Application of 4-aminoquinazoline in female mosquito reproduction prevention and control

Abstract

The invention discloses application of 4-aminoquinazoline in female mosquito reproduction control, which is characterized in that 4-aminoquinazoline powder is dissolved in a dimethyl sulfoxide (DMSO) solvent and then is proportionally mixed with a glucose solution, and ovarian development of female mosquitoes is inhibited by a method of feeding female adult mosquitoes in mouths, so that the egg laying amount of the female mosquitoes is reduced. The new application of the existing drug 4-aminoquinazoline is developed, the quantity of mosquito populations is controlled by directly inhibiting the reproductive potential energy of female adult mosquitoes, and a new solution is provided for the current situation that current mosquito chemical insecticides are generally resistant in China.

Description

Technical Field

[0001] The invention belongs to the technical field of mosquito vector control, and particularly relates to application of 4-aminoquinazoline in female mosquito reproduction control. Background Art

[0002] Mosquitoes are blood-sucking insects that can mix the pathogens they carry into their saliva and infect the host during the blood-sucking process, spreading a variety of mosquito-borne diseases such as malaria, dengue fever, Zika virus disease, chikungunya fever, Japanese encephalitis, and filariasis. Mosquito-borne diseases account for about 17% of all infectious diseases in the world, killing more than 700,000 people each year, seriously threatening human health and posing a huge challenge to global public health. my country has a vast territory, a suitable climate, rich vegetation, a wide variety of mosquito species, and a high population density. Mosquito-borne diseases may break out at any time, so it is urgent to strengthen the prevention and control of mosquito-borne diseases.

[0003] Since most mosquito-borne diseases lack vaccines and targeted special drugs, mosquito control has become an important means of controlling mosquito-borne diseases. At present, mosquito control strategies mainly include the following four methods: 1. Environmental control: by transforming the land, water or vegetation environment to eliminate mosquito breeding grounds; 2. Physical control: mainly through the use of screen doors, screen windows, mosquito nets, mosquito hats and insect repellent nets to reduce the contact between hosts and mosquitoes; 3. Biological control: in recent years, Bacillus thuringiensis (Bt), Bacillus sphaericus (Bs) and Wolbachia are more common, but the cost is high and it is difficult to implement on a large scale; 4. Chemical control: by spraying chemical insecticides in space to eliminate indoor and outdoor mosquitoes. Currently, common chemical insecticides mainly include organophosphates, carbamates and pyrethroids. With the advantages of high efficiency, rapidity and convenience, chemical control has always been the main method in the comprehensive mosquito control strategy. The long-term and widespread use of traditional chemical insecticides has led to the emergence and development of mosquito resistance, so it is urgent to develop new insecticides that have no cross-resistance with traditional chemical insecticides. The high reproductive capacity of female mosquitoes is conducive to maintaining the population and thus playing the role of efficient disease vectors. Therefore, reducing the reproductive potential of female mosquitoes is considered to be a mosquito control strategy with broad application prospects.

[0004] 4-aminoquinazoline is an off-white powdered organic compound that is slightly soluble in water and soluble in organic solvents. Its preparation method is to reduce nitroquinazoline with a reducing agent (such as sodium sulfite, sodium bisulfite or sulfoxide), and then obtain it by crystallization purification. The structure of 4-aminoquinazoline derivatives is similar to chloroquine, with few side effects, and can be used to treat and prevent malaria. The exact mechanism of its antimalarial effect is still unclear. It may be that 4-aminoquinoline antimalarial drugs accumulate continuously in the acidic organelles of malarial parasites, and inhibit the growth of malarial parasites by maintaining a high concentration; or 4-aminoquinoline antimalarial drugs will prevent cellular functions such as RNA synthesis and DNA replication, thereby leading to the death of malarial parasites. Nowadays, many new derivatives have been developed with 4-aminoquinazoline as the core, which can inhibit the activity of kinases such as vascular endothelial growth factor receptor-2 (VEGFR-2) tyrosine kinase and phosphatidylinositol-3 (PI3Ka), and have been widely used in the treatment of tumor targets such as lung cancer, breast cancer, colon cancer, and prostate cancer.

[0005] The search did not find any application of 4-aminoquinazoline in the field of mosquito control. Summary of the invention

[0006] In view of the shortcomings of the existing technical problems, the purpose of the present invention is to provide an application of 4-aminoquinazoline in female mosquito reproductive control. The present invention proposes a new use of 4-aminoquinazoline, provides a new strategy for existing mosquito vector control, and helps to solve the problem of drug resistance of traditional chemical insecticides.

[0007] The technical solution adopted by the present invention to solve the technical problem is:

[0008] In a first aspect, the present invention protects the use of 4-aminoquinazoline in mosquito control.

[0009] In a second aspect, the present invention protects the use of 4-aminoquinazoline in the preparation of drugs or insecticides for mosquito control.

[0010] In a specific embodiment, the molecular formula of the 4-aminoquinazoline is C8H7N3, and the structural formula is

[0011] In a specific embodiment, 4-aminoquinazoline in the drug or insecticide can inhibit the reproduction of female mosquitoes, specifically by inhibiting the development of the ovaries of female adult mosquitoes and reducing their egg-laying capacity.

[0012] In a specific embodiment, the mosquito is Culex pipiens of the genus Culex of the family Culicidae in the order Diptera.

[0013] In a specific embodiment, the drug or pesticide has 4-aminoquinazoline as an active ingredient, and the effective concentration is 25 to 400 mg / L.

[0014] In a specific embodiment, as the concentration of 4-aminoquinazoline increases, the inhibitory effect on the reproductive ability of female mosquitoes becomes more significant.

[0015] In a specific embodiment, the drug or insecticide is mixed with DMSO as a solvent and 1-10% preferably 5% glucose solution in a proportion, and is fed to the emerged female mosquitoes by oral feeding.

[0016] In a specific embodiment, the drug or insecticide can be prepared by the following method: weigh 4-aminoquinazoline off-white powder, add DMSO, and stir to dissolve to prepare a mosquito killer / insecticide.

[0017] In a specific embodiment, the mosquito repellent / insecticide uses 4-aminoquinazoline as the main active ingredient, and DMSO is used as a solvent to prepare 4-aminoquinazoline into a 10 mg / mL stock solution, which is then diluted with 5% glucose solution to a solution with a concentration of 25 to 400 mg / L when used, such as 25 mg / L, 50 mg / L, 100 mg / L, 200 mg / L, 400 mg / L solutions, etc.

[0018] Among them, the preparation of 10 mg / mL 4-aminoquinazoline stock solution: weigh 50 mg of 4-aminoquinazoline powder and put it into a pre-sterilized beaker, add DMSO to the beaker and place it on a magnetic stirrer to stir and dissolve for 30 minutes, and finally make the volume to 5 mL to prepare 5 mL of 10 mg / mL stock solution, which is divided into aliquots and frozen for later use.

[0019] Subsequently, the 10 mg / mL 4-aminoquinazoline stock solution was diluted to different working concentrations using a high-pressure sterilized 5% glucose solution as a solvent.

[0020] In a specific embodiment, the drug or insecticide also includes other drugs or agents for controlling mosquitoes or other related active ingredients.

[0021] In a second aspect, the present invention protects a method for controlling mosquitoes, wherein 4-aminoquinazoline or a drug or insecticide containing 4-aminoquinazoline is applied to a mosquito-borne disease epidemic area by way of sugar bait.

[0022] Beneficial effects: Compared with the prior art, the present invention has the following advantages:

[0023] (1) The present invention uses the existing drug 4-aminoquinazoline for mosquito control, thus opening up a new use for the drug.

[0024] (2) Sugar water is an important food for the growth and development of adult mosquitoes and the reproduction of offspring. The sugar bait of the present invention is made of 4-aminoquinazoline, DMSO and glucose water. The raw materials of the sugar bait formula are simple and easy to obtain, the preparation process is simple, and the bait attracts mosquitoes to eat.

[0025] (3) The medicament prepared from 4-aminoquinazoline, DMSO and glucose solution of the present invention, 400 mg / L, 200 mg / L, 100 mg / L, 50 mg / L and 25 mg / L, can have a good inhibitory effect on mosquito reproduction. At the same time, with the increase of the concentration used, the ovarian volume of female mosquitoes is significantly reduced, the number of follicles in the ovaries is significantly reduced, and the egg laying amount decreases more significantly, but there is no significant difference in the egg hatching rate.

[0026] (4) The 4-aminoquinazoline of the present invention reduces the reproduction of female mosquitoes and thus suppresses the population, thereby solving the problem of drug resistance of traditional chemical mosquito killers and can be used as an auxiliary agent and adjuvant of a new type of composite mosquito killer or other mosquito killers. BRIEF DESCRIPTION OF THE DRAWINGS

[0027] Figure 1 The effects of different concentrations of 4-aminoquinazoline on the ovarian development of female mosquitoes before egg laying (48h PBM) are shown in Figure 2. Figure 1 Figure A is the WT group. Figure 1 Figure B is the Control group. Figure 1 Figure C shows the group with a 4-aminoquinazoline concentration of 25 mg / L. Figure 1 Figure D shows the group with a 4-aminoquinazoline concentration of 50 mg / L. Figure 1 Figure E shows the group with a 4-aminoquinazoline concentration of 100 mg / L. Figure 1 Figure F shows the group with a 4-aminoquinazoline concentration of 200 mg / L. Figure 1 Figure G shows the group with 4-aminoquinazoline concentration of 400 mg / L (N=10, *P<0.05, **P<0.01, ***P<0.001, ****P<0.0001).

[0028] (Magnification 2.5x10).

[0029] Figure 2 The effect of different concentrations of 4-aminoquinazoline on the number of follicles in female mosquitoes before egg laying (48h PBM). (N=10,

[0030] *P<0.05, **P<0.01, ***P<0.001, ***P<0.001, ****P<0.0001).

[0031] Figure 3 The effects of different concentrations of 4-aminoquinazoline on the egg raft morphology of female mosquitoes are shown in Figure 2. Figure 3Figure A is the WT group. Figure 3 Figure B is the Control group. Figure 3 Figure C shows the group with 4-aminoquinazoline concentration of 25 mg / L. Figure 3 Figure D shows the group with a 4-aminoquinazoline concentration of 50 mg / L. Figure 3 Figure E shows the group with a 4-aminoquinazoline concentration of 100 mg / L. Figure 3 Figure F shows the group with a 4-aminoquinazoline concentration of 200 mg / L. Figure 3 Figure G shows the group with 4-aminoquinazoline concentration of 400 mg / L (N=30, *P<0.05, **P<0.01, ***P<0.001, ****P<0.0001). (Magnification 2.5x10).

[0032] Figure 4 The effect of different concentrations of 4-aminoquinazoline on the egg-laying capacity of female mosquitoes. (N=30, *P<0.05, **P<0.01,

[0033] ***P<0.001, ****P<0.0001).

[0034] Figure 5 The effect of different concentrations of 4-aminoquinazoline on the hatching rate of mosquito eggs (N=30, ns means no statistical difference, *P<0.05, **P<0.01, ***P<0.0001, ****P<0.0001). DETAILED DESCRIPTION

[0035] The technical scheme and technical effects of the present invention are further described and explained below in conjunction with specific embodiments, but the present invention is not limited to these specific embodiments. The reagents or instruments used without indicating the manufacturer are regarded as conventional products that can be purchased on the market.

[0036] 1. Mosquitoes and feed:

[0037] The Culex pipiens used in the present invention was collected in Tangkou County, Shandong Province, and then subcultured in this laboratory, and was not exposed to any pesticides or toxic substances before the experiment. The breeding conditions are as follows: temperature 28-30°C, relative humidity 50%-70%, and daily light-dark ratio of 14h:10h.

[0038] Larvae from 1 to 4 instars were fed with high-pressure mouse feed powder, and adult mosquitoes were fed with 5% glucose solution.

[0039] After mating, adult male and female mosquitoes were passaged by sucking fresh blood from ICR mice.

[0040] 2. Preparation of 4-aminoquinazoline mother solution

[0041] (1) 4-Aminoquinazoline (CAS: 15018-66-3), alias: quinazoline-4-amine, 4-quinazolineamine, molecular formula: C8H7N3, molecular weight: 145.16, melting point: 274-275℃, boiling point: 328.3℃, storage conditions: 2-8℃, properties: off-white crystalline powder.

[0042] (2) Dimethyl sulfoxide: molecular weight: 78.13, stored at room temperature.

[0043] (3) 5% glucose solution: Weigh 50 g of glucose powder and dissolve it in sterile water to make up to 1000 mL. Sterilize the prepared 5% glucose solution by high pressure.

[0044] Weigh 100 mg of 4-aminoquinazoline powder (Macklin, Q841658), dissolve it in DMSO (Sigma, 472301), mix it thoroughly, and dilute it to 10 mL to prepare a 10 mg / mL stock solution, which is then diluted with 5% glucose solution to different concentrations for oral feeding to female mosquitoes.

[0045] 3. Determine the effect of 4-aminoquinazoline on mosquito reproductive ability at different concentrations

[0046] Take 250 μL of the above 4-aminoquinazoline mother solution and add 6.00 mL of 5% glucose solution to prepare a 400 mg / L working solution.

[0047] Take 250 μL of the above 4-aminoquinazoline mother solution and add 12.25 mL of 5% glucose solution to prepare a 200 mg / L working solution.

[0048] Take 250 μL of the above 4-aminoquinazoline mother solution and add 24.75 mL of 5% glucose solution to prepare a 100 mg / L working solution.

[0049] Take 250 μL of the above 4-aminoquinazoline mother solution and add 49.75 mL of 5% glucose solution to prepare a 50 mg / L working solution.

[0050] Take 250 μL of the above 4-aminoquinazoline mother solution and add 99.75 mL of 5% glucose solution to prepare a 25 mg / L working solution.

[0051] Adult female mosquitoes at 0 hours of eclosion were orally fed with the above working solution until they laid eggs. The WT group was a blank control group orally fed with 5% glucose only, the Control group was an experimental control group of a mixture of dimethyl sulfoxide (DMSO) and 5% glucose solution, and the groups treated with different concentrations of 4-aminoquinazoline were experimental groups of a mixture of 4-aminoquinazoline, DMSO and 5% glucose solution in different proportions.

[0052] The ovaries of female mosquitoes 48 h after blood feeding (48 h PBM) were dissected, and the changes in ovarian morphology of each group were observed under an OLMPUS phase-contrast inverted microscope, and the number of follicles was counted (N=10).

[0053] Three days after emergence, after the male and female mosquitoes fully mated, they were given a blood meal, and the egg laying rate of female mosquitoes in each group was observed 72 hours after blood feeding (72h PBM).

[0054] The number of mosquito eggs in each egg raft was counted under an OLMPUS phase-contrast inverted microscope (N=30). The egg rafts were returned to the laboratory for normal rearing. When the larvae hatched, the first-instar larvae were counted and the hatching rate was calculated (N=30).

[0055] The results are as follows:

[0056] Effects of 1.4-aminoquinazoline on the ovaries and follicles of female mosquitoes before egg laying

[0057] like Figure 1 As shown, compared with the WT group and the Control group, the ovarian volume of female mosquitoes in the experimental group was significantly reduced at 48h PBM.

[0058] like Figure 2 As shown, at 48h PBM, the number of follicles in female mosquitoes in the WT group was 196.1±10.2, the control group was 201.8±21.7, the 25mg / L treatment group was 135.7±13.9, the 50mg / L treatment group was 131.2±11.4, the 100mg / L treatment group was 108.3±16.8, the 200mg / L treatment group was 87.9±15.1, and the 400mg / L treatment group was 86.8±18.1 ( Figure 2 , ****P<0.0001, ***P<0.001, **P<0.01).

[0059] The above results suggest that 4-aminoquinazoline may reduce the reproductive capacity of female mosquitoes by inhibiting ovarian development, and the number of follicles in the ovaries is significantly reduced. It has a good inhibitory effect at low concentrations (25 mg / L), and the inhibitory effect on mosquito ovaries is significantly improved with the increase of 4-aminoquinazoline concentration.

[0060] 2. Effect of 4-aminoquinazoline on mosquito egg laying

[0061] like Figure 3 As shown, compared with the WT group and the Control group, the raft morphology of the experimental group was significantly reduced.

[0062] like Figure 4As shown, at 72h PBM, the number of eggs laid by female mosquitoes in the WT group was 194.1±24.6, the control group was 190.83±23.3, the 25mg / L treatment group was 121.4±23.7, the 50mg / L treatment group was 105.8±28.2, the 100mg / L treatment group was 75.5±28.9, the 200mg / L treatment group was 63.5±26.2, and the 400mg / L treatment group was 62±19.6 ( Figure 4 , ****P<0.0001, ***P<0.001, **P<0.01).

[0063] The above results suggest that the egg-laying capacity of female mosquitoes decreased after treatment with 4-aminoquinazoline, and as the concentration of 4-aminoquinazoline increased, the egg-laying capacity of female mosquitoes decreased more significantly.

[0064] Effects of 3,4-aminoquinazoline on the hatching rate of mosquito eggs

[0065] like Figure 5 As shown, the hatching rate of female mosquitoes in the WT group was 94.4% ± 4.4%, the hatching rate of the control group was 94.3% ± 4.6%, the hatching rate of the 25 mg / L treatment group was 94.2% ± 5.2%, the hatching rate of the 50 mg / L treatment group was 95.1% ± 4.2%, the hatching rate of the 100 mg / L treatment group was 95.2% ± 3.0%, the hatching rate of the 200 mg / L treatment group was 95.7% ± 3.8%, and the hatching rate of the 400 mg / L treatment group was 88.9% ± 2.5% ( Figure 5 ).

[0066] The above results suggest that the hatching rate of mosquito eggs is not affected after treatment with 4-aminoquinazoline.

[0067] The protection content of the present invention is not limited to the above embodiments. Without departing from the spirit and scope of the inventive concept, changes and advantages that can be thought of by those skilled in the art are included in the present invention and are protected by the attached claims.

Claims

1. Application of 4-aminoquinazoline in mosquito control. 2.4-aminoquinazoline in the preparation of mosquito control drugs or insecticides.

3. The use according to claim 1 or 2, characterized in that: The molecular formula of 4-aminoquinazoline is C8H7N3, and its structural formula is 4. The use according to claim 2, characterized in that: The 4-aminoquinazoline in the medicine or insecticide can inhibit the ovarian development of female adult mosquitoes and reduce their egg-laying amount.

5. The use according to claim 2, characterized in that: The mosquito is Culex pipiens of the genus Culex of the family Culicidae in the order Diptera.

6. The use according to claim 2, characterized in that: The medicine or insecticide takes 4-aminoquinazoline as an effective ingredient, and the effective concentration is 25-400 mg / L.

7. The use according to claim 6, characterized in that: As the concentration of 4-aminoquinazoline increases, the reproductive ability of female mosquitoes is more significantly inhibited.

8. The use according to claim 6, characterized in that: The medicine or insecticide uses DMSO as solvent, is mixed with 1-10% glucose solution in proportion, and is fed to the emerged female mosquitoes by oral feeding.

9. The use according to claim 6, characterized in that: The medicine or insecticide also includes other medicines or agents for controlling mosquitoes or other related active ingredients.

10. A method for controlling mosquitoes, characterized in that: 4-aminoquinazoline or drugs or insecticides containing 4-aminoquinazoline are applied to mosquito-borne disease epidemic areas in the form of sugar baits.

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