阅读说明：本技术 10-十一烯酰氨基甲基丙酸乙酯及其制备方法和在驱蚊中的应用 (10-undecylenoyl amino ethyl propionate, preparation method thereof and application thereof in mosquito repelling ) 是由 吴华 戴建青 陈大嵩 黄鸿 郑基焕 于 2021-01-22 设计创作，主要内容包括：本发明公开了10-十一烯酰氨基甲基丙酸乙酯及其制备方法和在驱蚊中的应用。化合物10-十一烯酰氨基甲基丙酸乙酯,其结构式如式I所示。该化合物合成工艺路线简单,原材料易得,毒性低,无过敏,无刺激性,使用安全；具有良好的驱蚊效果,质量分数为10％的10-十一烯酰氨基甲基丙酸乙酯有效保护时间为8.3±1.0h,达到《GB/T13917.9-2009农药登记用卫生杀虫剂室内药效试验及评价第9部分：驱避剂》A级标准,具有进一步研究和扩大使用价值。(The invention discloses 10-undecylenoyl ethyl aminomethylpropionate, a preparation method thereof and application thereof in mosquito repelling. The compound 10-undecylenoyl ethyl aminomethylpropionate has a structural formula shown in formula I. The compound has the advantages of simple synthesis process route, easily obtained raw materials, low toxicity, no allergy, no irritation and safe use; the mosquito repellent agent has a good mosquito repellent effect, the effective protection time of 10-undecenoyl ethyl aminomethylpropionate with the mass fraction of 10% is 8.3 +/-1.0 h, and the mosquito repellent agent reaches the part 9 of GB/T13917.9-2009 pesticide registration sanitary insecticide indoor efficacy test and evaluation: the repellent has further research and expanded use value according to the A-grade standard.)

1. The compound 10-undecylenoyl ethyl aminomethylpropionate is characterized in that the structural formula is shown as a formula I:

2. the method for producing ethyl 10-undecenoylaminomethylpropionate according to claim 1, wherein 10-undecenoyl chloride and ethyl 3- (methylamino) propionate are used as raw materials, and the ethyl 10-undecenoylaminomethylpropionate is produced by condensation reaction in an organic solvent, separation and purification.

3. The method for preparing ethyl 10-undecenoylaminomethylpropionate according to claim 2, comprising the steps of:

a. dropwise adding ethyl 3- (methylamino) propionate into a mixed solution of 10-undecylenoyl chloride and an organic solvent while stirring, and then continuously stirring and reacting for 0.5-2h after the dropwise adding of the ethyl 3- (methylamino) propionate is finished, wherein the reaction temperature is 30-40 ℃, so as to obtain a solution after reaction;

b. adding a sodium hydroxide aqueous solution into the solution after the reaction, fully stirring, separating liquid, extracting an organic solvent layer, washing the organic solvent layer by using the sodium hydroxide aqueous solution, then washing the organic solvent layer by using a hydrochloric acid solution and water in sequence, and drying by using anhydrous sodium sulfate to obtain an organic solvent solution; evaporating the organic solvent in the organic solvent solution under reduced pressure to obtain a crude extract of 10-undecylenoyl ethyl methyl propionate;

c. and (3) carrying out acid-base washing on the crude extract of the 10-undecylenoyl ethyl methyl propionate, extracting by using an organic solvent, and evaporating under reduced pressure to obtain purified 10-undecylenoyl ethyl methyl propionate.

4. The method of claim 3, wherein the organic solvent is diethyl ether or n-hexane.

5. The method for preparing ethyl 10-undecenoylaminomethylpropionate according to claim 3, wherein the mass-to-volume ratio of 10-undecenoyl chloride to diethyl ether in step (a) is 1:1 to 3g/mL, and the mass ratio of 10-undecenoyl chloride to ethyl 3- (methylamino) propionate is 1.5 to 2.5: 1.

6. The method according to claim 3, wherein the aqueous sodium hydroxide solution in step (b) is a 5% by weight aqueous sodium hydroxide solution, the volume ratio of diethyl ether to the aqueous sodium hydroxide solution is 4:5, and the hydrochloric acid solution is a 10% by volume hydrochloric acid solution.

7. A mosquito repellent is characterized by containing 10-undecylenoyl ethyl methyl propionate with effective dose.

8. The mosquito repellent according to claim 7, wherein the mosquito repellent is in the form of an emulsion, an ointment, a tincture, a spray or an aerosol.

9. The use of the ethyl 10-undecenoylaminomethylpropionate of claim 1 for repelling mosquitoes.

10. The use of ethyl 10-undecenoylaminomethylpropionate for repelling mosquitoes according to claim 9, wherein said mosquitoes are aedes albopictus.

The technical field is as follows:

the invention belongs to the technical field of mosquito repelling, and particularly relates to 10-undecylenoyl ethyl methyl propionate, a preparation method thereof and application thereof in mosquito repelling.

Background art:

DEET has been reported as an effective repellent in the 50 s, and is currently used as a standard repellent in most countries and regions, being the most widely used repellent. It has been found that DEET has some disadvantages in use. If certain toxic and side effects exist, the symptoms of erythema, urticaria and the like can be caused to the user; the fabric has certain solubility to chemical fiber clothes; has certain permeability and irritation to skin. Even neurotoxicity may result, and mosquito resistance to DEET has now been found to be a potential source. Therefore, researches for searching new varieties of repellents and overcoming the existing defects are continuously carried out.

The invention content is as follows:

the invention solves the problems in the prior art, and provides 10-undecylenoyl amino methyl ethyl propionate, a preparation method and application thereof in mosquito repelling.

The invention aims to provide a compound 10-undecylenoyl ethyl aminomethylpropionate, which has a structural formula shown in a formula I:

the Chinese name of the compound is: ethyl 10-undecenoylaminomethylpropionate, alternative name of Chinese: 3- (N-10-undecylenoyl-N-methyl) -aminopropionic acid ethyl ester.

The invention also provides a preparation method of the 10-undecylenoyl ethyl methyl propionate, which comprises the steps of carrying out condensation reaction on 10-undecylenoyl chloride and ethyl 3- (methylamino) propionate serving as raw materials in an organic solvent, and separating and purifying to obtain the 10-undecylenoyl ethyl methyl propionate.

According to the proton theory of acid and base, active chlorine (Cl)^-) Readily and proton (H)⁺) The reaction produced hydrochloric acid (HCl). Providing chlorine (Cl) by 10-undecenoyl chloride^-) 3- (methylamino) propanoic acid ethyl ester donates a proton (H)⁺) To produce hydrochloric acid (HCl) and Ethyl 10-undecenoylaminomethylpropionate (Ethyl methyl 10-undecylaminopropionate). The chemical reaction equation is as follows:

preferably, the preparation method of the 10-undecenoyl ethyl aminomethylpropionate specifically comprises the following steps:

a. dropwise adding ethyl 3- (methylamino) propionate into the mixed solution of 10-undecylenoyl chloride and diethyl ether while stirring, and then continuously stirring and reacting for 0.5-2h after the dropwise adding of the ethyl 3- (methylamino) propionate is finished, wherein the reaction temperature is 30-40 ℃, so as to obtain a solution after reaction;

b. adding a sodium hydroxide aqueous solution into the solution after the reaction, fully stirring, separating liquid, extracting an organic solvent layer, washing the organic solvent layer by using the sodium hydroxide aqueous solution, then washing the organic solvent layer by using a hydrochloric acid solution and water in sequence, and drying by using anhydrous sodium sulfate to obtain an organic solvent solution; evaporating the organic solvent in the organic solvent solution under reduced pressure to obtain a crude extract of 10-undecylenoyl ethyl methyl propionate;

c. and (3) carrying out acid-base washing on the crude extract of the 10-undecylenoyl ethyl methyl propionate, extracting by using an organic solvent, and evaporating under reduced pressure to obtain purified 10-undecylenoyl ethyl methyl propionate.

More preferably, the organic solvent is diethyl ether or n-hexane.

Further preferably, the mass-to-volume ratio of the 10-undecylenoyl chloride to the diethyl ether in the step (a) is 1:1-3g/mL, and the mass ratio of the 10-undecylenoyl chloride to the ethyl 3- (methylamino) propionate is 1.5-2.5: 1.

More preferably, the sodium hydroxide aqueous solution in step (b) is a 5% by weight sodium hydroxide aqueous solution, the volume ratio of the diethyl ether to the sodium hydroxide aqueous solution is 4:5, and the hydrochloric acid solution is a 10% by volume hydrochloric acid solution.

The third purpose of the invention is to protect a mosquito repellent, which contains an effective amount of 10-undecylenoyl ethyl aminomethylpropionate. Preferably, the mosquito repellent contains 10 mass percent of 10-undecylenoyl ethyl methyl propionate.

Preferably, the dosage form of the mosquito repellent is emulsion, paste, tincture, spray or aerosol.

The fourth purpose of the invention is to protect the application of the 10-undecylenoyl ethyl methyl propionate in repelling mosquitoes.

Preferably, the mosquito is aedes albopictus.

Compared with the prior art, the invention has the following advantages:

the 10-undecylenoyl ethyl aminomethylpropionate has the advantages of good mosquito repellent effect, low toxicity, no allergy to skin, no irritation, safe use of compounds, uncomplicated synthesis process route, easily available raw materials, and further research and expanded use values.

Description of the drawings:

FIG. 1 is an infrared spectrum of ethyl 10-undecenoylaminomethylpropionate prepared in example 1;

FIG. 2 is a GC/MS mass spectrum of ethyl 10-undecenoylaminomethylpropionate prepared in example 1;

FIG. 3 is a nuclear magnetic resonance spectrum of ethyl 10-undecenoylaminomethylpropionate prepared in example 1.

The specific implementation mode is as follows:

the following examples are further illustrative of the present invention and are not intended to be limiting thereof.

Unless otherwise specified, the experimental materials and reagents in the invention are all conventional commercial products in the technical field.

Example 1:

synthesis method of compound

Preparation of ethyl 10-undecenoylaminomethylpropionate: in a 500mL three-necked reaction flask equipped with a stirrer, a dropping funnel and a thermometer, 40g of 10-undecenoyl chloride and 80mL of diethyl ether were charged. 20g of ethyl 3- (methylamino) propionate is slowly dropped into a dropping funnel, the speed of dropping the ethyl 3- (methylamino) propionate (about 0.5g/min) is controlled by paying attention to the boiling condition, the reaction process is allowed to be smooth, and the reaction temperature is 30-40 ℃. After the dripping is finished, stirring is continued for 1 h. Then, 100mL of a 5% aqueous solution of sodium hydroxide was added to the reaction mixture, and the mixture was stirred sufficiently for 1 hour to complete the reaction. Then, the ether layer was extracted by liquid separation. The ether layer was washed once more with 5% aqueous sodium hydroxide. The ether layer was then washed once with 10% by weight of each of a hydrochloric acid solution and water. Drying over anhydrous sodium sulfate yielded 48mL of ether layer. Evaporation of 48mL of the ether layer under reduced pressure gave 41.235g of the final product, ethyl 10-undecenoylaminomethylpropionate, in 90.87% yield. The 41.235g sample was washed with 10% by mass hydrochloric acid solution and 5% by mass sodium hydroxide aqueous solution in this order, and then extracted with ether and evaporated under reduced pressure to give 95% purity. The finally obtained 10-undecenoyl aminomethyl ethyl propionate sample is a light yellow oily liquid, is insoluble in water and is soluble in a common organic solvent.

Structural identification of compounds

The obtained purified 10-undecenoyl ethyl aminomethylpropionate compound is subjected to compound structure identification and analysis by infrared spectroscopy, GC/MS mass spectrometry and nuclear magnetic resonance. The inspection basis is as follows: general rule of infrared spectroscopic analysis GB/T6040-2002; general rule of organic mass spectrometry JY/T003-; the general rule of superconducting pulse Fourier transform nuclear magnetic resonance spectroscopy JY/T007-.

The structure identification results are as follows:

infrared spectrum analysis: infrared spectroscopic analysis (FIG. 1), IR, v/cm, was conducted on a sample of ethyl 10-undecenoylaminomethylpropionate^-1: v is stretching, δ is bending, β is in-plane bending, γ is out-of-plane bending, ω is out-of-plane rocking, s is symmetric vibration, and as is asymmetric vibration. IR, v/cm^-1：3 077(C＝Cν_C-H)，2 978(-CH₃ν_asC-H)，2 927(-CH₂-ν_asC-H)，2 855(-CH₂-ν_sC-H)，1 735(-COO-ν_C＝O)，1 645(N-C＝Oν_C＝O)，1 465(-CH₃δ_C-H)，1 406(-CH₂-δ_C-H)，1 375(-CH₃δ_C-H)，1 184，1 046(ν_C-O-C)，995(γ_＝CH)，910(γ_＝CH2)，724(-CH₂-ω_C-H). According to the expected structure of the compound.

GC/MS mass spectrometry: GC/MS mass spectrometry of a sample of ethyl 10-undecenoylaminomethylpropionate revealed in FIG. 2 that EI-MS, m/z,%: according to the expected structure of the compound.

Nuclear magnetic resonance spectrum analysis: the NMR spectrum analysis of a sample of ethyl 10-undecenoylaminomethylpropionate was carried out, and as can be seen from FIG. 3, the hydrogen spectrum¹H NMR(300MHz，CDCl₃)δ：1.917-2.260(m，16H，-CH₂-CH₂-CH₂-CH₂-CH₂-CH₂-CH₂-CH₂-)，4.816-5.762(m，3H，CH₂＝CH-)，2.839、2.962(m，3H，)，2.468-3.576(m，4H，-CH₂-CH₂-)，4.036-4.087(m，2H，-O-CH₂-)，1.220(m，3H，-CH₃) (ii) a Carbon spectrum¹³C NMR(75MHz，CDCl₃)δ：173.554(1C，)，28.800-34.012(8C，-CH₂-CH₂-CH₂-CH₂-CH₂-CH₂-CH₂-CH₂-)，138.989，114.040(2C，CH₂＝CH-)，36.322(2C，)，44.428、45.474、32.56(2C，-CH₂-CH₂-)，172.055(1C，)，60.483、60.859(1C，-O-CH₂-)，14.059(1C，-CH₃). According to the expected structure of the compound.

Therefore, the compound 10-undecenoylaminomethylpropionic acid ethyl ester has a chemical structure shown in the formula I:

example 2

The same as example 1, except that: and a step a, continuously stirring and reacting for 0.5h, wherein the mass-volume ratio of 10-undecylenoyl chloride to diethyl ether is 1:1g/mL, and the mass ratio of 10-undecylenoyl chloride to ethyl 3- (methylamino) propionate is 1.5: 1.

Example 3

The same as example 1, except that: and a step a, continuously stirring and reacting for 2 hours, wherein the mass-volume ratio of 10-undecylenoyl chloride to diethyl ether is 1:3g/mL, and the mass ratio of 10-undecylenoyl chloride to ethyl 3- (methylamino) propionate is 2.5: 1.

Example 4

The same as example 1, except that: the organic solvent was n-hexane, and the amount of n-hexane used was the same as that of ether in example 1.

Test example 1

The compound prepared in example 1 was tested as follows:

1. toxicity testing of Compounds

The acute oral toxicity of the ethyl 10-undecylenoylaminomethylpropionate to mice is tested according to a test method provided in GB15193.3-2014 national food safety standard acute oral toxicity test. Upper-lower limit test: the test subjects were given to 1 animal at a dose of 2000mg/kg body weight and if the animals died within 48h, the test was performed as a standard test. If the animal survived within 48h, taking another 4 animals to administer the test substance with the same dose, such as 3 out of 5 animals die, and performing formal test; if more than 3 animals survive, the test is terminated, and the test subject LD is obtained₅₀>2000mg/kg body weight.

The compound toxicity test results are as follows:

the acute oral toxicity of the 10-undecylenic acylaminomethylpropionic acid ethyl ester on the white mice is determined to be female mouse LD₅₀>2000mg/kg, male mouse is LD₅₀>2000 mg/kg. According to the national food safety standard (GB15193.3-2014) of China, the rat oral administration LD₅₀The toxicity is 501-5000 mg/kg. Therefore, the 10-undecylenoyl amino methyl ethyl propionate is low in toxicity.

During the indoor and field repelling experiments by referring to the GB/T13917.9-2009 test method, the hand skin and the leg skin of the tested persons (24 persons) have no drug allergy phenomenon and no irritation.

2. Laboratory mosquito repellent effect of compounds

The laboratory mosquito repellent effect of the 10-undecylenoyl ethyl aminomethylpropionate on Aedes albopictus is as follows: preparing 10-undecylenoyl methyl ethyl propionate into ethanol solutions with the mass fractions of 2.5%, 5.0%, 7.5% and 10.0% respectively by using absolute ethanol, preparing butyl ethyl acetamido propionate (RI3535, mosquito repellent ester) into an ethanol solution with the mass fraction of 10%, preparing Diethyltoluamide (DEET) into an ethanol solution with the mass fraction of 10%, and using the absolute ethanol as a reference for later use.

Referring to GB/T13917.9-2009 indoor pesticide effect test and evaluation part 9 of sanitary pesticides for pesticide registration: experiments were performed using the test methods provided in repellents. Selecting 4 or more testers with qualified offensive power (half of male and female, and should not drink before and during the testTea or coffee, should not use products containing essences), and the skin area of 50mm x 50mm is drawn on the back of each hand, wherein the skin area of one hand is 1.5mg/cm²(paste repellent) or 1.5. mu.L/cm²The dosage of the (liquid repellent) is uniformly coated on the repellent to be tested, the skin with the thickness of 40mm multiplied by 40mm is exposed, the rest part is closely shielded, and the other hand is a blank control (namely coated with 1.5 mu L/cm)²Absolute ethanol of (ii). Smearing repellent for 2h, putting hands into mosquito cage with qualified attack power for 2min, and observing whether there is mosquito coming before and stopping and sucking blood. And then testing every 1h, and judging that the repellent fails as long as one mosquito sucks blood in front of the mosquito. The effective protection time (h) of the repellent was recorded. The contrast test is firstly carried out by the contrast hand each time, the test insects with qualified attack force can be continuously tested, and the test insects with unqualified attack force need to be replaced by qualified test insects for testing.

The mosquito repelling result in the laboratory is as follows:

the repellent effect of the ethyl 10-undecylenoylaminomethylpropionate on the aedes albopictus is increased along with the increase of the concentration, and the effective protection time and the concentration show linear positive correlation. The effective protection time of 10-undecylenoyl ethyl aminomethylpropionate with the mass fraction of 10% is 8.3 +/-1.0 h, and reaches the part 9 of GB/T13917.9-2009 pesticide registration sanitary insecticide indoor efficacy test and evaluation: repellents "level a standard. The effective protection time of the control drugs of the butyl ethyl levamisole (RI3535, anophelifuge) and the DEET at the same concentration is 6.0 +/-0.8 h and 5.8 +/-1.0 h respectively (Table 1).

TABLE 1 mosquito repellent Effect of different concentrations of ethyl 10-undecenoylaminomethylpropionate

Table 1 Repellent effects of Ethyl methyl 10-undecylaminopropionate at different concentrations

The data in the table are mean numbers ± standard deviation (n ═ 4); values are the means ± SD (n ═ 4).

3. On-site mosquito repellent effect of the compounds

The on-site mosquito repelling effect of the 10-undecylenoyl ethyl aminomethylpropionate on Aedes albopictus is as follows:

the field mosquito repelling test is carried out in 5 months in 2020, and in the mosquito activity time, in the institute of animal research of the academy of sciences of Guangdong province. The trees beside the roads in the yard are mainly prepared from bauhinia variegata, kapok, mango trees, magnolia alba, syzygium samarangense and ficus auriculata, green belts around the building are mainly planted with calamus marjoris and yueli, and a fishpond is arranged outside the enclosure wall. The test is divided into 4 groups, each group comprises 5 persons, the test medicine group is 10 mass percent of 10-undecylenoyl ethyl methyl propionate ethanol solution, the reference medicine group 1 is 10 mass percent of butyl ethyl acetamido propionate (RI3535, mosquito repellent ester) ethanol solution, the reference medicine group 2 is 10 mass percent of DEET ethanol solution, and the blank reference group is absolute ethyl alcohol. Test samples are randomly distributed to subjects, the subjects wear long clothes and trousers, the left leg and the trouser leg are rolled up, and 3mL of the test samples are uniformly coated on the parts below the knee joint and above the ankle joint of the left leg. The medicine applying part is kept exposed all the time. Standing for 15min every 1h to a place where mosquitoes are dense in an outdoor green belt tree, observing the mosquito sting condition at any time, and recording the number of the stinging mosquitoes and the sting time. The effective protection time (h) and the effective protection rate (%) were calculated. The calculation formula of the effective protection rate is as follows:

the field mosquito repelling result is as follows:

the ethanol solution of 10-undecylenic acylamino ethyl propionate with the mass fraction of 10% is applied to protect the human body on site, no people are bitten by mosquitoes in 5h, 6h and 7h, and the protection rate is 100%; 2 persons are bitten in the 8 th hour, the protection rate is 92.13%, 5 persons are bitten in the 9 th hour, and the protection rate is 78.65%.

The effective protection rate of the general repellent is more than or equal to 80 percent and is considered to be effective, the effective protection rate is less than 80 percent and is considered to be no protection effect, the protection rate of 8 hours on site is 92.13 percent, so the effective protection time of 8 hours on site is also effective, and the effective protection time of 10 percent of 10-undecylenoyl ethyl methyl propionate on site to human bodies is 8 hours. Compared with an ethanol solution of 10% of ethyl butylacetamidopropionate (RI3535, anophelifuge) and an ethanol solution of 10% of DEET, no person is bitten by mosquitoes in the 5 th hour, and the protection rate is 100%; 4 persons are bitten in the 6 th hour, the protection rates are respectively 90.11 percent and 89.01 percent, and the 7 th hour 5 persons are bitten, and the protection rates are respectively 57.30 percent and 56.18 percent; the placebo ethanol group 5 was bitten (table 2).

TABLE 2 on-site mosquito repellent effect of 10-undecenoylaminomethylpropionic acid Ethyl ester at 10% by mass fraction

Table 2 Field repellent effect of 10％Ethyl methyl 10-undecylaminopropionate

Note: the average sting number of the mosquitoes in the blank control ethanol group is 16.2-18.2 per 15 min; the average number of moles in The blank controlled alcohol group waters is 16.2-18.2/15 min;

the data in the table are mean numbers ± standard deviation (n ═ 5); values are the means ± SD (n ═ 5).

The above embodiments are only for the purpose of helping understanding the technical solution of the present invention and the core idea thereof, and it should be noted that those skilled in the art can make several improvements and modifications to the present invention without departing from the principle of the present invention, and these improvements and modifications also fall within the protection scope of the claims of the present invention.