Application of compound fluoropropylazole in preparation of tick-killing drugs
The tick-killing drug prepared by using the compound flupropazole solves the problems of insufficient lethality and reproductive inhibition of existing drugs against ticks, achieving a highly efficient and safe tick-killing effect, and is suitable for various formulations and scenarios.
Patent Information
- Application Number
- CN202511894104.6
- Authority / Receiving Office
- CN · China
- Patent Type
- Applications(China)
- Current Assignee / Owner
- Filing Date
- 2025-12-16
- Publication Date
- 2026-01-16
AI Technical Summary
Existing tick-killing drugs such as flurana are insufficient in their lethality and reproductive inhibition capabilities against certain tick species at low concentrations. Furthermore, with the increasing resistance of parasites to these drugs, the development of new drugs with stronger killing activity and lower usage concentrations has become a key research focus.
Using the compound flupropazole as the active ingredient, a tick-killing drug suitable for veterinary or agricultural use is prepared, including injections, oral formulations, topical formulations, and sustained-release or controlled-release formulations. The concentration of flupropazole is 1~100ppm, preferably 5~30ppm, for killing Haemaphysalis longicornis, and can be used in combination with other insecticides.
Fluprozil has a 100% lethality rate and significantly delayed oviposition ability against Haemaphysalis longhorn at low concentrations. It is also more effective at killing ticks than existing drugs. The formulation is flexible and adaptable to different scenarios, has high safety, and is suitable for livestock and laboratory animals.
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Abstract
Description
Technical Field
[0001] This invention relates to the field of pharmaceutical technology, specifically to the application of the compound flupropionazole in the preparation of tick-killing drugs. Background Technology
[0002] The chemical formula of fluoropropanezide is C 15 H6Cl2F8N4 is a newly synthesized phenylpyrazole compound by researchers from the Veterinary Drug Innovation and Resistance Team at the Lanzhou Institute of Animal Husbandry and Veterinary Medicine, Chinese Academy of Agricultural Sciences. The mechanism of action of phenylpyrazole insecticides mainly involves interfering with γ-aminobutyric acid (GABA)-gated chloride ion channels in the insect nervous system, leading to uncontrolled neurological excitability and death in pests. Representative drugs in this class include fipronil, but long-term use has resulted in resistance issues, necessitating the development of novel insecticides with unique mechanisms of action that remain effective against resistant insect strains. In existing technologies, isoxazoline compounds (such as fleroranar) have been widely used as GABA receptor antagonists for the control of ectoparasites in pets such as dogs and cats. Fluoranarar has advantages such as a broad insecticidal spectrum, long-lasting effect, and high safety. While its mechanism of action is similar to that of phenylpyrazoles, there are differences in molecular structure, receptor binding characteristics, and resistance spectrum.
[0003] However, although fluranal, as a commercially available product (Bevido), is highly effective in controlling fleas and ticks in dogs and cats, there is still room for improvement in its lethality against certain tick species (such as Haemaphysalis longicornis) at low concentrations and its ability to inhibit tick reproduction. Especially in practical applications, with the increasing resistance of parasites, developing new drugs with stronger killing activity, lower application concentrations, and the ability to effectively disrupt the tick life cycle has become a key focus of current research.
[0004] Fluprozil, a new member of the phenylpyrazole class, has shown superior tick-killing activity and a significant inhibitory effect on tick oviposition behavior compared to fleranal in preliminary experiments. Experiments indicate that, at the same concentration, fluprozil has a higher mortality rate against Haemaphysalis longicornis and significantly delays its oviposition time, suggesting a unique advantage in controlling tick population reproduction.
[0005] Currently, research on the application of fluprofenazole in tick-killing agents is still in its preliminary stages, and its specific mechanism of action, pharmacodynamic properties, safety, and systematic formulation development require further investigation. Therefore, developing the application of fluprofenazole in the preparation of tick-killing agents will not only help enrich the existing range of insecticides but also provide a new technical approach to solving current resistance problems and improving control efficacy. Summary of the Invention
[0006] The purpose of this invention is to provide the application of the compound flupropionazole in the preparation of tick-killing drugs, so as to solve the problems mentioned in the background art.
[0007] To achieve the above objectives, the present invention provides the following technical solution: the use of the compound fluprozil in the preparation of a drug for the prevention or treatment of parasitic infections caused by ticks, wherein the drug is used to kill Haemaphysalis longicornis, and the chemical formula of the compound fluprozil is C. 15 H6Cl2F8N4.
[0008] Preferably, the drug comprises a therapeutically effective amount of fluprofenazole as the active ingredient, and a pharmaceutically acceptable carrier or excipient.
[0009] Preferably, the drug is formulated into a dosage form suitable for veterinary or agricultural use, the dosage form being selected from at least one of injections, oral preparations, topical preparations, sustained-release or controlled-release preparations.
[0010] Preferably, the oral preparations are tablets, chewable tablets, granules, and powders, and the topical preparations include drops, sprays, lotions, powders, and gels.
[0011] Preferably, the effective concentration of flupropazole in the drug is 1~100ppm, preferably 5~30ppm, which has a significant killing effect and reproductive inhibition ability on Haemaphysalis longicornis within this concentration range.
[0012] Preferably, the flupropanediol is used in combination with other insecticides to form a compound formulation, wherein the other insecticides are selected from at least one of organophosphates, pyrethroids, and isoxazoline compounds.
[0013] Preferably, the method for preparing the drug includes the following steps:
[0014] Mix flupropazole with a suitable carrier or diluent;
[0015] Add auxiliary materials;
[0016] The desired dosage form is prepared using conventional pharmaceutical processes.
[0017] Preferably, the excipients are selected from at least one of adhesives, disintegrants, lubricants, and stabilizers;
[0018] The carrier is selected from at least one of dimethyl sulfoxide (DMSO), propylene glycol, vegetable oil, starch, lactose, and microcrystalline cellulose.
[0019] Preferably, the mass percentage of flupropazole in the drug is 0.1% to 10%, more preferably 0.5% to 5%.
[0020] Preferably, the drug is suitable for livestock and laboratory animals.
[0021] Compared with existing technologies, the beneficial effects of this invention are as follows: This invention uses an in vitro immersion method to systematically evaluate the efficacy of fluprofenazole against Haemaphysalis longicornis. The results show that at a concentration of 5 ppm, the mortality rate reaches 100% within 48 hours, and at a concentration of 1 ppm, it can significantly delay oviposition and inhibit reproduction, and its efficacy is superior to that of the positive control drug fleranal. This invention is highly effective, rapidly and thoroughly killing ticks at low concentrations; it can inhibit reproduction and block the tick life cycle; it has high safety and is mammal-friendly; it has resistance management potential and can control resistant tick strains; the formulation is flexible and can be made into various dosage forms to adapt to different scenarios. Attached Figure Description
[0022] Figure 1 For the identification of adult ticks (female, starving state);
[0023] Figure 2 The tick mortality rate (24h, 48h) after treatment with the compound flupropanezide.
[0024] Figure 3 The tick mortality rate (24h, 48h) after treatment with fluranal was used as the positive control group. Detailed Implementation
[0025] The following embodiments, in conjunction with the accompanying drawings, facilitate a better understanding of the present invention, but do not limit the scope of the invention. Unless otherwise specified, the experimental methods used in the following embodiments are conventional methods. Unless otherwise specified, the experimental materials used in the following embodiments are commercially available. The compound flupropanezole was prepared by researchers from the Veterinary Drug Innovation and Resistance Team of the Lanzhou Institute of Animal Husbandry and Veterinary Medicine, Chinese Academy of Agricultural Sciences.
[0026] The ticks used in this experiment were all collected from sheep at a sheep farm in Xiangle Town, Ning County, Qingyang City, Gansu Province. They were initially identified as Haemaphysalis longicornis ticks and were cultured for three consecutive generations on female New Zealand White rabbits (4 months old, SPF grade, dorsal parasites) until their traits were stable (Experimental Animal Production License No.: SCXK(Gan)2020-0002). All subcultures were conducted at the Animal Laboratory of the Lanzhou Institute of Animal Husbandry and Veterinary Medicine, Chinese Academy of Agricultural Sciences (Animal Experiment Welfare and Ethics Number: 2025-24). The tick experiments were conducted in a constant temperature and humidity chamber at 27±1℃ and 70% humidity.
[0027] The experimental group drug was flufenoxuron, synthesized by members of the Veterinary Drug Innovation and Resistance Team at the Lanzhou Institute of Animal Husbandry and Veterinary Medicine, Chinese Academy of Agricultural Sciences, in powder form. The control group drug was fluranarane (powder, batch number 23M2014HA1, manufactured by Intervet International GmbH, Austria). Dimethyl sulfoxide (DMSO, brand: Solarbio, catalog number: D8371-100ml) was purchased from Beijing Solarbio Technology Co., Ltd., and was diluted 1000 times with sterile water as a solvent according to the drug dosage ratio during the experiment. The LED-type fixed artificial climate chamber (constant temperature and humidity incubator) was purchased from Ningbo Southeast Instrument Co., Ltd., model: RDN-260B.
[0028] 1.1 Reproduction of ticks
[0029] Tick passage and culture were conducted at the Lanzhou Institute of Animal Husbandry and Veterinary Medicine, Chinese Academy of Agricultural Sciences. Experimental animals—New Zealand White rabbits (SPF grade, female, 3 months old)—were purchased from the Animal Center of the Lanzhou Veterinary Research Institute, Chinese Academy of Agricultural Sciences. Ticks were collected from Ning County, Qingyang City, Gansu Province. Morphological identification of the ticks was performed before the experiment; all ticks in this batch belonged to the Haemaphysalis longicornis species. Before the experiment, the fur on the backs of the experimental animals was shaved. A gel was prepared and used with a cloth cover to adhere to the backs of the animals, and ticks were inoculated. After the ticks engorged, laid eggs, and hatched, new experimental animals were replaced, and the process of engorgement, egg laying, and hatching was repeated three times until the ticks were free of pathogen infection and their condition was stable before the experiment began. Ticks were fed to the experimental animals on their backs when engorged, and ticks not engorged were cultured in an indoor incubator at 25°C and 70% humidity.
[0030] 1.11 Identification of ticks
[0031] Under a stereomicroscope, tick species were differentiated based on biological characteristics such as the dorsal features of female Haemaphysalis longicornis ticks, the location of the genital opening and spiracular plates, and the length-to-width ratio of the pseudobulb base. Preliminary identification was performed based on morphological characteristics, referring to the *Atlas of Common Medical Ticks and Mites* and *Systematic Taxonomy of Ticks*. Based on the morphological identification results, DNA was extracted from 11 randomly selected ticks, and PCR amplification was performed using primers for the tick cytochrome C oxidase subunit I (COXI / COI) gene. The PCR products were confirmed using a 1% nucleic acid gel, and positive products were sent to the company for sequencing.
[0032] 1.12 PCR identification results
[0033] PCR amplification results showed that the COI gene amplified tick DNA and produced 10 positive bands. Sequencing analysis revealed that the ticks were closely related to the reported Haemaphysalis longicornis Nianxian and Baiyin strains, but less related to Dermacentor silvarum, Hyalomma rufipes, Hyalomma asiaticum, and Rhipicephalus microplus. Therefore, the ticks were identified as Haemaphysalis longicornis.
[0034] Example 1
[0035] 1.2 Pre-screening by compound concentration
[0036] This case study used the model tick species *Haemaphysalis longicornis* (female) as the research subject, employing high concentrations of compounds for pre-screening to narrow down the range of effective tick-killing concentrations. Fluproazole (powder) was diluted 1000-fold with dimethyl sulfoxide (DMSO) to create a stock solution. This stock solution was then diluted sequentially to four concentrations: 100 ppm, 50 ppm, 10 ppm, and 5 ppm (1000 ppm = 1 mg / mL), designated as the experimental groups. The positive control group (flurane, powder) was set up with the same concentration gradient as the experimental groups (100 ppm, 50 ppm, 10 ppm, 5 ppm). The negative control group was a 10 ppm dilution (DMSO). Each group consisted of 10 engorged adult ticks, with three parallel biological replicates for each compound concentration group. Ticks were treated using an in vitro immersion method, immersing for 5 minutes. After immersion, the ticks were blotted dry with filter paper and placed in an incubator at 27–28°C, maintaining 70% relative humidity. After 24 hours, the tick mortality status was observed and recorded.
[0037] Example 2
[0038] 1.3 Compound treatment of ticks experiment
[0039] Based on Example 1, the drug concentration range for the experimental group and the positive control group was further narrowed. The experimental group had four drug concentration gradients: 1 ppm, 5 ppm, 10 ppm, 20 ppm, and 30 ppm (1000 ppm = 1 mg / mL). The positive control group had five concentration gradients: 1 ppm, 5 ppm, 10 ppm, 20 ppm, and 30 ppm. One negative control group (DMSO, 0.1 ppm) was included. Six engorged adult ticks were used in each group, and three parallel biological replicates were set up for each drug concentration. Ticks were treated using an in vitro immersion method, immersing for 5 minutes. After immersion, the drug solution on the surface of the ticks was blotted dry with filter paper, and the ticks were placed in an incubator at 27–28°C, maintaining 70% relative humidity. Tick survival rates were observed and calculated after 24 and 48 hours of the experiment.
[0040] Estimated using the BLISS method (Probit analysis):
[0041] 48-hour LC50 of flupropiconazole against Haemaphysalis longicornis 50 = 0.80ppm.
[0042] Example 3
[0043] 1.4 Effects of compounds on tick oviposition time
[0044] Based on Example 2, the effects of the compound flupropiconazole and the positive control group fleranal on tick oviposition time at the lowest insecticidal concentration were compared. The flupropiconazole experiment consisted of an experimental group (1 ppm), a negative control group (DMSO, 0.1 ppm), and a blank control group (distilled water). The fleranal experiment consisted of experimental group 1 (10 ppm), experimental group 2 (5 ppm), experimental group 3 (1 ppm), a negative control group (DMSO, 0.1 ppm), and a blank control group (distilled water). Six engorged adult ticks were used in each group, and three parallel biological replicates were set up for each drug concentration. Ticks were treated using an in vitro immersion method, immersing for 5 minutes. After immersion, the tick surface was blotted dry with filter paper, and the ticks were placed in an incubator at 27–28°C and 70% relative humidity for 10 consecutive days to observe and record changes in tick oviposition time.
[0045] 1.5 Statistical Indicators and Data Processing
[0046] Tick mortality rate (%) = (Number of dead ticks / Number of experimental ticks) x 100%. Mortality rate evaluation criteria: ticks not crawling, limbs extended and not retracted, body shriveled and shrunken, exoskeleton dull, surface dark brown or black, overall stiff and inelastic. Oviposition time evaluation criteria: Comparison of the start of oviposition time under the same environmental conditions and different drug treatments. Data were processed using EXCEL software, and one-way ANOVA was performed using SPPS 19.0 software. The median lethal dose (LD50) was calculated using the BLISS method.
[0047] 2. Results
[0048] 2.1 Identification of tick species
[0049] Based on the dorsal characteristics of female Haemaphysalis longicornis ticks: The body color is reddish-brown or grayish-brown. After engorgement, it becomes oval, measuring (2.52~3.01) mm × (1.57~1.75) mm. Adults can swell to the size of a mung bean or soybean after feeding. The punctures on the scutum are medium-sized, evenly distributed, and relatively dense. The genital opening is located on the anterior half of the ventral surface, usually at the level of the coxae of legs II and III. The spiracles are oval, located posterolaterally to the coxae of leg IV. The pseudocephalic base is rectangular, twice as wide as long, with a prominent, triangular, strong basal projection. The outer edges of the palps protrude moderately outwards, forming an angular shape. The female scutum is sub-circular, wide in the middle, with an arc-shaped or slightly wavy edge, measuring (0.95~1.15) mm × (0.82~1.16) mm. The marginal bulge is narrow, long, and clearly defined, located on the posterior edge of the scutum. Based on these characteristics, after microscopic observation and analysis... Figure 1 The ticks were identified as Haemaphysalis longicornis.
[0050] 2.2 Results of Pre-screening Tests on Compound Concentration in the Experimental Group
[0051] The results of the pre-screening test of compound concentration in the experimental group showed that when flupropazole was diluted with 10 ppm of DMSO, the insecticidal rate reached 80% at 5 ppm, and the insecticidal rate reached 100% when the concentration was ≥10 ppm (Table 1).
[0052] In the positive control group, the insecticidal rate of fluranal was 40% when the concentration was ≥50 ppm, and the insecticidal rate could reach 100% when the concentration was ≥50 ppm (Table 2).
[0053] Preliminary experimental results showed that at the same low concentrations (5 ppm, 10 ppm), flupropiconazole was more effective than flenazone in killing ticks.
[0054] 2.3 Results of the compound treatment experiment on ticks
[0055] The results of the compound treatment test on ticks showed that when flupropazole was diluted with 0.1 ppm DMSO and treated with ticks for 24 hours (Table 3), the tick mortality rate reached over 60% at a concentration of 5 ppm, over 70% at a concentration of 10 ppm, and 100% of ticks were killed at concentrations above 20 ppm. Figure 2 After 48 hours of application (Table 4), the tick mortality rate was 61% at a concentration of 1 ppm, and 100% tick mortality was achieved at concentrations above 5 ppm. Figure 2 ).
[0056] When fluranal was diluted with 0.1 ppm DMSO and treated on ticks for 24 hours (Table 5), the mortality rate of ticks at a concentration of 30 ppm was 61%, while the mortality rate decreased to 44% at a concentration of 20 ppm. Figure 3 When applied to ticks for 48 hours (Table 6), the mortality rate was 100% at a concentration of 30 ppm, 61% at 20 ppm, and decreased to 44% at 10 ppm. Figure 3 ).
[0057] The final results show that, within the same treatment time and at the same concentration, flupropiconazole is more effective than flerostat in killing ticks. At 24 hours, a concentration of 20 ppm of flupropiconazole showed the best tick-killing effect; at 48 hours, a concentration of 5 ppm of flupropiconazole showed the best tick-killing effect.
[0058] 2.4 Effects of compounds on tick oviposition time
[0059] At a concentration of 1 ppm, flupropiconazole caused ticks to begin laying eggs on the fourth day, with only four ticks laying eggs. In the control group, egg laying began on the third day, peaking around the seventh day (Table 7). At a concentration of 1 ppm, flupropiconazole caused ticks to begin laying eggs on the second day, peaking between the seventh and tenth days (Table 8). At a concentration of 10 ppm, flupropiconazole caused ticks to begin laying eggs on the third day, a effect comparable to that of flupropiconazole at 1 ppm. This indicates that flupropiconazole at low concentrations significantly delays tick egg laying.
[0060] Table 1 Pre-screening of compound concentrations in the experimental group
[0061] concentration Survivor count Death count mortality rate(%) 100ppm (30 pieces) 0 30 100% 50ppm (30 pieces) 0 30 100% 10ppm (30 pieces) 0 30 100% 5ppm (30 pieces) 0 30 80% Negative control group (10 ppm, DMSO, 30 animals) 30 0 0
[0062] Table 2 Screening of fluranarine concentration in the positive control group
[0063] concentration Survivor count Death count mortality rate(%) 100ppm (30 pieces) 0 30 100% 50ppm (30 pieces) 0 30 100% 10ppm (30 pieces) 18 12 40% 5ppm (30 pieces) 30 0 0 Negative control group (10 ppm, DMSO, 30 animals) 30 0 0
[0064] Table 3. Tick mortality rate (24h) after treatment with the compound flupropazole
[0065] concentration Survivor count Death count mortality rate(%) 30ppm (18 pieces) 0 18 100% 20ppm (18 pieces) 0 18 100% 10ppm (18 pieces) 4 14 77.8% 5ppm (18 pieces) 7 11 61.1% 1 ppm (18 pieces) 12 6 33.3% Negative control group (0.1 ppm, DMSO, 18 animals) 18 0 0 Blank control group (distilled water, 18 animals) 18 0 0
[0066] Table 4. Tick mortality rate (48h) after treatment with the compound flupropazole
[0067] concentration Survivor count Death count mortality rate(%) 30ppm (18 pieces) 0 18 100% 20ppm (18 pieces) 0 18 100% 10ppm (18 pieces) 0 18 100% 5ppm (18 pieces) 0 18 100% 1 ppm (18 pieces) 7 11 61.1% Negative control group (0.1 ppm, DMSO, 18 animals) 18 0 0 Blank control group (distilled water, 18 animals) 18 0 0
[0068] Table 5. Tick mortality rate (24h) after fluranal treatment in the positive control group.
[0069] concentration Survivor count Death count mortality rate(%) 30ppm (18 pieces) 7 11 61.1% 20ppm (18 pieces) 10 8 44.4% 10ppm (18 pieces) 13 5 27.8% 5ppm (18 pieces) 16 2 11.1% 1 ppm (18 pieces) 17 1 5.56% Negative control group (0.1 ppm, DMSO, 18 animals) 18 0 0 Blank control group (distilled water, 18 animals) 18 0 0
[0070] Table 6. Tick mortality rate after fluranal treatment in the positive control group (48h)
[0071] concentration Survivor count Death count mortality rate(%) 30ppm (18 pieces) 0 18 100% 20ppm (18 pieces) 8 11 61.1% 10ppm (18 pieces) 10 8 44.4% 5ppm (18 pieces) 12 6 33.3% 1 ppm (18 pieces) 13 5 27.8% Negative control group (0.1 ppm, DMSO, 18 animals) 18 0 0 Blank control group (distilled water, 18 animals) 18 0 0
[0072] Table 7. Oviposition time of ticks after treatment with the compound flupropazole
[0073]
[0074] Although embodiments of the invention have been shown and described, it will be understood by those skilled in the art that various changes, modifications, substitutions and alterations can be made to these embodiments without departing from the principles and spirit of the invention, the scope of which is defined by the appended claims and their equivalents.
Claims
1. Use of the compound fluazaindazol for the preparation of a medicament for the prevention or treatment of parasitic infections caused by ticks, characterized in that, The medicine is used for killing Haemaphysalis longicornis.
2. Use of the compound fluazaindazoles according to claim 1 for the preparation of a veterinary drug, characterized by the fact that: The medicine comprises a therapeutically effective amount of fluazaindolyl as an active ingredient, and a pharmaceutically acceptable carrier or adjuvant.
3. Use of the compound fluazaindazoles according to claim 1 for the preparation of a veterinary drug, characterized in that: The medicine is prepared into a dosage form suitable for veterinary or agricultural use, which is at least one selected from injection, oral preparation, external preparation, sustained-release or controlled-release preparation.
4. Use of the compound fluazaindazoles according to claim 3 for the preparation of a veterinary drug, characterized by the fact that: The oral preparation is tablet, chewable tablet, granule and powder, and the external preparation includes drop, spray, lotion, powder and gel.
5. Use of the compound fluazaindazoles according to claim 1 for the preparation of a veterinary drug, characterized by the fact that: The effective concentration of fluazaindolyl in the medicine is 1-100 ppm.
6. Use of the compound fluazaindazoles according to claim 1 for the preparation of a veterinary drug, characterized in that: The fluazaindolyl is used in combination with other insecticides to form a compound preparation, which is at least one selected from organophosphorus, pyrethroid and isoxazoline.
7. Use of the compound fluazaindazoles according to claim 1 for the preparation of a veterinary drug, characterized by the fact that: The preparation method of the medicine comprises the following steps: mixing fluazaindolyl with a suitable carrier or diluent; adding an adjuvant; preparing a required dosage form through a conventional preparation process.
8. The use of compound fluazaindolyl in the preparation of a medicine for killing ticks according to claim 7, characterized in that: the adjuvant is at least one selected from binding agent, disintegrating agent, lubricant and stabilizer; the carrier is at least one selected from dimethyl sulfoxide (DMSO), propylene glycol, vegetable oil, starch, lactose and microcrystalline cellulose.
9. Use of the compound fluazaindazoles according to claim 7 for the preparation of a veterinary drug, characterized by the fact that: The mass percentage of fluazaindolyl in the medicine is 0.1%-10%.
10. Use of the compound fluazaindazoles according to claim 1 for the preparation of a veterinary drug, characterized by the fact that: The medicine is suitable for domestic animals and experimental animals.