CN113817008B - Preparation method and application of novel succinyl hexadecanoic macrolide - Google Patents

Preparation method and application of novel succinyl hexadecanoic macrolide Download PDF

Info

Publication number
CN113817008B
CN113817008B CN202110798419.6A CN202110798419A CN113817008B CN 113817008 B CN113817008 B CN 113817008B CN 202110798419 A CN202110798419 A CN 202110798419A CN 113817008 B CN113817008 B CN 113817008B
Authority
CN
China
Prior art keywords
compound
culturing
column chromatography
culture medium
ivermectin
Prior art date
Legal status (The legal status is an assumption and is not a legal conclusion. Google has not performed a legal analysis and makes no representation as to the accuracy of the status listed.)
Active
Application number
CN202110798419.6A
Other languages
Chinese (zh)
Other versions
CN113817008A (en
Inventor
王继栋
葛海霞
张绍勇
齐欢
张立钦
Current Assignee (The listed assignees may be inaccurate. Google has not performed a legal analysis and makes no representation or warranty as to the accuracy of the list.)
Huzhou University
Original Assignee
Huzhou University
Priority date (The priority date is an assumption and is not a legal conclusion. Google has not performed a legal analysis and makes no representation as to the accuracy of the date listed.)
Filing date
Publication date
Application filed by Huzhou University filed Critical Huzhou University
Priority to CN202110798419.6A priority Critical patent/CN113817008B/en
Publication of CN113817008A publication Critical patent/CN113817008A/en
Application granted granted Critical
Publication of CN113817008B publication Critical patent/CN113817008B/en
Active legal-status Critical Current
Anticipated expiration legal-status Critical

Links

Images

Classifications

    • C—CHEMISTRY; METALLURGY
    • C07—ORGANIC CHEMISTRY
    • C07H—SUGARS; DERIVATIVES THEREOF; NUCLEOSIDES; NUCLEOTIDES; NUCLEIC ACIDS
    • C07H17/00—Compounds containing heterocyclic radicals directly attached to hetero atoms of saccharide radicals
    • C07H17/04—Heterocyclic radicals containing only oxygen as ring hetero atoms
    • C07H17/08—Hetero rings containing eight or more ring members, e.g. erythromycins
    • A—HUMAN NECESSITIES
    • A01—AGRICULTURE; FORESTRY; ANIMAL HUSBANDRY; HUNTING; TRAPPING; FISHING
    • A01N—PRESERVATION OF BODIES OF HUMANS OR ANIMALS OR PLANTS OR PARTS THEREOF; BIOCIDES, e.g. AS DISINFECTANTS, AS PESTICIDES OR AS HERBICIDES; PEST REPELLANTS OR ATTRACTANTS; PLANT GROWTH REGULATORS
    • A01N43/00—Biocides, pest repellants or attractants, or plant growth regulators containing heterocyclic compounds
    • A01N43/90—Biocides, pest repellants or attractants, or plant growth regulators containing heterocyclic compounds having two or more relevant hetero rings, condensed among themselves or with a common carbocyclic ring system
    • C—CHEMISTRY; METALLURGY
    • C07—ORGANIC CHEMISTRY
    • C07H—SUGARS; DERIVATIVES THEREOF; NUCLEOSIDES; NUCLEOTIDES; NUCLEIC ACIDS
    • C07H1/00—Processes for the preparation of sugar derivatives
    • C07H1/06—Separation; Purification
    • C—CHEMISTRY; METALLURGY
    • C12—BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
    • C12N—MICROORGANISMS OR ENZYMES; COMPOSITIONS THEREOF; PROPAGATING, PRESERVING, OR MAINTAINING MICROORGANISMS; MUTATION OR GENETIC ENGINEERING; CULTURE MEDIA
    • C12N1/00—Microorganisms, e.g. protozoa; Compositions thereof; Processes of propagating, maintaining or preserving microorganisms or compositions thereof; Processes of preparing or isolating a composition containing a microorganism; Culture media therefor
    • C12N1/20—Bacteria; Culture media therefor
    • C—CHEMISTRY; METALLURGY
    • C12—BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
    • C12P—FERMENTATION OR ENZYME-USING PROCESSES TO SYNTHESISE A DESIRED CHEMICAL COMPOUND OR COMPOSITION OR TO SEPARATE OPTICAL ISOMERS FROM A RACEMIC MIXTURE
    • C12P19/00—Preparation of compounds containing saccharide radicals
    • C12P19/44—Preparation of O-glycosides, e.g. glucosides
    • C12P19/60—Preparation of O-glycosides, e.g. glucosides having an oxygen of the saccharide radical directly bound to a non-saccharide heterocyclic ring or a condensed ring system containing a non-saccharide heterocyclic ring, e.g. coumermycin, novobiocin
    • C12P19/62—Preparation of O-glycosides, e.g. glucosides having an oxygen of the saccharide radical directly bound to a non-saccharide heterocyclic ring or a condensed ring system containing a non-saccharide heterocyclic ring, e.g. coumermycin, novobiocin the hetero ring having eight or more ring members and only oxygen as ring hetero atoms, e.g. erythromycin, spiramycin, nystatin
    • Y—GENERAL TAGGING OF NEW TECHNOLOGICAL DEVELOPMENTS; GENERAL TAGGING OF CROSS-SECTIONAL TECHNOLOGIES SPANNING OVER SEVERAL SECTIONS OF THE IPC; TECHNICAL SUBJECTS COVERED BY FORMER USPC CROSS-REFERENCE ART COLLECTIONS [XRACs] AND DIGESTS
    • Y02—TECHNOLOGIES OR APPLICATIONS FOR MITIGATION OR ADAPTATION AGAINST CLIMATE CHANGE
    • Y02A—TECHNOLOGIES FOR ADAPTATION TO CLIMATE CHANGE
    • Y02A50/00—TECHNOLOGIES FOR ADAPTATION TO CLIMATE CHANGE in human health protection, e.g. against extreme weather
    • Y02A50/30—Against vector-borne diseases, e.g. mosquito-borne, fly-borne, tick-borne or waterborne diseases whose impact is exacerbated by climate change

Landscapes

  • Chemical & Material Sciences (AREA)
  • Life Sciences & Earth Sciences (AREA)
  • Organic Chemistry (AREA)
  • Engineering & Computer Science (AREA)
  • Health & Medical Sciences (AREA)
  • Biotechnology (AREA)
  • General Health & Medical Sciences (AREA)
  • Genetics & Genomics (AREA)
  • Zoology (AREA)
  • Wood Science & Technology (AREA)
  • Biochemistry (AREA)
  • Bioinformatics & Cheminformatics (AREA)
  • Molecular Biology (AREA)
  • Microbiology (AREA)
  • General Engineering & Computer Science (AREA)
  • Biomedical Technology (AREA)
  • Dentistry (AREA)
  • Tropical Medicine & Parasitology (AREA)
  • Medicinal Chemistry (AREA)
  • Agronomy & Crop Science (AREA)
  • Pest Control & Pesticides (AREA)
  • Plant Pathology (AREA)
  • Virology (AREA)
  • Environmental Sciences (AREA)
  • Chemical Kinetics & Catalysis (AREA)
  • General Chemical & Material Sciences (AREA)
  • Preparation Of Compounds By Using Micro-Organisms (AREA)
  • Saccharide Compounds (AREA)
  • Agricultural Chemicals And Associated Chemicals (AREA)
  • Pharmaceuticals Containing Other Organic And Inorganic Compounds (AREA)

Abstract

The invention relates to a novel macrolide compound 4 '-O- (6-O-succinyl-glucosyl) -25-methyl ivermectin and 4' -O- (6-O-succinyl-glucosyl) -25-ethyl ivermectin, a microbial conversion preparation method thereof and application thereof in preparing medicaments for preventing and controlling agricultural and forestry pests and mites.

Description

Preparation method and application of novel succinyl hexadecanoic macrolide
Technical Field
The invention belongs to the technical field of biology, and particularly relates to a method for preparing 4 '-O- (6-O-succinyl-glucosyl) -25-methyl ivermectin and 4' -O- (6-O-succinyl-glucosyl) -25-ethyl ivermectin by microbial transformation and application thereof in preparing medicaments for preventing and controlling agricultural and forestry pests and mites.
Background
25-methyl ivermectin and 25-ethyl ivermectin are novel sixteen-membered macrolide antibiotics, and have the characteristics of broad spectrum, high efficiency, low toxicity, small dosage, safer use and the like, and the chemical structural formulas of the antibiotics are as follows:
Figure BDA0003163632400000011
compared with avermectin, ivermectin and milbemycin, the active compound has the effect of dispelling and killing internal and external parasitic diseases such as nematodes, mites, ticks, lice, flies and maggots of animals such as pigs, cattle, sheep, horses, rabbits and poultry, and has a good prospect.
Microbial transformation is to modify a complex substrate by microbial cells, that is, to catalyze a reaction on a specific site (group) of the substrate by using a certain or a certain series of enzymes generated in the metabolic process of the microorganism, so that the molecular structure of the substrate is changed into another similar compound. The microbial transformation has the advantages of directional reaction, single product, less side reaction, rapid biomass accumulation, short transformation time, high expression efficiency of invertase, mild reaction condition, safety, environmental protection and the like, and is a convenient means for carrying out structural modification on natural active compounds.
According to the invention, the bacillus subtilis strain Bacillus subtilis ATCC 6633 is utilized to convert 25-methyl/25-ethyl ivermectins, so that 2 novel sixteen-membered macrolide active compounds with novel structures are obtained, the activity of the novel sixteen-membered macrolide active compounds is equivalent to that of a precursor compound 25-methyl/25-ethyl ivermectins, but the crystallinity of the novel sixteen-membered macrolide active compounds is better than that of the precursor compound, and the novel sixteen-membered macrolide active compounds are favorable for quality control of subsequent products and reduction of production cost. In addition, the structure of compounds 1-2 has a carboxyl group compared to the parent compound, indicating that water solubility is increased, which is advantageous for development as a water-based insecticide.
Disclosure of Invention
1. A compound having the structural formula:
Figure BDA0003163632400000031
wherein r=ch 3 Or CH (CH) 2 CH 3 。
Wherein R is 1 ＝CH 3 Which is compound 1 of the formula:
Figure BDA0003163632400000041
or alternatively
R 1 ＝CH 2 CH 3 Which is compound 2 of the formula:
Figure BDA0003163632400000051
the invention provides a preparation method of a compound 1-2, which comprises the following steps:
1) Preparing a penicillium griseofulvum seed solution: taking a bacillus subtilis strain Bacillus subtilis with the deposit number of ATCC 6633, streaking and inoculating on a PDA solid culture medium, culturing for 5-7 days in a constant temperature incubator at 20-28 ℃, transferring the activated strain into a soybean meal liquid culture medium, and culturing for 40 hours in a horizontal shaking table at 20-28 ℃ and 250rpm to obtain seed liquid;
2) Preparation of Compounds 1-2: transferring 1mL of seed solution into fresh soybean meal liquid culture medium, culturing at 180rpm/min and 28 ℃ for 24 hours, adding 1mL of 25-methyl/25-ethyl ivermectins (with concentration of 10 mg/mL) dissolved by organic solvent into each bottle, and culturing under the same condition for 96 hours to obtain fermentation liquor; extracting the fermentation liquor by using an extractant, recovering the extractant, and then obtaining a pure product of the compound 1-2 through chromatographic separation.
As a preferable technical scheme, the preparation method of the liquid culture medium in the step 1) comprises the following steps: taking 30g of glucose, 5g of yeast extract, 5g of NaCl and K 2 HPO 4 5g, distilled water is fixed to 1000ml, pH is regulated to 7.0-7.2 by 1mol/l NaOH, 50ml is subpackaged per bottle by adopting a 250ml triangular flask, 0.25-0.30g of soybean meal is weighed and placed into the triangular flask, bandaged, and sterilized for 20min under the conditions of 121 ℃ and 0.15 MPa.
As a preferable technical scheme, the organic solvent in the step 2) comprises one or more of methanol, ethanol and DMSO.
As a preferable technical scheme, the extractant in the step 2) comprises one or more of ethyl acetate, n-butanol, isobutyl acetate, diethyl ether, petroleum ether, dichloromethane or chloroform.
As a preferred embodiment, the purification of step 2) comprises reversed-phase high performance liquid chromatography.
As a preferred embodiment, the purification in step 2) further comprises silica gel column chromatography.
The present invention further provides pesticidal compositions comprising compound 1 and/or compound 2 as described above, said compositions further comprising one or more conventional carriers and/or diluents. The pesticide composition can be in the form of water dispersible granules, emulsifiable concentrates, aqueous suspensions, oil suspensions, microemulsions or tablets.
The invention further provides application of the compound 1-2 in preparing medicines for preventing and controlling crop diseases and insect pests. The crop diseases and insect pests comprise nematodes and red spiders.
Drawings
FIG. 1 high resolution mass spectrum of compound 1 obtained in example 1;
FIG. 2 shows the hydrogen spectrum of compound 1 obtained in example 1;
FIG. 3 carbon spectrum of Compound 1 obtained in example 1;
FIG. 4 DEPT135 spectrum of compound 1 obtained in example 1;
FIG. 5 Compound 1 obtained in example 1 1 H- 1 H COSY carbon profile;
FIG. 6 HMQC spectrum of Compound 1 obtained in example 1;
FIG. 7 is a HMBC spectrum of Compound 1 obtained in example 1;
FIG. 8 high resolution mass spectrum of compound 2 obtained in example 2;
FIG. 9 shows the hydrogen spectrum of compound 2 obtained in example 2;
FIG. 10 carbon spectrum of compound 2 obtained in example 2;
FIG. 11 DEPT135 spectrum of compound 2 obtained in example 2;
FIG. 12 Compound 2 obtained in example 2 1 H- 1 H COSY carbon profile;
FIG. 13 HMQC spectrum of Compound 2 obtained in example 2;
FIG. 14 HMBC spectra of compound 2 obtained in example 2.
Detailed Description
The invention is illustrated below with specific examples, it being understood that the examples are intended to illustrate the invention and not to limit it, the scope and core content of which are defined by the claims.
Example 1
1) Taking a bacillus subtilis strain Bacillus subtilis with the deposit number of ATCC 6633, streaking and inoculating on a PDA solid culture medium, culturing for 5-7 days in a constant temperature incubator with the temperature of 20-28 ℃, transferring the activated strain into a soybean meal liquid culture medium, culturing for 40 hours in a horizontal shaking table with the temperature of 20-28 ℃ and the speed of 250rpm to obtain seed liquid, transferring 1mL of the seed liquid into 100mL of a fresh soybean meal liquid culture medium per bottle, culturing for 24 hours at 180rpm and the temperature of 28 ℃, respectively adding 25-methyl ivermectin (with the concentration of 10 mg/mL) dissolved by 1mL of an organic solvent into each bottle, and culturing for 96 hours under the same condition to obtain a fermentation broth.
2) Fermenting and culturing to obtain 2L fermentation liquor according to the method, extracting the fermentation liquor with equal volume of ethyl acetate for three times to obtain ethyl acetate extract liquid, concentrating the extract liquid to dryness under the condition of reducing pressure at 50 ℃ to obtain 4.5g oily substance.
The obtained oily substance was subjected to column chromatography on a silica gel column (particle size 100 to 200 mesh) with chloroform: gradient elution with methanol=100:0-60:40 (V/V) was performed and detected by thin layer identification (TLC) to give 3 fractions. Component 3 was subjected to gel (Sephadex LH-20) column chromatography to give component 3-1, which was then further purified using semi-preparative column chromatography to give pure 4"-O- (6-O-succinyl-glucosyl) -25-methyl ivermectin (1).
Wherein the thin layer identification method comprises the following steps: the converted sample and blank control point are spread on a silica gel G thin layer plate under the condition of developing agent of chloroform-methanol (8:2), taken out for airing after running, observed under an ultraviolet lamp of 254nm, developed with concentrated sulfuric acid, and the conversion result is inspected.
Semi-preparative column chromatography conditions: mobile phase: methanol-water (containing thousandth of acetic acid) (90:10); column C18,9.4 x 250mm; detection wavelength: 244nm; flow rate: 1.5mL/min; column temperature: 30 ℃; the sample injection amount is as follows: 50uL. The peak with retention time of 14.35min was collected to give 4"-O- (6-O-succinyl-glucosyl) -25-methyl ivermectin (1, 14.5 mg).
3) Structural identification of compound 1.
The structure of compound 1 was determined by 1D and 2D NMR, MS, etc. spectroscopy as follows:
compound identification involves the following instrumentation:
the nuclear magnetic resonance spectrum is measured by a superconducting nuclear magnetic resonance apparatus (ZHONGKE-400) of the middle Ke oxford company;
the mass spectrum and the high-resolution mass spectrum are measured by a Q-TOF Micro LC-MS-MS mass spectrometer of Waters company;
the UV spectrum was measured using a Varian Cary 300Bio spectrophotometer spectrometer from Varian company;
specific data for the compounds are as follows:
structure of compound 1:
Figure BDA0003163632400000091
property of white crystalline powder;
solubility, namely, the water-insoluble organic matter is easy to dissolve in chloroform, acetone, methanol and ethanol;
molecular formula C 55 H 82 O 22 ；
HRESI-MS m/z:1117.5198[M+Na] + (calculated value: C 55 H 82 O 22 Na,1117.5195)；
UVλ max (EtOH)nm(logε):245(4.45)；
IR v max cm -1 ：3381,2963,1724,1680,1448,1384,1261,1091；
Hydrogen spectrum @ 1 H NMR) and carbon spectrum [ ] 13 C NMR) data are shown in table 1.
Example 2
1) Taking a bacillus subtilis strain Bacillus subtilis with the deposit number of ATCC 6633, streaking and inoculating on a PDA solid culture medium, culturing for 5-7 days in a constant temperature incubator with the temperature of 20-28 ℃, transferring the activated strain into a soybean meal liquid culture medium, culturing for 40 hours in a horizontal shaking table with the temperature of 20-28 ℃ and the speed of 250rpm to obtain seed liquid, transferring 1mL of the seed liquid into 100mL of a fresh soybean meal liquid culture medium per bottle, culturing for 24 hours at 180rpm and the temperature of 28 ℃, respectively adding 1mL of 25-methyl ivermectin (with the concentration of 10 mg/mL) dissolved by an organic solvent into each bottle, and culturing for 96 hours under the same condition to obtain fermentation liquor.
2) Fermenting and culturing to obtain 2L fermentation liquor according to the method, extracting the fermentation liquor with equal volume of ethyl acetate for three times to obtain ethyl acetate extract liquid, concentrating the extract liquid to dryness under the condition of reducing pressure at 50 ℃ to obtain 5.3g oily substance.
The obtained oily substance was subjected to column chromatography on a silica gel column (particle size 100 to 200 mesh) with chloroform: gradient elution with methanol=100:0-60:40 (V/V) was performed and detected by thin layer identification (TLC) to give 3 fractions. Component 3 was subjected to gel (Sephadex LH-20) column chromatography to give component 3-1, which was then further purified using semi-preparative column chromatography to give pure and 4"-O- (6-O-succinyl-glucosyl) -25-ethyl ivermectin (2, 25.8 mg).
Wherein the thin layer identification method comprises the following steps: the converted sample and blank control point are spread on a silica gel G thin layer plate under the condition of developing agent of chloroform-methanol (8:2), taken out for airing after running, observed under an ultraviolet lamp of 254nm, developed with concentrated sulfuric acid, and the conversion result is inspected.
Semi-preparative column chromatography conditions: mobile phase: methanol-water (containing thousandth of acetic acid) (90:10); column C18,9.4 x 250mm; detection wavelength: 244nm; flow rate: 1.5mL/min; column temperature: 30 ℃; the sample injection amount is as follows: 50uL. The peak with retention time of 14.63min was collected to give 4"-O- (6-O-succinyl-glucosyl) -25-ethyl ivermectin (2).
3) Structure identification of compound 2.
Structure of compound 2:
Figure BDA0003163632400000111
property of white crystalline powder;
solubility, namely, the water-insoluble organic matter is easy to dissolve in chloroform, acetone, methanol and ethanol;
molecular formula C 56 H 84 O 22 ；
HRESI-MS m/z 1131.5356 (calculated value: C) 56 H 84 O 22 Na,1131.5352)；
UVλ max (EtOH)nm(logε):245(4.06)；
IR v max cm -1 ：3466,2931,1719,1451,1380,1120,1066,993；
Hydrogen spectrum @ 1 H NMR) and carbon spectrum [ ] 13 C NMR) data are shown in tables 1 and 2.
TABLE 1 Compounds 1-2 in CD 3 Nuclear magnetic data in the hydrogen spectrum (400 MHz) and carbon spectrum (100 MHz) in OD
Figure BDA0003163632400000112
Figure BDA0003163632400000121
Figure BDA0003163632400000131
Example 3
Biological Activity of Compounds 1-2 against Tetranychus cinnabarinus
Test organism: tetranychus cinnabarinus (Tetranychus cinnabarinus): under the condition of artificial climate chamber [ (26+ -1) DEGC, RH (70+ -5)%, H/D14], inoculating on broad bean seedling for culturing.
The experimental method comprises the following steps: adopts leaf disc insect immersing method: and selecting adult mites which are fed indoors and have consistent physiological states. Selecting broad bean leaves with consistent growth, making leaf discs with the diameter of 2cm by using a puncher, placing the leaf backs of the leaf discs on absorbent cotton in the center of a plastic dish, picking up 3 leaf discs of each dish by using a small-sized writing brush, inoculating mites on the leaf discs, adding a proper amount of water into 30 heads of each leaf disc, and placing the leaf discs in a culture room with the temperature of 26+/-1 ℃ and the illumination intensity of 3000-4500 lx, 14h/d and RH of 50-75%. After 2 hours, the number of mites is checked under a stereoscopic microscope, and the number of mites on each leaf dish is not less than 20. Dissolving a sample to be tested in a small amount of acetone, diluting with water by taking alkylphenol ethoxylates as an emulsifying agent, preparing medicaments with mass concentrations of 0.005, 0.01, 0.02, 0.04 and 0.08mg/L, placing the medicaments in a beaker, clamping blades by forceps, sequentially soaking the medicaments from low concentration to high concentration for 5 seconds, treating female mites by using distilled water in comparison, wherein each mass concentration is treated, and repeating each treatment for 3 times. And (3) after the medicines on the leaves are dried, placing the treated leaf discs in a climatic chamber with the photoperiod of (26+/-1) DEG C and 14 hours for culturing for 24 hours, and adding a small amount of water into a culture dish for moisturizing. Mites are very active after the drug infusion, slow down the activity after 5-8 hours after treatment, and the insects are static after 12-24 hours. Death criterion: when the test is performed, the mite body is touched by the writing brush, and the dead person is judged to be dead. The test results are shown in Table 2
Table 2: activity of Compounds 1-2 against Tetranychus cinnabarinus
Figure BDA0003163632400000141
Example 4
Activity assay of Compounds 1-2 on pine wood nematodes
Test organism: takes pine wood nematodes as insect test agents
The test method comprises the following steps: inoculating prepared pine wood nematodes to PDA culture medium full of Botrytis cinerea, culturing in a constant temperature incubator at 25deg.C for 7 days, picking out the culture medium containing pine wood nematodes, soaking in distilled water for 24 hr, filtering to separate out nematodes, and preparing into suspension of 2500 head/mL for use.
The method comprises the steps of dissolving a sample to be detected in a small amount of acetone by adopting an immersion liquid method, diluting with water by taking alkylphenol ethoxylates as an emulsifying agent, and respectively preparing the mixture into 5 mass concentrations of 1, 2, 5, 10 and 20mg/L. Taking 10 mu L of the test agent and 90 mu L of pine wood nematode suspension in a 96-well culture plate, taking the emulsifiable concentrate without the sample to be tested as a control, and culturing in a culture box at 25 ℃. Each mass concentration was 1 treatment, each treatment was repeated 3 times, and after 24 hours microscopic examination, the survival number and death number of pine wood nematodes were counted.
Nematodes moving or in the shape of "S", waves, curls, and spirals are considered living worms, while nematodes that are motionless and in the shape of "C", J ", stiff, and have no light-shielding properties in the body wall are considered dead worms. Mortality and corrective mortality were calculated separately and virulence regression equation, LC, was calculated using software SPSS 22.0 50 Values, correlation coefficients, and 95% confidence limits. Wherein die outRate = (number of dead nematodes/number of nematodes tested) ×100%; corrected mortality = (treatment mortality-control mortality)/(1-control mortality) ×100%. The test results are shown in Table 3.
Table 3: activity of Compounds 1-2 against pine wood nematodes
Figure BDA0003163632400000151
The invention uses bacillus subtilis Bacillus subtilis ATCC 6633 to convert 25-methyl/25-ethyl ivermectins, and can prepare 2 novel sixteen-membered macrolide compounds: 4"-O- (6-O-succinyl-glucosyl) -25-methyl ivermectin (1) and 4" -O- (6-O-succinyl-glucosyl) -25-ethyl ivermectin (2). The 2 compounds have good effects in preventing and controlling agricultural and forestry pests or mites.
The use of the compounds 1-2 according to the present invention has been described by way of specific examples, and those skilled in the art can, with reference to the present disclosure, suitably modify the starting materials, process conditions, etc. to achieve corresponding other objects, all of which do not depart from the spirit of the invention, and all similar substitutions and modifications will be apparent to those skilled in the art and are intended to be included within the scope of the present invention.
The above-described embodiments are merely preferred embodiments of the present invention, and the present invention is not limited in any way, and other variations and modifications may be made without departing from the technical aspects set forth in the claims.

Claims (6)

1. A macrolide compound has the following general structure:
Figure FDA0004171880860000011
wherein r=ch 3 Or CH (CH) 2 CH 3。
2. The macrolide compound according to claim 1, having the structure:
Figure FDA0004171880860000012
3. the macrolide compound according to claim 1, having the structure:
Figure FDA0004171880860000021
4. a process for preparing a compound according to claims 1-3, characterized in that the process comprises the steps of:
1) Preparing a penicillium griseofulvum seed solution: taking a bacillus subtilis strain Bacillus subtilis with the deposit number of ATCC 6633, streaking and inoculating on a PDA solid culture medium, culturing for 5-7 days in a constant temperature incubator at 20-28 ℃, transferring the activated strain into a soybean meal liquid culture medium, and culturing for 40 hours in a horizontal shaking table at 20-28 ℃ and 250rpm to obtain seed liquid;
2) Preparation of the compound: transferring 1mL of seed liquid into fresh soybean meal liquid culture medium, culturing for 24 hours at the temperature of 28 ℃ at 180rpm/min, adding 1mL of 25-methyl ivermectin or 25-ethyl ivermectin dissolved by organic solvent into each bottle, and culturing for 96 hours under the same condition at the concentration of 10mg/mL to obtain fermentation liquor;
3) Fermenting and culturing to obtain 2L fermentation liquor according to the method, extracting the fermentation liquor with equal volume of ethyl acetate for three times to obtain ethyl acetate extract liquor, concentrating the extract liquor to dryness under the condition of reducing pressure at 50 ℃ to obtain oily substances; subjecting the oily substance to column chromatography on silica gel column with particle size of 100-200 meshes, gradient eluting with chloroform-methanol with volume ratio of 100:0-60:40, and identifying and detecting by TLC thin layer to obtain 3 components; subjecting the component 3 to Sephadex LH-20 gel column chromatography to obtain a component 3-1, and further purifying by semi-preparative column chromatography to obtain pure compound 1 or 2, wherein the semi-preparative column chromatography conditions are as follows: the mobile phase is 90:10Methanol-water, wherein the water contains one thousandth of acetic acid; column C18,9.4 x 250mm; detection wavelength: 244nm; flow rate: 1.5mL/min; column temperature: 30 ℃; the sample injection amount is as follows: 50uL, collecting a peak with a retention time of 14.35min to obtain a compound 1, and collecting a peak with a retention time of 14.63min to obtain a compound 2, wherein the compound 1 is:
Figure FDA0004171880860000022
compound 2 is->
Figure FDA0004171880860000023
5. A pharmaceutical composition comprising one or more of the compounds of claims 1-3, together with one or more conventional carriers and/or diluents.
6. Use of a compound according to any one of claims 1 to 3 or a pharmaceutical composition according to claim 5 in the manufacture of a medicament for controlling crop diseases and insect pests, which are nematodes or red arachnids.
CN202110798419.6A 2021-07-15 2021-07-15 Preparation method and application of novel succinyl hexadecanoic macrolide Active CN113817008B (en)

Priority Applications (1)

Application Number Priority Date Filing Date Title
CN202110798419.6A CN113817008B (en) 2021-07-15 2021-07-15 Preparation method and application of novel succinyl hexadecanoic macrolide

Applications Claiming Priority (1)

Application Number Priority Date Filing Date Title
CN202110798419.6A CN113817008B (en) 2021-07-15 2021-07-15 Preparation method and application of novel succinyl hexadecanoic macrolide

Publications (2)

Publication Number Publication Date
CN113817008A CN113817008A (en) 2021-12-21
CN113817008B true CN113817008B (en) 2023-05-23

Family

ID=78912681

Family Applications (1)

Application Number Title Priority Date Filing Date
CN202110798419.6A Active CN113817008B (en) 2021-07-15 2021-07-15 Preparation method and application of novel succinyl hexadecanoic macrolide

Country Status (1)

Country Link
CN (1) CN113817008B (en)

Families Citing this family (1)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
CN114931144A (en) * 2022-06-21 2022-08-23 台州科技职业学院 Macrolide compound and application thereof in prevention and treatment of agricultural and forestry pests and pest mites

Citations (1)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
CN1738828A (en) * 2002-12-20 2006-02-22 辛根塔参与股份公司 Avermectin b1 and avermectin b1 monosaccharide derivatives having an alkoxymethyl substituent in the 4''- or 4'-position

Family Cites Families (9)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
US5070015A (en) * 1990-10-15 1991-12-03 Merck & Co., Inc. Novel avermectins are produced from known microorganisms
GB9300883D0 (en) * 1993-01-18 1993-03-10 Pfizer Ltd Antiparasitic agents
ATE296832T1 (en) * 1996-09-04 2005-06-15 Abbott Lab 6-O-SUBSTITUTED KETOLIDES WITH ANTIBACTERIAL ACTION
DE19913534A1 (en) * 1999-03-25 2000-10-05 Bayer Ag Avermectin derivatives
EP1632498A1 (en) * 2004-09-03 2006-03-08 Syngenta Participations AG Derivatives of avermectin, avermectin monosaccharide and avermectin aglycone
US8426460B2 (en) * 2008-12-04 2013-04-23 Merial Limited Dimeric avermectin and milbemycin derivatives
CN106046083A (en) * 2015-06-10 2016-10-26 北京维德维康生物技术有限公司 Avermectin artificial antigen and preparation method and application thereof
CN106167815A (en) * 2016-05-30 2016-11-30 浙江农林大学 The Preparation method and use of it dimension streptozotocin derivative
CN107417774B (en) * 2017-09-21 2021-04-09 福建省农业科学院农业生物资源研究所 Ivermectin-coupled Bt insecticidal toxin and application thereof

Patent Citations (1)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
CN1738828A (en) * 2002-12-20 2006-02-22 辛根塔参与股份公司 Avermectin b1 and avermectin b1 monosaccharide derivatives having an alkoxymethyl substituent in the 4''- or 4'-position

Also Published As

Publication number Publication date
CN113817008A (en) 2021-12-21

Similar Documents

Publication Publication Date Title
RU2139874C1 (en) Polycyclic compounds, method and strain for their preparing, antiparasitic composition and method of supression of parasitic infections
DE69402308T2 (en) NEW A83543 CONNECTIONS AND METHOD FOR THEIR PRODUCTION
JPH0335796A (en) Cyclic depsipeptide substance, its production and anthelmintics containing the same
CN103740606B (en) Plant raw streptomycete and produce antibiotic promise and irrigate method and the application of nest mycin
CN106167815A (en) The Preparation method and use of it dimension streptozotocin derivative
CN113817008B (en) Preparation method and application of novel succinyl hexadecanoic macrolide
CN113461757B (en) Preparation method and application of novel sixteen-membered macrolide
CN103242338B (en) Macrolide compound as well as preparation method and application thereof
CN104098585B (en) Mibemycin analogue, its preparation method and application
FI94262B (en) Process for preparing macrolide against parasites
CN111153797B (en) Active substance for killing nematode and its preparing process and application
CN112795617A (en) Marine fungus secondary metabolite and preparation and application thereof
CN107722032B (en) Anhydride compound with prodenia litura poisoning activity and preparation method and application thereof
CN111909166B (en) Two new milbemycins, preparation method and application thereof
WO2012077783A1 (en) Novel depsipeptide pf1442 substance and process for production thereof
CN106912489A (en) The preparation method and applications of AVM analog derivative
CN117903265B (en) Macrocyclic peptide compound acalitide and preparation method and application thereof
CN1265704C (en) Bacteriostatic chemical-yijunmeisu and its application
CN105462859B (en) It is a kind of for preventing and treating the branch acremonium bacterial strain of cigarette aphid
CN113200995A (en) Compound Sterigmatocystin, preparation method thereof and application thereof in pesticide
EP0026485B1 (en) Herbicoline, process for its preparation and compositions containing it
JPH0665246A (en) Novel substance pf 1101a substance, pf 1101b substance, their production and use
CN117964631A (en) Indole diketopiperazine compound and preparation method and application thereof
JPS63227590A (en) Antibiotic milbemycin compound and production thereof
CN112010870A (en) Novel sixteen-membered macrolide compound, preparation method and application thereof

Legal Events

Date Code Title Description
PB01 Publication
PB01 Publication
SE01 Entry into force of request for substantive examination
SE01 Entry into force of request for substantive examination
GR01 Patent grant
GR01 Patent grant