Application of 4-aminoquinazoline in the reproductive control of female mosquitoes

Abstract

This invention discloses the application of 4-aminoquinazoline in the reproductive control of female mosquitoes. The invention involves dissolving 4-aminoquinazoline powder in dimethyl sulfoxide (DMSO), then mixing it with glucose solution in a specific ratio. This mixture is then orally fed to female adult mosquitoes to inhibit ovarian development and reduce egg production. This invention expands the application of the existing drug 4-aminoquinazoline by directly inhibiting the reproductive potential of female adult mosquitoes, thereby controlling mosquito populations and providing a new solution to the current widespread resistance to chemical insecticides in mosquitoes in my country.

Description

Technical Field

[0001] This invention belongs to the field of mosquito vector control technology, specifically relating to the application of 4-aminoquinazoline in the reproductive control of female mosquitoes. Background Technology

[0002] Mosquitoes are blood-sucking insects that, during their blood-feeding process, can inject pathogens they carry into their saliva and infect their hosts, transmitting a variety of mosquito-borne diseases such as malaria, dengue fever, Zika virus disease, chikungunya, Japanese encephalitis, and filariasis. Mosquito-borne diseases account for approximately 17% of all infectious diseases worldwide, claiming more than 700,000 lives annually, seriously threatening human health and posing a significant challenge to global public health. my country, with its vast territory, suitable climate, and abundant vegetation, has a diverse range of mosquito species and high mosquito population densities, making outbreaks of mosquito-borne diseases a constant possibility. Therefore, strengthening mosquito-borne disease prevention and control is urgently needed.

[0003] Since most mosquito-borne diseases lack vaccines and specific effective drugs, mosquito vector control has become a crucial means of controlling these diseases. Currently, mosquito vector control strategies mainly include the following four methods: 1. Environmental control: eliminating mosquito breeding grounds by modifying land, water bodies, or vegetation; 2. Physical control: reducing contact between hosts and mosquitoes through methods such as using screen doors and windows, mosquito nets, mosquito hats, and insect repellent nets; 3. Biological control: Bacillus thuringiensis (Bt), Bacillus sphaeroides (Bs), and Wolbachia have become more common in recent years, but these methods are costly and difficult to implement on a large scale; 4. Chemical control: eliminating indoor and outdoor mosquitoes through space spraying of chemical insecticides. Common chemical insecticides currently include organophosphates, carbamates, and pyrethroids. Due to their high efficiency, speed, and ease of use, chemical control has always been a major method in integrated mosquito control strategies. The long-term and widespread use of traditional chemical insecticides has led to the emergence and development of mosquito resistance, thus necessitating the development of novel insecticides that do not exhibit cross-resistance with traditional chemical insecticides. The high reproductive capacity of female mosquitoes is beneficial for maintaining population numbers and thus enabling them to function as highly effective disease vectors; therefore, reducing the reproductive potential of female mosquitoes is considered a promising mosquito control strategy.

[0004] 4-Aminoquinazoline is a grayish-white powdery organic compound, slightly soluble in water but soluble in organic solvents. It is prepared by reducing nitroquinazoline with a reducing agent (such as sodium sulfite, sodium bisulfite, or sulfoxide), followed by purification through crystallization. 4-Aminoquinazoline derivatives have structures similar to chloroquine, fewer side effects, and can be used to treat and prevent malaria. The exact mechanism of its antimalarial effect is unclear; it may be that 4-aminoquinazoline antimalarials accumulate in the acidic organelles of Plasmodium, maintaining a high concentration and thus inhibiting the growth of the parasite; or 4-aminoquinazoline antimalarials may inhibit cellular functions such as RNA synthesis and DNA replication, leading to the death of the parasite. Currently, many new derivatives based on 4-aminoquinazoline have been developed, inhibiting the activity of vascular endothelial growth factor receptor-2 (VEGFR-2) tyrosine kinase and phosphatidylinositol-3 (PI3Ka) kinases, and are widely used in targeted therapy for tumors such as lung cancer, breast cancer, colon cancer, and prostate cancer.

[0005] No application of 4-aminoquinazoline in mosquito control was found in the search. Summary of the Invention

[0006] To address the shortcomings of existing technologies, the present invention aims to provide the application of 4-aminoquinazoline in the reproductive control of female mosquitoes. This invention proposes a new use for 4-aminoquinazoline, offering a novel strategy for existing mosquito vector control and helping to solve the problem of resistance to traditional chemical insecticides.

[0007] The technical solution adopted by this invention to solve its technical problem is:

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

[0009] Secondly, this invention protects the use of 4-aminoquinazoline in the preparation of medicaments or insecticides for mosquito control.

[0010] In a specific implementation scheme, the molecular formula of the 4-aminoquinazoline is C8H7N3, and its structural formula is [not specified].

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

[0012] In the specific implementation plan, the mosquito is Culex pipiens pallens, belonging to the Culicidae family of the order Diptera.

[0013] In a specific implementation plan, the drug or insecticide uses 4-aminoquinazoline as the active ingredient, with an effective concentration of 25–400 mg / L.

[0014] In the specific implementation plan, the inhibition of female mosquito reproductive capacity becomes more significant as the concentration of 4-aminoquinazoline increases.

[0015] In a specific implementation plan, the drug or insecticide is mixed with DMSO as a solvent and then mixed with 1-10% (preferably 5%) of glucose solution in a proportional manner, and fed orally to female mosquitoes after they have emerged.

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

[0017] In a specific implementation scheme, the mosquito repellent / insecticide uses 4-aminoquinazoline as the main active ingredient. A stock solution of 4-aminoquinazoline at a concentration of 10 mg / mL is prepared using DMSO as the solvent. Before use, it is further diluted with 5% glucose solution to a concentration of 25–400 mg / L, specifically solutions of 25 mg / L, 50 mg / L, 100 mg / L, 200 mg / L, and 400 mg / L.

[0018] The preparation of the 10 mg / mL 4-aminoquinazoline stock solution is as follows: Weigh 50 mg of 4-aminoquinazoline powder into a pre-sterilized beaker, add DMSO to the beaker, and then stir on a magnetic stirrer for 30 min to dissolve. Finally, bring the volume to 5 mL to prepare 5 mL of 10 mg / mL stock solution, which is then dispensed and frozen for later use.

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

[0020] In specific implementation schemes, the drug or insecticide may also include other drugs or reagents or other related active ingredients for controlling mosquitoes.

[0021] Secondly, the present invention protects a method for mosquito control, wherein the method uses 4-aminoquinazoline or a drug or insecticide containing 4-aminoquinazoline to be applied to mosquito-borne disease endemic areas via sugar bait.

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

[0023] (1) This invention uses the existing drug 4-aminoquinazoline for mosquito control, opening up new applications for this drug.

[0024] (2) Sugar water is an important food for adult mosquitoes to grow, develop, and reproduce. This invention is made from 4-aminoquinazoline, DMSO and glucose water. The raw materials for this sugar bait formula are simple and readily available, the preparation process is simple, and it attracts mosquitoes to feed on it.

[0025] (3) The agent prepared by 4-aminoquinazoline, DMSO and glucose solution of the present invention can produce a good inhibitory effect on mosquito reproduction at concentrations of 400mg / L, 200mg / L, 100mg / L, 50mg / L and 25mg / L. At the same time, as the concentration increases, the ovarian volume of female mosquitoes decreases significantly, the number of follicles in the ovary decreases significantly, and the number of eggs laid decreases more significantly, but the hatching rate of eggs does not differ significantly.

[0026] (4) The 4-aminoquinazoline of the present invention inhibits the population size by reducing the reproduction of female mosquitoes, thus solving the problem of drug resistance of traditional chemical mosquito killers. It can be used as an adjuvant and adjuvant for new compound mosquito killers or other mosquito killers. Attached Figure Description

[0027] Figure 1 The effects of different concentrations of 4-aminoquinazoline on ovarian development in female mosquitoes before oviposition (48 h PBM) were investigated. Figure 1 Figure A in the diagram represents group WT. Figure 1 In the diagram, B represents the Control group. Figure 1 Figure C in the diagram represents the group with a 4-aminoquinazoline concentration of 25 mg / L. Figure 1 Figure D represents the group with a 4-aminoquinazoline concentration of 50 mg / L. Figure 1 Figure E in the diagram represents the group with a 4-aminoquinazoline concentration of 100 mg / L. Figure 1 Figure F in the diagram represents the group with a 4-aminoquinazoline concentration of 200 mg / L. Figure 1 The G figure represents the group with a 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 female mosquito follicles before oviposition (48 h 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 morphology of female mosquito egg rafts were investigated. Figure 3Figure A in the diagram represents group WT. Figure 3 In the diagram, B represents the Control group. Figure 3 Figure C in the diagram represents the group with a 4-aminoquinazoline concentration of 25 mg / L. Figure 3 Figure D represents the group with a 4-aminoquinazoline concentration of 50 mg / L. Figure 3 Figure E in the diagram represents the group with a 4-aminoquinazoline concentration of 100 mg / L. Figure 3 Figure F in the diagram represents the group with a 4-aminoquinazoline concentration of 200 mg / L. Figure 3 The G figure in the figure represents the group with a 4-aminoquinazoline concentration of 400 mg / L. (N = 30, *P < 0.05, **P < 0.01, ***P < 0.001, ****P < 0.0001). (Magnification 2.5 x 10).

[0032] Figure 4 The effect of different concentrations of 4-aminoquinazoline on oviposition in 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 mosquito egg hatching rate. (N=30, ns represents no statistical difference, *P<0.05, **P<0.01, ***P<0.0001, ****P<0.0001). Detailed Implementation

[0035] The technical solutions and effects of the present invention will be further described and explained below with reference to specific embodiments, but the present invention is not limited to these specific embodiments. Reagents or instruments used without a specified manufacturer are considered to be conventional products that can be purchased on the market.

[0036] 1. Mosquitoes and their food:

[0037] The Culex pipiens pallens used in this invention were collected from Tangkou County, Shandong Province, and subsequently bred in our laboratory. Prior to the experiment, the mosquitoes had not been exposed to any insecticides or toxic substances. Breeding conditions: temperature 28–30℃, relative humidity 50%–70%, and a daily light-dark ratio of 14h:10h.

[0038] Larvae in their first to fourth instar were fed high-pressure ground rodent food powder, while adult mosquitoes were fed a 5% glucose solution.

[0039] Adult male and female mosquitoes were passaged by feeding on fresh blood from ICR strain mice after mating.

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

[0041] (1) 4-Aminoquinazoline (CAS: 15018-66-3), also known as 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: grayish-white crystalline powder.

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

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

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

[0045] 3. Determining the effect of different concentrations of 4-aminoquinazoline on mosquito reproductive capacity.

[0046] Take 250 μL of the above 4-aminoquinazoline stock 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 stock 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 stock 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 stock 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 stock solution and add 99.75 mL of 5% glucose solution to prepare a 25 mg / L working solution.

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

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

[0053] Three days after emergence, male and female mosquitoes were fully mated and fed blood. The number of eggs laid by female mosquitoes in each group was observed 72 hours after 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 then returned to the laboratory for normal rearing until the larvae hatched. The number of first-instar larvae was counted, and the hatching rate was calculated (N=30).

[0055] Here are the results:

[0056] Effects of 1,4-aminoquinazoline on the ovaries and follicles of female mosquitoes before oviposition.

[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, in the control group 201.8±21.7, in the 25mg / L treatment group 135.7±13.9, in the 50mg / L treatment group 131.2±11.4, in the 100mg / L treatment group 108.3±16.8, in the 200mg / L treatment group 87.9±15.1, and in the 400mg / L treatment group 86.8±18.1. Figure 2 , ****P<0.0001, ***P<0.001, **P<0.01).

[0059] The above results suggest that 4-aminoquinazoline reduces female mosquito fertility by inhibiting ovarian development, resulting in a significant reduction in the number of follicles in the ovaries. A good inhibitory effect was observed at a low concentration (25 mg / L), and the inhibitory effect on mosquito ovaries was significantly enhanced with increasing concentrations of 4-aminoquinazoline.

[0060] Effects of 2,4-aminoquinazoline on mosquito oviposition

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

[0062] like Figure 4As shown, at 72h PBM, the oviposition rate of 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 oviposition rate of female mosquitoes decreased after treatment with 4-aminoquinazoline, and the decrease was more pronounced with increasing concentration of 4-aminoquinazoline.

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

[0065] like Figure 5 As shown, the hatching rate of female mosquitoes in the WT group was 94.4% ± 4.4%, the hatching rate in the control group was 94.3% ± 4.6%, the hatching rate in the 25 mg / L treatment group was 94.2% ± 5.2%, the hatching rate in the 50 mg / L treatment group was 95.1% ± 4.2%, the hatching rate in the 100 mg / L treatment group was 95.2% ± 3.0%, the hatching rate in the 200 mg / L treatment group was 95.7% ± 3.8%, and the hatching rate in 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 was not affected by 4-aminoquinazoline treatment.

[0067] The scope of protection of this invention is not limited to the above embodiments. Variations and advantages that can be conceived by those skilled in the art without departing from the spirit and scope of the inventive concept are included in this invention and are protected by the appended claims.

Claims

The application of 1,4-aminoquinazoline in mosquito control is characterized by, The 4-aminoquinazoline inhibits female mosquito reproduction. The application of 2,4-aminoquinazoline in the preparation of mosquito control drugs, characterized in that, The 4-aminoquinazoline inhibits female mosquito reproduction.

3. The application according to claim 2, characterized in that, The drug contains 4-aminoquinazoline, which can inhibit ovarian development in female adult mosquitoes and reduce their egg production.

4. The application according to claim 2, characterized in that, The mosquito in question is Culex pipiens pallens, belonging to the Culicidae family, Diptera order.

5. The application according to claim 2, characterized in that, The drug uses 4-aminoquinazoline as its active ingredient, with an effective concentration of 25~400 mg / L.

6. The application according to claim 5, characterized in that, The drug is prepared by mixing DMSO with a 1-10% glucose solution and then feeding it orally to newly emerged female mosquitoes.

7. The application according to claim 5, characterized in that, The drug also includes other drugs for controlling mosquitoes.

8. A method for mosquito control, characterized in that, The mosquito control method involves applying 4-aminoquinazoline or drugs containing 4-aminoquinazoline to mosquito-borne disease endemic areas via sugar bait, thereby inhibiting the reproduction of female mosquitoes.

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