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Application of quinoline compound broxaldine in inhibition of toxoplasma gondii activity

A technology of broxadin and compounds, applied in the application field of quinoline compound broxadin inhibiting the activity of Toxoplasma gondii

Pending Publication Date: 2021-07-06
LANZHOU INST OF ANIMAL SCI & VETERINARY PHARMA OF CAAS
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

[0005] Broxaldine, a quinoline compound, can be used in combination with dibromohydroxyquine to treat leprosy and giardiasis, but no studies have shown that broxaldine has an active inhibitory effect on Toxoplasma gondii

Method used

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  • Application of quinoline compound broxaldine in inhibition of toxoplasma gondii activity
  • Application of quinoline compound broxaldine in inhibition of toxoplasma gondii activity
  • Application of quinoline compound broxaldine in inhibition of toxoplasma gondii activity

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0028] A kind of quinoline compound broxadin is applied to the inhibition of Toxoplasma gondii RH-2F tachyzoites, and is verified by the following methods:

[0029] Use biochemiluminescent method to measure the inhibitory effect of broxadin to Toxoplasma gondii RH-2F tachyzoites, comprising the following steps:

[0030] Toxoplasma gondii type I virulent strain RH-2F expressing β-galactosidase was selected for growth inhibition assay. Harvest fresh and vigorous RH-2F tachyzoites from Vero cells, count them with a hemocytometer, and adjust the concentration of RH-2F to 1×105 / mL. Each concentration of broxadin was prepared in this way, and added to a white 96-well plate, 0.25% DMSO was used as the control group, and the group containing only the culture medium was used as the blank group. Add Beta- As for the detection reagent, after 30 minutes of action, the luminescence value was detected in a photometer to calculate the survival rate of RH-2F.

[0031] result:

[0032] Th...

Embodiment 2

[0034] The anti-invasion application of a quinoline compound broxadin to Toxoplasma gondii RH is verified by the following method: use indirect immunofluorescence to measure the anti-invasion effect of broxadin on toxoplasma gondii RH, comprising the following steps:

[0035] 1 × 105 / mL RH tachyzoites containing various concentrations of active compounds were inoculated in monolayer Vero cells after being treated in an incubator for 1 h, 0.25% DMSO was used as a negative control group, and 10 μg / mL pyrimethamine (PYR ) as a positive control. After 2 hours of invasion, try to wash away the non-invading tachyzoites with PBS. After fixation, antigen retrieval and blocking, the extracellular T. gondii was labeled with mouse anti-T. gondii monoclonal antibody, and the secondary antibody was goat anti-mouse IgG H&L (Alexa 647), after permeabilization with 0.2% TritonX-100, the rabbit anti-toxoplasma polyclonal antibody was selected to label the intracellular Toxoplasma gondii, and...

Embodiment 3

[0040] The anti-proliferation application of a quinoline compound broxadin to RH or Pru ​​in Toxoplasma gondii cells is verified by the following method, using Giemsa staining to measure the anti-proliferation effect of broxadin on intracellular RH and Pru, including the following steps :

[0041] Inoculate Vero cells in a 6-well plate. When the cell monolayer is formed, inoculate RH and Pru tachyzoites. The inoculation amount of RH tachyzoites is 2×105 / well, and the inoculation amount of Pru tachyzoites is 3×105 / well. After culturing for 8 hours, discard the culture medium, thoroughly wash with PBS to remove uninfected parasites, add broxadin 4 μg / mL medium for incubation, and set up negative control group (Toxoplasma gondii RH) and blank control group (cell ) and positive control group (pyrimethamine 10 μg / mL). After incubation for 24h, 48h and 72h, observe the growth status of the parasites and the state of the cells, take them out, stain them with Giemsa, and take picture...

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Abstract

The invention relates to the technical field of toxoplasmosis, and discloses application of quinoline compound broxaldine in inhibition of toxoplasma gondii activity, application of broxaldine to an inhibitor of toxoplasma gondii RH-2F tachyzoites, application of broxaldine to an anti-invasion preparation of toxoplasma gondii RH, application of broxaldine to an anti-proliferation preparation of RH or Pru in toxoplasma gondii cells and application of broxaldine to drugs for treating toxoplasmosis. The quinoline compound broxaldine disclosed by the invention not only has a remarkable inhibition effect on a toxoplasma gondii I type virulent strain RH, but also has a remarkable inhibition effect on intracellular proliferation of a toxoplasma gondii II type attenuated strain Pru, has a certain treatment effect on acute toxoplasmosis caused by RH and Pru under a certain dosage, and can be used for remarkably inhibiting weight loss of diseased mice, delaying the disease time and reducing the death rate.

Description

technical field [0001] The invention relates to the technical field of toxoplasmosis, in particular to an application of a quinoline compound broxadin to inhibit the activity of toxoplasma gondii. Background technique [0002] Toxoplasma belongs to the subphylum Apicomplexa, class Sporozoasida, subclass Coccidiasina, order Eucoccidiorida, suborder Eimeriina, family Toxoplasmatidae , Toxoplasma. The genus Toxoplasma has only one species, Toxoplasma gondii, commonly known as Toxoplasma gondii. However, according to different regions, different hosts, different virulence, life history and development time, etc., it can be divided into three different genotypes, type I (RH and GT-1 strains), type II (ME49 and PRU strains) and type III (CEP and VEG strains). The three genotype strains have significantly different virulence phenotypes, among which the type I strain is a virulent strain with a high lethality rate, mainly for acute infection; while the type II and III strains are...

Claims

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Application Information

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IPC IPC(8): A61K31/438A61K47/44A61P33/02
CPCA61K31/438A61K47/44A61P33/02
Inventor 张继瑜邱燕华王玮玮翟斌涛周绪正杨枭荣李冰程富胜魏小娟尚小飞
Owner LANZHOU INST OF ANIMAL SCI & VETERINARY PHARMA OF CAAS
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