Application of PLK4 inhibitor Centrinone in the preparation of drugs for treating echinococcosis

Abstract

The present invention discloses an application of a PLK4 inhibitor Centrinone in the preparation of a drug for treating echinococcosis. The PLK4 inhibitor Centrinone can be used as an active ingredient, alone or in combination, and provides a new option for preparing a drug for treating echinococcosis. The PLK4 inhibitor Centrinone of the present invention eliminates the proliferative cells of the cysts of Echinococcus granulosus and prevents the growth and development of the cysts. The PLK4 inhibitor Centrinone of the present invention, combined with selinexor, gemcitabine and 5-fluorouracil, or combined with selinexor and MALAT1-N-12, has a significant inhibitory effect on the growth of Echinococcus granulosus cysts, and can be used to prepare a drug for treating echinococcosis.

Description

Technical Field

[0001] The present invention belongs to the field of medical technology and relates to the use of a PLK4 inhibitor Centrinone in the preparation of a drug for treating echinococcosis. Background Art

[0002] Hydatid disease is a serious zoonotic parasitic disease caused by the tapeworm Echinococcus in humans or host animals. It is distributed worldwide, with my country being a high-incidence area. Echinococcosis includes cystic echinococcosis (caused by Echinococcus granulosus) and alveolar echinococcosis (caused by Echinococcus multilocularis). Cystic echinococcosis (CE) is the predominant form in my country. Currently, treatment options for CE primarily include surgery and medication. Surgical treatment has significant limitations, with cyst rupture and subsequent infection a common complication. Currently, the most widely used anti-CE drugs are the benzimidazoles, including albendazole and mebendazole. However, these drugs have low intestinal absorption, poor medication compliance, and only inhibit parasite growth rather than completely eliminate it. This results in a significant number of patients requiring long-term, high-dose medications, which still result in a high postoperative recurrence rate. Furthermore, research has revealed that the β-tubulin target of these drugs is highly similar between the parasite and the host, leading to severe adverse drug reactions in multiple systemic systems. Therefore, it is of great significance to find or develop new drugs for the treatment of echinococcosis with significant efficacy and few side effects.

[0003] Centrinone is a reversible Polo-like kinase 4 (PLK4) inhibitor that can destroy the activity of cancer cell centriole and inhibit cancer cell growth and proliferation. 26 H 25 F2N7O6S2, with a molecular weight of 633.65, is used to treat breast cancer, liver cancer, melanoma, etc., but there are no reports of Centrinone being used to treat cystic echinococcosis.

[0004] Selinexor is an orally available XPO1 inhibitor that causes tumor suppressor proteins to accumulate in the cell nucleus, which will restart and amplify their tumor suppressor function, leading to selective apoptosis of cancer cells without significantly affecting normal cells. 17 H 11 F6N7O, with a molecular weight of 443.31, is primarily used in multiple myeloma research, but there are no reports of selinexor being used to treat cystic echinococcosis.

[0005] MALAT1-N-1 is a potent and specific Malat1 inhibitor. MALAT1-N-1 regulates Malat1 downstream genes in a dose-dependent manner. Its molecular formula is C 19 H 21N3O2, molecular weight 323.39. MALAT1-N-1 has been reported to be used to treat abdominal aortic aneurysms, but there are no reports of MALAT1-N-1 being used to treat cystic echinococcosis.

[0006] Gemcitabine is a new cytosine nucleoside derivative with the chemical formula C9H 11 F2N3O4, with a molecular weight of 263.198. Once in the body, it is activated by deoxycytidine kinase and metabolized by cytidine deaminase. Clinically, gemcitabine is effective against a variety of solid tumors. 5-Fluorouracil (5-fluorouracil), with a chemical formula of C4H3FN2O2 and a molecular weight of 130.077, is a pyrimidine analog and an antimetabolite with some efficacy against various tumors, including gastrointestinal tumors, breast cancer, ovarian cancer, choriocarcinoma, cervical cancer, liver cancer, and bladder cancer. Gemcitabine and 5-fluorouracil can reduce the number of circulating MDSCs and promote a more robust anti-tumor immune response. However, these drugs are not specific for MDSCs and affect all rapidly proliferating cells, including anti-tumor T cells. Combination therapy with gemcitabine can significantly improve the performance of patients with pancreatic cancer. Currently, gemcitabine and 5-fluorouracil have not been used to treat cystic echinococcosis. Summary of the Invention

[0007] Purpose of the Invention: The present invention provides a method for preparing a drug for treating echinococcosis using a PLK4 inhibitor, Centrinone. Centrinone eliminates proliferative cells in Echinococcus granulosus vesicles and inhibits their growth and development. Therefore, Centrinone can be used alone or in combination with other drugs to prepare a drug for treating echinococcosis.

[0008] Technical solution: The purpose of the present invention is achieved through the following technical solution:

[0009] The present invention provides an application of a PLK4 inhibitor Centrinone in the preparation of a medicine for treating echinococcosis.

[0010] The PLK4 inhibitor Centrinone can be used as the sole active ingredient of the drug, and its structural formula is as follows:

[0011] .

[0012] The PLK4 inhibitor Centrinone can be used in combination with other compounds.

[0013] The present invention also provides the use of a combination of a PLK4 inhibitor Centrinone, selinexor, gemcitabine and 5-fluorouracil in the preparation of a drug for treating echinococcosis.

[0014] The present invention also provides the use of a combination of a PLK4 inhibitor Centrinone, selinexor and MALAT1-N-1 in the preparation of a drug for treating echinococcosis.

[0015] The medicine further includes a pharmaceutically acceptable carrier or excipient.

[0016] The excipients include one or more of antioxidants, emulsifiers, diluents, preservatives, solubilizers, disintegrants, wetting agents, adhesives or lubricants.

[0017] The dosage form of the medicine is granules, tablets, capsules, suspensions, oral solutions, injections or infusions.

[0018] The medicaments of the present invention can be administered in various known ways, such as orally or by injection. The medicaments of the present invention can be administered alone or in combination with other drugs. Oral compositions can be in any orally acceptable dosage form, including but not limited to granules, tablets, capsules, and suspensions.

[0019] Sterile injectable compositions can be formulated using suitable dispersing agents or wetting agents and suspending agents according to techniques known in the art. Pharmaceutically acceptable carriers and solvents that can be used include water, sodium chloride solution, and the like.

[0020] The drug of the present invention can be prepared into common preparations, and can also be prepared into sustained-release preparations, controlled-release preparations, targeted preparations and various microparticle drug delivery systems.

[0021] The actual dosage level of the active ingredient in the medicament of the present invention can be varied to obtain an amount of the active ingredient that is effective to achieve the desired therapeutic response for a particular patient, composition, and mode of administration, without being toxic to the patient. The selected dosage level depends on a variety of factors, including the route of administration, time of administration, rate of excretion, duration of treatment, other drugs, compounds, and / or materials used in combination with the PLK4 inhibitor Centrinone, the age, sex, weight, general health, and past medical history of the patient being treated, and similar factors well known in the medical arts.

[0022] Polo-like kinase 4 (PLK4) and nuclear export protein 1 (XPO1) play an important role in the proliferation and differentiation of germinal layer cells of Echinococcus granulosus vesicles. Centrinone and selinexor, as targeted inhibitors of PLK4 and XPO1, respectively, can eliminate the proliferative cells of Echinococcus granulosus vesicles and prevent the growth and development of vesicles.

[0023] The structural formula of selinexor is as follows:

[0024]

[0025] Malat1 inhibition is a hallmark of CD4+ T cell activation, and Malat1 expression is positively correlated with Maf expression. Overexpression of Malat1 leads to an imbalance between Th1 and Th2 immune responses in mice with cystic echinococcosis, resulting in immune disequilibrium. Malat1 inhibition in activated CD4+ T cells is a key determinant of optimal immunity against chronic infection. MALAT1-N-1, a potent and specific Malat1 inhibitor, effectively promotes CD4+ T cell activation and prevents immune disequilibrium, effectively addressing immune escape in cystic echinococcosis.

[0026] The structural formula of MALAT1-N-1 is as follows:

[0027]

[0028] MDSCs are a major cause of immunosuppression in mice infected with cystic echinococcosis. Gemcitabine and 5-fluorouracil can functionally inactivate MDSCs by targeting their immunosuppressive mechanisms, thereby preventing their suppressive effects on the immune system of CE mice.

[0029] The structural formula of gemcitabine is as follows:

[0030]

[0031] The structural formula of 5-fluorouracil is as follows:

[0032] .

[0033] Beneficial effects:

[0034] The PLK4 inhibitor Centrinone described in the present invention can be used as an active ingredient, alone or in combination with other drugs, to treat echinococcosis. This opens up new uses for Centrinone and provides a new option for the preparation of hydatid disease treatment drugs. The PLK4 inhibitor Centrinone described in the present invention eliminates proliferative cells in Echinococcus granulosus vesicles and prevents the growth and development of vesicles. Therefore, the PLK4 inhibitor Centrinone can be used alone or in combination with other drugs to prepare hydatid disease treatment drugs.

[0035] The PLK4 inhibitor Centrinone of the present invention, combined with selinexor, gemcitabine and 5-fluorouracil, or combined with selinexor and MALAT1-N-1, has a significant inhibitory effect on the growth of Echinococcus granulosus vesicles and can be used to prepare a drug for treating echinococcosis.

[0036] The present invention discloses for the first time the application of Centrinone, Selinexor, MALAT1-N-1, gemcitabine and 5-fluorouracil in treating cystic echinococcosis, expands the application of these drugs, and develops new drugs for treating cystic echinococcosis. BRIEF DESCRIPTION OF THE DRAWINGS

[0037] Figure 1 This is a bar graph of the wet weight of capsules in each group after 15 days of drug treatment;

[0038] Figure 2 This is a bar graph of the inhibition results of each group after 15 days of drug treatment;

[0039] Figure 3 This is the vesicle status diagram of the control group after administration of CMC-Na for 15 days;

[0040] Figure 4 The vesicle status diagrams after 15 days of treatment with Centrinone, Selinexor, MALAT1-N-1, gemcitabine, and 5-fluorouracil, respectively;

[0041] Figure 5 This is the vesicle status diagram after 15 days of treatment with the positive drug albendazole;

[0042] Figure 6 This is a diagram of the vesicle status of combination drug groups 1 and 2 after 15 days of treatment. DETAILED DESCRIPTION

[0043] The technical solution of the present invention is described in detail below through specific embodiments, but the protection scope of the present invention is not limited to the embodiments.

[0044] If no specific techniques or conditions are specified in the examples, the experiments were carried out according to the techniques or conditions described in the literature in the field or according to the product instructions. If no manufacturer is specified for the reagents or instruments used, they are all conventional products that can be purchased through regular channels.

[0045] The experimental methods in the following examples are conventional methods unless otherwise specified. The experimental materials used in the following examples are commercially available products unless otherwise specified.

[0046] Example 1 Effects of Centrinone, Selinexor, MALAT1-N-1, Gemcitabine, and 5-Fluorouracil on Echinococcus granulosus Vesicles in Mice

[0047] Centrinone, selinexor, MALAT1-N-1, gemcitabine, and 5-fluorouracil were purchased from Shanghai Aladdin Biochemical Technology Co., Ltd.

[0048] Experimental animals: 7-8 week-old Kunming mice weighing approximately 25 g.

[0049] Experimental methods:

[0050] Untreat group: Administer 0.4% CMC-Na aqueous solution at a volume of 0.1 mL / 10 g × mouse body weight as the control group.

[0051] Treatment group: Centrinone was dissolved in 0.4% CMC-Na in a mortar and ground, and then orally administered to each mouse at a daily dose of 20 mg / kg / day;

[0052] Selinexol was dissolved in 0.4% CMC-Na in a mortar and ground, and then orally administered to each mouse at a daily drug intake of 15 mg / kg / day;

[0053] MALAT1-N-1 was dissolved in 0.4% CMC-Na and ground in a mortar, and then orally administered to each mouse at a daily drug intake of 15 mg / kg / day;

[0054] Gemcitabine was dissolved in 0.4% CMC-Na in a mortar and ground, and then orally administered to each mouse at a daily drug intake of 40 mg / kg / day;

[0055] 5-Fluorouracil was dissolved in DMSO solvent and administered by intraperitoneal injection at a daily drug intake of 50 mg / kg / day per mouse.

[0056] After 15 days of continuous drug administration, the mice in different drug administration groups were euthanized, the cysts were collected from the abdominal cavity, the wet weight of the cysts was measured, and the inhibition rate of the drug on the cysts was calculated.

[0057] Inhibition rate = (blank group cyst wet weight - treatment group cyst wet weight) / blank group cyst wet weight * 100%.

[0058] The experimental results are shown in Table 1.

[0059] The effects of different drugs on Echinococcus granulosus vesicles can be observed by comparing the wet weight of vesicles in the Untreat group and different drug groups. The vesicles in the Untreat group were intact and numerous (see Figure 3 ); while the number of vesicles in the mice in the drug-treated group was smaller and their volume was significantly reduced (see Figure 4 Compared with the Untreat group, the wet weight of vesicles in the mice in several drug-treated groups decreased to varying degrees (see Figure 1 , Figure 2 This result indicates that Centrinone, selinexor, MALAT1-N-1, gemcitabine and 5-fluorouracil administered alone have a significant inhibitory effect on the growth of Echinococcus granulosus cysts.

[0060] Example 2 Effects of the Positive Drug Albendazole (ABZ) on Echinococcus granulosus Vesicles in Mice

[0061] Experimental animals: 7-8 week-old Kunming mice weighing approximately 25 g.

[0062] Experimental methods:

[0063] Albendazole was dissolved in 0.4% CMC-Na in a mortar and ground, and then orally administered to each mouse at a daily dose of 50 mg / kg / day.

[0064] After 15 days of continuous dosing, mice in the treatment group were euthanized, and cysts were collected from the peritoneal cavity. The wet weight of the cysts was measured, and the inhibition rate of drug-induced cyst formation was calculated. Inhibition rate = (cyst wet weight of blank group - cyst wet weight of treatment group) / cyst wet weight of blank group * 100%. The experimental results are shown in Table 1.

[0065] The effects of different drugs on Echinococcus granulosus vesicles can be observed by comparing the wet weight of vesicles in the Untreat group and the Albendazole group. The vesicles in the Untreat group were intact and numerous (see Figure 3 In the albendazole group, the number of vesicles in the mice was smaller, the vesicle walls were thinner, and the volume was significantly reduced (see Figure 5 Compared with the Untreat group, the wet weight of vesicles in the albendazole group was significantly reduced (see Figure 1 , Figure 2 This result indicates that albendazole has a significant inhibitory effect on the growth of Echinococcus granulosus cysts.

[0066] Example 3 Effects of Centrinone, Selinexor, and the Combination of Gemcitabine and 5-Fluorouracil (Combination Group 1) on Echinococcus granulosus Vesicles in Mice

[0067] Experimental animals: 7-8 week-old Kunming mice weighing approximately 25 g.

[0068] Experimental methods:

[0069] The mice in the drug administration group were sequentially treated with Centrinone, selinexor, and a combination of gemcitabine and 5-fluorouracil according to the method in Example 1. After 15 days of continuous administration, the mice were euthanized, and the cysts were collected from the abdominal cavity. The wet weight of the cysts was measured, and the inhibition rate of the drugs on the cysts was calculated. The inhibition rate = (blank group cyst wet weight - treatment group cyst wet weight) / blank group cyst wet weight * 100%.

[0070] Finally, SPSS 20.0 was used for statistical analysis. Data were expressed as mean ± standard deviation. One-way ANOVA was used to analyze the differences between groups. A p value less than 0.05 was considered statistically significant. The experimental results are shown in Table 1.

[0071] The effect of combined drug therapy on Echinococcus granulosus vesicles was observed by comparing the wet weight of vesicles in the Untreat group with those in the Centrinone, Selinexor, and Gemcitabine and 5-fluorouracil combined groups. The vesicles in the Untreat group were intact and numerous, whereas the vesicles in the Centrinone, Selinexor, and Gemcitabine and 5-fluorouracil combined groups were few in number, significantly reduced in size, and even showed a certain degree of calcification (see Figure 6 The wet weight of vesicles in the combined drug group was significantly reduced (see Figure 1 , Figure 2 The results showed that Centrinone, selinexor, and the combination of gemcitabine and 5-fluorouracil had a significant inhibitory effect on the growth of Echinococcus granulosus cysts.

[0072] Example 4 Effects of Centrinone, Selinexor, and MALAT1-N-1 Combination (Combination Group 2) on Echinococcus granulosus Vesicles in Mice

[0073] Experimental animals: 7-8 week-old Kunming mice weighing approximately 25 g.

[0074] Experimental methods:

[0075] Centrinone, selinexor, and MALAT1-N-1 were administered to mice according to the administration method in Example 1. After 15 days of continuous administration, the mice were euthanized, and the cysts were collected from the abdominal cavity. The wet weight of the cysts was measured, and the inhibition rate of the drugs on the cysts was calculated. The inhibition rate = (blank group cyst wet weight - treatment group cyst wet weight) / blank group cyst wet weight * 100%.

[0076] The experimental results are shown in Table 1.

[0077] The effect of combined drug treatment on Echinococcus granulosus vesicles was observed by comparing the wet weight of vesicles in the Untreat group and the Centrinone, Selinexor and MALAT1-N-1 combined groups. The vesicles in the Untreat group were intact and numerous (see Figure 3 ); while the number of vesicles in the mice of the combined drug group was very small and the volume was significantly reduced (see Figure 6 The wet weight of vesicles in the combined drug group was significantly reduced. This result indicates that the combined administration of Centrinone, selinexor and MALAT1-N-1 has a significant inhibitory effect on the growth of Echinococcus granulosus cysts, but the inhibitory effect is slightly lower than that of the combined drug group 1 (see Figure 1 , Figure 2 ).

[0078] As shown in Table 1 and Figure 1As shown in the figure, after 3 weeks of treatment with various drugs, the wet weight of cysts in mice in each group was significantly reduced compared with the blank group (4.3255±2.7955). The wet weight of cysts in mice in combination group 1 (0.7344±0.2273) and combination group 2 (0.9334±0.3273) decreased significantly, and the cysts in the mice almost disappeared (p=0.019, p=0.032). At the same time, the comparison between combination groups 1 and 2 and the albendazole group showed that the therapeutic effect of combination group 1 (inhibition rate reached 83.0%) was significantly better than that of combination group 2 (inhibition rate reached 78.4%) (p=0.021, p=0.032). Figure 2 In addition, the wet weight of cysts in mice in the albendazole, centrinone (2.3375±0.8558), selinexor (3.1375±1.2564), MALAT1-N-1 (1.9765±0.7264), gemcitabine (1.4436±0.6345), and 5-fluorouracil (1.0233±0.4273) groups also decreased significantly. These drugs alone can also slow down the growth of cysts, but the effect is not as good as that in the combination group.

[0079] Table 1 Wet weight of cysts and inhibition rate of mice in each group after 3 weeks of treatment

[0080]

[0081] Centrinone, selinexor, MALAT1-N-1, gemcitabine, and 5-fluorouracil, as well as their combination, can be used to develop anti-echinococcosis drugs. In vivo pharmacodynamic data suggest that these drugs are highly effective in treating echinococcosis and can serve as targets for screening new drugs for Echinococcus granulosus echinococcosis. The present invention provides a new approach for the treatment of cystic echinococcosis.

[0082] As described above, although the present invention has been shown and described with reference to specific preferred embodiments, it should not be construed as limiting the present invention itself. Various changes may be made to it in form and detail without departing from the spirit and scope of the present invention as defined in the appended claims.

Claims

1. Application of PLK4 inhibitor Centrinone in the preparation of drugs for the treatment of echinococcosis.

2. The use according to claim 1, characterized in that: The PLK4 inhibitor Centrinone is the only active ingredient of the drug, and its structural formula is as follows: 。 3. Application of PLK4 inhibitors Centrinone, selinexor, gemcitabine and 5-fluorouracil in combination for the preparation of drugs for the treatment of echinococcosis.

4. Application of the PLK4 inhibitor Centrinone, Selinexor and MALAT1-N-1 in combination for the preparation of a drug for the treatment of echinococcosis, wherein the structural formula of MALAT1-N-1 is: 。 5. The use according to any one of claims 1 to 4, characterized in that: The drug also includes pharmaceutically acceptable excipients.

6. The use according to claim 5, characterized in that: The auxiliary materials include one or more of antioxidants, emulsifiers, diluents, preservatives, solubilizers, disintegrants, wetting agents, adhesives or lubricants.

7. The use according to any one of claims 1 to 4, characterized in that: The dosage form of the medicine is granules, tablets, capsules, suspensions, oral solutions, injections or infusions.

8. The use according to any one of claims 1 to 4, characterized in that: The PLK4 inhibitor Centrinone eliminates proliferative cells of Echinococcus granulosus vesicles and blocks the growth and development of vesicles.

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