Application of seltamides medicine in preparation of toxoplasmosis resisting medicine

Abstract

The invention belongs to the technical field of biological medicines, relates to a novel medicine application of a compound, and in particular relates to an application of a seltamides medicine in preparing a medicine for resisting toxoplasmosis. The at least one drug is a compound as shown in a formula I; the research of the invention shows that the seltamies not only can prolong the survival time of mice infected by toxoplasma virulent strains, but also can significantly reduce the death rate of mice infected by toxoplasma attenuated strains, that is, the seltamies have an obvious treatment effect on related diseases caused by the toxoplasma virulent strains or the toxoplasma attenuated strains. Therefore, the compound can be used for preparing medicines for treating toxoplasmosis.

Description

Technical Field

[0001] The present invention belongs to the field of biomedicine technology and relates to new drug uses of compounds, and specifically relates to the use of a statin-type drug in the preparation of an anti-toxoplasmosis drug. Background Art

[0002] The information disclosed in this background technology section is only intended to enhance the understanding of the overall background of the invention, and should not necessarily be regarded as an admission or any form of suggestion that the information constitutes the prior art already known to a person skilled in the art.

[0003] Toxoplasma gondii is an opportunistic pathogenic protozoan that can infect all warm-blooded animals, including humans. Malignant tumors can cause immunosuppression and increase the risk of toxoplasmosis. Therefore, there is an urgent need to develop drugs that are both anti-tumor and anti-toxoplasmic in clinical practice. Currently, pyrimethamine and sulfadiazine sodium are the standard treatments for toxoplasmosis. Although this treatment is effective, up to 80% of patients may relapse if treatment is not extended, and about 40% of patients may stop treatment due to severe toxic side effects. According to statistics, almost all treatments involve sulfonamides, which often cause allergic reactions and bone marrow suppression. In addition, these drugs are only effective for acute infections, ineffective for chronic infections, and have no inhibitory effect on tumor cells. Therefore, it is necessary to provide a new drug for toxoplasmosis. Summary of the invention

[0004] In order to overcome the deficiencies of the prior art, the purpose of the present invention is to provide an application of a statin-type drug in the preparation of an anti-toxoplasmosis drug. Studies of the present invention show that statin-type drugs can not only prolong the survival time of mice infected with a strong strain of Toxoplasma gondii, but also significantly reduce the mortality rate of mice infected with a weak strain of Toxoplasma gondii, that is, statin-type drugs have obvious therapeutic effects on related diseases caused by a strong strain of Toxoplasma gondii or a weak strain of Toxoplasma gondii.

[0005] Based on the above research results, the present invention provides the following technical solutions:

[0006] A use of a statin-type drug in the preparation of an anti-toxoplasmosis drug, wherein the statin-type drug is a compound represented by formula I;

[0007]

[0008] In some embodiments, the anti-toxoplasmosis drug is used to treat toxoplasmosis caused by infection with the virulent strain RH or the weak strain ME49 of Toxoplasma gondii.

[0009] In some embodiments, the anti-toxoplasmosis drug is used to treat toxoplasmosis caused by Toxoplasma gondii infecting human foreskin fibroblasts (HFF).

[0010] In some embodiments, the dosage of the anti-toxoplasmosis drug is: 8 to 12 mg of the statin drug per kilogram of body weight per day.

[0011] In some embodiments, the active ingredient in the anti-toxoplasmosis drug is a compound of Formula I or a pharmaceutically acceptable salt, hydrate, or solvate thereof.

[0012] The pharmaceutically acceptable salts of the present invention refer to salts formed by the compound of formula I and non-toxic inorganic or organic acids. The inorganic acid may be hydrochloric acid, sulfuric acid, phosphoric acid, etc.; the organic acid may be acetic acid, propionic acid, lactic acid, oxalic acid, maleic acid, fumaric acid, malic acid, etc.

[0013] In some embodiments, the anti-toxoplasmosis drug further comprises a pharmaceutically acceptable excipient.

[0014] The pharmaceutically acceptable excipients described in the present invention refer to excipients that are pharmacologically and / or physiologically compatible with the subject and the active ingredient, including but not limited to: binders, fillers, disintegrants, pH adjusters, surfactants, adjuvants, diluents, preservatives, etc.

[0015] In some embodiments, the anti-toxoplasmosis drug is in the form of tablets, capsules, granules, drops, pills, etc.

[0016] In some embodiments, the anti-toxoplasmosis drug is administered orally, by gavage, or by subcutaneous injection.

[0017] In some embodiments, the anti-toxoplasmosis drug is administered to a human or non-human mammal. The non-human mammal of the present invention may be a mouse, a rat, a rabbit, a dog, a dog, etc.

[0018] The beneficial effects of the present invention are:

[0019] The present invention provides a new pharmaceutical use of an anti-toxoplasmosis drug. In vitro cell experiments have shown that the statin class of drugs has low toxicity to human foreskin fibroblasts, wherein the statin class of drugs has no toxicity to human foreskin fibroblasts below 0.1 μM, and the half inhibition concentration of the growth of human foreskin fibroblasts is 67.61 μM; at the same time, the statin class of drugs has a significant inhibitory effect on the Toxoplasma RH-GFP strain in human foreskin fibroblasts, and the half inhibition concentration is 10.73 nM. In vivo animal experiments have shown that the statin class of drugs can prolong the survival time of mice infected with the strong strain RH of Toxoplasma gondii, and can significantly improve the survival rate of mice infected with the weak strain ME49 of Toxoplasma gondii. Therefore, the statin class of drugs can be used for the preparation of drugs in the field of anti-toxoplasmosis, and similar drugs can be developed on this basis. BRIEF DESCRIPTION OF THE DRAWINGS

[0020] The accompanying drawings in the specification, which constitute a part of the present invention, are used to provide a further understanding of the present invention. The exemplary embodiments of the present invention and their descriptions are used to explain the present invention and do not constitute improper limitations on the present invention.

[0021] Figure 1 This is the cytotoxicity curve of the statin-type drugs on HFF in Example 1 of the present invention;

[0022] Figure 2 This is the inhibition curve of the statin-type drugs on Toxoplasma gondii in HFF cells in Example 2 of the present invention;

[0023] Figure 3 This is the survival curve of the statin-type drugs in Example 3 of the present invention on mice infected with the virulent strain RH Toxoplasma gondii;

[0024] Figure 4 This is the survival curve of the statin-type drugs in Example 3 of the present invention on mice infected with the attenuated strain ME49 of Toxoplasma gondii. DETAILED DESCRIPTION

[0025] In order to enable those skilled in the art to more clearly understand the technical solution of the present invention, the technical solution of the present invention will be described in detail below in conjunction with specific embodiments.

[0026] In the following examples, the statin drugs used are all compounds represented by Formula I;

[0027]

[0028] Example 1: Cytotoxicity test of statin drugs

[0029] 1. Material Description

[0030] Human foreskin fibroblasts (HFF), statins (MedChemExpress), CCK-8 kit (Biyuntian), dimethyl sulfoxide (DMSO) (Solebol), 96-well cell culture plates (Corning), DMEM medium (Gibco) and fetal bovine serum (FBS) (Gibco).

[0031] 2. Experimental Methods

[0032] HFF cell lines were cultured at 2-5 × 10 3The density of each well was inoculated in a 96-well microplate and cultured for 24 hours. After adding different concentrations of compounds (0.001μM, 0.01μM, 0.1μM, 1μM, 10μM, 100μM, 1000μM) to treat various cell lines for 48 hours, the CCK-8 kit was used to detect cell viability. The specific operation is as follows: Add 10μL CCK-8 reagent to each well to be tested and incubate in an incubator for 1-2 hours, and then use a microplate reader to measure the absorbance value (OD) at a wavelength of 450nm. Inhibition rate % = 1-(OD sample-OD background) / (OD blank-OD background) × 100%; Use prism statistical software to calculate IC through the inhibition curve 50 value.

[0033] Results Figure 1 The concentration below 0.1 μM was not toxic to HFF cells, and its half inhibitory concentration (IC 50 ) is 67.61μM.

[0034] Example 2: Experiment on the inhibition of intracellular Toxoplasma by statins

[0035] 1. Material Description

[0036] Human foreskin fibroblasts (HFF), statins (MedChemExpress), RH-GFP strain (donated by Shanxi Agricultural University), 24-well cell culture plates (Corning), DMEM medium (Gibco) and fetal bovine serum (FBS) (Gibco).

[0037] 2. Experimental Methods

[0038] Purified RH-GFP tachyzoites were inoculated into HFF cells at an MOI of 5. After 3 h, the extracellular parasites were washed off with PBS. Stastatin drugs were added at final concentrations of 1, 2, 4, 6, 8, 10, 25, and 50 nM, respectively. After 24 h, images were acquired using a fluorescence microscope (Carl Zeiss AG, Germany). The fluorescence area was counted using Image J software.

[0039] Results Figure 2 The EC50 of statins against RH-GFP tachyzoites of Toxoplasma gondii 50 ) is 10.73nM.

[0040] Example 3: Stastatin drugs have anti-Toxoplasma activity in vivo

[0041] 1. Material Description

[0042] C57BL / 6 mice, RH and ME49 insect strains (donated by Shanxi Agricultural University), statins (MedChemExpress), Tween-80 (Solebao), PEG-300 (Solebao), distilled water, DMSO (Biyuntian)

[0043] Solvent preparation: 10% DMSO, 40% PEG300, 5% Tween-80 and distilled water in volume percentage are mixed in sequence and each time the solvent is added, it must be mixed thoroughly before the next solvent is added.

[0044] 2. Experimental Methods

[0045] To evaluate the protective effect of statins against acute infection, mice in the infection group were intraperitoneally injected with 100 virulent RH tachyzoites or 500 virulent ME49 tachyzoites in sterile PBS. The control group was mock infected with sterile PBS. Treatment began 24 hours after infection, and the infected mice were divided into a solvent group (10% DMSO, 40% PEG-300, 5% Tween 80 and ddH2O, ig) and a statin group (10 mg / kg, 10% DMSO, 40% PEG-300, 5% Tween 80 and ddH2O, ig). The drug was administered once a day for 7 consecutive days. The mice were observed at 7 am, 12 noon and 7 pm, and the survival rate at each time point was recorded.

[0046] Results Figure 3 and Figure 4 .like Figure 3 As shown in the figure, all mice in the solvent + 100RH group died on the 10th day after infection. All mice in the statin + 100RH group died on the 12th day after infection. This indicates that the statin treatment group prolonged the survival time of mice. Figure 4 As shown, all mice in the solvent + 500ME49 group died on the 27th day after infection. 60% of mice in the statin + 500ME49 group survived, indicating that statins have a good protective effect on mice.

[0047] The above description is only a preferred embodiment of the present invention and is not intended to limit the present invention. For those skilled in the art, the present invention may have various modifications and variations. Any modification, equivalent replacement, improvement, etc. made within the spirit and principle of the present invention shall be included in the protection scope of the present invention.

Claims

1. Use of a statin-type drug in the preparation of an anti-toxoplasmosis drug, wherein the statin-type drug is a compound represented by formula I; 2. The use according to claim 1, characterized in that: The anti-toxoplasmosis drug is used for treating toxoplasmosis caused by infection with the strong strain RH or the weak strain ME49 of Toxoplasma gondii.

3. The use according to claim 1, characterized in that: The anti-toxoplasmosis drug is used for treating toxoplasmosis caused by Toxoplasma gondii infecting human foreskin fibroblasts.

4. The use according to claim 1, characterized in that: The dosage of the anti-toxoplasmosis drug is: 8 to 12 mg of the statin drug per kilogram of body weight per day.

5. The use according to claim 1, characterized in that: The active ingredient in the anti-toxoplasmosis drug is the compound shown in Formula I or a pharmaceutically acceptable salt, hydrate or solvate thereof.

6. The use according to claim 1, characterized in that: The anti-toxoplasmosis drug also includes pharmaceutically acceptable excipients.

7. The use according to claim 6, characterized in that: Pharmaceutically acceptable excipients include one or more of binders, fillers, disintegrants, pH adjusters, surfactants, adjuvants, diluents, and preservatives.

8. The use according to claim 1, characterized in that: The dosage form of the anti-toxoplasmosis drug is tablets, capsules, granules, drops or pills.

9. The use according to claim 1, characterized in that: The anti-toxoplasmosis drug is administered orally, by gavage or by subcutaneous injection.

10. The use according to claim 1, characterized in that: The anti-toxoplasmosis drug is administered to humans or non-human mammals.

Images