Application of glutamate transporter agonist in preparation of ivermectin-resistant drug for treating parasites

Abstract

The invention relates to application of a glutamate transporter agonist in preparation of an ivermectin drug-resistant drug for treating parasites, and belongs to the field of drug resistance. The glutamic acid transporter agonist reduces the content of glutamic acid in the body by enhancing the function of the glutamic acid transporter. The ceftriaxone sodium is used for treating the ubr-1 mutant, the content of glutamic acid in the ubr-1 mutant is reduced, expression of a chloride ion channel controlled by glutamic acid is increased, and therefore IVM drug resistance caused by UBR-1 protein mutation is relieved. The invention finds that the curative effect of the IVM on parasitic diseases is effectively recovered, the service life of the medicine is prolonged, and the use cost of the IVM is reduced.

Description

Technical Field

[0001] The present invention relates to the field of drug resistance, and more specifically, to a method for alleviating ivermectin resistance of parasites, and in particular to the use of a glutamate transporter agonist in the preparation of an ivermectin-resistant drug for treating parasites. Background Art

[0002] Ivermectin (IVM) is a macrolide derivative and a chemically modified derivative of the naturally occurring avermectin B1. 22 ,C 23 -Dihydroavermectin B 1a and 20% C 22 ,C 23 -Dihydroavermectin B 1b It is composed of 100 kinds of antibiotics and has a strong lethal effect on many pathogenic parasitic nematodes. IVM was first introduced into the animal parasitic disease pharmaceutical market in 1981. As the use of IVM has increased year by year, IVM has become a very popular animal therapeutic pharmaceutical product in the late 1980s. Not only that, IVM also has a huge application in the human medical and health market. Since the launch of the "IVM Donation Program" in 1987, more than 1.4 billion approvals for the control and elimination of onchocerciasis and 1.2 billion approvals for the control and elimination of lymphatic filariasis have been approved. Most of the diseases caused by onchocerciasis and lymphatic filariasis in poor countries in the world have been eliminated, which has made a significant contribution to human public health.

[0003] The receptor of IVM in parasites is glutamate-gated chloride channel (GluCl). IVM can activate GluCl in the parasite, causing a large influx of chloride ions, causing neuronal hyperpolarization, leading to paralysis of body wall muscles, inhibition of movement and feeding, and death of the parasite. However, as IVM is used more and more frequently and in greater quantities to treat parasitic diseases, parasites are becoming more and more resistant to IVM, and there are more and more cases of parasitic nematodes being resistant to IVM. Summary of the invention

[0004] In order to better solve the resistance of parasites to ivermectin (Ivermectin, referred to as IVM) and restore the therapeutic effect of IVM, the present invention studies the use of glutamate transporter agonists to enhance the function of glutamate transporters to reduce the glutamate content in the body, thereby increasing the expression of glutamate-gated chloride ion channels, thereby alleviating the ivermectin resistance of parasites.

[0005] According to the purpose of the present invention, there is provided the use of a glutamate transporter agonist in the preparation of an ivermectin-resistant drug for treating parasites.

[0006] Preferably, the glutamate transporter agonist is ceftriaxone sodium (Cef for short).

[0007] Preferably, the concentration of the glutamate transporter agonist is less than or equal to 50 μg / mL.

[0008] Preferably, the concentration of the glutamate transporter agonist is greater than or equal to 0.05 μg / mL.

[0009] Preferably, the glutamate transporter agonist is used to enhance the function of the glutamate transporter to reduce the glutamate content in the body, thereby increasing the expression of glutamate-gated chloride ion channels, thereby alleviating the ivermectin resistance of the parasite.

[0010] Preferably, the ivermectin resistance of the parasite is resistance caused by UBR-1 protein mutation.

[0011] Preferably, the parasite is Caenorhabditis elegans.

[0012] In general, the above technical solution conceived by the present invention has the following technical advantages compared with the prior art:

[0013] (1) The glutamate transporter agonist of the present invention reduces the glutamate content in the body by enhancing the function of the glutamate transporter. The present invention treats ubr-1 with Cef to reduce the glutamate content in the body and increase the expression of glutamate-gated chloride ion channels, thereby alleviating IVM resistance caused by UBR-1 protein mutation.

[0014] (2) In the present invention, under the background of 5 ng / mL IVM treatment, the present invention used different concentrations of Cef to treat ubr-1. As the Cef concentration gradually increased, the survival rate of ubr-1 gradually decreased, and when the Cef concentration reached 50 μg / mL, the survival rate of ubr-1 dropped to the lowest, which was equivalent to the survival rate of WT. This shows that Cef can effectively alleviate the ivermectin resistance caused by UBR-1 protein mutation.

[0015] (3) The present invention has effectively restored the efficacy of IVM against parasitic diseases, extended the useful life of the drug and reduced the cost of using IVM. BRIEF DESCRIPTION OF THE DRAWINGS

[0016] Figure 1:Survival rate, body length, pharyngeal velocity and movement speed of WT and ubr-1 treated with different concentrations of IVM. A represents the survival rate of WT and ubr-1 after nematodes were treated with 0ng / mL and 5ng / mL IVM; B represents the body length of WT and ubr-1 after nematodes were treated with 0ng / mL and 5ng / mL IVM; C represents the pharyngeal velocity of WT and ubr-1 after nematodes were treated with 0ng / mL and 5ng / mL IVM; D represents the movement speed of WT and ubr-1 after nematodes were treated with 0ng / mL and 5ng / mL IVM.

[0017] Figure 2 :GluCl fluorescence expression diagram. The expression levels of three GluCl proteins AVR-14, AVR-15, and GLC-1 in C.elegans were decreased in ubr-1.

[0018] Figure 3 :The survival rate, body length, pharyngeal velocity and movement speed of ubr-1 treated with different concentrations of Cef. A and B represent the survival rate of ubr-1 treated with different concentrations of Cef under 5 ng / mL IVM treatment; C represents the survival rate of WT and ubr-1; D represents the body length of WT and ubr-1; E represents the pharyngeal velocity of WT and ubr-1; F represents the movement speed of WT and ubr-1. The concentration of IVM used in C, D, E and F is 5 ng / mL, and the concentration of Cef used is 50 μg / mL. "-" indicates no drug added, and "+" indicates drug added.

[0019] Figure 4 : GluCl fluorescence expression diagram. The expression level of GluCl protein AVR-15 in C.elegans was increased in ubr-1 after Cef treatment. DETAILED DESCRIPTION

[0020] In order to make the purpose, technical solutions and advantages of the present invention more clearly understood, the present invention is further described in detail below in conjunction with the accompanying drawings and embodiments. It should be understood that the specific embodiments described herein are only used to explain the present invention and are not intended to limit the present invention. In addition, the technical features involved in the various embodiments of the present invention described below can be combined with each other as long as they do not conflict with each other.

[0021] The present invention utilizes the high conservation of GluCl between Caenorhabditis elegans (C. elegans for short) and parasites, and explores new drug strategies by studying the IVM resistance of Caenorhabditis elegans.

[0022] The wild-type nematode WT and the mutant nematode ubr-1 were treated with 0 ng / mL and 5 ng / mL of ivermectin, respectively.

[0023] Example 1

[0024] Survival test (eg Figure 1 (A) There was no significant difference in the survival rates of WT and ubr-1 in the 0 ng / mL IVM treatment group; however, in the 5 ng / mL IVM treatment group, the survival rate of WT was decreased compared with that of ubr-1.

[0025] Example 2

[0026] Body length test (such as Figure 1 (B) There was no significant difference in body length between WT and ubr-1 in the 0 ng / mL IVM treatment group; whereas in the 5 ng / mL IVM treatment group, the body length of WT decreased compared with that of ubr-1.

[0027] Example 3

[0028] Pharyngeal pump rate test (eg Figure 1 C), there was no significant difference in pharyngeal pumping rate between WT and ubr-1 in the 0 ng / mL IVM treatment group; while in the 5 ng / mL IVM treatment group, the pharyngeal pumping rate of WT was decreased compared with that of ubr-1.

[0029] Example 4

[0030] Movement speed test (such as Figure 1 D), there was no significant difference in the movement speed between WT and ubr-1 in the 0 ng / mL IVM treatment group; while in the 5 ng / mL IVM treatment group, the movement speed of WT decreased compared with ubr-1, indicating that ubr-1 nematodes have a certain resistance to IVM.

[0031] The reason for drug resistance is that the glutamate content in ubr-1 increases, resulting in a decrease in the expression level of GluCl in the body (such as Figure 2 ), thereby generating resistance. Ceftriaxone sodium is a glutamate transporter agonist, which mainly enhances the function of glutamate transporters to reduce the glutamate content in the body. The present invention uses Cef to treat ubr-1, reduce the glutamate content in the body, increase the expression of glutamate-gated chloride ion channels, and thus alleviate IVM resistance caused by UBR-1 protein mutation.

[0032] In the context of 5 ng / mL IVM treatment, the present invention uses different concentrations of Cef to treat ubr-1. As the Cef concentration gradually increases (e.g. Figure 3 A in Figure 3In B), the survival rate of ubr-1 gradually decreased, and when the Cef concentration reached 50 μg / mL, the survival rate of ubr-1 dropped to the lowest, which was equivalent to that of WT. Then, 5 ng / mL IVM and 50 μg / mL Cef were used to treat wild-type nematodes WT and mutant nematodes ubr-1, respectively.

[0033] Example 5

[0034] Survival test (eg Figure 3 (C), The survival rate of ubr-1 worms treated with both IVM and Cef was reduced compared with that of ubr-1 worms treated with IVM alone.

[0035] Example 6

[0036] Body length test (such as Figure 3 D), The body length of ubr-1 worms treated with both IVM and Cef was reduced compared with that of ubr-1 worms treated with IVM alone.

[0037] Example 7

[0038] Pharyngeal pump rate test (eg Figure 3 (E), Pharyngeal pumping rate was decreased in ubr-1 worms treated with both IVM and Cef compared to ubr-1 worms treated with IVM alone.

[0039] Example 8

[0040] Movement speed test (such as Figure 3 F) shows that the movement speed of ubr-1 nematodes treated with both IVM and Cef decreased compared with that of ubr-1 nematodes treated with IVM alone, indicating that Cef can effectively alleviate the ivermectin resistance caused by UBR-1 protein mutation.

[0041] In order to further illustrate the relationship between Cef and GluCl, the present invention used 50 μg / mL Cef to treat ubr-1 nematodes to observe the expression level of GluCl. Figure 4 After Cef treatment, the expression level of GluCl protein AVR-15 in ubr-1 increased, indicating that the alleviation of ivermectin resistance in ubr-1 nematodes is achieved by increasing the expression of GluCl and thus providing ivermectin sensitivity.

[0042] It will be easily understood by those skilled in the art that the above description is only a preferred embodiment of the present invention and is not intended to limit the present invention. Any modifications, equivalent substitutions and improvements made within the spirit and principles of the present invention should be included in the protection scope of the present invention.

Claims

1. Application of glutamate transporter agonists in the preparation of ivermectin-resistant drugs for treating parasites.

2. The use according to claim 1, characterized in that The glutamate transporter agonist is ceftriaxone sodium.

3. The use according to claim 1 or 2, characterized in that: The concentration of the glutamate transporter agonist is less than or equal to 50 μg / mL.

4. The use according to claim 1 or 2, characterized in that: The concentration of the glutamate transporter agonist is greater than or equal to 0.05 μg / mL.

5. The use according to claim 1, characterized in that The glutamate transporter agonist is used to enhance the function of the glutamate transporter to reduce the glutamate content in the body, thereby increasing the expression of glutamate-gated chloride ion channels, thereby alleviating the ivermectin resistance of parasites.

6. The use according to claim 1, characterized in that The ivermectin resistance of the parasite is caused by UBR-1 protein mutation.

7. The use according to claim 1, characterized in that The parasite is Caenorhabditis elegans.

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