mainly comprises surgical treatment and auxiliary treatment by an anti-echinococcosis medicament. Some patients are seriously injured due to factors such as parasitic positions and sizes of the hydatid, complete excision of the external bursa cannot be realized, the postoperative recurrence rate is high, the operation treatment cost is high, and the drug therapy becomes the best treatment means. The research of anti-tumor, anti-infection and Chinese herbal medicine extract and other medicines for treating echinococcosis has made certain progress, but still more effective and safe medicines for clinical treatment are needed. It is composed ofMi nitazoxanide (D)Nitazoxanide，NTZ) The anti-parasite spectrum is wide, and research reports show that the anti-parasite compound has the effect of killing echinococcus granulosus in vitro, and the effect is obvious, but the related reports in vivo are few. Benzimidazole medicines (such as ABZ and the like) recommended by the world health organization for treating echinococcosis have the problems of difficult dissolution, low bioavailability, side effects caused by long-term taking and the like, and the medicines for efficiently treating echinococcosis are still lacked so far, so that a new medicine target and a new medicine molecule are searched, and the method has important significance for preventing and treating echinococcosis.

In the field of the study of parasites,Hemer Setc. find andE. granulosusthe relationship between parents and sources is very closeE. multilocularisIn the field of diabetes, an insulin receptor signaling pathway exists for regulating glucose uptake and transport, and the pathway is proved to be an important anti-echinococcosis drug target. Researches find that the insulin receptor signal path has an important role in the interaction between parasites and hosts, and the downstream signal path is activated to regulate the glucose metabolism of organisms by sensing the insulin signal molecules of the hosts in the parasitic environment so as to be beneficial to successful parasitism.

IGF-1R is the main membrane receptor mediating the physiological or glucose uptake function of IGF-1, has high homology with IR, has structural similarity as high as 84%, and can activate AKT in tumor cells, further activate PI3K/AKT signal pathway, and regulate cell growth, differentiation, apoptosis, tumor angiogenesis and metastasis. BMS536924 is a novel small molecule inhibitor targeting IGF-1R, and can significantly reduce the volume of tumor in tumor research. However, BMS536924 (insulin-like growth factor 1 receptor inhibitor) has not been reported as a drug for treating cystic echinococcosis at present.

Disclosure of Invention

The invention provides an application of an insulin-like growth factor 1 receptor inhibitor BMS536924 in preparation of a medicament for treating cystic echinococcosis, overcomes the defects of the prior art, and can supplement and solve the problem that no related report that the insulin-like growth factor 1 receptor inhibitor BMS536924 is used as a medicament for treating the cystic echinococcosis exists at present; meanwhile, the benzimidazole medicine has the problems of poor intestinal absorption, long-term administration and easy generation of drug resistance, so that the treatment effect is unsatisfactory.

The technical scheme of the invention is realized by the following measures: an application of insulin-like growth factor 1 receptor inhibitor BMS536924 in preparing the medicines for treating cystic echinococcosis is disclosed.

The following is further optimization or/and improvement of the technical scheme of the invention:

the above further comprises albendazole, insulin-like growth factor 1 receptor inhibitor BMS536924 and albendazole in combination.

The mass ratio of the insulin-like growth factor 1 receptor inhibitor BMS536924 to albendazole is 1-2: 1.

The cystic echinococcosis is a disease caused by echinococcus granulosus infection.

The Echinococcus granulosus is selected from the group consisting of: echinococcus granulosus larvae, Echinococcus granulosus embryonic cells, Echinococcus granulosus germinal layer, or a combination thereof.

The invention discloses the application of an insulin-like growth factor 1 receptor inhibitor BMS536924 in preparing a medicament for treating cystic echinococcosis for the first time; the survival rate detection of the in vitro metacercaria and the vesicle and the pharmacodynamics experiment data in the body of the mouse indicate that the insulin-like growth factor 1 receptor inhibitor BMS536924 (BMS 536924) is a high-efficiency anti-echinococcosis drug molecule. In vitro experiment results show that the BMS536924 can inhibit the growth and survival of the metacercaria and destroy the ultrastructure of the metacercaria, the activity of the vesicle is inhibited after in vitro administration, germinal cells in a germinal layer of the vesicle are reduced, and the BMS536924 and the ABZSO medicament have a synergistic effect when being used together, the medicinal effect is higher than that of the BMS536924 and the ABZSO medicament which are independently administered, the survival rate of the vesicle is obviously reduced, and the ultrastructure of the vesicle is destroyed; meanwhile, the dosage of the combined albendazole sulfoxide of the insulin-like growth factor 1 receptor inhibitor BMS536924 is reduced by 1/4 to 1/2 compared with the dosage of the albendazole sulfoxide alone, so that the dosage and the drug resistance problem are greatly reduced, and a good treatment effect is achieved.

Drawings

FIG. 1 is a diagram showing the survival rate of Echinococcus granulosus metacercaria in vitro intervention of BMS 536924.

FIG. 2 is a scanning electron micrograph of Echinococcus granulosus metacercaria in the DMSO group.

FIG. 3 is a scanning electron micrograph of Echinococcus granulosus metacercaria at 15. mu. mol/L ABZSO group.

FIG. 4 is a scanning electron micrograph of 100. mu. mol/L BMS536924 group Echinococcus granulosus metacercaria.

FIG. 5 is a scanning electron micrograph of 50. mu. mol/L BMS536924 group Echinococcus granulosus metacercaria.

FIG. 6 is a scanning electron micrograph of 25. mu. mol/L BMS536924 group Echinococcus granulosus metacercaria.

FIG. 7 is a graph of the survival rate of Echinococcus granulosus vesicles in vitro with combination of BMS536924, ABZSO, BMS536924 and ABZSO.

FIG. 8 is a scanning electron micrograph of Echinococcus granulosus vesicles from the set of DMSO groups.

FIG. 9 is a scanning electron micrograph of Echinococcus granulosus vesicles paired with 15. mu. mol/L ABZSO.

FIG. 10 is a scanning electron micrograph of 100. mu. mol/L BMS536924 group intervening Echinococcus granulosus vesicles.

FIG. 11 is a scanning electron micrograph of a 50. mu. mol/L BMS536924 group of intervening Echinococcus granulosus vesicles.

FIG. 12 is a scanning electron micrograph of 25. mu. mol/L BMS536924 group intervening Echinococcus granulosus vesicles.

FIG. 13 is a projection electron micrograph of Echinococcus granulosus vesicles from the solvent set.

FIG. 1450 mg/kg ABZ set projection electron micrograph of Echinococcus granulosus vesicles.

FIG. 15 is a projection electron micrograph of Echinococcus granulosus vesicles prepared from BMS536924 high dose set.

FIG. 16 is a projection electron micrograph of Echinococcus granulosus vesicles prepared from the dose set of BMS 536924.

FIG. 17 is a projection electron micrograph of Echinococcus granulosus vesicles prepared from BMS536924 low dose set.

Detailed Description

The present invention is not limited by the following examples, and specific embodiments may be determined according to the technical solutions and practical situations of the present invention. The various chemical reagents and chemical articles mentioned in the invention are all the chemical reagents and chemical articles which are well known and commonly used in the prior art, unless otherwise specified; the percentages in the invention are mass percentages unless otherwise specified; the solution in the present invention is an aqueous solution of water as a solvent, for example, a hydrochloric acid solution is an aqueous hydrochloric acid solution, unless otherwise specified.

Example 1, the use of insulin-like growth factor 1 receptor inhibitor BMS536924 for the preparation of a medicament for the treatment of cystic echinococcosis.

Example 2, which is an optimization of the above examples, further comprises albendazole, insulin-like growth factor 1 receptor inhibitor BMS536924 and albendazole in combination.

Example 3, optimized for the above example, the insulin-like growth factor 1 receptor inhibitor BMS536924 and albendazole are in a mass ratio of 1 to 2: 1. Specifically, the mass ratio of the insulin-like growth factor 1 receptor inhibitor BMS536924 to albendazole sulfoxide in the in vitro test of Echinococcus granulosus metacercaria and vesicles is 1-8: 1, and the mass ratio of the insulin-like growth factor 1 receptor inhibitor BMS536924 to albendazole in the in vivo test of a mouse is 1-2: 1.

Example 4, optimized for the above example, cystic echinococcosis is a disease caused by echinococcus granulosus infection.

Example 5 as an optimization of the above examples, echinococcus granulosus is selected from the group consisting of: echinococcus granulosus larvae, Echinococcus granulosus embryonic cells, Echinococcus granulosus germinal layer, or a combination thereof.

The application of the insulin-like growth factor 1 receptor inhibitor BMS536924 in preparing the medicine for treating the cystic echinococcosis is tested as follows:

in vitro experiment of insulin-like growth factor 1 receptor inhibitor BMS536924 (BMS 536924 for short)

1.1 BMS536924 experiment for in vitro intervention of Procoenuruses

1.1.1 Experimental purposes: explore the survival rate of the Echinococcus granulosus prototheca in vitro intervention at each concentration with the experimental period of 4 days

1.1.2 Experimental groups (overview)

The experiment is divided into 9 groups, namely a BMS536924 intervention group 1, a BMS536924 intervention group 2, a BMS536924 intervention group 3, a BMS536924 intervention group 4, a BMS536924 intervention group 5, a BMS536924 intervention group 6, an ABZSO group, a Negative group (Negative Control) and a DMSO group; each group is provided with 3 multiple holes; the experimental period was set for 4 days.

1.1.3 mother liquor preparation

Weighing 4.7996mg BMS536924, dissolving in 1mL DMSO, and uniformly mixing to prepare (10 mM) mother solution I; taking 500 mu L of mother liquor, adding 500 mu L of DMSO to obtain mother liquor II; taking 500 mu L of mother liquor, adding 500 mu L of DMSO to obtain mother liquor; taking 500 mu L of mother liquor, adding 500 mu L of DMSO to obtain mother liquor; taking 500 microliter of mother liquor, adding 500 microliter of DMSO to obtain mother liquor; taking 500 microliter of mother liquor, adding 500 microliter of DMSO to obtain mother liquor; 3.97995mg of albendazole sulfoxide is weighed and dissolved in 1mL of DMSO, and the albendazole sulfoxide suspension mother solution (the concentration is 15 mM) is prepared by mixing uniformly.

1.1.4 preparation of Echinococcus granulosus culture solution

Adding fetal bovine serum and double antibodies into an RPMI1640 culture medium, and uniformly mixing to obtain an echinococcus granulosus culture solution, wherein the volume percentage of fetal bovine serum in the echinococcus granulosus culture solution is 25%, and the volume percentage of the double antibodies in the echinococcus granulosus culture solution is 1%.

1.1.5 obtaining Echinococcus granulosus metacercaria

Fresh goat liver infected with echinococcus granulosus was purchased from the Hualing slaughterhouse in Wuxyloji, Xinjiang, cyst fluid was extracted using a 50mL syringe under sterile conditions, the cyst wall was cut open with scissors and then the oocysts were peeled off with forceps and placed in a sterile petri dish, washing the cut capsule wall with normal saline, filtering with a screen to remove impurities, collecting metacercaria until the metacercaria naturally settles to the bottom of a centrifuge tube, washing with normal saline until the solution is clarified, discarding the supernatant, adding pepsin with the pH value adjusted to 2.0 by 1% HCI and the pH value adjusted to 50mL and 0.1%, digesting the supernatant in a constant-temperature water tank at 37 ℃ for 30min, washing the protocercaria for 3 to 5 times by PBS containing 1% penicillin and streptomycin (double antibody), at least 5min each time, filtering the protocercaria by using a 200 mu m screen, removing impurities, uniformly mixing the washed protocercaria, dyeing the protocercaria by using a 1% eosin solution for 10s, and then using a counter to count the total number of the protocercaria and die the protocercaria under an optical microscope.Counting the number of the metacercaria, adding a part of metacercaria with the survival rate of more than 95 percent into a culture medium (1640 culture medium, double antibody and fetal calf serum) to culture in an incubator at 37 ℃, freezing the other part of metacercaria to a refrigerator at-80 ℃ for subsequent extraction of metacercaria DNA, and performing PCR amplification and sequence sequencing on metacercaria used in the research, wherein the species of metacercaria used in the research isE. granulosus s.s.。

1.1.6 configurations of BMS536924 intervention group 1, BMS536924 intervention group 2, BMS536924 intervention group 3, BMS536924 intervention group 4, BMS536924 intervention group 5, BMS536924 intervention group 6, ABZSO group, negative group and DMSO group

The echinococcus granulosus protocoenurus is prepared in groups: mu.L of mother solution (i) is added into 198 mu.L of echinococcus granulosus culture solution and mixed evenly to obtain BMS536924 dry preparation group 1 (100 mu mol/L BMS 536924); adding 2 mu L of mother solution (namely BMS 536924) into 198 mu L of Echinococcus granulosus culture solution, and uniformly mixing to obtain BMS536924 dry pre-group 2 (50 mu mol/L BMS 536924); adding 2 mu L of mother solution to 198 mu L of Echinococcus granulosus culture solution, and mixing to obtain BMS536924 dry pre-group 3 (25 mu mol/L BMS 536924); adding 2 mu L of mother liquor (4) into 198 mu L of Echinococcus granulosus culture solution, and mixing to obtain BMS536924 dry pre-group 4 (12.5 mu mol/L BMS 536924); adding 2 muL of mother liquor into 198 muL of Echinococcus granulosus culture solution, and mixing to obtain BMS536924 dry pre-group 5 (6.25 mumol/L BMS 536924); adding 2 mu L of mother solution to 198 mu L of echinococcus granulosus culture solution, and mixing to obtain BMS536924 dry pre-group 6 (3.125 mu mol/L BMS 536924); adding 2 mu L (concentration is 1.5 mM) albendazole sulfoxide suspension into 198 mu L echinococcus granulosus culture solution, and mixing uniformly to obtain albendazole sulfoxide group (ABZSO group) (15 mu mol/L ABZSO); obtaining a negative group by taking 200 mu L of echinococcus granulosus culture solution; mu.L of Echinococcus granulosus culture solution was added to 2. mu.L of DMSO and mixed to obtain a DMSO group.

1.1.7 Experimental procedures

Taking the metacercaria cultured in vitro adaptively for 2 days, paving the metacercaria in a 96-well plate according to the number of 250-300 per well, and arranging 3 multiple wells in each group. Then, BMS536924 dry preparation group 1 (100. mu. mol/L BMS 536924), BMS536924 dry preparation group 2 (50. mu. mol/L BMS 536924), BMS536924 dry preparation group 3 (25. mu. mol/L BMS 536924), BMS536924 dry preparation group 4 (12.5. mu. mol/L BMS 536924), BMS536924 dry preparation group 5 (6.25. mu. mol/L BMS 536924), BMS536924 dry preparation group 6 (3.125. mu. mol/L BMS 536924), ABZSO group (15. mu. mol/L ABZSO), negative group and DMSO group were added and mixed well.

The in vitro intervention survival rate of Echinococcus granulosus metacercaria after day 4 is shown in the figure 1; the scanning electron micrograph of Echinococcus granulosus metacercaria by DMSO group after day 4 is shown in FIG. 2; the scanning electron micrograph of the ABZSO group on Echinococcus granulosus metacercaria after day 4 is shown in FIG. 3; the scanning electron micrograph of Echinococcus granulosus metacercaria in BMS536924 dried cohort 1 after day 4 is shown in FIG. 4; the scanning electron micrograph of Echinococcus granulosus metacercaria in BMS536924 dried cohort 2 after day 4 is shown in FIG. 5; the scanning electron micrograph of Echinococcus granulosus metacercaria in BMS536924 stem control group 3 after day 4 is shown in FIG. 6; it can be seen from fig. 1 to fig. 6 that after the in vitro intervention for 4 days, the abazso group metacercaria cephalic hooks partially disappear, the sucking discs are invaginated, calcium particles on the worm body disappear, and germinal cells in the vesicle germinal layer are reduced; the protozoon and the vesicle in the BMS536924 intervention group are all dead, the electron microscope result shows that the BMS536924 intervenes the shrinkage of the parasite body and damages the parasite body, the germinal cells in the vesicle germinal layer are obviously reduced, the germinal cell number in the vesicle germinal layer is in positive correlation with the BMS536924 administration concentration, which shows that the insulin-like growth factor 1 receptor inhibitor BMS536924 is a good external pesticide for echinococcus granulosus, and the effect is better than that of albendazole sulfoxide.

In vitro experiment of insulin-like growth factor 1 receptor inhibitor BMS536924 (BMS 536924 for short) combined with albendazole sulfoxide

1.1 BMS536924 Albendazole sulfoxide Combined in vitro experiment

1.1.1 Experimental purposes: researches the survival rate of the vesicle in vitro intervention by combining BMS536924 and albendazole sulfoxide

1.1.2 experimental groups (overview):

the experiment is divided into 8 groups, namely a BMS536924 dry preparation group (25 mu mol/L BMS536924, 50 mu mol/L BMS536924, 100 mu mol/L BMS 536924), an albendazole sulfoxide group (15 mu mol/L ABZSO), a BMS536924 combined albendazole sulfoxide group (25 mu mol/L BMS536924+15 mu mol/L ABZSO, 50 mu mol/L BMS536924+15 mu mol/L ABZSO, 100 mu mol/L BMS536924+15 mu mol/L ABZSO) and a solvent group (DMSO); the metacercaria is paved in a 96-well plate, each group is provided with 3 multiple wells, and the experimental period is set for 4 days.

1.1.3 BMS536924 mother liquor preparation

Accurately weighing 3.97995mg of albendazole sulfoxide, dissolving in 1mL of DMSO, and uniformly mixing to prepare (the concentration is 15 mM) albendazole sulfoxide suspension; 4.7996mg of BMS536924 were dissolved in 1mL of DMSO and mixed well to prepare a 10mM BMS536924 suspension.

1.1.4 preparation of in vitro vesicle culture solution

Prepare 600 mL vesicle culture medium, need 54 mL 5% yeast, 8.4 mL 30% glucose, 150 mL fetal bovine serum, 6 mL double antibody, 390 mL 1640 medium mixed well, placed at 4 ℃.

1.1.5 Echinococcus granulosus vesicle acquisition

Injecting 2000 heads and 100 mu L of fresh metacercaria into each mouse, feeding the mice in an animal feeding center of Xinjiang medical university, after 9 months of modeling, removing necks of the mice to kill the mice, dissecting and taking out vesicles and putting the vesicles into a culture dish containing normal saline, stripping tissues of the mice, cleaning the tissues of the mice by using the normal saline for 3 to 5 times, putting the culture dish into an ultra-clean bench to clean the tissues by using 0.9 percent of normal saline added with 1 percent of double antibody, discarding the vesicles with collapse and incomplete shape, and putting the vesicles with better activity into a culture bottle to be added with a mixed culture medium to be cultured in a cell culture box at 37 ℃.

1.1.6 different concentration BMS536924 dosing groups: 60 μ L (concentration: 0.25 mM, 0.5 mM, 1 mM) of BMS536924 suspension was added to 5.94 mL of vesicle culture medium, and mixed to obtain 25 μmol/L, 50 μmol/L, 100 μmol/L BMS536924 dry pre-groups; BMS536924 in combination with albendazole sulfoxide administration group: 60 mu L (with the concentration of 0.25 mM, 0.5 mM and 1 mM) of BMS536924 suspension is respectively added into 5.94 mL of vesicle culture solution, 60 mu L of 0.15 mM albendazole sulfoxide suspension is added into each well, and the mixture is mixed uniformly to obtain a combined administration group (25 mu mol/L BMS536924+15 mu mol/L ABZSO, 50 mu mol/L BMS536924+15 mu mol/L ZABO and 100 mu mol/L BMS536924+15 mu mol/L ABZSO); the DMSO groups were obtained by adding 60. mu.L of DMSO to 5.94 mL of vesicle culture medium, and mixing the resulting mixture.

1.1.7 Experimental procedures

Placing vesicles in 6-well plate, incubating, placing 15-20 vesicles in each well, adding 8 mL vesicle culture solution, mixing, and placing in 37 deg.C CO₂Culturing in a constant temperature incubator. After 4 days of administration, the culture solution is discarded, PBS is used for washing for three times, vesicles in each group are collected, 0.4% trypan blue is used for dyeing for 3 min, the PBS is used for washing away the dyeing solution, the dyeing condition of the vesicles is observed under an optical microscope, the number and the total number of dead vesicles are counted by taking vesicle collapse as a standard, the survival rate of the vesicles is calculated, and statistical analysis is carried out on the data.

The 2-day survival rate of the BMS536924, BMS536924 and ABZSO in vitro vesicle after combined administration is shown in figure 7, and the survival rate results of the vesicles after administration in figure 7 show that the combination of BMS536924, ABZSO and BMS536924 with ABZSO has different degrees of inhibition effects on vesicle growth, and the growth inhibition effect of the combination of BMS536924 and ABZSO on the vesicles is obviously better than that of the combination of BMS536924 and ABZSO when independently administered; after 4 days, a scanning electron microscope image of the Echinococcus granulosus vesicle by the DMSO group is shown in FIG. 8, and germinal cells on the inner surface of the vesicle are complete in shape and large in number; the scanning electron micrograph of the ABZSO group on the echinococcus granulosus vesicles after the 4 th day is shown in FIG. 9, and the germinal cell morphology is collapsed and the number is small; after day 4, the scanning electron micrograph of the echinococcus granulosus vesicle in BMS536924 stem group 1 is shown in fig. 10, and the number of germinal cells is significantly reduced compared with the solvent group; the scanning electron micrograph of the Echinococcus granulosus vesicles in BMS536924 stem control group 2 after day 4 is shown in FIG. 11, the germinal cell morphology is damaged to some extent, and the number of germinal cells is reduced compared with the solvent group; the scanning electron micrograph of the echinococcus granulosus vesicle in BMS536924 stem group 3 after day 4 is shown in fig. 12, and the destruction degree of the germinal cell morphological structure is weak. As can be seen from fig. 7 to 12, BMS536924, ABZSO and BMS536924 in combination with ABZSO exhibit different degrees of inhibition on vesicle growth, and the vesicle growth inhibition effect of the combination of BMS536924 and ABZSO is significantly better than that of BMS536924 alone and ABZSO alone, which indicates that the combination of insulin-like growth factor 1 receptor inhibitor BMS536924 and albendazole sulfoxide has better synergistic therapeutic effect. It was also found through experimental results that the survival rate of vesicles significantly decreased with the increase of the concentration of BMS536924 drug.

In vivo experiment of insulin-like growth factor 1 receptor inhibitor BMS536924 (BMS 536924 for short)

1. Pharmacodynamic study of BMS536924 on treatment of cystic echinococcosis in mice

1.1 purpose of experiment: obtain pharmacodynamic data of different dosages of BMS536924 after the treatment of cystic echinococcosis by the gastric lavage administration mode.

1.2 contents of the experiment

1.2.1 establishing a cystic echinococcosis mouse animal model:

(1) c57 mice (6 weeks old, 18g to 22 g) were acclimatized for one week, and then a cystic echinococcosis mouse model was established by intraperitoneal injection. Uniformly mixing clean original coenurses, sucking out 1-1.5 mL centrifuge tubes by using a pipette gun, adding 10 mu L of 1% eosin solution, dyeing for 10s, taking out 10 mu L of solution, dripping the 10 mu L of solution onto a glass slide, calculating the original coenurses with the survival rate of more than 95% by using a counter under an optical microscope, using the original coenurses for abdominal injection molding, injecting 200 mu L of physiological saline into each mouse, wherein the original coenurses contain 2000 original coenurses, and injecting 200 mu L of physiological saline into a blank control group.

(2) Molding time: the molding period of the cystic echinococcosis mouse animal model is 9 months.

(3) And (3) judging the success of molding: after 6 months of inoculation, the mice are subjected to ultrasonic B-ultrasonic detection molding, and the molding success is determined when the focus with the diameter exceeding about 0.5cm exists.

② mouse model: the intraperitoneal injection is divided into a solvent group, a positive drug group (ABZ group, namely albendazole group) and a treatment group (100 mg/kg BMS536924, 70 mg/kg BMS536924 and 35 mg/kg BMS 536924), and the total number of the groups is 5, and 5 to 8 groups are provided.

1.2.2 BMS536924 treatment of cystic echinococcosis mice:

(1) and selecting a mouse cystic echinococcosis animal model which is successfully modeled, and performing pharmacodynamic experiments.

(2) The experimental groups were 100 mg/kg BMS536924, 70 mg/kg BMS536924, 35 mg/kg BMS536924, positive drug group (ABZ group is albendazole group), solvent group (95% PEG400 and 5% DMSO mixed solution), oral gavage administration was adopted, administration volume was 0.1ml/10g, 1 time per day, administration was 30 days.

(3) The drugs were formulated at each concentration according to the test schedule: (same group is given according to different dosages with same concentration)

BMS536924 high dose group: 300 mg of BMS536924 yellow powder is precisely weighed and dissolved in 30 ml of 95% PEG400 and 5% DMSO mixed solution, and the dosage is 100 mg/kg.

Dose group in BMS 536924: 210 mg of BMS536924 yellow powder is precisely weighed and dissolved in 30 ml of 95% PEG400 and 5% DMSO mixed solution, and the dosage is 70 mg/kg.

BMS536924 low dose group: 105 mg of BMS536924 yellow powder is precisely weighed and dissolved in 30 ml of 95% PEG400 and 5% DMSO mixed solution, and the dosage is 35 mg/kg.

Positive drug group: 150 mg of ABZ is precisely weighed and dissolved in 30 ml of mixed solution of 95 percent PEG400 and 5 percent DMSO, thus obtaining the dosage of 50 mg/kg.

Solvent group: 95% PEG400 and 5% DMSO mixed solution

Storing the above 5 medicinal liquids in a refrigerator at 4 deg.C.

(4) After 30 days of intragastric administration, the cystic echinococcosis mice were killed by cervical dislocation, 2 vesicles in abdominal cavities of the mice were randomly selected from each experimental group and fixed in an electron microscope liquid, and the inner ultrastructure of the vesicles was observed by using a transmission electron microscope. The electron microscope image of the solvent group vesicles is shown in fig. 13, the electron microscope image of the ABZ group vesicles is shown in fig. 14, the electron microscope image of the BMS536924 high dose group vesicles is shown in fig. 15, the electron microscope image of the BMS536924 medium dose group vesicles is shown in fig. 16, and the electron microscope image of the BMS536924 low dose group vesicles is shown in fig. 17.

As can be seen from fig. 13 to 17, the vesicle grows layer structure in the solvent group is compact, and orderly arranged microvilli and nuclei with complete morphology can be clearly observed; the nuclear structure was destroyed in the 50 mg/kg ABZ group; the microvilli in the 100 mg/kg BMS536924 vesicle hair-growing layer is reduced, the nucleus with loose structure expands abnormally, the glycogen is reduced, and a large amount of vacuole structures appear; microvilli in 70 mg/kg BMS536924 vesicles become short, cell structures in a germinal layer are damaged, cell nuclei disappear, and lipid droplets appear in the germinal layer; glycogen accumulation and a large number of lipid droplets appear in the 35 mg/kg BMS536924 group vesicle hair-growing layer; the structural morphology of cells in the hair growth layer of the solvent group vesicle can be observed to be complete in a scanning electron microscope, the number of cells in the hair growth layer inside the vesicle in the 50 mg/kg ABZ group is reduced, and the number of hair growth cells in the hair growth layer of the vesicle is obviously reduced along with the increase of the drug dosage in the BMS536924 administration group. The insulin-like growth factor 1 receptor inhibitor BMS536924 can be used for treating mouse cystic echinococcosis well by oral administration and gastric lavage, and the effect is better than that of albendazole.

In conclusion, the invention discloses the application of an insulin-like growth factor 1 receptor inhibitor BMS536924 in preparing a medicament for treating the cystic echinococcosis for the first time; the survival rate detection of the in vitro metacercaria and the vesicle and the pharmacodynamics experiment data in the body of the mouse indicate that the insulin-like growth factor 1 receptor inhibitor BMS536924 (BMS 536924 for short) is a high-efficiency anti-echinococcosis drug molecule. In vitro experiment results show that the BMS536924 can inhibit the growth and survival of the metacercaria and destroy the ultrastructure of the metacercaria, the activity of the vesicle is inhibited after in vitro administration, germinal cells in a germinal layer of the vesicle are reduced, and the BMS536924 and the ABZSO medicament have a synergistic effect when being used together, the medicinal effect is higher than that of the BMS536924 and the ABZSO medicament which are independently administered, the survival rate of the vesicle is obviously reduced, and the ultrastructure of the vesicle is destroyed; meanwhile, the dosage of the combined albendazole serving as an insulin-like growth factor 1 receptor inhibitor BMS536924 is reduced by 1/4-1/2 compared with the dosage of the albendazole sulfoxide alone, so that the dosage and the drug resistance problem are greatly reduced, and a good treatment effect is achieved.

The technical characteristics form an embodiment of the invention, which has strong adaptability and implementation effect, and unnecessary technical characteristics can be increased or decreased according to actual needs to meet the requirements of different situations.

Claims

1. An application of insulin-like growth factor 1 receptor inhibitor BMS536924 in preparing the medicines for treating cystic echinococcosis is disclosed.

2. The use of insulin-like growth factor 1 receptor inhibitor BMS536924 as a medicament for the treatment of cystic echinococcosis according to claim 1, which further comprises albendazole, wherein insulin-like growth factor 1 receptor inhibitor BMS536924 is used in combination with albendazole.

3. The use of insulin-like growth factor 1 receptor inhibitor BMS536924 as claimed in claim 2 for the manufacture of a medicament for the treatment of cystic echinococcosis, characterized in that the mass ratio of insulin-like growth factor 1 receptor inhibitor BMS536924 to albendazole is 1 to 2: 1.

4. The use of the insulin-like growth factor 1 receptor inhibitor BMS536924 according to claim 1 or 2 or 3 for the manufacture of a medicament for the treatment of cystic echinococcosis, characterized in that cystic echinococcosis is a disease caused by echinococcus granulosus infection.

5. The use of the insulin-like growth factor 1 receptor inhibitor BMS536924 according to claim 4 for the preparation of a medicament for the treatment of cystic echinococcosis, characterized in that echinococcus granulosus is selected from the group consisting of: echinococcus granulosus larvae, Echinococcus granulosus embryonic cells, Echinococcus granulosus germinal layer, or a combination thereof.