Application of lenvatinib (E7080) in treatment of alveolar echinococcosis

Abstract

The invention discloses application of lenvatinib (E7080) in treatment of alveolar echinococcosis, and belongs to the field of biomedicine. The technical problem to be solved by the invention is how to treat and / or prevent alveolar echinococcosis. It is found that lenvatinib (E7080) can cause falling of a head hook of an original head section of echinococcus multilocularis, shrinkage of a body section, damage to structures such as a suction cup and microvillus, and the in-vivo pathogenicity is remarkably reduced; the structure of the vesicles in the middle polyester period can be destroyed, so that the hair growth layers of the vesicles shrink and are separated from the cuticle. It is shown that lenvatinib (E7080) has a good killing effect on echinococcus multilocularis and can be used for preparing the medicine for treating the alveolar echinococcosis.

Description

Technical Field

[0001] This invention belongs to the field of biomedicine, specifically relating to the application of lenvatinib (E7080) in the treatment of alveolar echinococcosis. Background Technology

[0002] Alveolar echinococcosis (AE) is caused by Echinococcus multilocularis (… Echinococcus multilocularis, E. multilocularis Echinococcosis (AE) is a significant zoonotic parasitic disease caused by infection, with a relatively high incidence in western China. The disease primarily affects the liver, exhibiting invasive, proliferative, and destructive malignant tumor-like characteristics, often referred to as "parasitic cancer," posing a major threat to human health. Currently, drug therapy remains the most widely used treatment for AE. However, albendazole (ABZ), the only clinically recommended drug for AE by the World Health Organization, primarily inhibits rather than kills the parasite. Furthermore, it suffers from high oral dosages, low bioavailability, long treatment cycles, and significant side effects, leading to poor patient adherence and a clinical cure rate of only 30%. Therefore, there is an urgent need to find new, reliable drugs for treating AE. In recent years, drug reuse and research into the potential therapeutic value of existing drugs in new indications have been considered effective solutions for treating various diseases, potentially offering a breakthrough in the current predicament of AE drug treatment.

[0003] Lenvatinib (E7080) is a novel multi-target tyrosine kinase inhibitor that effectively inhibits multiple tyrosine kinase receptors, such as vascular endothelial growth factor receptors (VEGFR 1-3) and fibroblast growth factor receptors (FGFR 1-4), thereby inhibiting angiogenesis and directly suppressing tumor cell proliferation. It has become a new standard of care for first-line treatment of unresectable advanced liver cancer. Furthermore, Lenvatinib (E7080) has been approved globally and in China for the treatment of various malignancies, including thyroid cancer, renal cell carcinoma, and endometrial cancer, demonstrating excellent efficacy. There are no reports of Lenvatinib (E7080) being used to treat alveolar echinococcosis. Summary of the Invention

[0004] The technical problem to be solved by this invention is the treatment and / or prevention of alveolar echinococcosis.

[0005] To address the aforementioned technical problems, this invention first provides the application of tyrosine kinase inhibitors in the preparation of products for the treatment and / or prevention of alveolar echinococcosis, or their application in the treatment and / or prevention of alveolar echinococcosis.

[0006] Specifically, the tyrosine kinase inhibitor is a multi-target tyrosine kinase inhibitor. Further, the tyrosine kinase inhibitor is Lenvatinib (E7080).

[0007] The use of a tyrosine kinase inhibitor in the preparation of a product for killing protoscolex or cysticercus of Echinococcus multilocularis, or in killing protoscolex or cysticercus of Echinococcus multilocularis, also falls within the protection scope of the present application.

[0008] The use of a tyrosine kinase inhibitor in the preparation of a product for inhibiting the growth or development of Echinococcus multilocularis, or in inhibiting the growth or development of Echinococcus multilocularis, also falls within the protection scope of the present application.

[0009] Specifically, the growth or development of Echinococcus multilocularis is the growth or development of protoscolex or cysticercus of Echinococcus multilocularis.

[0010] The use of a tyrosine kinase inhibitor in the preparation of a product for reducing the pathogenicity of Echinococcus multilocularis, or in reducing the pathogenicity of Echinococcus multilocularis, also falls within the protection scope of the present application.

[0011] Specifically, the pathogenicity of Echinococcus multilocularis is the pathogenicity of protoscolex of Echinococcus multilocularis.

[0012] The use of a tyrosine kinase inhibitor in the preparation of a product for inhibiting the infection of a human or an animal by Echinococcus multilocularis, or in inhibiting the infection of a human or an animal by Echinococcus multilocularis, also falls within the protection scope of the present application.

[0013] Specifically, the animal is a mammal. In an embodiment of the present application, the animal is a mouse.

[0014] The use of a tyrosine kinase inhibitor in the preparation of a product for inhibiting the metastasis of a hydatid disease lesion in a human or an animal, or in inhibiting the metastasis of a hydatid disease lesion in a human or an animal, also falls within the protection scope of the present application.

[0015] Specifically, the animal is a mammal. In an embodiment of the present application, the animal is a mouse.

[0016] Specifically, the metastasis of a hydatid disease lesion is the metastasis of a hydatid disease lesion to the liver.

[0017] The use of a tyrosine kinase inhibitor in the preparation of a product for inhibiting the growth of a hydatid disease lesion in a human or an animal, or in inhibiting the growth of a hydatid disease lesion in a human or an animal, also falls within the protection scope of the present application.

[0018] Specifically, the animal is a mammal. In an embodiment of the present application, the animal is a mouse.

[0019] In the above text, the tyrosine kinase inhibitor is a multi-target tyrosine kinase inhibitor. Further, the tyrosine kinase inhibitor is lenvatinib (E7080).

[0020] In this invention, the product may be a drug.

[0021] Experiments have shown that lenvatinib (E7080) can cause the loss of the protoscolex hooks, somatic segment shrinkage, and damage to structures such as suckers and microvilli in Echinococcus multilocularis, significantly reducing its pathogenicity in vivo. It can also disrupt the structure of vesicles in the mesotacarpal stage, leading to shrinkage of the vesicle germinal layer and separation from the cuticle. This indicates that lenvatinib (E7080) has a good killing effect on Echinococcus multilocularis and can be used as a drug for the treatment of alveolar echinococcosis. Attached Figure Description

[0022] Figure 1 Effects of different concentrations of Lenvatinib (E7080) on the protoscolex of Echinococcus multilocularis (methylene blue staining).

[0023] Figure 2 Effects of different concentrations of Lenvatinib (E7080) on the survival rate of protoscolex of Echinococcus multilocularis.

[0024] Figure 3 Scanning electron microscopy was used to observe the effects of Lenvatinib (E7080) (100 μM) intervention on the ultrastructure of the prostellar section of Echinococcus multilocularis. a, invaginated type; b, everted type; rostellar section (rc), sucker (sucker, su), body (bo), tegument (tg), microthrix (mt).

[0025] Figure 4 : The effect of Lenvatinib (E7080) (100μM) intervention on the morphology and structure of vesicles in the middle stage of Echinococcus multilocularis was observed by optical microscopy.

[0026] Figure 5 The effects of Lenvatinib (E7080) (100 μM) intervention on the ultrastructure of vesicles in the middle stage of Echinococcus multilocularis were observed using scanning electron microscopy. Laminated layer (LL) and germinal layer (GL) were also observed.

[0027] Figure 6Effect of Lenvatinib (E7080) intervention on the pathogenicity of Echinococcus multilocularis protoscolex infection in mice. A: Gross observation, with lesions circled; B: Gross observation of lesions; C: Lesion weight; D: Infection rate, the number to the left of the " / " indicates the number of infected mice, and the number to the right of the " / " indicates the total number of mice in the group; E: Liver metastasis rate, the number to the left of the " / " indicates the number of mice with liver metastasis, and the number to the right of the " / " indicates the total number of mice in the group. Detailed Implementation

[0028] The present invention will now be described in further detail with reference to specific embodiments. The given embodiments are merely illustrative of the invention and not intended to limit its scope. The embodiments provided below can serve as a guide for further improvements by those skilled in the art and do not constitute a limitation on the invention in any way.

[0029] Unless otherwise specified, the experimental methods used in the following examples are conventional methods, performed according to the techniques or conditions described in the literature in this field or according to the product instructions. Unless otherwise specified, the materials, reagents, instruments, etc., used in the following examples are commercially available.

[0030] The following examples used IBM SPSS Statistical 26.0 to perform statistical analysis on the data. The experimental results are expressed as mean ± standard deviation. One-way ANOVA was used for comparisons among multiple groups, and independent samples t-test was used for comparisons between two groups. P < 0.05 (*) indicates a significant difference, and P < 0.01 (**) indicates a highly significant difference.

[0031] Lenvatinib (E7080): APExBIO; Catalog No.: A2174; CAS: 417716-92-8.

[0032] Example 1: Application of Lenvatinib (E7080) in the preparation of a drug for treating alveolar echinococcosis This embodiment obtained data on the killing effect of Lenvatinib (E7080) on the protoscolex and vesicles of Echinococcus multilocularis cultured in vitro and in vivo infection data, and found that Lenvatinib (E7080) can be used as a drug for the preparation of treatment for alveolar echinococcosis. The specific experiments are as follows: 1. In vitro intervention with Lenvatinib (E7080) reduced the activity of the protoscolex of Echinococcus multilocularis. The prostomium of Echinococcus multilocularis was aseptically isolated and placed in a 96-well plate containing 200 μL of culture medium (1% penicillin-streptomycin (v / v) + 10% fetal bovine serum (v / v) + RPMI-1640 medium). Then, different concentrations of Lenvatinib (E7080) were added to the culture system for intervention (Lenvatinib was dissolved in DMSO to prepare a 10 mM stock solution). The experiment included a solvent control group (a 2% DMSO control group, i.e., adding the same volume of DMSO as the intervention group to the culture system, with a final concentration of 2%) and Lenvatinib (E7080) intervention groups with concentrations of 1 μM, 25 μM, 50 μM, 100 μM, and 200 μM, i.e., the final concentrations of Lenvatinib (E7080) in the culture system were 1 μM, 25 μM, 50 μM, 100 μM, and 200 μM, respectively). Protostomes were collected on days 1, 3, and 6 after intervention. The activity of protostomes was observed using methylene blue staining, and the survival rate was calculated. The ultrastructural changes of protostomes 6 days after Lenvatinib (E7080) (100 μM) intervention were observed using scanning electron microscopy.

[0033] The experimental results are shown in Figure 1 and Figure 2 Six days later, the survival rate of protostomes in the solvent control group was (79.09±5.57)%, while the survival rates of protostomes in the Lenvatinib (E7080) 1μM, 25μM, 50μM, 100μM, and 200μM intervention groups decreased to 77.26±7.74%, 42.16±16.18%, 29.66±9.88%, 3.77±1.05%, and 0%, respectively. The survival rates of protostomes in the 25μM, 50μM, 100μM, and 200μM intervention groups were significantly different from those in the solvent control group. F =47.59, P <0.0001).

[0034] Scanning electron microscopy revealed that after 6 days of intervention with a final concentration of 100 μM Lenvatinib (E7080) in the protoscolex, it caused shrinkage of the protoscolex segments, loss of head hooks, and damage to structures such as suckers and microvilli. Figure 3 This indicates that Lenvatinib (E7080) has a good killing effect on the protoscolex of Echinococcus multilocularis cultured in vitro.

[0035] 2. In vitro intervention with Lenvatinib (E7080) reduced the activity of mid-stage vesicles in Echinococcus multilocularis. Medium-stage vesicles of *Echinococcus multilocularis* cultured in vitro to a diameter of 2–3 mm were placed in 24-well plates containing 1 mL of culture medium (1% penicillin-streptomycin + 10% fetal bovine serum + RPMI-1640 medium). Lenvatinib (E7080) was then added to the culture system for intervention. The experiment included a Lenvatinib (E7080) intervention group (100 μM, i.e., the final concentration of Lenvatinib (E7080) in the culture system was set to 100 μM) and a solvent control group (final concentration of 1% DMSO, i.e., an equal volume of DMSO was added to the culture system, resulting in a final concentration of 1% (volume percentage)). After 9 days of intervention, the morphology and ultrastructure of the medium-stage vesicles were observed using optical microscopy and scanning electron microscopy.

[0036] The experimental results are shown in Figure 4 and Figure 5 Nine days after Lenvatinib (E7080) intervention in Echinococcus multilocularis vesicles, separation of the cuticle and germinal layer occurred in the intermediate stage vesicles, with the germinal layer turning black and shrunken into clumps. Figure 4 Scanning electron microscopy revealed that in the Lenvatinib (E7080) intervention group, the cuticle and germinal layer of the vesicles in the mesophase stage of Echinococcus multilocularis were separated, and the tissue structure of the germinal layer was destroyed. Figure 5 This indicates that Lenvatinib (E7080) has a significant killing effect on the middle stage vesicles of Echinococcus multilocularis cultured in vitro.

[0037] 3. Lenvatinib (E7080) intervention reduced the pathogenicity of Echinococcus multilocularis protoscolex infection in mice. Following the method described above for in vitro intervention of the protoscolex of Echinococcus multilocularis, a Lenvatinib (E7080) 100μM intervention group and a solvent control group were set up. Six days after in vitro intervention of the protoscolex, the protoscolex from both groups were intraperitoneally injected into female Balb / C mice (6-7 weeks old, weighing 15-18g) at a dose of 6500 protoscolex / mouse. Six mice were inoculated in each group. Mice and lesion samples were collected 24 weeks after inoculation to observe the ability of the protoscolex in the two groups to develop into vesicles in the middle tapetum stage, compare the lesion weight of the two groups of mice, and calculate the infection rate and metastasis rate.

[0038] Infection rate = (Number of mice infected with Echinococcus multilocularis / Total number of mice in the group) × 100%; Metastasis rate = Number of mice with liver metastases / Total number of mice in the group × 100%.

[0039] The experimental results are shown in Figure 6In the AE (Acute Acute Respiratory Syndrome) in mice, 24 weeks after inoculation, the infection rate in the solvent control group was 100%, and peritoneal vesicles and liver metastases were observed, with a metastasis rate of 100%. The infection rate in the Lenvatinib (E7080) intervention group was 83.3%, and the metastasis rate was 16.7%, significantly lower than that in the solvent control group. The lesion weight in the Lenvatinib (E7080) intervention group was 23.67±23.4 mg, significantly lower than the lesion weight of 4263.17±2835.6 mg in the solvent control group.

[0040] In summary, this invention discloses for the first time a novel multi-target tyrosine kinase inhibitor, lenvatinib (E7080), which is a novel drug for treating alveolar echinococcosis. Pharmacodynamic experimental data show that lenvatinib (E7080) has a significant killing effect on the protoscolex of Echinococcus multilocularis. In vivo models confirm that the pathogenicity of the protoscolex is significantly reduced after drug intervention, and it can inhibit the growth of vesicles in the middle stage of the tapeworm, indicating that lenvatinib (E7080) can be used to treat alveolar echinococcosis.

[0041] The present invention has been described in detail above. For those skilled in the art, the invention can be practiced in a wide range of ways with equivalent parameters, concentrations, and conditions without departing from its spirit and scope, and without requiring unnecessary experiments. Although specific embodiments have been given, it should be understood that further modifications can be made to the invention. In summary, according to the principles of the invention, this application is intended to include any changes, uses, or improvements to the invention, including changes made using conventional techniques known in the art that depart from the scope disclosed herein. Some of the essential features can be applied within the scope of the following appended claims.

Claims

1. The use of tyrosine kinase inhibitors in the preparation of products for the treatment and / or prevention of alveolar echinococcosis, or in the treatment and / or prevention of alveolar echinococcosis.

2. The application according to claim 1, characterized in that: The tyrosine kinase inhibitor is Lenvatinib (E7080).

3. The use of tyrosine kinase inhibitors in the preparation of products that kill Echinococcus multilocularis protoscolex or vesicles, or in the killing of Echinococcus multilocularis protoscolex or vesicles.

4. The use of tyrosine kinase inhibitors in the preparation of products that inhibit the growth or development of Echinococcus multilocularis, or in the inhibition of the growth or development of Echinococcus multilocularis.

5. The application of tyrosine kinase inhibitors in the preparation of products that reduce the pathogenicity of Echinococcus multilocularis, or in the reduction of the pathogenicity of Echinococcus multilocularis.

6. The use of tyrosine kinase inhibitors in the preparation of products that inhibit human or animal infection with Echinococcus multilocularis, or in the inhibition of human or animal infection with Echinococcus multilocularis.

7. The use of tyrosine kinase inhibitors in the preparation of products that inhibit the metastasis of alveolar echinococcosis lesions in humans or animals, or in the inhibition of the metastasis of alveolar echinococcosis lesions in humans or animals.

8. The use of tyrosine kinase inhibitors in the preparation of products that inhibit the growth of alveolar echinococcosis lesions in humans or animals, or in the use of tyrosine kinase inhibitors in the inhibition of the growth of alveolar echinococcosis lesions in humans or animals.

9. The application according to any one of claims 3-8, characterized in that: The tyrosine kinase inhibitor is Lenvatinib (E7080).

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