Pesticide active compound or parent compound as well as preparation method and application of pesticide active compound or parent compound

Abstract

The invention relates to a raw pesticide or a parent pesticide as well as a preparation method and application of the raw pesticide or the parent pesticide. The raw medicine or the parent medicine contains at least two kinds of materials from 1-N glycosyl chain ridinyl-2-amino-(L-lysyl) n-L-lysine-2-deoxygulosamine, and the raw medicine or the parent medicine contains at least two kinds of materials from 1-N glycosyl chain ridinyl-2-amino-(L-lysyl) The total content of the 1-N glycosyl lindinyl-2-amino-(L-lysyl) n-L-lysine-2-deoxygulosamine is more than 70% in terms of the total mass of the raw medicine or the parent medicine; wherein n represents the number of L-lysyl and is any integer from 0 to 4. The raw pesticide or the parent pesticide of the pesticide contains 1-N glycosyl-lindinyl-2-amino-(L-lysyl) n-L-lysine-2-deoxygulosamine with relatively high content, and has a better effect in the aspects of stability of effective components, stability of antibacterial ability and the like compared with the raw pesticide or the parent pesticide with relatively low content.

Description

Technical Field

[0001] The invention belongs to the technical field of agriculture, and specifically relates to a pesticide technical or mother drug and a preparation method and application thereof. Background Art

[0002] 1-N-Glycosyl-2-amino-(L-lysyl) n -L-lysine-2-deoxygulosamine (where n represents the number of L-lysyl groups and can be any integer between 0 and 4) has excellent bactericidal properties and can resist Gram-positive bacteria, Gram-negative bacteria, mycobacteria, yeasts and filamentous fungi. It is an agricultural antibiotic with good application prospects.

[0003] 1-N-Glycosyl-2-amino-(L-lysyl) n -L-Lysine-2-deoxygulosamine Currently, there is only one technical or mother drug product registered in my country, and its active ingredient content is 12%. The active ingredient content is low and the product quality is poor.

[0004] CN113896753A discloses a method for efficiently separating the agricultural antibiotic from the fermentation broth, by combining ceramic membrane filtration and cationic resin exchange adsorption and other methods to obtain 1-N-glycosidyl-2-amino-(L-lysyl) n -L-lysine-2-deoxygulosamine content of 25-65% of the original drug, its color and purity are greatly improved compared with the 12-24% products on the market. However, the applicant found in the experiment that the original drug obtained by CN113896753A degraded rapidly during storage and the antibacterial activity decreased significantly. In order to better meet the needs of actual production, it is urgent to develop a 1-N-glycosidic acid chain 2-amino-(L-lysyl) n -L-lysine-2-deoxygulosamine original drug or parent drug. Summary of the invention

[0005] In order to solve the above technical problems, according to one aspect of the present application, a pesticide raw material or a raw material is provided, wherein the pesticide raw material or the raw material contains 1-N-glycosidyl-2-amino-(L-lysyl) n - at least 2 of L-lysine-2-deoxygulosamine;

[0006] Based on the total mass of the original drug or the parent drug, the 1-N-glycosidyl-2-amino-(L-lysyl) n -The total content of L-lysine-2-deoxygulosamine is more than 70%;

[0007] Wherein n represents the number of L-lysyl groups, and can be any integer between 0 and 4.

[0008] Optionally, based on the total mass of the original drug or the mother drug, the total content of the 1-N-glycosidyl-2-amino-(L-lysyl) nL-lysine-2-deoxygulosamine is 70%-94%.

[0009] Optionally, based on the total mass of the original drug or the mother drug, the total content of the 1-N-glycosidyl-2-amino-(L-lysyl) nL-lysine-2-deoxygulosamine is 70%-90%.

[0010] Optionally, the original drug or mother drug contains 1-N-glycosidyl-2-amino-(L-lysyl) n - At least 3 of L-lysine-2-deoxygulosamine.

[0011] Optionally, the original drug or mother drug contains 1-N-glycosidyl-2-amino-(L-lysyl) n - At least 4 of L-lysine-2-deoxygulosamine.

[0012] Optionally, the original drug or mother drug contains 1-N-glycosidyl-2-amino-(L-lysyl) n - At least 5 of L-lysine-2-deoxygulosamine.

[0013] According to another aspect of the present application, a method for preparing the above-mentioned pesticide technical or concentrate is provided, comprising the following steps: a fermentation step and a purification step.

[0014] According to another aspect of the present application, there is provided the use of the above-mentioned pesticide technical or bulk drug or the pesticide technical or bulk drug obtained according to the above-mentioned method in the preparation of a fungicide.

[0015] The positive and progressive effects of the present invention are:

[0016] The pesticide technical or bulk drug provided in this application, based on the total mass of the technical or bulk drug, contains 1-N-glycosidyl-2-amino-(L-lysyl) n -L-lysine-2-deoxygulosamine has a total content of more than 70%, and its stability in terms of active ingredient content and antibacterial effect is better than that of the original drug or mother drug with a lower content, which is more conducive to extending the shelf life of the original drug or mother drug. n The total content of -L-lysine-2-deoxygulosamine is controlled within the range of 70%-94%, which can better ensure that the hygroscopicity of the original drug or the mother drug is weak, and at the same time can greatly reduce the degradation rate of its active ingredients, and can better maintain its antibacterial ability, and has better comprehensive performance in terms of processing performance, storage convenience and storage stability. BRIEF DESCRIPTION OF THE DRAWINGS

[0017] Figure 1 This is the liquid phase spectrum of the pesticide technical or mother drug prepared in Example 2, wherein the darker color in the data table is 1-N-glycosidyl-2-amino-(L-lysyl) n -L-lysine-2-deoxygulosamine, the retention time n is 0, 1, 2, 3, and 4 from small to large;

[0018] Figure 2 This is the liquid phase spectrum of the pesticide technical or mother drug prepared in Example 3, wherein the darker color in the data table is 1-N-glycosidyl-2-amino-(L-lysyl) n -L-lysine-2-deoxygulosamine, the retention time n is 0, 1, 2, 3, and 4 from small to large;

[0019] Figure 3 This is the liquid phase spectrum of the pesticide technical or mother drug prepared in Example 5, wherein the darker color in the data table is 1-N-glycosidyl-2-amino-(L-lysyl) n -L-lysine-2-deoxygulosamine, the retention time n is 0, 1, 2, 3, and 4 from small to large;

[0020] Figure 4 This is the liquid phase spectrum of the pesticide technical or mother drug prepared in Example 6, wherein the darker color in the data table is 1-N-glycosidyl-2-amino-(L-lysyl) n -L-lysine-2-deoxygulosamine, the retention time n is 0, 1, 2, 3, and 4 from small to large;

[0021] Figure 5 It is a diagram of moisture absorption of samples of Example 2, Example 3, Example 5 and Example 6;

[0022] Figure 6 The antibacterial effect diagram of the samples of Example 2, Example 3, Example 5 and Example 6 is shown in FIG. Figure 6 A is the antibacterial effect of the sample not treated at 60℃ / 25% RH. Figure 6 B is the antibacterial effect of the sample treated under 60°C / 25% RH conditions. DETAILED DESCRIPTION

[0023] For the purpose of the present invention, unless otherwise specified, the terms used in this application have the following meanings:

[0024] "Active ingredient" refers to the ingredient in pesticide products that has toxic and killing activity against diseases, insects, weeds, etc. In this application, it specifically refers to "1-N-glycosidyl-2-amino-(L-lysyl) n-L-lysine-2-deoxygulosamine".

[0025] "1-N-glycosidyl-2-amino-(L-lysyl) n -L-lysine-2-deoxygulosamine" refers to the BF component of Zhongshengmycin. When n=0, it is the Zhongshengmycin F component, when n=1, it is the Zhongshengmycin E component, when n=2, it is the Zhongshengmycin D component, when n=3, it is the Zhongshengmycin C component, and when n=4, it is the Zhongshengmycin B component.

[0026] "Technical product" refers to the product obtained in the production process consisting of active ingredients and related impurities. A small amount of additives may be added if necessary.

[0027] "Material drug" refers to the product obtained in the production process consisting of active ingredients and related impurities, which may contain small amounts of necessary additives and appropriate diluents.

[0028] In this application, the original drug or parent drug contains 1-N-glycosidyl-2-amino-(L-lysyl) n -L-lysine-2-deoxygulosamine, the rest are different from 1-N-glycosidic chain 2-amino-(L-lysyl) n -L-lysine-2-deoxygulosamine and other substances, which can be extracted from the fermentation broth except 1-N-glycosidyl-2-amino-(L-lysyl) n -Other substances other than L-lysine-2-deoxygulosamine (such as related impurities) may also be other additionally added substances (such as additives or diluents).

[0029] Based on the total mass of the original drug or the parent drug, the 1-N-glycosidyl-2-amino-(L-lysyl) n The "total content" in "total content of L-lysine-2-deoxygulosamine" means that the original drug or parent drug contains at least two 1-N-glycosidyl-2-amino-(L-lysyl) n -L-lysine-2-deoxygulosamine, wherein n is any integer from 0 to 4, and all 1-N-glycosidic chain lidinyl-2-amino-(L-lysyl) n -The percentage of the total mass of L-lysine-2-deoxygulosamine to the total mass of the original drug or parent drug.

[0030] For example, the original drug or parent drug contains a total of 2 1-N-glycosidic chain 2-amino-(L-lysyl) n -L-lysine-2-deoxygulosamine, n is 0 and 1 respectively, the total mass of the original drug or parent drug is 100g, these two 1-N-glycosidic chain liridyl-2-amino-(L-lysyl)n -L-lysine-2-deoxygulosamine has a total mass of 70 g, then, based on the total mass of the original drug or mother drug, the 1-N-glycosidyl-2-amino-(L-lysyl) n -L-lysine-2-deoxygulosamine has a total content of 70%. It contains two 1-N-glycosidic chain 2-amino-(L-lysyl) n -L-lysine-2-deoxygulosamine original drug or mother drug also includes but is not limited to the following scenarios: n is 0 and 2 respectively; or n is 0 and 3 respectively; or n is 0 and 4 respectively; or n is 1 and 2 respectively; or n is 1 and 3 respectively; or n is 2 and 3 respectively; or n is 2 and 4 respectively; or n is 3 and 4 respectively; or other combinations.

[0031] Alternatively, for example, the original drug or parent drug contains a total of three 1-N-glycosidic chain lysine-2-amino-(L-lysyl) n -L-lysine-2-deoxygulosamine, n is 0, 1, 2, the total mass of the original drug or parent drug is 100g, these three 1-N-glycosidic chain liridyl-2-amino-(L-lysyl) n -L-lysine-2-deoxygulosamine has a total mass of 80 g, then, based on the total mass of the original drug or mother drug, the 1-N-glycosidyl-2-amino-(L-lysyl) n -L-lysine-2-deoxygulosamine has a total content of 80%. It contains three 1-N-glycosidic chain 2-amino-(L-lysyl) n -L-lysine-2-deoxygulosamine original drug or mother drug also includes but is not limited to the following scenarios: n is 0, 1, 3 respectively; or n is 0, 1, 4 respectively; or n is 1, 2, 3 respectively; or n is 1, 3, 4 respectively; or other combinations.

[0032] Or, for example, the original drug or parent drug contains a total of four 1-N-glycosidic chain 2-amino-(L-lysyl) n -L-lysine-2-deoxygulosamine, n is 0, 1, 2, 3, the total mass of the original drug or parent drug is 100g, these four 1-N-glycosidic chain 2-amino-(L-lysyl) n -L-lysine-2-deoxygulosamine has a total mass of 90 g, then, based on the total mass of the original drug or mother drug, the 1-N-glycosidyl-2-amino-(L-lysyl) n -L-lysine-2-deoxygulosamine has a total content of 90%. It contains three 1-N-glycosidic chain 2-amino-(L-lysyl) n-L-lysine-2-deoxygulosamine original drug or mother drug also includes but is not limited to the following scenarios: n is 0, 1, 2, 4 respectively; or n is 0, 2, 3, 4 respectively; or n is 1, 2, 3, 4 respectively; or other combinations.

[0033] Alternatively, for example, the original drug or parent drug contains a total of 5 1-N-glycosidic chain 2-amino-(L-lysyl) n -L-lysine-2-deoxygulosamine, n is 0, 1, 2, 3, 4, the total mass of the original drug or parent drug is 100g, these five 1-N-glycosidic chain liridyl-2-amino-(L-lysyl) n -L-lysine-2-deoxygulosamine has a total mass of 90 g, then, based on the total mass of the original drug or mother drug, the 1-N-glycosidyl-2-amino-(L-lysyl) n -The total content of L-lysine-2-deoxygulosamine is 90%.

[0034] In a specific embodiment, the pesticide technical or stock drug provided by the present application has a total active ingredient content of 70% or more. Compared with pesticide technical or stock drugs with lower active ingredient content, such as technical or stock drugs with a total active ingredient content of 60%, the technical or stock drug provided by the present application has slower degradation of active ingredients during storage, and the antibacterial activity is better maintained.

[0035] Furthermore, the inventors of the present application have found that 1-N-glycosidyl-2-amino-(L-lysyl) nL-lysine-2-deoxygulosamine is a hygroscopic compound, especially at high content, its hygroscopicity is strong, for example, when the content is above 96%. Therefore, the present application preferably provides a raw material or mother drug with a total active ingredient content of 70%-94%, and this content range can ensure that the raw material or mother drug has weak hygroscopicity, and can also effectively reduce the degradation rate of its active ingredients, and can better maintain its antibacterial ability, etc. Further preferably, the total content of active ingredients in the original drug or mother drug is a range value consisting of any two of 70%, 71%, 72%, 73%, 74%, 75%, 76%, 77%, 78%, 79%, 80%, 81%, 82%, 83%, 84%, 85%, 86%, 87%, 88%, 88.5%, 89%, 89.5%, 90%, 90.5%, 91%, 91.5%, 92%, 93%, 94%. Further preferably, the total content of active ingredients in the original drug or mother drug is any one of 70%-94%, 70%-90%, 70%-85%, 70%-83%, 70%-80%, 70%-75%.

[0036] In a specific embodiment, the preparation method of the pesticide technical or concentrate provided in the present application comprises: a fermentation step and a purification step.

[0037] In a specific embodiment, the strain used for fermentation is not limited, and the effective ingredient 1-N-glycosidyl-2-amino-(L-lysyl) n -L-lysine-2-deoxygulosamine. Preferably, it is Streptomyces lavendulae var. hainanensis.

[0038] The present application does not limit the method for purifying pesticide technical or mother drug from fermentation broth, as long as the total content of the active ingredient obtained is within the scope of the present application. Preferably, the purification step includes: a fermentation broth bacterial separation step; a resin adsorption and analysis step; and / or a concentration step. The bacterial separation includes but is not limited to centrifugation, filtration, etc. The resin is preferably an ion exchange resin, such as 724 weakly acidic cation exchange resin; the analytical solution is preferably an acidic analytical solution, such as a hydrochloric acid solution. The concentration includes but is not limited to nanofiltration concentration, etc. Specific embodiment:

[0040] The reagents and solvents used in the following examples are commercially available or prepared by conventional methods known to those skilled in the art. All commercially available reagents and solvents were not further purified. The experimental methods in the following examples without specifying specific conditions were performed according to conventional methods and conditions.

[0041] The sample testing in the examples was carried out according to the following method.

[0042] 1. Determination of active ingredient content in pesticide technical or parent drug

[0043] (1) Preparation of sample solution:

[0044] Weigh 0.025 g of the sample to be tested, place it in a 50 mL volumetric flask, dilute to the scale with water, shake well, and filter.

[0045] (2) Test conditions:

[0046] Chromatographic column: 250 mm × 4.6 mm (id) stainless steel column, filled with C18, 5 μm filler.

[0047] Mobile phase: Gradient elution conditions are shown in Table 1.

[0048] Flow rate: 1.0mL / min.

[0049] Column temperature: room temperature (temperature variation should not exceed 2°C).

[0050] Detection wavelength: 200nm.

[0051] Injection volume: 10 μL.

[0052] Table 1 Gradient elution conditions

[0053] Time / min Acetonitrile Mixed salt solution 0 85 15 2 85 15 10 78 22 20 78 22 22 85 15 30 85 15

[0054] The above operating parameters are typical and can be appropriately adjusted according to the characteristics of different instruments in order to obtain the best results. The retention time of each component of the active ingredient is confirmed using the standard of the corresponding component.

[0055] (3) Calculation of results

[0056] The content of active ingredients was calculated by area normalization method.

[0057] 2. Determination of the degradation rate of active ingredients in pesticide technical or parent drug

[0058] (1) Preparation of sample solution:

[0059] Same as the above “Determination of the content of active ingredients in pesticide technical or bulk drug”.

[0060] (2) Test conditions:

[0061] Same as the above “Determination of the content of active ingredients in pesticide technical or bulk drug”.

[0062] (3) Calculation of results

[0063] The result is calculated according to the following formula:

[0064] M=(A1-A2) / A1×100%

[0065] Where:

[0066] M - degradation rate of active ingredients;

[0067] A1 - the total peak area of ​​the active ingredients of the sample not treated at 60℃ / 25%RH;

[0068] A2 - Total peak area of ​​active ingredients in samples treated at 60°C / 25% RH.

[0069] Preparation Example

[0070] Example 1 Preparation of pesticide technical or parent drug

[0071] 1. Preparation of fermentation broth

[0072] The fermentation broth is obtained by fermenting the Hainan variant of Streptomyces lilacinus in a culture medium containing glucose, corn starch, corn flour, soybean flour, inorganic salts, etc. The fermentation process is provided by Shaanxi Micro Biotechnology Co., Ltd., the applicant of CN113896753A (the company is a subsidiary of Yuanda Industrial Holdings Co., Ltd., the applicant of this application).

[0073] 2. Purification

[0074] According to steps 1)-4) of Example 3 of CN113896753A, the fermentation liquid is filtered through a ceramic membrane, and the materials of the high unit clear liquid tank and the low unit clear liquid tank are collected respectively. The materials of the low unit tank are adsorbed and analyzed by LX-731-X1 modified resin, and the materials of the high unit tank and the eluent are mixed, concentrated and dried to obtain the pesticide technical or mother drug. It is determined that the obtained pesticide technical or mother drug contains 1-N-glycosidyl-2-amino-(L-lysyl) n -L-lysine-2-deoxygulosamine, n is 0, 1, 2, 3, 4 respectively, and the total content of each component is 48.8% based on the total mass of the original drug or the mother drug.

[0075] Example 2 Preparation of pesticide technical or parent drug

[0076] 1. Preparation of fermentation broth

[0077] Same as Example 1.

[0078] 2. Purification

[0079] According to steps 1)-4) of Example 1 of CN113896753A, the fermentation liquid is filtered through a ceramic membrane, and the materials of the high unit clear liquid tank and the low unit clear liquid tank are collected respectively. The materials of the low unit tank are adsorbed and analyzed by LX-731-X1 modified resin, and the analyzed liquid is concentrated and dried to obtain the pesticide technical or mother drug. It is determined that the obtained pesticide technical or mother drug contains 1-N-glycosidyl-2-amino-(L-lysyl) n -L-lysine-2-deoxygulosamine, n is 0, 1, 2, 3, 4, respectively. Based on the total mass of the original drug or parent drug, the total content of each component is 60.0%. The liquid chromatogram is shown in Figure 1 shown.

[0080] Example 3 Preparation of pesticide technical or parent drug

[0081] 1. Preparation of fermentation broth

[0082] Same as Example 1.

[0083] 2. Purification

[0084] (1) Acidification of the fermentation broth: acidifying the fermentation broth to a pH of 3.5;

[0085] (2) Bacteria isolation: The acidified material is centrifuged and the resulting clear liquid is filtered twice through an ultrafiltration membrane;

[0086] (3) Resin adsorption and analysis: Use 724 weakly acidic cation exchange resin to adsorb the filtrate, rinse with water until the effluent is colorless, and analyze with 5% hydrochloric acid. After discarding the 0.5 BV effluent, start collecting samples and stop collecting samples after 5 BV;

[0087] (4) Nanofiltration concentration: The analytical solution is concentrated by nanofiltration;

[0088] (5) Drying: The concentrated solution is spray dried to obtain the pesticide technical or the mother drug. The obtained pesticide technical or the mother drug is tested to contain 1-N-glycosidyl-2-amino-(L-lysyl) n -L-lysine-2-deoxygulosamine, based on the total mass of the original drug or parent drug, n is 0, 1, 2, 3, 4, and the total content of each component is 72.9%. The liquid chromatogram is shown in Figure 2 shown.

[0089] Example 4 Preparation of pesticide technical or parent drug

[0090] 1. Preparation of fermentation broth

[0091] Same as Example 1.

[0092] 2. Purification

[0093] (1) Acidification of the fermentation broth: acidifying the fermentation broth to a pH of 3.5;

[0094] (2) Bacteria isolation: The acidified material is centrifuged and the resulting clear liquid is filtered twice through an ultrafiltration membrane;

[0095] (3) Resin adsorption and analysis: Use 724 weakly acidic cation exchange resin to adsorb the filtrate, rinse with water until the effluent is colorless, and analyze with 5% hydrochloric acid. After discarding the 0.5BV effluent of the analytical solution, start collecting samples and stop collecting samples after 5BV; collect the analytical solution and repeat the above resin adsorption process once more;

[0096] (4) Nanofiltration concentration: The analytical solution is concentrated by nanofiltration;

[0097] (5) Drying: The concentrated solution is spray dried to obtain the pesticide technical or the mother drug. The obtained pesticide technical or the mother drug is tested to contain 1-N-glycosidyl-2-amino-(L-lysyl) n -L-lysine-2-deoxygulosamine, n is 0, 1, 2, 3, 4 respectively, and the total content of each component is 82.3% based on the total mass of the original drug or the mother drug.

[0098] Example 5 Preparation of pesticide technical or parent drug

[0099] 1. Preparation of fermentation broth

[0100] Same as Example 1.

[0101] 2. Purification

[0102] (1) Acidification of the fermentation broth: acidifying the fermentation broth to a pH of 3.5;

[0103] (2) Bacteria isolation: The acidified material is centrifuged and the resulting clear liquid is filtered through an ultrafiltration membrane three times;

[0104] (3) Resin adsorption and analysis: Use 724 weakly acidic cation exchange resin to adsorb the filtrate, rinse with water until the effluent is colorless, and analyze with 5% hydrochloric acid. After discarding the 1BV effluent of the analytical solution, start collecting samples and stop collecting samples after the 4BV; collect the analytical solution and repeat the above resin adsorption process once more;

[0105] (4) Nanofiltration concentration: The analytical solution is concentrated by nanofiltration;

[0106] (5) Drying: The concentrated solution is spray dried to obtain the pesticide technical or the mother drug. The obtained pesticide technical or the mother drug is tested to contain 1-N-glycosidyl-2-amino-(L-lysyl) n -L-lysine-2-deoxygulosamine, n is 0, 1, 2, 3, 4, respectively. Based on the total mass of the original drug or parent drug, the total content of each component is 89.2%. The liquid chromatogram is shown in Figure 3 shown.

[0107] Example 6 Preparation of pesticide technical or parent drug

[0108] 1. Preparation of fermentation broth

[0109] Same as Example 1.

[0110] 2. Purification

[0111] (1) Acidification of the fermentation broth: acidifying the fermentation broth to a pH of 3.5;

[0112] (2) Bacteria isolation: The acidified material is centrifuged and the resulting clear liquid is filtered through an ultrafiltration membrane three times;

[0113] (3) Resin adsorption and analysis: Use 724 weakly acidic cation exchange resin to adsorb the filtrate, rinse with water until the effluent is colorless, and analyze with 5% hydrochloric acid. After discarding the 1BV effluent of the analytical solution, start collecting samples and stop collecting samples after the 4BV; collect the analytical solution and repeat the above resin adsorption process twice;

[0114] (4) Nanofiltration concentration: The analytical solution is concentrated by nanofiltration;

[0115] (5) Drying: The concentrated solution is spray-dried to obtain the pesticide technical or the raw material. The obtained pesticide technical or the raw material is tested to contain 1-N-glycosidic acid chain 2-amino-(L-lysyl) n -L-lysine-2-deoxygulosamine, n is 0, 1, 2, 3, 4, respectively. Based on the total mass of the original drug or parent drug, the total content of each component is 96.2%. The liquid chromatogram is shown in Figure 4 shown.

[0116] Effect embodiment:

[0117] The mother drugs or original drugs of Example 2 (active ingredient content 60.0%), Example 3 (active ingredient content 72.9%), Example 5 (active ingredient content 89.2%), and Example 6 (active ingredient content 96.2%) were placed at 60°C / 25%RH for 30 days, their moisture absorption was observed, and the degradation rate of their active ingredients and changes in their antibacterial effects were detected.

[0118] 1. Moisture absorption

[0119] After being placed under 60°C / 25% RH conditions for 30 days, the moisture absorption of Example 2, Example 3, Example 5, and Example 6 is as follows: Figure 5 As shown. As can be seen from the figure, the hygroscopicity of the samples of Example 2, Example 3, and Example 5 is relatively weak. In Example 2 and Example 3, only a very small amount of samples are slightly moistened in a circle outside the sample pile, and in Example 5, a small amount of samples are moistened in a circle outside the sample pile and on the surface. However, most of the samples of these three embodiments are still in powder form. The wet part is removed with a glass rod, and the remaining powdered samples still have very good fluffiness and can be used normally for subsequent preparation processing. Most of the samples of Example 6 have become pasty and are very easy to stick to the glass rod. This kind of hygroscopic original drug or mother drug is very easy to stick to the machine or conveyor during the preparation processing, which is inconvenient to operate, causing processing difficulties, and also poses a huge challenge to the storage conditions. It can be seen that the sample of Example 6 has a significantly enhanced hygroscopicity due to the excessively high content of active ingredients, poor processing performance, and more stringent storage conditions. 1-N-Aglycone chain lidyl-2-amino-(L-lysyl) n The hygroscopicity of -L-lysine-2-deoxygulosamine may be due to the presence of more hydroxyl groups in its molecular structure.

[0120] 2. Degradation rate of active ingredients

[0121] Samples to be tested:

[0122] The samples of Example 2, Example 3, Example 5 and Example 6 that were not treated under 60°C / 25% RH conditions;

[0123] The samples of Example 2, Example 3, Example 5 and Example 6 (dried) were placed under the conditions of 60°C / 25% RH for 30 days.

[0124] The results of the degradation rate determination of the active ingredients of each sample are shown in Table 2.

[0125] Table 2 Degradation rate of each sample

[0126] Technical / main drug samples Degradation rate % Example 2 7.69％ Example 3 4.03％ Example 5 3.27％ Example 6 6.88％

[0127] It can be seen from the above results that the samples of Example 2 and Example 6 have an effective component degradation rate of more than 5% after being placed under 60°C / 25% RH for 30 days, which is difficult to meet the production needs. Among them, the sample of Example 2 has the largest degradation rate, which may be due to its low effective component content and contains more other impurities, affecting the stability of the effective component; followed by the sample of Example 6. It can be seen from the above hygroscopicity results that the effective component content of Example 6 is too high and the hygroscopicity is strong. After being placed under 60°C / 25% RH for 30 days, most of the sample has become a paste. The high water content environment may be an important reason for the degradation of its effective component. The effective component degradation rates of Examples 3 and 5 are relatively small, below 5%, and their effective components are relatively stable, which is more in line with the needs of actual production.

[0128] 3. Determination of antibacterial effect

[0129] (1) Sample to be tested

[0130] The samples of Example 2, Example 3, Example 5 and Example 6 which were not treated under 60°C / 25% RH conditions were prepared into the following solutions for use:

[0131] Take 0.1 g of the sample of Example 2 and dissolve it in 1.2 L of sterile water to obtain a solution with an active ingredient concentration of 50 μg / mL, named Solution 1 (front);

[0132] Take 0.1 g of the sample of Example 3 and dissolve it in 1.458 L of sterile water to obtain a solution with an active ingredient concentration of 50 μg / mL, named Solution 2 (front);

[0133] Take 0.1 g of the sample of Example 5 and dissolve it in 1.784 L of sterile water to obtain a solution with an active ingredient concentration of 50 μg / mL, named Solution 3 (front);

[0134] Take 0.1 g of the sample of Example 6 and dissolve it in 1.924 L of sterile water to obtain a solution with an active ingredient concentration of 50 μg / mL, which is named Solution 4 (front).

[0135] The samples of Example 2, Example 3, Example 5 and Example 6 (dried) which were placed under 60°C / 25% RH for 30 days were diluted by the same multiple as the corresponding samples which were not treated under 60°C / 25% RH to prepare the following solutions for use:

[0136] 0.1 g of the sample of Example 2 was dissolved in 1.2 L of sterile water, and the resulting solution was named Solution 1 (latter);

[0137] 0.1 g of the sample of Example 3 was taken and dissolved in 1.458 L of sterile water. The obtained solution was named Solution 2 (latter);

[0138] 0.1 g of the sample of Example 5 was taken and dissolved in 1.784 L of sterile water. The obtained solution was named Solution 3 (latter).

[0139] 0.1 g of the sample of Example 6 was taken and dissolved in 1.924 L of sterile water. The obtained solution was named Solution 4 (latter).

[0140] (2) Preparation of bacterial layer

[0141] Take 200 mL of the culture medium that has been melted and cooled to 48°C to 50°C, add 1 mL of 1×10 -8 The suspension of citrus canker bacteria at CFU / mL was shaken evenly, poured into a culture dish, and allowed to stand to solidify to obtain a bacterial layer.

[0142] (3) Addition of medicine

[0143] Take a culture dish containing a bacterial layer, add 4 sterile filter papers, and drop 10 μL of solution 1 (front), solution 2 (front), solution 3 (front), and solution 4 (front) on the filter papers respectively. Repeat 3 times.

[0144] Take another culture dish containing a bacterial layer, add 4 sterile filter papers, and drop 10 μL of solution 1 (after), solution 2 (after), solution 3 (after), and solution 4 (after) on the filter papers respectively. Repeat 3 times.

[0145] The above culture dishes were cultured in a 28°C incubator for 48 h.

[0146] (4) Investigation of antibacterial effect

[0147] After the incubation, the diameter of the inhibition zone was measured by the cross method and the average value was calculated. The results are shown in Table 3 and Figure 6 As shown, Figure 6 A is the antibacterial zone image of the sample not treated at 60℃ / 25% RH. Figure 6 B is a picture of the inhibition zone of the sample after being placed under 60°C / 25% RH conditions for 30 days.

[0148] Table 3 Antibacterial effect of each sample

[0149]

[0150] It can be seen from the above results that the antibacterial abilities of the samples of Example 2, Example 3, Example 5 and Example 6 are almost the same when they are not treated under 60°C / 25% RH conditions. After being treated under 60°C / 25% RH conditions, the average diameter of the antibacterial zone of the samples of Example 2 and Example 6 is reduced by more than 5mm, and the stability of the antibacterial ability is poor. Among them, the antibacterial ability of Example 2 is most significantly reduced, and the average diameter of the antibacterial zone is reduced from 27.3mm to 21.2mm, which is reduced by 6.1mm; followed by the sample of Example 6, the average diameter of the antibacterial zone is reduced from 28.7mm to 23.3mm, which is reduced by 5.4mm. The antibacterial ability of the samples of Example 3 and Example 5 is well maintained, and the average diameter of the antibacterial zone is reduced from 27.9mm to 24.1mm, which is only reduced by 3.8mm, and from 28.2mm to 24.8mm, which is only reduced by 3.4mm.

[0151] Based on the above experimental results, the original drug or mother drug (active ingredient content of 60.0%) of Example 2 has weak hygroscopicity, which is convenient for subsequent processing and storage, but it performs the worst in terms of maintaining the active ingredient and maintaining the antibacterial effect. This may be due to the fact that the content of its active ingredient 1-N-glycosidyl-2-amino-(L-lysyl) nL-lysine-2-deoxygulosamine is too low, the content of other impurities is high, and the stability of the active ingredient is greatly affected by other impurities and is easily degraded. Therefore, only by increasing the content of 1-N-glycosidyl-2-amino-(L-lysyl) nL-lysine-2-deoxygulosamine to more than 70% can the stability of the active ingredient and antibacterial ability be better maintained to meet the needs of actual production.

[0152] Further, although the original drug or mother drug of Example 6 (with an active ingredient content of up to 96.2%) is less affected by other impurities, due to its high content and strong hygroscopicity, it not only affects the subsequent preparation processing technology and causes storage difficulties, but also causes the active ingredient to be easily degraded. Therefore, it also performs poorly in terms of maintaining the active ingredient and maintaining the antibacterial effect. It can be seen that controlling the content of 1-N-glycosidyl-2-amino-(L-lysyl) nL-lysine-2-deoxygulosamine within the range of 70%-94% can not only ensure that it has low hygroscopicity, but also greatly reduce the degradation rate of its active ingredient and maintain its antibacterial ability, thereby further improving the comprehensive performance of the original drug or mother drug.

[0153] The above embodiments are preferred implementation modes of the present invention, but the implementation modes of the present invention are not limited to the above embodiments. Any other changes, modifications, substitutions, combinations, and simplifications that do not deviate from the spirit and principles of the present invention should be equivalent replacement methods and are included in the protection scope of the present invention.

Claims

1. A pesticide technical or bulk drug, characterized in that: The pesticide technical or mother drug contains 1-N-glycosidyl-2-amino-(L-lysyl) n - at least 2 of L-lysine-2-deoxygulosamine; Based on the total mass of the pesticide original or parent drug, the 1-N-glycosidyl-2-amino-(L-lysyl) n -The total content of L-lysine-2-deoxygulosamine is more than 70%; Wherein n represents the number of L-lysyl groups, and can be any integer between 0 and 4.

2. The pesticide technical or bulk drug according to claim 1, characterized in that: Based on the total mass of the pesticide original or parent drug, the 1-N-glycosidyl-2-amino-(L-lysyl) n -The total content of L-lysine-2-deoxygulosamine is 70%-96.2%.

3. The pesticide technical or bulk drug according to claim 1, characterized in that: Based on the total mass of the pesticide original or parent drug, the 1-N-glycosidyl-2-amino-(L-lysyl) n -The total content of L-lysine-2-deoxygulosamine is 72%-90%.

4. The pesticide technical or bulk drug according to claim 1, characterized in that: Based on the total mass of the pesticide original or parent drug, the 1-N-glycosidyl-2-amino-(L-lysyl) n -The total content of L-lysine-2-deoxygulosamine is 75%-90%.

5. The pesticide technical or bulk drug according to claim 1, characterized in that: Based on the total mass of the pesticide original or parent drug, the 1-N-glycosidyl-2-amino-(L-lysyl) n -The total content of L-lysine-2-deoxygulosamine is 80%-90%; Preferably, based on the total mass of the pesticide original or parent drug, the 1-N-glycosidyl-2-amino-(L-lysyl) n -The total content of L-lysine-2-deoxygulosamine is 70%-85%; Preferably, based on the total mass of the pesticide original or parent drug, the 1-N-glycosidyl-2-amino-(L-lysyl) n -The total content of L-lysine-2-deoxygulosamine is 70%-80%.

6. Use of the pesticide technical or concentrate according to any one of claims 1 to 5 in the preparation of pesticide preparations or pesticide compositions.

7. A pesticide preparation or a pesticide composition, characterized in that: The pesticide preparation or pesticide composition contains the pesticide technical or concentrate according to any one of claims 1 to 5.

8. The pesticide preparation or pesticide composition according to claim 7, characterized in that: The pesticide preparation or pesticide composition is in the form of an aqueous solution or a soluble solution.

9. Use of the pesticide technical or concentrate according to any one of claims 1 to 5, or the pesticide preparation or pesticide composition according to any one of claims 7 to 8 in the preparation of a fungicide.

10. Use of the pesticide technical or concentrate according to any one of claims 1 to 5, or the pesticide preparation or pesticide composition according to any one of claims 7 to 8 in sterilization.

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