CN110269859B - Application of quinoxaline-1, 4-dioxide compound in resisting toxoplasma gondii infection - Google Patents
Application of quinoxaline-1, 4-dioxide compound in resisting toxoplasma gondii infection Download PDFInfo
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- A61K31/395—Heterocyclic compounds having nitrogen as a ring hetero atom, e.g. guanethidine or rifamycins
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- A61P31/00—Antiinfectives, i.e. antibiotics, antiseptics, chemotherapeutics
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Abstract
The invention relates to application of quinoxaline-1, 4-dioxide compounds in resisting Toxoplasma gondii infection, in particular to novel quinoxaline-1, 4-dioxide compounds, especially geometric isomers and pharmaceutically acceptable solvates and/or hydrates thereof, and application of a pharmaceutical composition containing the compounds in resisting Toxoplasma gondii (Toxoplasma gondii) infection. Experiments prove that the compound has the functions of protecting cells infected by Toxoplasma gondii and inhibiting replication and reproduction of Toxoplasma gondii, and has potential effects on resisting Toxoplasma gondii infection.
Description
Technical Field
The invention relates to a novel quinoxaline-1, 4-dioxide compound, in particular to a geometric isomer thereof, a pharmaceutically acceptable solvate and/or hydrate thereof, and a pharmaceutical composition containing the compound, and application of the compound in resisting Toxoplasma gondii (Toxoplasma gondii) infection.
Background
Quinoxaline-1, 4-dioxide compounds (formula I) were originally developed for use as antibacterial agents. In the early research of quinoxaline-1, 4-dioxide compounds, strong and efficient in-vitro activity against Treponema hyodysenteriae (Treponema hyodysenteriae) is widely concerned. To date, the compounds olaquindox, carba and the like are the main drugs for resisting the pathogens. For years, the quinoxaline-1, 4-dioxide compound is still highly valued by people, and a plurality of scholars continuously carry out various structural modifications on the quinoxaline compound, but most structural modifications retain the basic chemical skeleton of the quinoxaline-1, 4-dioxide compound. Therefore, the basic chemical framework of the quinoxaline-1, 4-dioxide compound has very important and irreplaceable functions. Our research is therefore also a search for new biological activities and uses based on this basic chemical framework.
At present, quinoxaline-1, 4-dioxide compounds commonly used in veterinary clinical practice in China comprise quinocetone, mequindox, olaquindox and the like. Mequindox (mequindox) and Quinocetone (QCT) belong to artificially synthesized quinoxaline 1, 4-dioxide drugs, are novel veterinary drugs developed by the Lanzhou veterinary drug research institute of the Chinese academy of agricultural sciences, are mainly used as novel quinoxaline antibacterial growth promoters, and are widely applied to animal growth promotion in the breeding industry in China to improve the conversion rate of animal feed. However, the application of the medicine in resisting Toxoplasma gondii is not reported.
Toxoplasma gondii (academic name: Toxoplasma gondii) is a protozoan of the order Coccidia, also known as Toxoplasma gondii or Toxoplasma gondii, and is a parasitic organism of the genus Toxoplasma. Toxoplasma is a worldwide parasitic protozoa, and its intermediate hosts are extensive, including reptiles, fish, insects, birds, mammals and other animals and humans; the final host is only the cat and the feline. Toxoplasmosis (toxoplasmosis) is a disease caused by toxoplasma which causes serious zoonosis. Toxoplasma infection in early-pregnancy women can lead to abortion, premature birth and even stillbirth, and can also cause newborn deformity and fetal eye complications through placenta. In people with immune deficiency or hypoimmunity (such as AIDS and tumor patients), Toxoplasma gondii can cause serious clinical symptoms (such as Toxoplasma gondii encephalopathy) and even death. In immunocompromised hosts, toxoplasma often forms a chronic infection causing irreversible central nervous system and vision damage. Toxoplasma, a protozoan of an opportunistic intracellular parasite, primarily elicits a cellular immune response in the host.
At present, the therapeutic drugs for toxoplasmosis are mainly sulfonamides, wherein sulfadiazine, sulfamethoxydiazine, sulfamethoxazole and synergist dimethoxybenzylamine and trimethoprim are used more frequently. Different degrees of sulfonamide resistant toxoplasma have been generated clinically today. In addition, macrolide antibiotics such as roxithromycin, spiramycin and azithromycin and lincomycin antibiotics such as clindamycin and milomycin also have effects of preventing and treating toxoplasmosis to different degrees. However, the medicaments have the defects of long treatment time, difficulty in thoroughly killing the insects, easy relapse of animals, large side effect and the like.
The quinoxaline-1, 4-dioxide compound of the invention belongs to quinoxaline compounds, and is different from the existing toxoplasmosis treatment drugs on the market in terms of chemical structure. And the existing research shows that the chemical structure of the sulfonamides is similar to that of p-aminobenzoic acid (PABA), and can compete with the PABA for dihydrofolate synthetase to influence the synthesis of dihydrofolate, so that the growth and the reproduction of bacteria, toxoplasma and the like are inhibited; macrolide antibiotics, which are the general term for a class of antibacterial drugs having a 12-16 carbon lactone ring in their molecular structure, act by inhibiting bacterial protein synthesis by blocking peptidyl transferase activity in the 50s ribosome of the pathogen. The action mechanism of the quinoxaline-1, 4-dioxide compound is still unclear at present, and the existing research shows that the compound can release 1, 4-dioxygen groups on the parent nucleus in a pathogen, so that a large number of free radicals are generated in the pathogen cell, the pathogen generates an oxidative stress reaction, and the oxidative reaction further causes the oxidation of macromolecular substances such as protein, nucleic acid and the like in the pathogen cell to lose activity so as to generate the biological activity of the compound.
Disclosure of Invention
The invention discovers that the quinoxaline-1, 4-dioxide compound (formula I) has the functions of protecting toxoplasma gondii infected cells and inhibiting the replication and reproduction of toxoplasma gondii through creative research, and has potential effect on resisting toxoplasma gondii infection. Further preferably, 2 compounds have remarkable effects and good effects, and can effectively reduce the number of live toxoplasma gondii, reduce cytopathy caused by toxoplasma gondii infection and reduce the level of the number of tachyzoites of the toxoplasma gondii at the concentration of mu g/mL.
Specifically, the invention provides a quinoxaline-1, 4-dioxide compound with a structure shown in a formula I, a geometric isomer thereof, and a pharmaceutically acceptable solvate or hydrate thereof:
wherein R1, R2, R3 may optionally be selected independently from each other from: h (hydrogen), OH (hydroxyl), Cl (chlorine), NO2(nitro), -OCH3(methoxy), -NC2H5(N, N-dimethylamino).
In addition, the invention also relates to a pharmaceutical composition containing the quinoxaline-1, 4-dioxide compound (formula I) and pharmaceutically acceptable solvates or hydrates thereof and a pharmaceutically acceptable carrier. The pharmaceutical composition can be used in various ways, such as oral tablets, capsules, powders, oral liquids, injections and transdermal preparations. Pharmaceutically acceptable carriers include diluents, fillers, disintegrants, wetting agents, lubricants, colorants, flavoring agents, or other conventional additives, according to conventional pharmaceutical practice. Typical pharmaceutically acceptable carriers include, for example, microcrystalline cellulose, starch, cross-linked polyvidones, povidone, polyvinylpyrrolidone, maltitol, citric acid, sodium dodecyl sulfate or magnesium stearate, and the like.
Another aspect of the invention relates to pharmaceutical compositions comprising at least one pharmaceutically acceptable carrier for a compound of the invention. The pharmaceutical composition may be prepared in various forms according to different administration routes.
Also provided is the use of a compound of formula (I) for combating Toxoplasma gondii (Toxoplasma gondii) infection in order to protect an animal from Toxoplasma attack.
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FIG. 1 is an in vitro observation of the proliferation inhibition of Toxoplasma gondii tachyzoite by drug screening, and the proliferation status of Toxoplasma gondii after 48h of partial drug action under a fluorescence microscope;
FIG. 2 is a graph showing the proliferation state of Toxoplasma gondii after DL-QCT (No. 4) acts on Toxoplasma gondii tachyzoite in vitro and DL-QCT acts for 48 hours under a fluorescence microscope;
FIG. 3 is a graph showing the proliferation state of Toxoplasma gondii in vitro by JQ-QCT (No. 8) on Toxoplasma gondii tachyzoite and 48h after JQ-QCT action under a fluorescence microscope
Detailed Description
The following examples are illustrative of preferred embodiments of the present invention and are not to be construed as limiting the invention in any way.
The starting materials or reagents used in the examples are, unless otherwise specified, commercially available.
The experimental procedures, in which specific conditions are not noted in the following examples, are generally carried out under conventional conditions or conditions recommended by the manufacturers.
The Toxoplasma gondii standard strength strain (RH strain) used in the embodiment has biological characteristics representing the characteristics of Toxoplasma gondii in China.
In the examples, the lytic agent is dimethyl sulfoxide, which has the function of increasing solubility and permeability of the drug and has no toxic effect on experimental cells or animals.
The structural formula, the name or the number of the quinoxaline-1, 4-dioxide compound (formula I) related in each embodiment of the invention are shown in the table 1.
TABLE 1 substituent, name code or numbering of quinoxaline-1, 4-dioxide compound (formula I)
Example 1: preparation of quinoxaline-1, 4-dioxide compound for inhibiting toxoplasma gondii proliferation in Vero cells
1. Synthesis of benzofurazan monoxide
In a 250ml four-neck flask, about 0.2mol of o-nitroaniline (molecular weight: 138), 0.28mol of KOH are dissolved in 250ml of 95% ethanol, the mixture is stirred until the o-nitroaniline is dissolved, 250ml of NaClO solution is dropwise added under ice bath, yellow precipitate is gradually precipitated, the reaction is monitored by TLC, and a developing agent PE: EA (petroleum ether: ethyl acetate) ═ 5: 1. After 8h, after the reaction is finished, adding water for filtration, washing with water to obtain yellow precipitate, recrystallizing with 70% ethanol to obtain 22.64g of product with the yield of 82.35%, wherein the reaction formula is
Electrospray full-scan mass spectrometry, excimer [ M + H ]]+ m/z is 137. mp: 68.8-70.5 ℃.
2. Synthesis of 2-acetyl-3-methylquinoxaline-1, 4-dioxide (mequindox, MAQO)
Adding 0.05mol of benzofurazan into a 250ml four-mouth bottle, then adding 0.12mol of acetylacetone (molecular weight: 100), stirring at room temperature until the mixture is dissolved, carrying out water bath at room temperature, dropwise adding 22ml of triethylamine (molecular weight 101.19), heating to 40-45 ℃ for several minutes, stirring to gradually separate out yellow crystals (or standing until the mixture is naturally cooled and completely reacts for 10 hours to obtain a large amount of yellow crystals), monitoring the reaction by TLC, wherein a developing agent is PE: EA (petroleum ether: ethyl acetate) ═ 1: 1, carrying out suction filtration, washing by a small amount of ethanol, recrystallizing by 70% of ethanol to obtain 6.5g of product, the yield is 60%, and the reaction formula is that
Electrospray full-scan mass spectrometry, excimer [ M + H ]]+ m/z is 219. Namely mequindox (molecular weight: 218.21).
3. Synthesis of Compounds 1-9
In a 250ml four-necked flask,respectively heating and stirring 15-25 mmol of benzaldehyde, p-hydroxybenzaldehyde, o-hydroxybenzaldehyde, m-nitrobenzaldehyde, p-methoxybenzaldehyde, p-chlorobenzaldehyde, p-dimethylaminobenzaldehyde and the like, dissolving the above materials in 20-50 ml of absolute ethyl alcohol or methanol, then respectively adding 2.2g (about 10mmol) of 2-acetyl-3-methyl quinoxaline-1, 4-dioxide at 50-55 ℃, stirring until the mixture is dissolved, controlling the temperature to be 50-55 ℃, dropwise adding 2-10 ml of diethylamine, stirring at 50-55 ℃ for overnight after the addition is finished, gradually changing the color to yellow, brown or reddish brown and gradually precipitating with precipitates, carrying out monitoring and tracking reaction by Thin Layer Chromatography (TLC), adding water, stirring, filtering, washing with a small amount of ethanol to remove excessive p-hydroxybenzaldehyde, Recrystallizing reaction substrates such as m-nitrobenzaldehyde, solvents and impurities with 95% ethanol, and naturally drying to obtain compounds 1-9 respectively, namely: p-hydroxyquinocetone (DQ-QCT), o-hydroxyquinocetone (LQ-QCT), m-hydroxyquinocetone (JQ-QCT), m-nitroquinocetone (JX-QCT), p-nitroquinocetone (DX-QCT), p-methoxyquinocetone (DKY-QCT), p-chloroquinocetone (DL-QCT) and p-dimethylaminoquinocetone (DJA-QCT), wherein the reaction formula is
Wherein R is hydrogen, p-hydroxyl, o-hydroxyl, m-nitro, p-methoxyl, p-chloro and p-dimethylamino.
4. Identification of Compounds 1-9
The synthesized compounds respectively take DMSO as a solvent, perform hydrogen spectrum analysis on a Bruker 400MHz type nuclear magnetic resonance instrument, perform infrared spectrum analysis in a KBr tablet sample preparation mode, perform ultraviolet spectrum scanning analysis by taking chromatographic grade methanol as a solvent, and perform electrospray ionization source (ESI) mass spectrum analysis by taking acetonitrile as a solvent. Meanwhile, the content of each drug is measured by adopting a high performance liquid chromatography, and the content of all tested compounds is ensured to be more than 98%.
Example 2: activity identification for inhibiting toxoplasma gondii proliferation in Vero cells
The implementation steps are as follows:
1. preparation of a drug stock solution:
50mg of each of the above drugs (quinoxaline-1, 4-dioxide compound (formula I), No. 1 to 9) was dissolved in 10mL of dimethyl sulfoxide (DMSO) to obtain a stock solution of 5mg/mL, and the stock solution was stored at 4 ℃. The working solution of the drug is diluted by 10 percent of FBS DMEM medium before use, and is subjected to filtration sterilization, so that the final concentration of the drug contains DMSO (dimethyl sulfoxide) not more than 1.0 percent, and the working solution of the drug is used for toxicity test of Vero cells and inhibitory effect test of Toxoplasma gondii proliferation.
2. Vero cells:
vero cells were kept in an animal parasitology-intensive open laboratory in Shanghai veterinary research institute of agricultural rural area, China academy of agricultural sciences. At 37 ℃ with 5% CO2Culturing in an incubator, culturing in 10% FBS DMEM medium, digesting with pancreatin, transferring 2-3 generations until cells are stable, and counting with a cell counter and adjusting the number for later use. Fetal bovine serum, DMEM medium, diabody (penicillin, streptomycin antibiotic) were purchased from Gibco, usa, and 24-well cell culture plates and cell culture bottles were purchased from Corning, usa.
3. Toxoplasma tachyzoite:
toxoplasma RH/GFP strain tachyzoites were given by the university of livestock products, national zone of Japan, and were subcultured in Vero cells. The Toxoplasma gondii RH/GFP strain tachyzoite is a worm body expressing green fluorescent protein, and because the green fluorescent protein gene is transfected into the worm body and integrated into the genome of the worm body, the Toxoplasma gondii can be expressed along with the replication of the worm body, and the Toxoplasma gondii strain tachyzoite is not lost after passage, so that the Toxoplasma gondii strain tachyzoite is easier to observe after infection.
4. Cytotoxicity test:
the MTT method was used to divide the experiment into three groups, a blank control group (no Vero cells and drug added), a test control group (Vero cells only added), and a drug test group (Vero cells added and drug at various concentrations). Taking 96-well plate, adding 10% FBS DMEM medium (DMSO is not more than 1.0%) containing drugs with final concentration of 50, 20, 10, 5, 2.5, 1.0 μ g/mL when the cell growth is more than 80%, and adding 5% CO at 37 deg.C2After 24 hours of culture in an incubator, 20 mu L of MTT working solution with the concentration of 5mg/mL is added into each hole, after 4 hours, the supernatant is discarded, 200 mu L of DMSO is added for 0.5 hour,the microplate reader reads the OD at a wavelength of 570nm (average of 5 replicate wells). The drug concentration in which the cell survival rate is higher than 80% is selected for the research of the inhibition effect of the proliferation of the anti-toxoplasma tachyzoite.
5. And (3) worm body culture:
culturing Vero cell and toxoplasma in 5% CO237 ℃ CO2An incubator. 8% fetal bovine serum and double antibody are added into the culture solution. When the cells grow to be more than 80% of confluency, the cells are digested by pancreatin, collected and passed to a new cell culture bottle, and after 48 hours of culture, the growth condition of the cells is observed under a mirror. The worm bodies are inoculated to cells paved in a 70% cell culture bottle, after the cells are cultured for 72 hours, the worm bodies and the cells are scraped by a cell scraper, a needle of a 26G (0.45#) syringe repeatedly sucks and blows 3 times to lyse the cells, a 5 mu m needle filter filters and removes cell debris to purify the worm bodies, the worm bodies are centrifuged at 2000rpm for 10min, the worm body sediment is resuspended by a DMEM medium containing 10% FBS, and the green fluorescent spot worm bodies are observed and counted by a fluorescence microscope.
6. Toxoplasma gondii proliferation resistance test:
vero cells were passaged into 24-well cell culture plates, 800 Vero cells per well in a volume of 1mL, and after 24h of culture, 2000 purified RH/GFP Toxoplasma gondii tachyzoites in a volume of 100. mu.L were added per well. After the two are cultured for 2h, the culture solution in each well is aspirated, and the Vero extracellular tachyzoites are removed by washing twice with the cell culture solution. Adding prepared drug working solution, and setting drug group, positive control group (PBS group) and cell blank control group. The drug groups were infected with somatic insect cells and given the corresponding drugs, with final concentrations of the drugs selected at 12, 3 and 0.6 μ g/mL (DMSO not more than 1.0%): the positive control group is infected worm somatic cells which are not dosed but are given PBS with corresponding volume; the cell blank group was a cell culture that was not administered either drug or somatic infection. Each experimental group was set up in 3 duplicate wells and the experiment was repeated three times. Observing the proliferation condition and morphological change of the toxoplasma gondii tachyzoite at 48h and 72h after administration under a fluorescence microscope, counting the number of live insects according to the green fluorescence points (one fluorescence point represents one insect body because the insect body proliferates in an exponential mode, and one fluorescence point is counted as one insect body during counting), and analyzing the reduction rate of the drug to the toxoplasma gondii:
the reduction rate is (average infection number of positive control group polypide-average infection number of drug group polypide) ÷ average infection number of positive control group polypide is multiplied by 100%
7. Statistical analysis:
the counts were expressed as mean. + -. standard deviation (ā. + -.s), the variances were compared and a t-test was performed, and P < 0.05 indicated that the differences were statistically significant.
8. Results
Cytotoxicity
The MTT method detects that each drug has certain toxicity to Vero cells in a soluble range. The drug concentrations of the drugs 1 to 9 for the cell viability of more than 80% are mostly about 10 mug/mL. Therefore, 12. mu.g/mL was selected as the highest drug concentration for the anti-Toxoplasma test to perform the efficacy test.
In vitro observation of drug screening for inhibition of toxoplasma tachyzoite proliferation
After administration for 48h and 72h, the proliferation and morphological change of toxoplasma tachyzoites in each experimental group were observed by an inverted fluorescence microscope. After 48h, the change is obvious, and the positive control group (PBS group) Toxoplasma gondii tachyzoite can be seen under a fluorescence microscope to proliferate in Vero cells in a large amount, and the shape is approximately normal. The PBS group toxoplasma tachyzoites proliferate actively and are represented as a plurality of fluorescent spots; compared with the prior art, the Toxoplasma gondii tachyzoite in Vero cells can be seen under each low-concentration drug group mirror, and the Toxoplasma gondii tachyzoite has normal shape and good activity. But with the increase of the concentration of the medicine, the toxoplasma tachyzoite density intensity of each administration group is weakened and reduced to different degrees; meanwhile, a phenomenon of drug crystal precipitation (mainly quinoxaline mother nucleus dideoxy compounds with numbers 1-9) is seen in certain high-concentration drug groups, which indicates that the drugs have poor solubility in a toxoplasma culture environment, and further observation shows that the compounds have poor growth inhibition effect on the toxoplasma. 72h further observation shows that in the drug group with the inhibiting effect in 48h observation, the toxoplasma tachyzoite density intensity is further weakened, and a large number of dead insect bodies without activity appear along with the continuous effect of the drugs and the consumption of nutrient substances. The results are shown in FIG. 1.
Drug counts for Toxoplasma inhibition
The live insect counts were performed on each drug group (compounds numbered 1-9 and 2-methylquinoxaline-1, 4-dioxy, olaquindox) with significant inhibitory effect, and the results are shown in table 2. And 48h, compared with a positive control group, the number of the live insects in a plurality of drug groups is reduced, and the number of the live insects is significant along with the increase of the concentration. Wherein, compared with other medicines, the number of the live insects in the medicine groups of the numbers 4 and 8 is obviously reduced at the medicine dose of 0.6 mu g/mL. The drug doses of numbers 1,4 and 8 are all significantly reduced above 3 mug/mL. And 72h, the number of live insects in each experimental group is obviously reduced, and a large number of dead insects appear. The drugs numbered 1,4 and 8 are indicated to be better able to inhibit toxoplasma tachyzoite proliferation in vitro at a certain dose for continued validation studies.
TABLE 2 Toxoplasma results in Vero cells against each of the screened drugs
Note: each group was provided with 3 replicates, 3 replicates. P < 0.05 compared to PBS group; denotes P < 0.01.
And (4) conclusion:
the experiments show that the synthesized quinoxaline-1, 4-dioxide compound (formula I, number 1-9) and quinamine alcohol have certain effect of inhibiting the reproduction of toxoplasma gondii in vitro, but the parent nucleus deoxyribose compound of the quinoxaline-1, 4-dioxide compound (formula I, number 1-9) has poor solubility and almost no insect resistance effect. The mother nucleus 1, 4-dioxy of the quinoxaline-1, 4-dioxy compound is an important functional group for resisting toxoplasma gondii, and the effect of resisting toxoplasma gondii can be increased through structural modification, so that the DL-QCT and JQ-QCT compounds are screened to have better toxoplasma gondii resisting effect, but the effect of the DL-QCT and JQ-QCT compounds needs to be further researched and confirmed.
Example 3: experiment for inhibiting toxoplasma gondii proliferation in vitro by using DL-QCT and JQ-QCT
The experimental preparation and the specific operation were as described in example 2, except that the relevant experiments were performed for DL-QCT, JQ-QCT (numbers 4, 8, respectively).
1. Toxoplasma gondii proliferation resistance test:
vero cells were passaged into 24-well cell culture plates, 800 Vero cells per well in a volume of 1mL, and after 24h of culture, 2000 purified RH/GFP Toxoplasma gondii tachyzoites in a volume of 100. mu.L were added per well. After the two are cultured for 2h, the culture solution in each well is aspirated, and the Vero extracellular tachyzoites are removed by washing twice with the cell culture solution. Adding prepared medicinal working solution, and setting administration group and blank control group (PBS group). Selecting drugs with final concentrations of 0.5, 1, 2 and 4 mug/mL (DMSO is not more than 1.0 percent) respectively; the blank control group was given only the corresponding volume of PBS. Each experimental group was set up in 3 duplicate wells and the experiment was repeated three times. Observing the proliferation condition and the morphological change of the toxoplasma tachyzoite at 48h and 72h after the administration under a fluorescence microscope, counting the number of live insects according to green fluorescence points (one fluorescence point represents one insect body because the insect body proliferates in an exponential mode, and one fluorescence point is counted as one insect body during counting), and analyzing the insect reduction rate of the drug to the toxoplasma:
the reduction rate is (average infection number of control polypide-average infection number of test polypide) ÷ average infection number of control polypide multiplied by 100%
2. Statistical analysis:
the counts were expressed as mean. + -. standard deviation (ā. + -.s), the variances were compared and a t-test was performed, and P < 0.05 indicated that the differences were statistically significant.
3. Results
In vitro observation of Toxoplasma tachyzoite proliferation of each group
After administration for 48h and 72h, the proliferation and morphological change of toxoplasma tachyzoites in each experimental group were observed by an inverted fluorescence microscope. After 48h, the change is obvious, the toxoplasma tachyzoite of the blank control group is greatly increased in Vero cells under a fluorescence microscope, and the shape is approximately normal. The toxoplasma tachyzoite in the blank control group proliferates actively and is represented as a plurality of fluorescent spots; compared with the Vero cell, toxoplasma tachyzoite in Vero cells can be seen in each drug group with the concentration of 0.5 mu g/mL under a microscope, and the Vero cell is normal in shape and good in activity. But with the increase of the concentration of the medicine, the density and the intensity of the toxoplasma tachyzoite of each administration group are weakened and reduced to different degrees; among them, the effect is most remarkable in DL-QCT. And 72h, the density and the strength of toxoplasma tachyzoites of each experimental group are further weakened, and a large number of dead insects without activity appear along with the continuous action of the medicine and the consumption of nutrient substances. The results are shown in FIGS. 2 and 3.
The drug count the inhibitory effect on Toxoplasma gondii
The number of live insects in each drug experiment group was counted and shown in Table 3. 48h, compared with the non-administration group, the number of the live insects in each administration group is reduced, and the live insects are significant along with the increase of the concentration. Wherein, the number of the living insects in the group administered with the DL-QCT medicament is remarkably reduced by more than 2 mug/mL of medicament dose. And 72h, the number of live insects in each experimental group is obviously reduced, and a large number of dead insects appear. It is shown that both the drugs Nos. 4 and 8 are better able to inhibit Toxoplasma tachyzoite proliferation in vitro at a certain dosage.
TABLE 3 counting results of DL-QCT, JQ-QCT against Toxoplasma gondii in vitro
Note: each group was provided with 3 replicates, 3 replicates. P < 0.05 compared to PBS group; denotes P < 0.01.
And (4) conclusion:
through repeated verification tests of in vitro Toxoplasma gondii propagation, the DL-QCT and DQ-QCT have good effects of inhibiting Toxoplasma gondii propagation in vitro, but the insect-resistant effect of the DL-QCT is optimal.
Claims (2)
1. The application of the quinoxaline-1, 4-dioxide compound with the general formula I in preparing the medicine for resisting Toxoplasma gondii infection is provided, the quinoxaline-1, 4-dioxide compound with the general formula I can effectively inhibit the number of invading cells of Toxoplasma gondii (Toxoplasma gondii) and effectively reduce the cytopathic capability of Toxoplasma gondii, thereby having the effect of resisting Toxoplasma gondii infection,
wherein the content of the first and second substances,
r1 is H (hydrogen), R2 is H (hydrogen), R3 is Cl (chloro);
r1 is H (hydrogen), R2 is OH (hydroxyl), and R3 is H (hydrogen).
2. The use of claim 1, wherein the medicament further comprises a pharmaceutically acceptable carrier.
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