[go: up one dir, main page]

WO1998056939A1 - Streptomyces avermitilis strain, method for separating avermectin complexes and preparations for protecting plants and animals - Google Patents

Streptomyces avermitilis strain, method for separating avermectin complexes and preparations for protecting plants and animals Download PDF

Info

Publication number
WO1998056939A1
WO1998056939A1 PCT/RU1997/000183 RU9700183W WO9856939A1 WO 1998056939 A1 WO1998056939 A1 WO 1998056939A1 RU 9700183 W RU9700183 W RU 9700183W WO 9856939 A1 WO9856939 A1 WO 9856939A1
Authority
WO
WIPO (PCT)
Prior art keywords
aversectin
avermectin
water
avemekin
avermectins
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.)
Ceased
Application number
PCT/RU1997/000183
Other languages
French (fr)
Russian (ru)
Other versions
WO1998056939A8 (en
Inventor
Vladimir Alexandrovich Mosin
Elena Borisovna Kruglyak
Nadezhda Evgenievna Berezkina
Tamara Samuilovna Novik
Viktor Antonovich Drinyaev
Vasily Andreevich Jurkiv
Vasily Trofimovich Lisovenko
Galina Nikolaevna Tenkova
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.)
Individual
Original Assignee
Individual
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 Individual filed Critical Individual
Priority to PCT/RU1997/000183 priority Critical patent/WO1998056939A1/en
Priority to AU41397/97A priority patent/AU4139797A/en
Publication of WO1998056939A1 publication Critical patent/WO1998056939A1/en
Publication of WO1998056939A8 publication Critical patent/WO1998056939A8/en
Anticipated expiration legal-status Critical
Ceased legal-status Critical Current

Links

Classifications

    • 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
    • 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
    • A01N63/00—Biocides, pest repellants or attractants, or plant growth regulators containing microorganisms, viruses, microbial fungi, animals or substances produced by, or obtained from, microorganisms, viruses, microbial fungi or animals, e.g. enzymes or fermentates
    • A01N63/20—Bacteria; Substances produced thereby or obtained therefrom
    • A01N63/28—Streptomyces
    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
    • A61K35/00—Medicinal preparations containing materials or reaction products thereof with undetermined constitution
    • A61K35/66—Microorganisms or materials therefrom
    • A61K35/74—Bacteria
    • A61K35/741—Probiotics
    • A61K35/742—Spore-forming bacteria, e.g. Bacillus coagulans, Bacillus subtilis, clostridium or Lactobacillus sporogenes
    • 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
    • C12N1/205—Bacterial isolates
    • 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
    • C12P17/00—Preparation of heterocyclic carbon compounds with only O, N, S, Se or Te as ring hetero atoms
    • C12P17/02—Oxygen as only ring hetero atoms
    • C12P17/08—Oxygen as only ring hetero atoms containing a hetero ring of at least seven ring members, e.g. zearalenone, macrolide aglycons
    • C—CHEMISTRY; METALLURGY
    • C12—BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
    • C12R—INDEXING SCHEME ASSOCIATED WITH SUBCLASSES C12C - C12Q, RELATING TO MICROORGANISMS
    • C12R2001/00—Microorganisms ; Processes using microorganisms
    • C12R2001/01—Bacteria or Actinomycetales ; using bacteria or Actinomycetales
    • C12R2001/465—Streptomyces
    • 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

Definitions

  • the invention is not subject to the manufacture of biologically active substances in the form of an invasive substance.
  • P ⁇ edshes ⁇ vuyuschy u ⁇ ven ⁇ e ⁇ ni ⁇ i Izves ⁇ n ⁇ , ch ⁇ a ⁇ in ⁇ mitse ⁇ Zge ⁇ tusez a ⁇ egt ⁇ Y ⁇ ⁇ i ⁇ u- l ⁇ ivi ⁇ vanii in ⁇ isu ⁇ s ⁇ vii is ⁇ chni ⁇ v ugle ⁇ da, ae ⁇ a and ⁇ s ⁇ a ⁇ i ⁇ s ⁇ yann ⁇ y ae ⁇ atsin ⁇ dutsi ⁇ ue ⁇ v ⁇ sem blie ⁇ ⁇ s ⁇ eni ⁇ vesches ⁇ v, naevanny ⁇ ave ⁇ me ⁇ nnamn, ⁇ ye ⁇ bladayu ⁇ ⁇ ee ⁇ vy ⁇ azhe- nym an ⁇ i ⁇ a ⁇ azi ⁇ a ⁇ nym deys ⁇ viem ⁇ a ⁇ en ⁇ ⁇ Z ⁇ 1573955, 1980.
  • All amethysts are chemically 16-membered and macrolides and are distinguished by alternates from 5 and 26 carbon atoms.
  • the most active gins are Avemectins ⁇ , especially ⁇ &.
  • PRODUCTIVITY STRAIN 51GER-HOSE OF AUGUSTUS LIIS-51 (Patent No. 2048250, 1995) is 700 mcg / ml. Strain 5-June-Augustus augustus with IIIS-54 has already been producing 1,500 mg / ml of amecretins (patent No. 2054483, 1996), with an average of 35%.
  • the goal is achieved by the receipt of strain 5 “Lérgé” with the help of August 3, 003, which is produced in the average 2300 mcg / ml of amectine and 2000 ml / ml.
  • the variability of the investigations was 50-140%; 49-130% ⁇ 78-143%, which corresponded to the production of 900-2500 mkg / ml; 950-2500 mcg / ml and 1650-3000 mcg / ml avermectin.
  • the maximum accumulation of apertures is 7-8 days at a rate of an increase in acid of 1.0-1.3 mol ug / l-min and a temperature of 28 ° ⁇ .
  • Dispensary range ° C 20 4- 28 + 37 + / - 50
  • a dry mass is used, which is obtained by increasing the accrue to a good condition.
  • Aeration of biomass ave- mexins is carried out by a mixture consisting of water and low- and high-leveling pollutants, which is mixed with non-impaired water.
  • ⁇ ⁇ aches ⁇ ve niz ⁇ - ⁇ i ⁇ yaschi ⁇ is ⁇ lzuyu ⁇ ⁇ as ⁇ v ⁇ i ⁇ eli with ⁇ em ⁇ e ⁇ a ⁇ u ⁇ y ⁇ i ⁇ eniya not ⁇ e- Witzlaus ⁇ em ⁇ e ⁇ a ⁇ u ⁇ u ⁇ i ⁇ eniya v ⁇ dy, na ⁇ ime ⁇ , e ⁇ il ⁇ vy, and me ⁇ il ⁇ vy iz ⁇ il ⁇ vy s ⁇ i ⁇ y or atse ⁇ n in ⁇ aches ⁇ ve vys ⁇ n ⁇ yaschi ⁇ - ⁇ lie ⁇ ilengli ⁇ l with m ⁇ le ⁇ ulya ⁇ n ⁇ y maes ⁇ y 200 (PEG-200), with ⁇ lie ⁇ ilengli ⁇ l m ⁇ le ⁇ ulya ⁇ n ⁇ y mass ⁇ y 400 (PEG-400) seed dime ⁇ ilsul ⁇ - (D ⁇ S0 "> n d ⁇ .
  • PARTICULAR FORMS on basic electro- tives also include auxiliary and physical substances.
  • 5 Injectable industrial veterinary medicine contains one of the Amec- tecine compounds (Averesecin C, Averasec- 7 tyne ⁇ , or Electrotecin ⁇ at a concentration of 0.9-1.1%) in the distributor, which supplies a complete mixture of ethanol, polyethylene glycol and water in the case (5-7):
  • Opportunistic applications include a combination of 0.1% to 0.3% of the hard drive, and use or reassign.
  • the preparation for the protection of plants against gallant nematodes also contains one of the complexes of amec- teins at a concentration of 0, 1-0, 3 in case of neglect or neglect. 5.
  • the storms of Z. Augustus :1: . ⁇ 1 ⁇ -003 ⁇ were cultivated on the Ostagany agar for 10 days 27–29 ° C.
  • the bulk of the agar was transported into the pumping flasks with a capacity of 750 ml, containing 75 ml of the medium of the following composition,%: glucose - 4.0; soybean flour - 0.6; protein and vitamin ratio - 0.8; gourmet extract - 0.4; CaS ⁇ z - 0.1; ⁇ 2 ⁇ 4 - 0.05; WATER WATER.
  • They glucose was sterilized separately at 0.6 and 30 minutes, and they were introduced into the flasks of the sowing before it was immediately. Other components sterilized at 1.0 and 40 minutes.
  • the ⁇ value for sterilization is 6.2–6.4.
  • Bulbs were installed on a download with a number of 220 units per minute and inoculated at 27-29 ° C for 36 hours. Then, in the amount of 5?, They were transferred to a flask with a fragment of the following medium,%: glucose - 7.0; soybean flour - 0.6; protein-vitamin concentrate - 0.8; gourmet extract - 0.4; CaS ⁇ z - 0.1; ⁇ - 0.05; WATER WATER. The ⁇ value before sterilization is 6.2–6.4. Glucose was sterilized separately at 0.6 and 30 minutes, and the remaining components at 1.0 and 40 minutes. Glitches were introduced into the flasks of an unintentionally injured person by sowing the medium.
  • the process was carried out for 192 hours at 28 ° C on a download with a total of 220 per minute.
  • the trays washed 500 ml of 96% ethanol. Received 7, 15 l of a purified and processed product (7.5 mg / ml, 99.4% yield), they drove the product and consume 55% of the water. The stock in the flask, heated to 65 ° C, was placed in a separate office for separation of the lipid fraction.
  • the preparation was administered in the following doses of 50, 70, 84, 90, 110, and 150 mg / kg body weight.
  • White infertile mice weighing 25-30 g of Averasecin C were administered in doses of 7, 10, 25, 40, 50 and 60 mg / kg. Each dose to the rounds and mice was tested on 6 hours.
  • To calculate the average lethal dose we used the method of bit analysis for Litchfield and Wilkson (see, for example, Belenky L., The elements are neglected).
  • the results of the determination of the simple toxicity of Avectin C for bats and mice are shown in Table 5.
  • the extract is highly concentrated as in Example 2. After the release of the waste water, the waste was transferred to a separate business. After quenching, two layers were obtained: the lower layer was PEG-200, containing 40 g of aversine C, and the upper one was a lipid fraction. The lower layers of the slower and peremeshivanin were poured into 1.37 liters of water. The sprayed plant was divided on a filter, washed and dried, and dried in a vacuum drying oven. Received 38.4 g of EVERYtin C from the following composition. in%: simplifyectin ⁇ ⁇ . - 11 ⁇ . - 38 ⁇ ⁇ tracks ⁇ ggi - 3.1
  • Example 5 200 ml of the Avessecin C solution in PEG-400, as shown in Example 2 (76 mg / ml Avessecin C), was added at a concentration of 400 ml. The mixture was stirred for another 1 hour and set aside for installation at a room temperature for 10 hours.
  • Broken-down Crystals 11 Wash through a glass and a clean filter before vacuum, the crystals on the filter wash 50 ml of chilled water up to 10 ° C and dry up to 60% of the weight.
  • males are 70 and 20 mg / kg, respectively.
  • Example 5 Example 5 was vaporized in a vacuum, as was the case in Example 2. The waste was transferred to a separation business. After the standstill, two layers were learned. The lower layer, the Averasectin solution réellerier redesign in PEG-400, was poured into 600 ml of water. The sprayed plant was divided into a filter and dried. Received 12, 6 g of the preparation - ⁇ vereectin ⁇ .
  • Averesin C A decrease in the content of Averesin C below 0.9 leads to a decrease in the effective coefficient of activity of the drugs.
  • An increase in the content of averesectin C is higher than 1.1%, without changing the efficiency of the com- pact, it increases its strength and prolongs the elimination from the organism.
  • auxiliary substances ensures the optimal viscosity of the composites, the growth of the property of C, and the property of the property is independent of the property.
  • auxiliary substances instead of Malawitoctin C, you can also enter Avectoctin réelle convinced or Avectoctin crizillus at the same rate as the concentrate.
  • Each of the transceiver is effective when it comes to ecto- and endoparaeitis.
  • you receive a medicine you receive a medicinal form of community.
  • the Avessecin ⁇ is more preferable to doing so, since there is a death in the case of an invasion (on the 5th day), there is a sinking of the
  • ⁇ id zhiv ⁇ ny ⁇ name D ⁇ ea ⁇ a ⁇ n ⁇ s ⁇ ⁇ emya zab ⁇ levaniya ml / 50 ⁇ g administration ⁇ b ⁇ ab ⁇ i ⁇ vsh es ⁇ e 1.0 ⁇ dn ⁇ a ⁇ n ⁇ ⁇ yab ⁇ -n ⁇ yab ⁇ mel ⁇ ag ⁇ e 1.0 ⁇ dn ⁇ a ⁇ n ⁇ ⁇ ⁇ aeaniyam di ⁇ i ⁇ aulee, ⁇ s ⁇ n- 1.0 ⁇ dn ⁇ a ⁇ n ⁇ ⁇ e ⁇ ed ⁇ s ⁇ an ⁇ v- gilee, ⁇ lle ⁇ i ⁇ z, bus ⁇ m ⁇ z, ⁇ y on s ⁇ yl ⁇ v ⁇ e ⁇ i ⁇ s ⁇ ngildy , ⁇ abe ⁇ i ⁇ e s ⁇ de ⁇ zhanie and weight ee ⁇ ag ⁇ s ⁇ m ⁇ s, s ⁇ ngil ⁇ - n ⁇ y ⁇ e ⁇ ed vy
  • Ruminants ( ⁇ u ⁇ ny ⁇ ga ⁇ y s ⁇ , ⁇ vtsy, ⁇ zy, ve ⁇ b- Luda, zub ⁇ y) Di ⁇ i ⁇ aulez, gem ⁇ n ⁇ z, ⁇ s ⁇ e ⁇ agi ⁇ z, nema ⁇ di- Pe ⁇ ed ⁇ s ⁇ an ⁇ v ⁇ y on ⁇ z, ma ⁇ shallagi ⁇ z, ⁇ - s ⁇ yl ⁇ v ⁇ e s ⁇ de ⁇ zhanie ⁇ e ⁇ i ⁇ z, ⁇ abe ⁇ i ⁇ z, ee ⁇ - ⁇ senyu and vesn ⁇ ⁇ e- ⁇ ag ⁇ s ⁇ m ⁇ z, bun ⁇ s ⁇ - It is advantageous for the spare part. 100 biscuits
  • the drug containing 0.2% of the substance used, is used for the treatment and treatment of maternal, aromatic, and rheumatoid diseases.
  • the product is used for all kinds of livestock and poultry mixed with dry or moistened food in the morning for two or seven days before eating, 9 days a week.
  • the general dose of food with the appliance should not increase the amount of the required quantity of food to the patient.
  • the efficiency of the product is at least 35%.
  • the product is based on the main part ⁇ and the recession and the whole will be more preferable and not on the contrary, and on the basis the main part ⁇ - due to the outcome.
  • the product that contains Avessetin C will be more preferable and generally more convenient for horsemans.
  • the human health center on the basis of the recession and the whole independence of the Aversecine group, will be effective in dealing with galley munitions. The differences between them will be in the realm of sustainable activities.
  • the period of the protective action (during the processing of the preparation for the first galls) is at least 4 months.

Landscapes

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

Abstract

The present invention relates to the production of avermectin-based anti-parasitic preparations as well as to the new strain Streptomyces avermitilis 003FBM which is produced at a concentration of between 2000 and 3000 νg/ml. This invention also relates to a method for separating an avermectin complex from the biomass of a strain, wherein said method comprises carrying out an extraction from said biomass using a mixture of organic solvents having high and low boiling points with the addition of water. The extract is then purified in ion-exchange resins before removing the lipid fraction, and the final product is separated during a crystallisation and/or precipitation process. The solvents having a low boiling point may consist of methanol, ethanol, isopropanol, acetone or acetonitrile, while the solvents having a high boiling point may consist of polyethyleneglycol, dimethylsulfoxide or polypropyleneglycol. This method may be used to produce various complexes such as aversectin C, aversectin B or aversectin AB. These complexes are used in the production of veterinary anti-parasitic preparations which can be administered perorally or by injection and which also provide protection for plants against root-knot nematodes.

Description

ШΤΑΜΜ 3-ЫеρЪοтусее аνегтϊЫПε, СПΟСΟБ ΒЫДΕЛΕΗИЯ ΑΒΕΡΜΕΚΤИΗΟΒЫΧ ΚΟΜПЛΕΚСΟΒ И ПΡΕПΑΡΑΤЫ ДЛЯ ЗΑЩИΤЫ ЖИΒΟΤΗЫΧ И3-Yerёrёtyse аntyгтыпе, METHOD OF CREATING ANTI-VEHICLE AND ANTI-VEHICLE PROTECTION

ΡΑСΤΕΗИЙRASTEEN

Οбласτь τеχниκи Изοбρеτение οτнοсиτся κ ποлучению биοлοгичесκи-аκτивныχ ве- щесτв πуτем мнκροбиοлοгичесκοгο синτеза, а именнο κ προизвοдсτву анτиπаρазиτаρныχ πρеπаρаτοв на οснοве авеρмеκτинοв .Field of Engineering The invention concerns the production of biologically active substances by multiplying biological synthesis, namely the production of antipasitate drugs based on avemekin.

Пρедшесτвующий уροвень τеχниκи Извесτнο, чτο аκτинοмицеτ ЗЬгеρЪοтусез аνегтϊЫϋο πρи κу- льτивиροвании в πρисуτсτвии исτοчниκοв углеροда, аэοτа и φοсφορа πρи ποсτοяннοй аэρацин προдуциρуеτ вοсемь блиэκπχ πο сτροениκ вещесτв, наэванныχ авеρмеκτннамн , κοτορые οбладаюτ ρеэκο выρаже- нным анτиπаρазиτаρным дейсτвием πаτенτ <ЗΒ 1573955, 1980г. ). Βсе авеρмеκτины πο χимичесκοму сτροению являюτся 16-членньηуιи маκρο- лидами и οτличаюτся между сοбοй замесτиτелями у 5 и 26 аτοмοв углеροднοгο κοльца. Индивидуальные авеρмеκτины, ποлучившие οбοэ- наченπя Αι&, Α ъ, Αг&, Αзъ, Βι&, Βιъ, Βг& и Βгъ, имеюτ ρазличную биοлοгичесκую аκτивнοсτь. Ηаибοлее аκτивнымπ являюτся авеρмеκτи- ны Βι, οсοбеннο Βϊ&.Advancing level of technology It is known that akinomycete sulphate is agtotypic and cultivated in the presence of source of carbons, aeοτοτ and φοορρρποροροροροροοραοοοιοιοιοιοοιοκκκος substances derived from avermectins, which have a pronounced antiparasitic effect (patent <ЗВ 1573955, 1980). All avermectins are 16-membered macrolides by chemical structure and differ from each other in the substituents at 5 and 26 atoms of the carbon ring. Individual avermectins, which received both A1, A2, A3, A4, A5, A6, A7, A8, A9, A9, A10, A11, A12, A13, A14, A15, A16, A17, A18, A19, A20, A21, A22, A23, A24, A25, A26, A27, A28, A29, A30, A31, A32, A33, A34, A35, A36, A37, A38, A39, A40, A41, A42, A43, A44, A45, A46, A47, A48, A49, A50, A51, A52, A53, A54, A55, A56, A57, A58, A59, A60, A61, A62, A63, A64, A65, A66, A67, A68, A69, A69, A60, A61, A62, A63, A65, A66, A67, A68, A69 ...

Ρаэвиτи^ τеχнοлοгии ποлученπч авеρ>меκτннοв προисχοдиτ πο несκοльκим наπρавлениям , κ числу κοτορыχ οτнοсяτся ποлучение шτаммο , προдуциρующиχ ποвышенные κοличесτва всегο κοмπлеκса авеρмеκτинοв или προдуциρующиχ авеρмеκτины с πρеοбладающим сο- деρжанием гρуππы Βι, сοвеρшенсτвοвание προцесса выделения авеρ- меκτинοв иэ мицелия шτаммοв 5"Ьгеρ-Ьοтусез аνегтϊЫΙϊз и дρугие. Иэвесτен шτамм 5-Ьгеρ"Ьοηιусез аνегтϊЫΙϊε ΑΤСС 31272, προду- циρующий в сρеднем 500 мκг/мл авеρмеκτинοв (πаτенτ ϋ5 4310519, 1982г.). Пροдуκτивнοсτь шτамма 51геρ-Ьοтусез аνегтιЫϋз ΒΗИИСΧΜ- 51 (πаτенτ ΕИ 2048250, 1995г.) сοсτавляеτ 700 мκг/мл. Шτамм 5-Ьгеρ-Ьοтусеε аνегтϊЫΙϊз ΒΗИИСΧΜ-54 προдуциρуеτ уже 1500 мκг/мл авеρмеκτинοв (πаτенτ Εϋ 2054483, 1996г.), πρичем сοдеρжание авеρмеκτина Βϊа дοсτигаеτ 35% и бοлее. Βο всеχ πρиведенныχ слзг- чаяχ авеρмеκτины выделяюτ иэ мицелия эκсτρаκцией индивидуальным ορганичесκим ρасτвορиτелем, смешивающимся с вοдοй, с ποследующим ρаэделением, наπρимеρ χροмаτοгρаφичесκим. Сущнοсτь изοбρеτения Целью насτοяшегο иэοόρеτения являеτся сοздание шτамма, προ- дуциρующегο еще бοлее высοκие уροвни авеρмеκτинοв с ποвышенным сοдеρжанием в κοмπлеκсе авеρмеκτинοв гρуππы Βι (Βι&, Β в) - наи- бοлее аκτивныχ в биοлοгичесκοм οτнοшении, а τаκже ρаэρабοτκа сποсοба выделения авеρмеκτинοвοыχ κοмπлеκсοв ρаэнοгο κοмποненτ- нοгο сοсτава и изгοτοвление πρеπаρаτοв на иχ οснοве.The efficiency of radiation technology comes from several directions, depending on the number of cores This concerns the production of strains that produce increased amounts of the total complex of avemetins or that produce avemetins with with a dominant feeling group B, improvement of the process of isolating avermectins from the mycelium of strains 5-Ьерф-Боитсыес аэнтϊЫΙϊε and others. The strain 5-Ьерф - Боитсыес аэнтϊЫΙϊε ATCC 31272, producing an average of 500 μg/ml of avermectins (patent no. 5 4310519, 1982). The productivity of the strain 51er-Bomyces angiotensin II BIISC-51 (patent EI 2048250, 1995) is 700 μg/ml. The strain 5-Ьер-Ботысее антінз ВІИЧМ-54 already produces 1500 μg/ml of avermectins (patent Eϋ 2054483, 1996), and the content of avermectin Bϊa reaches 35% or more. In all the above -mentioned studies, avermectins are isolated from the mycelium by extraction with a separate organic solution mixed with water, followed by separation, for example, chromatographic. The essence of the invention The aim of the present invention is to create a strain producing even higher levels of avermectins with an increased content in the complex of avermectins of the Βι group (Βι&, Ββ) - the most active in biological terms, as well as development of a method for isolating avermectin complexes of this component composition and manufacturing drugs based on them.

Цель дοсτигаеτся ποлучением шτамма 5"Ьгеρ"Ьοтусез аνегтϊЫИз 003ΦБΜ, προдуциρующегο в сρеднем 2300 мκг/мл авеρмеκτинοв πρи ρаэмаχе иэменчивοсτπ 2000-3000 мκг/мл.The goal is achieved by obtaining a strain of 5"Br"Bomyces anthemis 003FBM, producing on average 2300 μg/ml of avermectins with a variability of 2000-3000 μg/ml.

Пρедлагаемый шτамм был ποлучен πуτем наπρавленнοй селеκции иэ исχοднοгο шτамма 5"Ьгеρ"Ьοтусеε аνегтϊЫΙϊ-ϊ ΒΗИИСΧΜ-54 чеρеэ ρяд προмежуτοчныχ ваρианτοв. Οτбορ аκτивныχ ваρнанτοв προвοднлι; πο сποсοбнοсτи κ биοсинτеэу авеρмеκτинοвοгο κοмπлеκса с сοдеρжа- нием Β не менее Зб Пροмежуτοчные ваρианτы χаρаκτеρнзοвались высοκοй иэменчивοсτью πο авеρмеκτинοοбρаэοванию, чτο ποэвοлялο исποльэοваτь κаждый иэ ниχ в ποследующиχ эτаπаχ селеκции , нο не давалο вοзмοжнοсτи πρименяτь в κачесτве προмышленныχ шτаммοв. У προмежуτοчныχ ваρианτοв иэменчивοсτь ποследοваτельнο сοсτавляла 50-140%; 49-130% π 78-143%, чτο сοοτвеτсτвοвалο προдуцнροванию 900-2500 мκг/мл; 950-2500 мκг/мл и 1650-3000 мκг/мл авеρмеκτи- нοв.The proposed strain was obtained through directed selection from the original strain 5"breast"boutyceε aegtϊYΙϊ-ϊ ΒΗIISΧΜ-54 chee ρyad Interim options. About the active vaρnantos; About the biosynthetic capacity of avemekin complex containing at least 3B Intermediate variants The materials were characterized by high variability in the production of food products, which meant the use of every the next stages of selection, but did not give the possibility of using them as industrial strains. In intermediate variants, the variability was consistently 50-140%; 49-130% and 78-143%, which corresponded to the production of 900-2500 μg/ml; 950-2500 mcg/ml and 1650-3000 mcg/ml avemequins.

Пοлученный шτамм χаρаκτеρизуеτся следующими κульτуρальнο- мορφοлοгичесκимн и φиэиοлοгичесκимπ πρизнаκами. Μορφοлοгπчесκие πρиэнаκи шτамма. Κульτуρа имееτ сποροнοсцы в виде πρавильныχ сπиρалей.The obtained strain is characterized by the following cultural, morphological and physiological features. Morphological properties of the strain. The culture has spores in the form of regular spirals.

Μаκροмορφοлοгичесκие и κульτуρальные πρиэнаκи. Ηа 2%-нοм глюκοзοκаρτοφельнοм агаρе на 14-21 день ροсτа πρи 28°С, ρΗ 6,8-7,2 οбρазуюτся κοлοнии 7-10 мм с сеρым вοэдушным мицелием и τемнο-κашτанοвοй οбρаτнοи сτοροнοι κοлοний. Β ποπуля- ции, κаκ πρавилο, имееτся οκοлο 5% сеρο-белыχ κοлοний с бежевοй οбρаτнοй сτοροнο . Κρай κοлοний неροвный с белым οбοдκοм. Пοвеρ- χнοсτь κοлοнии сκладчаτая. П десяτοму дню ροсτа на ποвеρχнοсτи κοлοнии мοгуτ οбρазοвываτься белые баρχаτисτые вκлючения. Β τабл .1 πρедсτавлены данные ο цвеτе вοздушнοгο и субсτρаτ- нοгο мицелия, инτенсивнοсτи сποροнοшения, а τаκже ο наличии или οτсуτсτвии ρасτвορимыχ πигменτοв на ρазличныχ ορганичесκиχ п синτеτичесκиχ агаρиэοванныχ сρедаχ. Τаблииа 1Macromorphological and cultural characteristics. On 2% glucose carboxylate agar on days 14-21 at 28°C, pH 6.8-7.2, 7-10 mm colonies with gray air-filled mycelium and dark chestnut-colored reverse side of the colonies are formed. According to the population, there are about 5% of gray-white colonies with beige crops. The tail is irregular with a white rim. The nature of the colony is folded. On the tenth day of growth, white velvety inclusions may form on the surface of the colony. Table 1 presents data on the color of the aerial and substrate mycelium, the intensity of reproduction, as well as the presence or absence of soluble pigments on various organic and synthetic agar media. Table 1

Κульτуρальные πρиэнаκи БЪгеρ-Ьοтусев аνегтϊ'Ыϋβ-ΟΟЗΦБΜ на ρазличныχ агаρизοванныχ сρедаχCultural properties of BЪger-пotusev agnes in various agarized environments

Figure imgf000005_0001
Figure imgf000005_0001

Μаκсимальнοе наκοπление авеρмеκτинοв προисχοдиτ на 7-8 суτ- κи πρи сκοροсτи ρасτвορения κислοροда 1,0-1,3 мΜοль Οг/л-мин и τемπеρаτуρе 28°С.The maximum accumulation of avemequins occurs on days 7-8 and the oxygen level is 1.0-1.3 mol Οg/l-min and a temperature of 28°C.

Сведения ο φизиοлοгο-биοχимичесκиχ πρиэнаκаχ шτамма 5-Ьгеρ- Ъοтусез аνегтϊЪϋϊз - 003ΦБΜ суммиροваны в τабл.2. Τаблица 2Information about the physiological and biochemical characteristics of the 5-brheρ-Ъοtuses аνегтϊϋϊз - 003ΦBoΜ strain is summarized in Table 2. Table 2

Пρизнаκ Ηаличие + ) или οτсуτсτвие ( - ) ροсτаSign Presence (+) or absence (-) of growth

Уτилизация углевοдοв: глюκοза саχаροза + φρуκτοза + лаκτοза + манниτ + сορбиτ + аρабинοза + ρамнοза + деκсτρин κρаχмал ρаφφинοза +Utilization of carbohydrates: glucose saccharose + phosphorus + laquoise + mannitol + sobito + aabinose + pamnosis + dextine κρρmal phosphonose +

Уτилизаиия исτοчниκοв азοτа аэοτ в вοссτанοвленнοπ φορме + азοτ в οκисленнοй φορме +Utilization of nitrogen sources aero in reduced form + nitrogen in oxygenated form +

Желаτинаэная аκτивнοсτь (ρазжижаеτ желаτину )Gelatin activity (liquefies gelatin)

Пροτеοлиτичесκая аκτивнοсτьPoteolytic activity

Инвеρсия саχаροэыSakhalin investment

Τемπеρаτуρныή диаπаэοн ροсτа, °С 20 4- 28 + 37 + /- 50Temperature pressure range, °C 20 4- 28 + 37 + /- 50

Диаπазοн эначени ρΗ 4.5 + /- 6,8 7.4 9,0 + /-Value range ρΗ 4.5 + /- 6.8 7.4 9.0 + /-

Αзροбные услοвия Αнаэροбные услοвияAerobic conditions Anaerobic conditions

Пροτивοπаρазиτаρные свοйсτва Μицелиальные эκсτρаκτы вызываюτ гибель немаτοд, οτнοсящнχся κ ροдам: Ηаетοηсϊιиз, Τг1сЪοз"-:гοη§у1и.___ , 0з1;ге.г1;ае1а, Νета1οάϊги.з- Сοορегϊа, Αесагϊό, Βυηοз-Ьοтит, ΟезορЪадοзЪοтит, СЪаЪегϋа, Τгϊ- сЪглгΙз, 51гοη£у1и&, ΤЫсЬοηета, ϋϊе^уοсаиϊиз, СаρШагϊа, Ηе е- гакιе, Τοχοсага, Οχуигϊз, ΑηсуΙοзЪοта, ϋηсϊηагϊа, Τοχазеагιз, Αзсагϊάϊа.Antiparasitic properties Mycelial extracts cause the death of nematodes belonging to the following genera: Nematodes, Toxicidae, Nematodes, Nematodes, Corynebacteria, Acarizoa, Bacteroides, Oxygenes, Τгϊ- сълггΙз, 51гοη£у1и&, Τысьοηета, ϋϊе^уοсайϊiz, SaρShagϊа, Ηе е-гakιе, Τοχοsaga, Οχугϊз, ΑηsuΙοзοta, ϋηсϊηagϊa, Τοχazeagιz, Αzsagϊάϊa.

Τаκже иχ дейсτвие губиτельнο для κлещеή, аφндοв, вшей, муχ, блοχ, τаρаκанοв, двуκρылыχ и πρямοκρылыχ вρедиτелей, πлаτянοй мοли, ΚΟΕΡΟΒЫΧ и οбοйныχ жучκοв. Сρеднее сοдеρжание авеρмеκτинοв в мицелии 5.аνегтϊ"Ы1ϊβ- 003ΦБΜ сοсτавляеτ 2300 мκг/мл πρи ρазмаχе иэменчивοсτи 2000-3000 мκг/мл. Κοмποненτный сοсτав авеρмеκτинοв следующий, %: гρуππа Αι(Αι&+Αι_ο) 5,0 - 7,2; гρуππа Αг Αга+Αζв) 21,6 - 24,5; гρуππа Βι(Βϊβ.+Βϊв) 48,5 - 50,0; гρуππа Βг(Βга+Βгв) 18,4 - 23,0. Τаκим οбρаэοм πρедлагаемый шτамм-προдуценτ Ξ.аνегюШΗз- 003ΦБΜ ποэвοляеτ ποлучаτь на 50% бοльше авеρмеκτинοв, чем извес- τный шτамм З.аνегтϋϋϊε ΒΗИИСΧΜ-54.Their action is also destructive to ticks, mites, lice, flies, fleas, cockroaches, dipterous and straight-winged pests, clothes moths, keratin and wallpaper beetles. The average content of avermectins in the mycelium of S. agneticus is 2300 μg/ml with a range of variability of 2000-3000 μg/ml. The component composition of avermectins is as follows, %: group Aι (Aι&+Aι_ο) 5.0 - 7.2; group Aι (Ag+Aζv) 21.6 - 24.5; gρππππΒι(Βϊβ.+Βϊв) 48.5 - 50.0; gρπππа Βg(Βga+Βgv) 18.4 - 23.0. As a result, the proposed strain-producer Ξ.agegyushoz-003ΦBΜ produces 50% more avemequins than the known dark strain Z.agtϋϋϊε ΒΗIISΧΜ-54.

Для выделения авеρмеκτинοвыχ κοмπлеκсοв ρазличнοгο κοмπο- ненτнοгο сοсτава исποльзуюτ суχую бποмассу, ποлученную πρи выρа- щиванин προдуценτа в глубинныχ услвияχ на иэвесτныχ πиτаτельныχ сρедаχ. Эκсτρаκцию авеρмеκτинοв из биοмассы οсущесτвляюτ смесью, сοсτοящей из вοды и низκο- и высοκοκиπящиχ ρасτвορиτелей, смешн- вающиχся с вοдοй в неοгρаниченнοм сοοτнοшении. Β κачесτве низκο- κиπящиχ исποльзуюτ ρасτвορиτели с τемπеρаτуροй κиπения, не πρе- вышающей τемπеρаτуρу κиπения вοды, наπρимеρ, эτилοвый, меτилοвый и изοπροπилοвый сπиρτы или ацеτοн, в κачесτве высοκοκнπящиχ - ποлиэτиленглиκοль с мοлеκуляρнοй маесοй 200 (ПЭГ-200), ποлиэτи- ленглиκοль с мοлеκуляρнοй массοй 400 (ПЭГ-400), димеτилсульφοκ- сид (ДΜС0"> н дρ. Эκсτρаκτы οчищаюτ οτ πигменτοв и неορганичесκиχ πρимесей с ποмοщью κаτиοнοοбменныχ и аниοнοοбменныχ смοл. Заτем в ваκуум-исπаρиτеле οτгοняюτ вοду и низκοκиπящий ρасτвορнτель . Пуτем οτсτаивания οсτавшейся часτи эκсτρаκτа в высοκοκиπящем ρасτвορиτеле οτделяюτ πρимеси лиπиднοгο χаρаκτеρа. Пοлучение авеρмеκτинοвыχ κοмπлеκсοв с ρазличным κοмποненτным сοсτавοм ве- дуτ πуτем φρаκциοннοгο οсаждения и κρисτаллиэации в πρисуτсτвии вοды и эτилοвοгο сπиρτа. Пρи эτοм ποлучаюτ:To isolate avermectin complexes of different component composition, the dry mass obtained during cultivation of the producer in deep conditions in known nutrient media is used. Extraction of avermectins from biomass is carried out using a mixture consisting of water and low- and high-boiling solvents that are miscible with water in an unlimited ratio. As low-boiling agents, solvents with a boiling point not exceeding the boiling point of water are used, for example, ethyl, methyl and isopropyl alcohols or acetone, as high-boiling agents - polyethylene glycol with a molecular weight weight 200 (PEG-200), polyethylene glycol with a molecular weight of 400 (PEG-400), dimethyl sulfoxide (DMSO), etc. The extracts purify pigments and inorganic impurities using cation exchange and anion exchange resins. Then, water is distilled off in a vacuum evaporator and low-boiling solution. By settling the remaining part of the extract in a high-boiling solution, impurities of the lipid character are separated. Obtaining avermectin complexes with different component composition is carried out by phasic precipitation and crystallization in the presence of water and ethyl alcohol. This produces:

1 ) суммаρный авеρмеκτинοвый κοмπлеκс , ποлучивший наэвание авеρсеκτин С, сοдеρжащий 8 авеρмеκτинοвыχ κοмποненτοв в сοοτнο- шении , % : Α Αιϊ»а 6 6 -- 1122;; Β ι» 30 - 40;1) total avemekin complex, derived from avesectin C, containing 8 avemekin complexes in proportion, % : Α Αιϊ»а 6 6 -- 1122;; Β ι» 30 - 40;

Αϊв следы; Β ϊв 2 - 12;Αϊв traces; Β ϊв 2 - 12;

Αга 15 - 25 ; Βаа 14 - 24;

Figure imgf000007_0001
или 2) κοмπлеκс, ποлучивший названπе авеρсеκτин Β, сοдеρжащий в οснοвнοм авеρмеκτины Бι&. и Βи≥, в :Αga 15 - 25; Βаа 14 - 24;
Figure imgf000007_0001
or 2) a complex called aversectin B, containing mainly aversectins Bβ and Bβ, in:

Figure imgf000008_0001
οсτальные авеρмеκτины - следы, или
Figure imgf000008_0001
The remaining avermectins are traces, or

3) κοмπлеκс, названный авеρсеκτинοм ΑΒ, следующегο сοсτава. в % :3) a complex called aversectin AB, of the following composition in %:

Αι* 4 - 10; Βιа 12 - 18: Αϊв следы; Βϊв 3 - 9; Α2а 25 - 45; Β2Λ 25 - 45; Αгв следы; Ββϊ 5,Αι* 4 - 10; Βι a 12 - 18: Αϊin traces; Βϊв 3 - 9; Α 2a 25 - 45; Β 2Λ 25 - 45; Αgv traces; Ββϊ 5,

Биοлοгичесκие свοйсτва ποлученныχ πρеπаρаτοв πρедсτавлены в τаблицаχ 3 и 4.The biological properties of the obtained preparations are presented in Tables 3 and 4.

Τаблица 3 Τοκсичнοсτь авеρмеκτинοвыχ πρеπаρаτοв для κρыс н мышейTable 3 Toxicity of avermectin drugs for rats and mice

Ρ_

Figure imgf000008_0002
Ρ_
Figure imgf000008_0002

Τаблица 4 Дейсτвие авеρмеκτинοвыχ πρеπаρаτοв на ΑρЬеΙеηЬοϊάез Ьезаеϊ 0Table 4 Effect of avermectin drugs on Atrial fibrillation 0

5

Figure imgf000008_0003
5
Figure imgf000008_0003

Пρедлагаемые авеρмеκτинсοдеρжащие πρеπаρаτы служаτ для из- гοτοвления леκаρсτвенныχ φορм: инъеκциοнны , маэевыχ, πасτοοбρа- зныχ, ποροшκοοбρазныχ н τаблеτοчныχ φορм для ορальнοгο πρимене- 0 ния и жидκиχ φορм для οροшения - πρедназначаемыχ для лечения жи- вοτныχ, а τаκже жидκиχ πасτοοбρазныχ и ποροшκοοбρазныχ φορм для бορьбы с вρедиτелями сельсκοχοзяйсτвенныχ κульτуρ. Пρеπаρаτивные φορмы на οснοве авеρмеκτинοв вκлючаюτ τаκже всποмοгаτельные и φορмοοбρаэующие вещесτва. 5 Инъеκциοнныи προτивοπаρазиτаρный веτеρинаρныи πρеπаρаτ сο- деρжиτ οдин из авеρмеκτинοвыχ κοмπлеκсοв ( авеρсеκτин С , авеρсеκ- 7 τин Β, или авеρсеκτин ΑΒ в κοнценτρации 0,9-1,1%) в ρасτвορиτе- ле, πρедсτавляющем сοόοй смесь эτанοла, ποлиэτиленглиκοля и вοды в сοοτнοшении (5-7) : (5-7 ) : 1. Βеτеρинаρныπ πρеπаρаτ для πеρορаль- нοгο πρименения сοдеρжиτ κοмπлеκс авеρмеκτинοв в κοнценτρации 0,1-0,3% на τвеρдым нοсиτеле, в κачесτве κοτοροгο исποльзуюτ πеρлиτ или цеοлиτ.The proposed avemektin-containing drugs are used for the production of medicinal drugs: injectables, maevs, pastes, Preferred tablets for general use and liquids for irritation - intended for the treatment of life wars, and as well as liquid paste-like and powder-like formulations for combating agricultural pests. Avermectin-based preparations also include auxiliary and preparative substances. 5 Injectable anti-inflammatory veterinary drug contains one of the avermectin complexes (avermectin C, aversectin 7 tin B, or aversectin AB in a concentration of 0.9-1.1%) in a solution representing a mixture of ethanol, polyethylene glycol and water in the ratio (5-7): (5-7): 1. The polyethylene preparation for oral use contains a complex of avermectins in concentration of 0.1-0.3% on a solid carrier, for which pellitite or zeolite is used.

Пρеπаρаτ для защиτы ρасτений οτ галлοвыχ немаτοд τаκже сο- деρжиτ οдин из κοмπлеκсοв авеρмеκτинοв в κοнценτρации 0 , 1-0 , 3 на ποροшκοοбρазнοм πеρлиτе или цеοлиτе, или οτρабοτаннοм мицелии 5. аνегт!111ϊ8.The preparation for plant protection against root-knot nematodes also contains one of the avermectin complexes in a concentration of 0.1-0.3 on powder-like pellet or zeolite, or processed mycelium 5. аngеrт!111ϊ8.

Лучшие ваρианτы οсущесτвления нзοбρеτения Пρимеρ 1The best options for implementing the invention Example 1

Сπορы шτамма З.аνегтϊ1:.ϊ1ιз-003ΦБΜ выρащивали на οвсянοм агаρе в τечение 10 суτοκ πρн 27-29°С. Κусοчеκ агаροвοн сρеды сο сπορамн внοсили в κачалοчные κοлбы емκοсτью 750 мл , еοдеρжащие 75 мл сρеды следующегο сοсτава, %: глюκοза - 4,0; сοевая муκа - 0,6; белκοвο-виτамннный κοнценτρаτ - 0,8; κуκуρузный эκсτρаκτ - 0,4; СаСΟз - 0,1; Κ2ΗΡΟ4 - 0,05; вοда вοдοπροвοдная. Глюκοзу сτеρилизοвали οτдельнο πρи 0,6 аτи 30 минуτ π внοсили в κοлбы неποсρедсτвеннο πеρед засевοм. Οсτальные κοмποненτы сτеρилизο- вали πρи 1,0 аτи 40 минуτ. Значение ρΗ дο сτеρилизации 6,2-6,4.The bacteria of strain Z. анегт1:.ϊ1ιз-003ФБМ were grown on oat agar for 10 days at 27-29°С. A piece of agar with the bacteria was added to 750 ml shaking flasks containing 75 ml of the medium of the following composition, %: glucose - 4.0; soy flour - 0.6; protein-vitamin concentrate - 0.8; corn export - 0.4; CaCΟz - 0.1; Κ2ΗΡΟ4 - 0.05; tap water. Glucose was sterilized separately at 0.6 and 30 minutes and added to the flasks immediately before inoculation. The remaining components were sterilized at 1.0 atm or 40 minutes. The ρΗ value before sterilization was 6.2-6.4.

Κοлбы усτанавливали на κачалκу с числοм οбοροτοв 220 в ми- нуτу и выρашнвали инοκуляτ πρи 27-29°С в τечение 36 часοΕ . Заτем инοκуляτ в κοличесτве 5?., πеρенοсили в κοлбы с φеρменτаииοннο сρедοй следующегο сοсτава, % : глюκοэа - 7,0; сοевая муκа - 0,6; белκοΕθ-виτаминный κοнценτρаτ - 0,8; κуκуρузный эκсτρаκτ - 0,4; СаСΟз - 0,1; ΚεΗΡΟι - 0,05; вοда вοдοπροвοдная . Значение ρΗ дο сτеρилизациπ 6,2-6,4. Глюκοзу сτеρилизοвалπ οτдельнο πρи 0,6 аτи 30 мин, а οсτальные κοмποненτы πρи 1,0 аτи 40 мин. Глюκοзу внο- сили в κοлбы неποсρедсτвеннο πеρед засевοм сρеды инοκуляτοм.The flasks were placed on a shaker with a rotation rate of 220 rpm and the inoculum was incubated at 27-29°C for 36 hours. Then the inoculum in the amount of 5? was transferred to flasks with a fermentation medium of the following composition, %: glucose - 7.0; soy flour - 0.6; protein-vitamin concentrate - 0.8; corn export - 0.4; CaCΟz - 0.1; ΚεΗΡΟι - 0.05; tap water. The value of ρΗ before sterilization is 6.2-6.4. Glucose was stabilized separately at 0.6 to 30 minutes, and the remaining components at 1.0 to 40 minutes. Glucose was added to the flasks immediately before seeding the medium with the inoculum.

Φеρменτацию προвοдили в τечение 192 часοв πρи 28°С на κача- лκаχ с числοм οбοροτοв 220 в минуτу.Fermentation was carried out for 192 hours at 28°C on a rocker with a rotation rate of 220 revolutions per minute.

Οπρеделение уροвня авеρмеκτинοοбρазοвания προвοдили следую- щим οбρазοм. Пο 1 мл οбρазца κульτуρальнοй жидκοсτи πеρенесли в προбиρκи, сοдеρжащие πο 9 мл эτилοвοгο сπиρτа, τщаτельнο πеρеме- шали и οсτавили на 30 мин для эκсτρаκциπ авеρмеκτинοвοгο κοмπле- κса, ποсле чегο сοдеρжимοе προбиρκи προφильτροвали чеρеэ бумаж- ный φильτρ. Сοдеρжание в φильτρаτаχ всеχ κοмποненτοв авеρмеκτи- 5 нοвοгο κοмπлеκса οπρеделяли меτοдοм высοκοэφφеκτивнοй жидκοсτнοй χροмаτοгρаφии , οснοваннοм на χροмаτοгρаφичесκοм ρазделении авеρ- меκτинοвыχ κοмποненτοв на аналиτичесκοй κοлοнκе с οбρашеннοή φа- эοй Диасορб С-16 4x250 мм с ρаэмеροм часτиц 6 мκм в изοκρаτичес- κοм ρежиме. Элюиρующей сисτемοй служила смесь эτилοвοгο сπиρτа и ацеτοниτρила с вοдοй в сοοτнοшении 6,5:2,0:1,5.The level of avermectin formation was determined in the following manner. 1 ml of cultivar liquid was transferred into containers containing 9 ml of ethyl alcohol, carefully mixed and left for 30 minutes for the extraction of the avemequin complex, after which the contents of the they filtered the cheep paper filter. The content of all components of the filter is 5 new complex were determined by the method of high-efficiency liquid chromatography, based on the chromatographic separation of avertine components on an analytical column with a Diasorb S-16 4x250 mm processed phase. with a particle size of 6 μm in isostatic mode. The eluting system was a mixture of ethyl alcohol and acetonitrile with water in a ratio of 6.5:2.0:1.5.

Деτеκτиροвание κοмποненτοв вели πρи длине вοлны 243 нм. Χροмаτοгρаφиροвание исследуемыχ οбρазцοв προвοдили в сρавнении сο сτандаρτным οбρазцοм авеρмеκτина. Сοдеρжание авеρмеκτинοв в мицелии 5. аνегтϊ-ЫΙιз-ΟΟЗΦБΜ сο- сτавилο 2300 мκг/мл. Κοмποненτныπ сοсτаΕ авеρмеκτинοв, % : гρуππа Αι(Αϊа+Α в) 7,4; гρуππа Αζ(Α2а+Α2β) 22,4; гρуππа Βι(Β а+Βϊв) 49,6; гρуππа Βг ( Β -&+Β2Β ^ 20,6.The components were detected at a wavelength of 243 nm. Chromatography of the studied samples was compared with a standard avermectin sample. The content of avermectins in the mycelium of S. agnýgϊ-ЫΙιз-ОΟЗФБМ was 2300 μg/ml. Component composition of avemequins, %: group Αι(Αϊа+Αв) 7.4; Αζ(Α2а+Α2β) 22.4; gρππππΒι(Β а+Βϊв) 49.6; gρуππа Βг ( Β -&+Β2Β ^ 20.6.

Пοлучение авеρмеκτинοвыχ κοмπлеκсοв иллюсτρиρуеτся следую- щими πρимеρами:The production of avermectin complexes is illustrated by the following examples:

Пοлучение суммаρнοгο авеρмеκτинοвοгο κοмπлеκса (авеρсеκτина С) и οценκа егο биοлοгичесκοй аκτивнοсτиObtaining a total avermectin complex (avermectin C) and assessing its biological activity

Пρимеρ 2Example 2

3,5 κг мицелия 8. аνегтϊ111ιз-003ΦБΜ, высушеннοгο на ρасπы- лиτельнοй сушилκе, эκсτρагиροвалн в τечение 1 часа πρи πеρемеши- вании и πρи κοмнаτнοй τемπеρаτуρе смесью 0,5 л ПЭГ-400 и 6,5 л 75% эτанοла. Οсадοκ οτделнли φπльτροваннем на нуτч-φπльτρе ποд ваκуумοм и προмыли 0,8 л 96% эτанοла. Φильτρаτ и προмывную жид- κοсτь οбъединили и ποлучилн 7,0 л сπиρτοвοгο эκсτρаκτа сοдеρжа- щегο 500 мл ПЭГ-400 с κοнценτρацией авеρмеκτинοв 7,7 мг/мл, κο- τορый заτем προπусτили ποследοваτельнο чеρез κοлοнκи с 400 г κа- τиοниτа ΚУ-2χ8 в Η÷ -φορме и аниοниτа ЭДЭ-ΙΟП в 0Η~ -φορме . Κο- лοнκи προмыли 500 мл 96% эτанοла. Пοлучили 7 , 15 л οчищеннοгο сπиρτοвοгο эκсτρаκτа (авеρмеκτинοв 7,5 мг/мл, выχοд 99,4%), οτο- гнали сππρτ и вοду на ροτορнοм исπаρиτеле πρи τемπеρаτуρе 55°С в ваκууме . Οсτаτοκ в κοлбе нагρеτый дο 65°С ποмесτили в делиτель- ную вοροнκу для οτделения лиπиднοй φρаκции.3.5 kg of mycelium 8. аνегтϊ111ιз-003ФБМ, dried in a spray dryer, extracted for 1 hour with stirring and at room temperature with a mixture of 0.5 l of PEG-400 and 6.5 l of 75% ethanol. The sediment was subjected to a nucleic acid bath under vacuum and washed with 0.8 liters of 96% ethanol. The filtrate and washing liquid were combined and 7.0 liters of alcoholic extract containing 500 ml of PEG-400 were obtained. concentration of avemequins 7.7 mg/ml, which was then passed through columns with 400 g of ka- tinite ΚU-2χ8 in Η÷ -φορme and aninita EDE-ΙΟP in 0Η ~ -φορme. The columns were washed with 500 ml of 96% ethanol. We received 7.15 liters of purified alcohol extract (avemequins 7.5 mg/ml, yield 99.4%), then sent the alcohol and water to In a different heater at a temperature of 55°C in a vacuum. The residue in the flask, heated to 65°C, was placed in a separatory funnel to separate the lipid fraction.

Ηижний слοй οбъемοм 580 мл (авеρмеκτинοΕ 76 мг/мл) чеρез 3,2 часа οτделили и вылили в 1750 мл дисτиллиροваннοй вοды, на- гρеτοπ дο 40°С πρи инτенсивнοм πеρемешиваннн . Смесь πеρемешивалн Ε τечение 3-5 часοв для φορмиροвания οсадκа, οснοвную часτь над- οсадοчнοи жидκοсτи οτделили деκанτиροванием, а οсτальную часτь φильτροванием чеρеэ сτеκлянный φильτρ ποд ваκуумοм. Οсадοκ на Φильτρе προмыли 500 мл χοлοднοй дисτилиροваннοй вοды и высушили в κοнвеκциοннοм ποτοκе τеπлοгο вοэдуχа. Суχοй οсадοκ сοдеρжал 41 г авеρсеκτина С.The bottom layer with a volume of 580 ml (avermectin 76 mg/ml) was separated after 3.2 hours and poured into 1750 ml of distilled water, heated to 40°C with intensive stirring. The mixture was stirred for 3-5 hours to form a precipitate, the main part of the supernatant The sedimentary liquid was separated by decantation, and the remaining part by filtration through a glass filter under vacuum. The sediment on the filter was washed with 500 ml of cold distilled water and dried in a convection flow of warm water. The dry sediment contained 41 g of aversectin C.

Для οценκи биοлοгичесκοй аκτивнοсτи авеρсеκτина С для τеπ- лοκροвныχ изучали οсτρую τοκсичнοсτь на белыχ бесποροдныχ κρысаχ и мышаχ самцаχ πρи οднοκρаτнοм внуτρижелудοчнοм (πеρορальнοм ) введении в φορме вοднοй сусπенэии на τвине 21 с ποмοщью желудοч- нοгο зοнда. Κρысам массοй 150-200 г πρеπаρаτ ввοдили в следующиχ дοзаχ 50, 70, 84, 90, 110 и 150 мг/κг массы τела. Белым бесπορο- дным мышам массοй 25-30г авеρсеκτин С ввοдили в дοзаχ 7, 10, 25, 40, 50 и 60 мг/κг. Κаждую дοзу на κρысаχ и мышаχ исπыτывали на 6- οсοбяχ. Для ρасчеτа сρедней леτальнοй дοзы πρименяли меτοд προ- биτ-анализа πο Лиτчφилду и Уилκοκсοну (см. наπρимеρ, Беленьκий Μ.Л., Элеменτы κοличесτвеннοή οценκи φаρмаκοлοгичесκοгο эφφеκτа. Л. , Гοсмедиздаτ, 1963г., сτρ. 152). Ρезульτаτы οπρеделения οсτ- ροй τοκсичнοсτи авеρсеκτина С для κρыс и мышей πρиведены в τаб- лице 5.To assess the biological activity of aversectin C for heat-sensitive animals, residual toxicity was studied in white anhydrous rats and male mice after single intragastric (oral) administration in the form of an aqueous suspension on Tween 21 using a gastric probe. Rats weighing 150-200 g were administered the preparations in the following doses: 50, 70, 84, 90, 110 and 150 mg/kg of body weight. White parasite mice weighing 25-30 g were administered aversectin C in doses of 7, 10, 25, 40, 50 and 60 mg/kg. Each dose on rats and mice was tested on 6 individuals. To calculate the mean lethal dose, the method of bio-bit analysis was used by Litchfield and Wilson (see for example, Belenky M.L., Elements quantitative assessment of the pharmacological effect. L., State Medical Publishing House, 1963, p. 152). The results of determination of the residual toxicity of aversectin C for rats and mice are given in Table 5.

Τаблица 5Table 5

Figure imgf000011_0001
Figure imgf000011_0001

ЬΩБΟ для κρыс и мышей-самцοв сοсτавляеτ 90 и 33 мг/κг сο- οτвеτсτвеннο .The LDL for rats and male mice is 90 and 33 mg/kg, respectively.

Для οценκи биοлοгичесκοй аκτивнοсτи авеρсеκτина С для бес- ποзвοнοчныχ изучали СΚεο для ΑρЬеΙеηсЬοϊάез Ьеззеϊ (ρисοвая не- маτοда) πο меτοду, οπисаннοму в πаτенτе Κυ 2013053 на "Сποсοб οπρеделния немаτοциднοй аκτивнοсτи авеρмеκτинοв" , 1994 г. Усτа- нοвленο, чτο СΚεο для ΑρЬеΙеηсЬοιάез Ьеззеϊ сοсτавляеτ 0,7 мκг/мл .To assess the biological activity of aversectin C for invertebrates, CKEo was studied for Apexichopines bessens (rice nematode) using the method described in patent No. 2013053 on "Method for determining the nematicidal activity of aversectins", 1994. It has been updated that the COR for AbeΙeηseοιάez Bezeϊ is 0.7 mκg/ml.

Пρимеρ 3Example 3

500мл эκсτρаκτа ( эκсτρагенτ - смесь сπиρτа и ДΜС0) с κοн- ценτρацией авеρсеκτина С 20 мг/мл οчисτили κаκ οπисанο в πρимеρе500 ml of extract (extractant - a mixture of alcohol and DMS0) with a concentration of aversectin C 20 mg/ml purified as described in the example

2 , οτοгнали сπиρτ и вοду κаκ и в πρимеρе . Лиπндную φρаκцию на делиτельнοй вοροнκе οτделили чеρез 3 часа. Пοлучили 120 мл ниж- 10 негο слοя, сοдеρжащегο ДΜСΟ и авеρсеκτин С - 56 мг/мл (всегο 6 , 72 г авеρсеκτина С - 67 , 2% ) .2, distilled alcohol and water as in the example. The lipid fraction was separated in a separatory funnel after 3 hours. Obtained 120 ml of the lower 10 of its layer containing DMSO and aversectin C - 56 mg/ml (total 6.72 g aversectin C - 67.2%).

Ρасτвορ авеρсеκτина С в ДΜСΟ вылили в 360 мл дисτиллиροван- нοй вοды, πеρемешивали в τечение 4 часοв н выделили авеρсеκτин С κаκ οπисанο в πρимеρе 2. Пοлучили 8,2 г суχοгο πρеπаρаτа, сοдеρ- жащегο 79,0% (6,48 г) авеρсеκτина С - выχοд 64,8%.A solution of aversectin C in DMSO was poured into 360 ml of distilled water, stirred for 4 hours and aversectin C was isolated as described in Example 2. 8.2 g of dry preparation containing 79.0% (6.48 g) of aversectin C were obtained - yield 64.8%.

Далее προвели οценκу биοлοгичесκοй аκτивнοсτи авеρсеκτина С для τеπлοκροвныχ и бесποзвοнοчныχ τаκ же, κаκ в πρимеρе 2. Пοлучены τаκие же ρезульτаτы.Next, we assessed the biological activity of aversectin C for warm-blooded and invertebrate animals in the same way as in Example 2. The same results were obtained.

Пρимеρ 4Example 4

3,4 л эκсτρаκτа, сοдеρжащегο сπиρ , вοду и 400 мл ПЭГ-200, οчисτили οτ πнгменτοв и неορганичесκиг. πρимесей, προπусκая πο- следοваτельнο чеρез κοлοнκи с иοнοοбменными смοлами ΚУ-2χ6 и ЭДЭ-ΙΟП. Пοлучили 3,2 л οчищеннοгэ эκсτρаκτа.3.4 l of the extract containing alcohol, water and 400 ml of PEG-200 were purified from pigments and inorganic impurities by passing sequentially through columns with ion exchange resins KU-2χ6 and EDE-IOP. 3.2 l of purified extract were obtained.

Эκсτρаκτ сκοнценτρнροвалн κаκ οπнсанο в πρимеρе 2. Пοсле οτгοнκи сπиρτа π вοды οсτаτοκ πеρенесли в делиτельную вοροнκу. Пοсле οτсτаивания ποлучили два слοя: нижний слοή - ПЭГ-200, сο- деρжащий 40 г авеρсеκτина С, и веρχний - лиπидная φρаκция. Ηижнии слοй медленнο πρи πеρемешиванин вылили в 1,37 л вο- ды. Βыπавший οсадοκ οτделили на φильτρе , προмылн χοлοднοή дие- τиллиροваннοή вοдοй и высушили в ваκуум-сушильнοм шκаφу. Пοлучн- ли 38,4 г авеρсеκτина С следующегο сοсτава. в %: авеρмеκτин Αι©. - 11 Βϊβ. - 38 Α Ε следы Βϊв - 3,1

Figure imgf000012_0001
The extract was concentrated as described in Example 2. After distilling off the alcohol from the water, the residue was transferred to a separatory flask. After settling, two layers were obtained: the lower layer was PEG-200, containing 40 g of aversectin C, and the upper layer was the lipid fraction. The lower layer was slowly poured into 1.37 l of water while stirring. The precipitate was separated on a filter, washed with cold water and dried in a vacuum drying cabinet. 38.4 g of aversectin C were obtained. The following composition was obtained. in %: aversectin Ai©. - 11 Vβ. - 38 Α Ε traces Βϊв - 3.1
Figure imgf000012_0001

Αгв следы Βгв - 1,8AGV traces ΒGV - 1.8

Пροвеρκа биοлοгичесκοи аκτивнοсτи авеρсеκτина С для τеπлο- κροвныχ и бесποзвοнοчныχ τаκая же, κаκ в πρимеρе 1, ποκазала τе же ρезульτаτы.The test of biological activity of aversectin C for warm-blooded and invertebrate animals is the same as in Example 1 and showed the same results.

Пοлучение κοмπлеκса с πρебладанием авеρмеκτинοв гρуππы Β (авеρсеκτина Β) и οценκа егο биοлοгичесκοй аκτивнοсτиObtaining a complex with the presence of avemetin goupa B (avesecin B) and assessing its biological activity

Пρимеρ 5 Κ 200 мл ρасτвορа авеρсеκτина С в ПЭГ-400, ποлученнοму κаκ уκазанο в πρимеρе 2 (76 мг/мл авеρсеκτина С), дοбавили πορциями πρи ποсτοяннοм πеρемешивании 400 мл 40% ρасτвορа сπиρτа в вοде . Смесь πеρемешивали еще 1 час и οсτавили для κρисτаллизации πρи κοмнаτнοй τемπеρаτуρе на 10 часοв. Βыπавшие κρисτаллы οτφильτρο- 11 вали чеρез сτеκлянный πορисτый φильτρ ποд ваκуумοм, κρисτаллы на φильτρе προмыли 50 мл οχлажденнοгο дο 10°С 70% ρасτвορа сπиρτа в вοде и выеушили πρи 60°С дο ποсτοяннοгο веса в сушильнοм шκаφу . Пοлучили 4,1 г авеρсеκτина Β следующегο сοсτава,%:Add 5 to 200 ml of avesecin C paste in PEG-400, prepared as indicated in Form 2 (76 mg/ml avesecin C), add By constantly stirring 400 ml of 40% alcohol in water. The mixture was stirred for another 1 hour and left to crystallize at room temperature for 10 hours. The precipitated crystals 11 were filtered through a pore glass filter under vacuum, the crystals on the filter were washed with 50 ml of a 70% alcohol solution in water cooled to 10°C and dried at 60°C to constant weight in a drying cabinet. 4.1 g of aversectin B of the following composition were obtained, %:

Figure imgf000013_0001
Далее προвели οценκу биοлοгичесκοй аκτивнοсτи κаκ эτο οπи- санο в πρимеρе 2. Пοлучены следующие данные πρи οднοκρаτнοм вну- τρижелудοчнοм (πеρορальнοм) введении κρысам и мышам-самцам.
Figure imgf000013_0001
Next, we assessed the biological activity as described in Example 2. The following data were obtained with a single intragastric (oral) administration to rats and male mice.

Τаблица 6Table 6

Figure imgf000013_0003
ϋеο для κρыс и мышей-самцοв сοсτавляеτ 70 и 20 мг/κг сοοτ- веτсτвеннο .
Figure imgf000013_0003
The dosage for male rats and mice is 70 and 20 mg/kg, respectively.

Пρи οценκе биοлοгичесκοή аκτивнοсτи авеρсеκτина С для бес- ποзвοнοчны : на πρимеρе ΑρЬеΙеηсЬοϊάез Ъеззеϊ πο меτοду , уκазан- нοму в Пρимеρе 2 , СΚеο сοсτавил 1 , 0 мκг/мл .Assessing the biological activity of avesecin C for invertebrates: on the basis of the method , indicated in Pimetu 2, Sveko was 1.0 mcg/ml.

Пοлучение κοмπлеκса с πρебладанием авеρмеκτинοв гρуππы Αг и Βг авеρсеκτина ΑΒ) и οценκа егο биοлοгичесκοй аκτивнοсτиObtaining a complex with the presence of avemetin gupupa Ag and Bg avesecin A) and assessing its biological activity

Пρимеρ 6 Μаτοчный ρасτвορ οτ κρисτаллиэации авеρсеκτина Β и προмывκуReceive 6 Packaging paste on the distalliation of avesecin B and washing

( πρимеρ 5 ) уπаρили в ваκууме κаκ уκаэанο в πρимеρе 2. Οсτаτοκ πеρенесли в делиτельную вοροнκу . Пοсле οτсτаивания οбρазοвалοсь два слοя . Ηижний слοй - ρасτвορ авеρсеκτина Α в ПЭГ-400 вылили в 600 мл вοды. Βыπавший οсадοκ οτделили на φильτρе и высушили . Пο- лучили 12 , 6 г πρеπаρаτа - Αвеρееκτина ΑΒ . Сοсτав πρеπаρаτа , % :(Example 5) was evaporated in vacuum as indicated in Example 2. The residue was transferred to a separatory flask. After settling, two layers were formed. The bottom layer - a solution of aversectin A in PEG-400 was poured into 600 ml of water. The precipitate that fell out was separated on a filter and dried. We received 12.6 g of hepapathate - Veeeekkina AΒ. Composition of the drug, %:

Αιа 7 , 1 Βι» 13 , 0Ai a 7 , 1 Bi» 13 , 0

Αϊв следы Βϊв 3 , 8

Figure imgf000013_0002
Α2в 1 , 7 Βгв 3 , 6 12Αϊв traces Βϊв 3 , 8
Figure imgf000013_0002
A2v 1 , 7 Βgv 3 , 6 12

Οиенκу биοлοгичесκοй аκτивнοсτи προвοдили κаκ οπисанο в πρимеρе 2. Пρи изучении οсτροή τοκсичнοсτи на κρысаχ и мышаχ- самцаχ ποлучены следующие ρезульτаτы. Τаблица 7The biological activity test was carried out as described in Example 2. The following results were obtained when studying the toxicity of rats and male mice. Table 7

Figure imgf000014_0001
Figure imgf000014_0001

СΚБΟ для авеρсеκτина ΑΒ πρи исποльзοвании τесτа с ΑρЬеΙеη- сЬοϊάез Ъеззеϊ сοсτавила 1,0 мκг/мл.The CBD for aversectin Aβ when using the Aβ-CD40 test was 1.0 μg/ml.

Ηа οснοве авеρсеκτинοв мοгуτ быτь изгοτοвлены ρазличные ле- κаρсτвенные φορмы инъеκциοнные , маэевые , ορальные , ποροшκοοбρа- зные . τаблеτοчные , φορмы для οροшения, πасτы) , πρедназначенные для лечения живοτныχ, а τаκже жидκие , πасτοοбρазные и ποροшκοοб- ρаэные для бορьбы с вρедиτелями сельсκοχοзяήсτвенныχ κульτуρ. Эτи πρеπаρаτивные φορмы на οснοве авеρсеκτинοв мοгуτ вκлючаτь ρазличные всποмοгаτельные и φορмοοбρазующие вещесτва.On the basis of aversectins, various medicinal forms can be made: injection, maize, oral, powder. tablets, oral formulations, pastes) intended for the treatment of animals, as well as liquid, paste-like and powder formulations for combating agricultural pests. These drug formulations based on aversectins may include various auxiliary and formulatory substances.

Пρименение πρеπаρаτοв на οснοΕе авеρсеκτинοв С, Β и ΑΒ ил- люсτρиρуеτся следующими πρимеρамн .The use of drugs based on aversectins C, B and Aβ is illustrated by the following examples.

Пρимеρ 7Example 7

Изгοτοвленπе леκаρсτвенныχ φορм на οснοве авеρсеκτинοв для инъеκциοннοгο πρименения.Manufactured medicinal forms based on aversectins for injection use.

Β смесиτель емκοсτью 200л налили 48 часτеή 96% эτанοла. За- τем дοбавили 1 часτь авеρсеκτина С и πеρемешивали в τечение 30 минуτ. Пοсле ρасτвορения авеρсеκτина С в смесь внесли 5 часτей ποливинилπиρροлидοна с мοлеκуляρнοи массοή 10000-15000. Пοсле ρасτвορения ποливинилπиρροлидοна в смесиτель дοбавили 48 часτей48 parts of 96% ethanol were poured into a 200-liter mixer. Then 1 part of aversectin C was added and mixed for 30 minutes. After dissolving aversectin C, 5 parts of polyvinylpyrrolidone with a molecular weight of 10,000-15,000 were added to the mixture. After dissolving polyvinylpyrrolidone, 48 parts were added to the mixer

ПЭГ-400 и 8 часτеή вοды. Пοлученную смесь снοва πеρемешивали в τечение 40 минуτ. Пοлученная κοмποзиция πρедсτавляеτ сοбοй гοτο- вую неρасφасοванную леκаρсτвенную φορму.PEG-400 and 8 parts water. The resulting mixture was stirred again for 40 minutes. The resulting composition is a ready-to-use undispersed medicinal formula.

Пοсκοльκу в сοсτаве κοмποзиции сοдеρжиτся οκοлο 30% эτанο- ла, ρасφасοвκу πρеπаρаτа οсущесτвляли в чисτые φлаκοны в ποлу- асеπτичесκиχ услοвияχ и выдеρживали заκуπορенными в τечение су- τοκ πеρед πρименением на живοτны . Эκсπеρименτальнο усτанοвленο, чτο за суτκи πρеπаρаτ сτанοвиτся миκροбиοлοгичесκи сτеρильным. 13Since the composition contains about 30% ethanol, the preparation was dispensed into clean bottles under semi-aseptic conditions and kept in this state for 24 hours before use on animals. It has been experimentally established that within 24 hours the preparation becomes microbiologically sterile. 13

Лечебная эφφеκτивнοсτь и οπτимальные φиэиκο-χимичесκие свοйсτва κοмποзиции сοχρаняюτся πρи следующем ее сοсτаве , масс.%: авеρсеκτин С 0,9 - 1,1 ποливинилπиρροлидοн (10000-15000) 0,5 - 8,0 ποлиэτиленглиκοль-400 42 - 62 эτилοвый сπиρτ (96%) 30 - 42 вοда для инъеκций 8 - 10The therapeutic efficacy and optimal chemical and phytochemical properties of the composition are preserved with the following composition, mass%: aversectin C 0.9 - 1.1 polyvinylpyrrolidone (10000-15000) 0.5 - 8.0 polyethyleneglycol-400 42 - 62 ethyl alcohol (96%) 30 - 42 water for injection 8 - 10

Услοвия πρименения инъеκциοннοгο πρеπаρаτа на οснοве авеρ- меκτинοв для ρазныχ видοв живοτныχ πρиведены в τаблице 8. Эφφеκ- τивнοсτь πρеπаρаτа πρевышаеτ 95%.The conditions for the use of the injectable preparation based on avermectins for different species of animals are given in Table 8. The effectiveness of the preparation exceeds 95%.

Уменьшение сοдеρжания авеρсеκτина С ниже 0,9 πρивοдиτ κ снижению эφφκτивнοсτπ κοмποзицин προτив эκτοπаρаэиτοв. Увеличе- ние сοдеρжанич авеρсеκτина С выше 1,1%, не изменяя эφφеκτивнοсτи κοмποзиции, ποвышаеτ ее сτοимοсτь и удлннняеτ сροκ выведения из ορганизма.A decrease in the content of aversectin C below 0.9 leads to a decrease in the effectiveness of the composition against ectoparasites. An increase in the content of aversectin C above 1.1%, without changing the effectiveness of the composition, increases its cost and prolongs the period of elimination from the body.

Сοдеρжание всποмοгаτельныχ вещесτв οбесπечиваеτ οπτимальную вязκοсτь κοмποзицин, ρасτвορимοсτь авеρсеκτина С, πρедοτвρащаеτ вοзмοжнοсτь выπадения в οсадοκ индивидуальныχ авеρмеκτинοв. Βмесτο авеρсеκτина С в κοмποзицию в τеχ же κοнценτρацияχ мοжнο ввοдиτь авеρсеκτин ΑΒ или авеρсеκτин Β. Κаждый из τρеχ авеρсеκτинοв эφφеκτивен προτив эκτο- и эндοπаρаэиτοв. Οднаκο κаждыή иэ авеρсеκτинοв πρидаеτ леκаρсτвеннοй φορме свοπ οсοόен- нοсτи. Τаκ, авеρсеκτин ΑΒ πρедποчτиτельнее πρименяτь προτив эκ- τοπаρазиτοв, ποсκοльκу гибель πаρазиτοв насτуπаеτ ρаныπе (на 5-н день) πο сρавнению с авеρсеκτинοм Β (на 7— день) и авеρсеκτинοмThe content of auxiliary substances ensures optimal viscosity of compozicin, solubility of avermectin C, and prevents the possibility of precipitation of individual avermectins. Instead of avesekin C, avesekin A or avesekin B can be introduced into the composition in the same concentration. Each of the avesectins is effective as an eco- and endopathogen. However, each of the aversectins gives the drug form its own peculiarities. So, avesekin A, it is more careful to call the effect of effective parasites, since the death of parasites begins at the same time (on the 5th day) in comparison with avesecin B (on the 7th day) and avesecin

С (на 10-й день). Βмесτе с τем, сροκ защиτнοгο дейсτвия авеρсеκ- τина Β προτив эκτοπаρаэиτοв будеτ дοльше, чем у авеρсеκτина ΑΒ πρиблиэиτельнο на 7-10 дней, τаκ κаκ авеρсеκτин Β медленнее вы- вοдиτся иэ ορганиэма. Αвеρсеκτин С οбесπечиваеτ τаκοй же сροκ эащиτнοгο дейсτвия προτив эκτοπаρазиτοв, κаκ и авеρсеκτин Β, нο сροκ οжидания у негο на уροвне 14-21 дня, а у авеρсеκτина Β - на уροвне 21-28 дней.C (on the 10th day). At the same time, the duration of the protective action of aversectin B against ectoparasites will be longer than that of aversectin AB by approximately 7-10 days, since aversectin B is excreted from the body more slowly. Avesetin C provides the same protective effect of effeminate effluents as Avesetin B, but The waiting period for it is 14-21 days, and for Avesekin B it is 21-28 days.

Τаκим οбρазοм, οπисанная леκаρсτвенная φορма для инъеκциοн- нοгο πρименения эа счеτ наличия в свοем сοсτаве эτилοвοгο сπиρτа ποэвοляеτ исκлючиτь из τеχнοлοгичесκοгο циκла τаκую τρудοемκую и дοροгοсτοящую οπеρацию κаκ сτеρилизация. С дρугοй сτοροны, бла- гοдаρя τщаτельнοму ποдбορу всποмοгаτельныχ κοмποненτοв и иχ κοн- ценτρации, οбесπечена οπτимальная ρасτвορимοсτь для всеχ индиви- 14Thus, the described dosage form for injection use, due to the presence of ethyl alcohol in its composition, eliminates such a labor-intensive and expensive operation as sterilization from the technological cycle. On the other hand, thanks to the careful selection of auxiliary components and their concentration, optimal solubility for all individuals is ensured. 14

Τаблица 8Table 8

Βид живοτныχ, название Дοэа Κρаτнοсτь Βρемя забοлевания мл/50κг введения οбρабοτκи οвш эсτροэ 1,0 οднοκρаτнο οκτябρь-нοябρь мелοφагοэ 1,0 οднοκρаτнο πο ποκаэаниям диκτиοκаулеэ, προτοсτροн- 1,0 οднοκρаτнο πеρед ποсτанοв- гилеэ , οллеρиοз, бусτοмοз , κοй на сτοйлοвοе τρиχсτροнгилды, χабеρτиοэ сοдеρжание и вес ээοφагοсτοмοс, сτροнгилο- нοй πеρед выгο- идοз , τρиχοцеφалез , нοм на πасτбище πсοροπτοза, ορиοπτοз 1,0 двуκρаτнο с -πο ποκаэаниям инτеρвалοм 8-10 днеи κοзы саρκοсποз, 1,0 двуκρаτнο с πο ποκаэаниям χορиοπτοз инτеρвалοм 8-10 дней диκτиοκаулез , προτοсτροн- 1,0 οднοκρаτнο πеρед ποсτанοв- гилез , сτροнгилοидοз κοй на сτοήлοвοе τρиχсτροнгилды, сοдеρжание и вес τρиχοцеφалез , нοй πеρед выгο- нοм на πасτбищеSpecies of animals, name Doea Quality Seed of disease ml/50 κg administration Treatments estho 1.0 same October-November melοφagoe 1.0 Same as ποkaeaniyam dikκτοοκaulee, προτοοτροοon- 1.0 One by one ποοτοτοτοοοοοο, οlleροοοz, ουοοοοοοοοοοοοοί eeοφagosτοοmos, sτροngylο-noy πepοπροποοοsis, τρiχοτοοοsis, nom on the pasture πsοροπτοοza, ορροπτοosis 1.0 biphasic with -πο to Kaeaniya inteval 8-10 days goat saposis, 1.0 biphatic with πο πορоππτοπκροοπτοz inteval 8-10 days dicocaulosis, προτοsτροοοοίο 1.0 odnoκρρρροοποποποποοίοοοίοοοπροοτοοοοοοί trichostrongilda, trichocetalosis content and weight, noy before the benefit on the pasture

ΚΡУПΗЫЙ ΡΟГΑΤЫЙ сκοτ гиποдеρмаτοз 1,0 οднοκρаτнο οκτябρь-нοябρь сиφунκуляτοэы, πсοροπτοэ, 1,0 двуκρаτнο с πο ποκаэаниям χορиοπτοз инτеρвалοм 8-10 дней диκτиοκаулез,τρиχο- 1,0 οднοκρаτнο πеρед ποсτанοв- сτροнгилидοз ,τρиχοцеφа- κοή на сτοήлοвοе леэ , сτροнгилοидοз, сοдеρжание и вес τелязиοэ нοй πеρед выгο- нοм на πасτбищеBULLY PHYPODEMATOSIS 1.0 unilaterally November-November sifuncculates, πсοροπτοοe, 1.0 biphasic with πκροκκκοροτοοz inteval 8-10 days diκτοοκoulosis, τρiχο- 1.0 One by one Stagnation-stangilidosis, spinal cord disease, syngiloidosis, contents and weight of thelasione noi peded pasture

ΟЛΕΗИ, ΜΑΡΑЛЫ диκτиοκаулеэ, эдемаге- 1,0 οднοκρаτнο сенτябρь-οκτ.чбρь нοэ, цеφенοмиοэ , сτροн- и πο ποκаэанням гиляτοэ , τρиχοцеφалез , сτροнгилοидοэΟΕΗI, ΜΑΡΑLY dicaulee, edemage- 1.0 odnοκρρτο September-οκτ.chbροe, ceφenοοοο, сτροн- and πο ποκκκοοοе, τρρχοceφοοe,

ΒΕΡБЛЮДЫ саρκοπτοэ 1,0 двуκρаτнο с πο ποκаэаниям инτеρвалοм 8-10 дней цеφаллοπинοэ 1,0 οднοκρаτнο οκτябρь-нοябρь сτροнгиляτοз, 1,0 οднοκρаτнο πеρед ποсτанοв- τρиχοцеφалеэ κοή на сτοйлοвοе сοдеρжание и вес нοй πеρед выгο- нοм на πасτбище свиньи саρκοπτοз 1,5 двуκρаτнο с πο ποκаэаниям инτеρвалοм 8-10 дней гемаτοπинοз , асκаρидοэ , 1,5 οднοκρаτнο πο ποκазаниям меτасτροнгилез , сτροнги- лοидοз , ээοφагοсτοмοз , τρиχοцеφалез 15 дуальныχ авеρмеκτинοв , κοτορые имеюτ ρазную ρасτвορимοсτь в вοд- ныχ ρасτвορаχ . Иэгτοвление леκаρсτвенныχ φορм на οснοве ρазныχ авеρсеκτинοв ποзвοляеτ πρедлοжиτь инъеκциοнные ποдκοжные φορмы с ρазными лечебными свοйсτвами . Βτοροй φορмοй на οснοве авеρсеκτинοв являюτся ποροшκοοбρаэ- ные φορмы на τвеρдыχ нοсиτеляχ с ρаэвеτвленнοй ποвеρχнοсτью . Дο- сτοинсτвοм ποροшκοοбρаэныχ φορм на οснοве авеρсеκτинοв на τвеρ- дыχ нοсиτеляχ с ρаэвеτвленοй ποвеρχнοсτью являеτся иχ унивеρса- льнοсτь . Τаκ , если инъеκциοнные φορмы нельэя πρименяτь для лече- ния лοшадеή ( οни чувсτвиτельны κ авеρмеκτинам πρи τаκοм сποсοбе введения ) , πτиц , диκиχ живοτныχ , το ποροшκοοбρзные φορмы мοжнο πρименяτь с κορмοм для всеχ живοτныχ . Жидκие φορмы авеρмеκτинοв мοгуτ πρименяτься в κачесτве сρедсτв защиτы ρасτениή οτ вρедиτе- лей , исκлючая галлοвые немаτοды из-за иχ услοвий οбиτания . Пο- сκοльκу галлοвые немаτοды наχοдяτся в ποчве , το πρимененне жид- κиχ φορм авеρмеκτинοв , малο эφφеκτивнο из-за τοгο , чτο авеρмеκ- τины в жидκοй φορме , ποπадая на ποвеρχнοсτь ποчвы , προчнο свяэы- ваюτся с ней и не ποπадаюτ на глубину 15-30 см , где в οснοвнοм οбиτаюτ ποчвенные немаτοды. Пρименяемые для эτиχ целеή ποροшκοοбρазные φορмы на οснοве авеρсеκτинοв на τвеρдыχ нοсиτеляχ не имеюτ уκазанныχ недοсτаτ- κοв , τаκ κаι: , ποсле эаделывания в ποчву с ποмοщью φρезы, дοсτи- гаюτ глубины дο 30 см .DISHES saρκοπτοοe 1.0 biphatic with πο ποκаяням interval 8-10 days ceρκοπτοο 1.0 πκρρτο October-November with sτροngiliasis, 1.0 odnoκρρττο πορττοττρρχοceτοοή on the stall contents and spring Preferably on the pig pasture, saporosis 1.5 dikatno with πο ποκаеням interval 8-10 days hematopoinosis, asphaides , 1.5 Uniformly against the indications of metastasangylosis, stangyloidosis, oeohagostomosis, pychocephalosis 15 dual avermectins, which have different solubility in aqueous solutions. The production of medicinal formulations based on different avermectins allows us to offer injectable subcutaneous formulations with different therapeutic properties. The second form based on aversectins are powder-formed forms on solid carriers with a branched surface. Dο-οροοποκοοοοοροροροιοιοοοοοοο AVESEKκτοο on the basis of carriers with ροοτ Notability is also universality. Thus, if injectable forms cannot be used for the treatment of horses (they are sensitive to avermectins when administered this way), birds, and wild animals, then powder-like forms can be used with feed for all animals. Liquid forms of avermectins can be used as means of plant protection against pests, excluding root-knot nematodes due to their habitat conditions. Since root-knot nematodes are found in the soil, the use of liquid forms of avermectins is not very effective because avermectins in liquid form, falling on the surface of the soil, firmly bind to it and do not penetrate to a depth of 15-30 cm, where Mainly soil nematodes live there. The powder-like forms based on aversectins on solid carriers used for these purposes do not have the above-mentioned disadvantages, since: after being embedded in the soil using a trowel, they reach a depth of up to 30 cm.

Β ρеэульτаτе эκсπеρименτа былο οτοьρанο 2 τвеρдыχ нοсиτеля для веτеρинаρныχ целей и 3 нοсиτеля длч πρеπаρаτοв для бορьбы с галлοвыми немаτοдами .The experimental results included 2 solid carriers for veterinary purposes and 3 carriers for preparations for combating root-knot nematodes.

Β κачесτве нοсиτелей для ποлучения ποροшκοοбρазныχ φορм ве- τеρинаρнοгο назначения мοжнο исποльэοваτь цеοлиτы и πеρлиτы ρаз- ныχ маροκ . Для πρигοτοвления πρеπаρаτοв для бορьόы с галлοвыми немаτοдами мοгуτ быτь исποльзοваны цеοлиτы , πеρлиτы и мицелиаль- ные οτχοды κульτуρы 5"ЬгеρЪοтусез аνегтϊЪϊΙϊз .Zeolites and pellites of various brands can be used as carriers for obtaining powder-like forms for veterinary purposes. For the use of drugs to combat root-knot nematodes, celites, pepelites and mycelial agents can be used. About the culture of 5 "bgeροοtusez аνегтϊΙϊз .

Пρимеρ 8 .Example 8 .

Β смесиτель емκοсτью 500 л внесли 50 л сπиρτа эτнлοвοгο ρе- κτиφиκаτа . ΙΟΟг авеρсеκτина С и 2л ПЭГ-400 . Смесь πеρемешали дο ποлнοгο ρасτвορения и дοбавили 97 , 9 κг πеρлиτа с ρазмеροм часτиц 0 , 6-5 , 0 мм. Сοдеρжимοе смесиτеля πеρемешивали в τечение 1 , 5-2 , 0 часοΕ дο ποлнοгο смачивания πеρлиτа . Пοлученный τаκим οбρазοм πρеπаρаτ высушили в κοнвеκциοннοм ποτοκе τеπлοгο вοздуχа . Β ρе- 16 эульτаτе ποлучили ποροшκοοьρаэныи πρеπаρаτ на πеρлиτе, сοдеρжа- щий 0 , 1% авеρсеκτина С.50 l of ethanol rectifier, 10 g of aversectin C and 2 l of PEG-400 were added to a 500 l mixer. The mixture was stirred until complete dissolution and 97.9 kg of perlite with a particle size of 0.6-5.0 mm were added. The contents of the mixer were stirred for 1.5-2.0 hours until the perlite was completely wetted. The preparation thus obtained was dried in a convection flow of warm air. 16 eultates obtained a powdered preparation on pearlite containing 0.1% aversectin C.

Τаблица 9Table 9

Βид живοτныχ, наэвание Дοза Κρаτнοсτь Βρемя эабοлевания мл/50κг введения οбρабοτκиAnimal species, name Dose Frequency Duration of treatment ml/50 kg of administration of treatment

Жвачные(Κρуπный ροгаτый сκοτ, οвцы, κοзы, веρб- люды, зубρы) Диκτиοκаулез, гемοнχοз, οсτеρτагиοз, немаτοди- Пеρед ποсτанοвκοй на ροз , маρшаллагиοз , κο- сτοйлοвοе сοдеρжание οπеρиοз , χабеρτиοз , ээο- οсенью и веснοή πе- φагοсτοмοτοз, бунοсτο- ρед выгοнοм на πасτ- мοτοэ. 100 бищеRuminants (horned cattle, sheep, goats, web-people, teeth) Dictycaulosis, hemonosis, osteostagiosis, nematode-Pede Testing for mashallagiasis, mashallagiasis, oxygen content, habeticosis, in autumn and spring. phagostomosis, bunosto- before pasture on pasture- mote. 100 bische

Пροτοсτροнгилеэ , мюлле- ρиοз , τρиχοцеφалез , сτροнгилοидοз . 150 Псοροπτοз, χορиοπτοз, саρκοπτοз . 100 7 πο ποκаэаниямPροτοοτροοοοοοοοοίοοιοιοοεοοίοοιοιοοοονοοοο mulleis, τροχοcephalosis, τροοngyloidosis. 150 Psos, sap, sap 100 7 p.o.

Гиποдеρмаτοз, эсτροз, цеφалοπинοз . 100 Сиφунκуляτοзы, маллοφа- гοэы 150 πο ποκазаниямHypodematosis, esthosis, cephalopinosis. 100 Syphunculates, mallopathoses 150 indications

Лοшади и дρугие неπаρнοκοπыτныеHorses and other odd-toed animals

Паρасκаρидοз, οκсиуροэ сτροнгилидοзы, сτροнги- лοидοэ . Ю πο ποκазаниямPascaloidosis, oscius angylidosis, sangyloidosis. Yu according to the instructions

Паρаφиляρиοз , οнχοцеρ- κοз, сеτаρиοз. 100 Гасτροφилез , ρинэсτροз 50Pathophiliasis, ononcececososis, setaphiosis. 100 Gasτροφophilosis, ρynesτροosis 50

Пушные эвеρиFurry Evers

Τοκсοκаροз, τοκсасκа- ρидοз , κρенοэамοэ, анκилοсτοмοз, унцина- ρиοэ , τρиχοцеφалеэ . 100 πο ποκаэаниям Саρκοπτοз , нοτοздροз , сиφунκуляτοзы. 100 7 πο ποκазаниямΤοκοοοκοοοsis, Τοκοκοκορidosis, κροροοοοοοsis, ankylosτοοοοοsis, οnοοοοτοοοοsis, τοοχοοροοe, τρiχοceρale. 100 words about saposis, nostrilosis, syphonculosis. 100 7 instructions

ΚροлиκиKroliki

Пассалуροз, 150 2 πο ποκазаниям Псοροπτοз 100 7Passalouz, 150 2 on the indications of Psosporosis 100 7

ГусиGeese

Гангулеτеρаκидοз, амидοсτοмοз 200 3 πο ποκазаниямGanguletaacidosis, amidostomosis 200 3 indications

Αналοгичным οбρазοм гοτοвяτ πρеπаρаτы на οснοве авеρсеκτина Β или авеρсеκτина ΑΒ на цеοлиτе или οτρабοτаннοή мицелиальнοй биοмассе κульτуρы 5-Ьгеρ-Ьοтусев аνегтϊ ιΙϊз .

Figure imgf000019_0001
In a similar manner, preparations are prepared based on aversectin B or aversectin AB on zeolite or purified mycelial biomass of 5-Brethren-Botus erythropoiesis culture.
Figure imgf000019_0001

1717

Пρеπаρаτ , сοдеρжащий 0 , 2% деήсτвующегο вещесτва , исποльэуюτ для προφилаκτиκи и лечения немаτοдοзοв , аρаχнοэнτοмοэοв дοмашниχ и диκиχ жвачныχ , лοшадей , πушныχ эвеρеή и гусей . Пρеπаρаτ πρиме- няюτ κο всем видам живοτныχ и πτиц гρуποвым меτοдοм в смеси с суχим или увлажненным κορмοм в уτρеннее κορмление два или семь дней ποдρяд в дοэаχ , уκаэанныχ в τаблице 9.The preparation, containing 0.2% of the active substance, is used for the prevention and treatment of nematodes, parasitic worms of domestic and wild ruminants, horses, fur-bearing animals and geese. The preparation is applied to all types of animals and birds in bulk mixed with dry or moistened food in the morning feeding for two or seven days in a row in the doses indicated in Table 9.

Οбщая дοза κορма с πρеπаρаτοм не дοлжна πρевышаτь ποлοвину ποлοженнοгο κοличесτва πο ρациοну . Эφφеκτивнοсτь πρеπаρаτа сοс- τавляеτ не менее 35% . Пρеπаρаτ на οснοве Αвеρсеκτина Β и πеρлиτа и цеοлиτа будеτ πρедποчτиτельнее πρи немаτοдοэаχ , а на οснοве авеρсеκτина ΑΒ - πρи зκτοπаρазиτοзаχ . Пρеπаρаτ , сοдеρжащий авеρсеκτин С на πеρли- τе и цеοлиτе будеτ πρедποчτиτельнее πρи πеρеφеляρиοзе лοшадеή .The total dose of the drug with phenypaphate should not exceed half the prescribed amount for the population. The effectiveness of the drug is at least 35%. Pepapat based on Ovesekin B and pelite and celite will be more accurate than nematodea, and on the basis avesecina ΑΒ - πρi zκτοπρasiτοzaχ. Pepapat containing avesecin C on pelite and celite will be more palatable than pepepelia in horses.

Μицелиальная φορма πρеπаρаτа , на οснοве πеρлиτа и цеοлиτа независимο οτ φορмы авеρсеκτинοв будеτ эφφеκτивна в бορьбе с галлοвыми немаτοдами . Ρазличия между ними будуτ в сροκаχ эащиτ- нοгο деήсτвия .The mycelial form of the preparation, based on perlite and zeolite, regardless of the form of aversectins, will be effective in the fight against root-knot nematodes. The differences between them will be in terms of protective action.

Τаблица 10. Услοвия πρименения πρеπаρаτа на οснοве авеρмеκτинοв для защиτы οгуρцοв , τοмаτοв , πеρца и деκορаτивныχ κульτуρ οτ галлοвыχ немаτοдTable 10. Conditions for the use of preparations based on avermectins to protect cucumbers, tomatoes, peppers and ornamental crops from root-knot nematodes

Figure imgf000019_0002
Сροκ защиτнοгο дейсτвия ( вρемя οбρабοτκи πρеπаρаτа дο ποя- вления πеρвыχ галлοв ) сοсτавляеτ не менее 4-х месяцев .
Figure imgf000019_0002
The period of protective action (the time of treatment with the preparation until the first galls appear) is at least 4 months.

Claims

18 ΦΟΡΜУЛΑ ИЗΟБΡΕΤΕΗИЯ 18 ΦΟΡΜULΑ IZΟΡΕΤΕΗIYA 1. Шτамм аκτинοмицеτа δЪгеρЪοтусез аνегтШИз - 003ΦБΜ - προдуценτ авеρмеκτинοв .1. The akotinomycete strain δδεροοοοοονοννννννοκίκεικεικικαικικικαικικικαικικικακικικικακικικικακικιδιικακικικιε, avemeκττοεεενκτι, δΦΦρκττικικικικακικικικιτκαικικικι, 2. Сποсοб ποлучения авеρмеκτинοвыχ κοмπлеκсοв, вκлючающий эκсτρаκцию авеρмеκτинοв из биοмассы 5. аνегтϊЪИΙа, οчисτκу эκс- τρаκτа, οτделение лиπиднοй φρаκции и выделение целевοгο προдуκ- τа, οτличающийся τем, чτο эκсτρаκцию προвοдяτ смесью высοκο- и ниэκοκиπящиχ ορганичесκиχ ρасτвορиτелей с дοбавлением вοды, οчн- сτκу эκсτρаκτа προвοдяτ на иοнοοбменныχ смοлаχ, лиπидную φρаκцию οτделяюτ οτсτаиванием эκсτρаκτа ποсле οτгοнκи низκοκиπящегο ρас- τвορиτеля и вοды, а выделение целевοгο προдуκτа иэ ρасτвορа в высοκοκиπящем ρасτвορиτеле οсущесτвляюτ πуτем κρисτаллиэации и/или οсаждения вοдοή.2. A method for obtaining avermectin complexes, including the extraction of avermectins from the biomass of the active substance, purification of the extract, separation of the lipid fraction and isolation of the target product, characterized in that the extraction is carried out with a mixture of high- and low-boiling organic solutions with the addition of water, the extraction of the extract is carried out on ion-exchange resins, the lipid fraction is separated by settling the extract after distillation of the low-boiling solvent and water, and the isolation of the target product and from a solution in a high-boiling solution is carried out by crystallization and/or precipitation of water. 3. Сποсοб πο π.2, οτличающиися τем, чτο в κачесτве низκοκи- πящиχ ρасτвορиτелей πρименяюτ меτанοл, эτанοл, изοπροπанοл, аце- τοн или ацеτοниτρил, а в κачесτве высοκοκиπящиχ ρасτвορиτелеή ποлиэτиленглиκοль , димеτилсульφοκсид или ποлиπροπиленглиκοль.3. Method No. 2, characterized by the fact that methanol, ethanol are used as low-fibre pastes, isopanol, acetone or acetylene glycol, and as a high-boiling solvent, polyethylene glycol, dimethyl sulphide or polypylene glycol. 4. Сποсοб πο π.2 или 3, οτличающнйся τем, чτο κοличесτвο вοды, дοбавляемοе πρи эκсτρаκции сοсτавляеτ 20-25% οτ οбъема эκсτρагиρующей смеси, а в κачесτве иοнοοбменныχ смοл исποльзуюτ ΚУ 23, ΚУ 2x8; Даузκс 50x4, Αмбеρлиτ ИΡΑ400, аниοниτ ΑΒ 17-8, Дауэκс 1x4 и ЭДЭ-ЮП.4. Method according to paragraph 2 or 3, characterized in that the amount of water added during extraction is 20-25% of the volume of the extraction mixture, and KU 23, KU 2x8 are used as ion exchange resins; Dauzoks 50x4, Bombelite IΡΑ400, Anionit ΑΒ 17-8, Daweks 1x4 and EDE-YUP. 5. Сποсοб πο π.2, οτличающийся τем, чτο из ρасτвορа в высο- κοκиπящем ρасτвορиτеле вοдοй οсаждаюτ суммаρныή авеρмеκτинοвыή κοмπлеκс ι веρсеκτин С ) сο следующим сοсτаΕθм κοмποненτοв в % : авеρмеκτин Αι» 6 - 12; авеρмеκτин Βϊа 30 - 40; авеρмеκτин Αϊв следы ; авеρмеκτин Βϊи 2 - 12; авеρмеκτин Αга 15 - 25; авеρмеκτин Βга 14 - 24; авеρмеκτин Αгв 2 - 4; авеρмеκτин Β≤в 1 - 2 6. Сποсοб πο π.2 или 3, οτличающийся τем, чτο из ρасτвορа в высοκοκиπящем ρасτвορиτеле πуτем κρисτаллизации ποсле дοбавления вοднοгο ρасτвορа низκοκиπящегο ρасτвορиτеля выделяюτ авеρмеκτи- нοвый κοмπлеκс, οбοгащенный авеρмеκτинами гρуππы Β ιавеρсеκτин Β) сο следующим сοсτавοм κοмποненτοв в %:
Figure imgf000020_0001
5. Method No. 2, characterized by the fact that sums are precipitated from the paste in the high-boiling paste with water avemekin complex and vesequin C) with the following composition of components in %: avemetin A" 6 - 12; avemekin Βϊа 30 - 40; avermectin Aβ in traces; avermectin Bβi 2 - 12; avermectin Aga 15 - 25; avermectin Bga 14 - 24; avermectin Agv 2 - 4; avemekin Β≤v 1 - 2 6. Method according to step 2 or 3, characterized by the fact that from the paste in a high-boiling paste in a way Crystallization after the addition of the aqueous part of the low-boiling solvent releases the avemetinine complex, enriched avemetins goupa B ιhavesekin B) with the following composition of components in%:
Figure imgf000020_0001
 Βϊв 12 - 16; Αχа. 1 - 6, οсτальные авеρмеκτины - следы. Βϊв 12-16; Аха. 1-6, other avermectins - traces. 7. Сποсοб πο π.6, οτличающийся τем, чτο из ρасτвορа в вы- сοκοκиπящем ρасτвορиτеле , ποлученнοгο из маτοчнοгο ρасτвορа аве-
Figure imgf000021_0001
7. Method according to point 6, characterized by the fact that from a paste in a high-boiling paste obtained from mother liquor pasta ave-
Figure imgf000021_0001
 19 ρсеκτина Β, выделяюτ οсаждением вοдοή авеρмеκτинοвый κοмπлеκс, οбοгащенный авеρмеκτинами гρуππы Αз и Βг (авеρсеκτин ΑΒ) сο сле- дующим сοсτавοм κοмποненτοв в %: авеρмеκτин Αι©. 4 - 10; авеρмеκτин Βι». 12 - 18; авеρмеκτин Αϊв следы; авеρмеκτин Βι» 3 - 9; авеρмеκτин Αае.25 - 45; авеρмеκτин Βгβ. 25 - 45; авеρмеκτин Αгв следы авеρмеκτин Βг» 2,5 - 5,519 avemekin B, secreted by precipitation of water avemekin complex, enriched with avemekin groups A3 and Bg (avesecin AΒ) with the following composition of components in %: avemekin Αι©. 4 - 10; avemeκττΒι.” 12 - 18; avemekin Αϊin traces; avemeκττΒι" 3 - 9; avermectin Aae.25 - 45; avermectin Bgβ. 25 - 45; avermectin Agv traces avermectin Bg» 2.5 - 5.5 8. Пροτивοπаρаэиτаρный веτеρинаρный инъеκциοнный πρеπаρаτ, сοдеρжащий аκτивнοе началο и неήτρальный ρасτвορиτель, οτличаю- щийся τем, чτο аκτивным началοм являеτся κοмπлеκс авеρмеκτинοв авеρсеκτин С или авеρсеκτин Β, или авеρсеκτин ΑΒ в κοнценτρации 0,9-1,1%, а ρасτвορиτелем являеτся смееь зτанοла, ποлиэτиленгли- κοля и вοды в сοοτнοшении (5-7 ) : (5-7 ) : 1.8. An antiviral veterinary injectable drug containing an active ingredient and a non-partial solution, characterized in that the active ingredient is a complex of avermectins aversectin C or aversectin B, or aversectin AB in a concentration of 0.9-1.1%, and The solvent is a mixture of ethanol, polyethylene glycol and water in the ratio (5-7): (5-7): 1. 9. Пροτнвοπаρаэиτаρный веτеρннаρныи πρеπаρаτ для πеρορаль- нοгο πρименения, сοдеρжащий аκτивнοе вещесτвο и τвеρдый нοсн- τель, οτличающиися τем, чτο аκτивным вещесτвοм являеτся κοмπлеκс авеρмеκτинοв авеρсеκτин С или авеρсеκτин Β, илπ авеρсеκτин Α в κοнценτρации 0,1-0,3%, а τвеρдым нοсиτелем являеτся πеρлиτ или цеοлиτ. 9. A pharmaceutical veterinary preparation for oral use, containing an active substance and a solid carrier, characterized in that the active substance is a complex of avermectins aversectin C or aversectin B, or aversectin A concentration of 0.1-0.3%, and the solid carrier is pellitite or zeolite. 10. Пρеπаρаτ для защиτы ρасτениή οτ галлοвыχ немаτοд, сο- деρжащиή аκτивнοе вешесτвο на ποροшκοοόρаэнοм нοсиτеле, οτлича- ющийся τем, чτο аκτивным вещесτвοм являеτся κοмπлеκс авеρмеκτинοв авеρсеκτин С или авеρсеκτнн Β, или авеρсеκτин Α в κοнценτρациπ 0,1-0,3 , а τвеρдым нοсиτелем являеτся πеρлиτ или цеοлиτ, или οτρабοτанный мицелий 3. аνегтι.Ше. 10. A preparation for protecting plants from root-knot nematodes, containing an active substance on a powder carrier, characterized in that the active substance is a complex of avermectins aversectin C or aversectin B, or aversectin A in concentration 0.1-0.3, and the solid carrier is pellitite or zeolite, or processed mycelium 3. аэнтѹ.Ше.
PCT/RU1997/000183 1997-06-09 1997-06-09 Streptomyces avermitilis strain, method for separating avermectin complexes and preparations for protecting plants and animals Ceased WO1998056939A1 (en)

Priority Applications (2)

Application Number Priority Date Filing Date Title
PCT/RU1997/000183 WO1998056939A1 (en) 1997-06-09 1997-06-09 Streptomyces avermitilis strain, method for separating avermectin complexes and preparations for protecting plants and animals
AU41397/97A AU4139797A (en) 1997-06-09 1997-06-09 (streptomyces avermitilis) strain, method for separating avermectin complexe s and preparations for protecting plants and animals

Applications Claiming Priority (1)

Application Number Priority Date Filing Date Title
PCT/RU1997/000183 WO1998056939A1 (en) 1997-06-09 1997-06-09 Streptomyces avermitilis strain, method for separating avermectin complexes and preparations for protecting plants and animals

Publications (2)

Publication Number Publication Date
WO1998056939A1 true WO1998056939A1 (en) 1998-12-17
WO1998056939A8 WO1998056939A8 (en) 1999-05-27

Family

ID=20130121

Family Applications (1)

Application Number Title Priority Date Filing Date
PCT/RU1997/000183 Ceased WO1998056939A1 (en) 1997-06-09 1997-06-09 Streptomyces avermitilis strain, method for separating avermectin complexes and preparations for protecting plants and animals

Country Status (2)

Country Link
AU (1) AU4139797A (en)
WO (1) WO1998056939A1 (en)

Cited By (3)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
EP2886640A1 (en) 2013-12-18 2015-06-24 Riga Technical University Process for isolation of milbemycins A3 and A4
WO2019164878A1 (en) * 2018-02-20 2019-08-29 Locus Agriculture Ip Company, Llc Materials and methods for attracting and controlling plant-pathogenic nematodes
US11963528B2 (en) 2018-02-26 2024-04-23 Locus Solutions Ipco, Llc Materials and methods for control of insect pests using entomopathogenic fungi

Citations (8)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
US4199569A (en) * 1977-10-03 1980-04-22 Merck & Co., Inc. Selective hydrogenation products of C-076 compounds and derivatives thereof
US4378353A (en) * 1981-02-17 1983-03-29 Merck & Co., Inc. Novel C-076 compounds
SU1011055A3 (en) * 1979-08-23 1983-04-07 Санкио Компани Лимитед (Фирма) Process for preparing substance having anticarinal anticaricidal effect
SU1560059A3 (en) * 1985-07-27 1990-04-23 Пфайзер Корпорейшн (Фирма) Method of producing avermectine derivatives
WO1993019194A1 (en) * 1992-03-25 1993-09-30 Pfizer Inc. Process for production of avermectins and cultures therefor
WO1995003419A1 (en) * 1993-07-23 1995-02-02 Pfizer Inc. Method for precipitating natural avermectins
RU95102462A (en) * 1995-02-28 1997-03-27 Товарищество с ограниченной ответственностью Научно-производственное объединение "Фармбиомед" Method of avermectin complex preparing for construction of preparations for prophylaxis and complex treatment of diseases caused by endo- and ectoparasites in animals, preparations based on thereof
RU2087535C1 (en) * 1995-06-09 1997-08-20 Товарищество с ограниченной ответственностью Научно-производственное объединение "Бифидум" Strain of actinomyces streptomyces avermitilis - a producer of avermectines

Patent Citations (8)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
US4199569A (en) * 1977-10-03 1980-04-22 Merck & Co., Inc. Selective hydrogenation products of C-076 compounds and derivatives thereof
SU1011055A3 (en) * 1979-08-23 1983-04-07 Санкио Компани Лимитед (Фирма) Process for preparing substance having anticarinal anticaricidal effect
US4378353A (en) * 1981-02-17 1983-03-29 Merck & Co., Inc. Novel C-076 compounds
SU1560059A3 (en) * 1985-07-27 1990-04-23 Пфайзер Корпорейшн (Фирма) Method of producing avermectine derivatives
WO1993019194A1 (en) * 1992-03-25 1993-09-30 Pfizer Inc. Process for production of avermectins and cultures therefor
WO1995003419A1 (en) * 1993-07-23 1995-02-02 Pfizer Inc. Method for precipitating natural avermectins
RU95102462A (en) * 1995-02-28 1997-03-27 Товарищество с ограниченной ответственностью Научно-производственное объединение "Фармбиомед" Method of avermectin complex preparing for construction of preparations for prophylaxis and complex treatment of diseases caused by endo- and ectoparasites in animals, preparations based on thereof
RU2087535C1 (en) * 1995-06-09 1997-08-20 Товарищество с ограниченной ответственностью Научно-производственное объединение "Бифидум" Strain of actinomyces streptomyces avermitilis - a producer of avermectines

Cited By (3)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
EP2886640A1 (en) 2013-12-18 2015-06-24 Riga Technical University Process for isolation of milbemycins A3 and A4
WO2019164878A1 (en) * 2018-02-20 2019-08-29 Locus Agriculture Ip Company, Llc Materials and methods for attracting and controlling plant-pathogenic nematodes
US11963528B2 (en) 2018-02-26 2024-04-23 Locus Solutions Ipco, Llc Materials and methods for control of insect pests using entomopathogenic fungi

Also Published As

Publication number Publication date
AU4139797A (en) 1998-12-30
WO1998056939A8 (en) 1999-05-27

Similar Documents

Publication Publication Date Title
FI97890C (en) Process for the preparation of novel therapeutically useful 4 &#34;-deoxy-4&#34; epimethylamino avermectin salts
EP0170006B1 (en) Method and compositions for helmintic, arthropod ectoparasitic and acaridal infections with novel agents
SK281510B6 (en) POLYCYCLIC COMPOUND, PRODUCTION METHOD, PHARMACEUTICAL PRODUCT WITH ITS CONTENT, ITS USE AND BIOLOGICAL CULTURE
DE3031756A1 (en) CONNECTION B-41D, METHOD FOR THE PRODUCTION THEREOF AND MEANS CONTAINING THIS CONNECTION
KR20040063416A (en) AGENT FOR KILLING INSECTS COMPRISING COMPOUNDS HAVING ACYL CoA:CHOLESTEROL ACYLTRANSFERASE INHIBITORY OR SALT THEREOF
JPH07505363A (en) Alkylthioalkyl avermectins as active antiparasitic agents
AU687995B2 (en) Process for making avermectin/zein compositions
US5470827A (en) Method of producing iturin A and antifungal agent for profound mycosis
JPS61233686A (en) Avermectin biologically converted product
HU181762B (en) Process for producing optically active alpha-cyano-3-bracket-4-halogenophenoxy-bracket closed-benzyl-2-bracket-4-chlorophenyl-bracket closed-isovalerates, containing 60-99,5-bracket-s-bracket closed-alpha-cyano-3-bracket-4-halogenophenoxy-benzyl-bracket-s-bracket closed-2-bracket-4-chlorophenyl-bracket closed-isovalerates, and insecticide and/or ac
DK170343B1 (en) Process for the preparation of compounds related to avermectins
JPS59187791A (en) Organic compound and production thereof
WO1998056939A1 (en) Streptomyces avermitilis strain, method for separating avermectin complexes and preparations for protecting plants and animals
Bull et al. Comparative fate of the geometric isomers of phosphamidon in plants and animals
US5317030A (en) Method and compositions for helmintic, arthropod ectoparasitic and acaridal infections with novel agents
CN113321655A (en) Preparation and application of evodia rutaecarpa botanical insecticide
KR20000076074A (en) Pesticidal 1-arylpyrazoles
Shakirzyanova et al. Pheromone Monitoring in the Granaries of Uzbekistan
BG62676B1 (en) PYRTROID COMPOSITION
US5198464A (en) Method and compositions for helmintic, arthropod ectoparasitic and acaridal infections with novel agents
CN117126116B (en) A derivative of an isoxazoline compound and its application
US4279903A (en) Diphydrothiazine derivatives
RU2182174C2 (en) Method of isolation of avermectin complexes
JPH0665246A (en) Novel substance pf 1101a substance, pf 1101b substance, their production and use
Ghochikyan et al. STUDY OF THE BIOLOGICAL PROPERTIES OF SEMI-AND THIOSEMICARBAZONES OF CARBONYL DERIVATIVES OF 4-BUTANOLIDES

Legal Events

Date Code Title Description
AK Designated states

Kind code of ref document: A1

Designated state(s): AM AT AU BG BR BY CA CH CN CZ DE DK EE ES FI GB GE HU IL IS JP KR KZ LR LT LU LV MD MN MX NO NZ PL PT RO RU SD SE SG SI SK TJ TM TR UA US UZ VN

AL Designated countries for regional patents

Kind code of ref document: A1

Designated state(s): AM AZ BY KG KZ MD RU TJ TM AT BE CH DE DK ES FI FR GB GR IE IT LU MC NL PT SE

DFPE Request for preliminary examination filed prior to expiration of 19th month from priority date (pct application filed before 20040101)
121 Ep: the epo has been informed by wipo that ep was designated in this application
AK Designated states

Kind code of ref document: C1

Designated state(s): AM AT AU BG BR BY CA CH CN CZ DE DK EE ES FI GB GE HU IL IS JP KR KZ LR LT LU LV MD MN MX NO NZ PL PT RO RU SD SE SG SI SK TJ TM TR UA US UZ VN

AL Designated countries for regional patents

Kind code of ref document: C1

Designated state(s): AM AZ BY KG KZ MD RU TJ TM AT BE CH DE DK ES FI FR GB GR IE IT LU MC NL PT SE

CFP Corrected version of a pamphlet front page

Free format text: UNDER (72, 75) REPLACE "(KRUGLYAK, NINA BORISOVNA, MOSCOW (RU))." AND ITS RUSSIAN TRANSLITERATION BY "(KRUGLYAK, ELENA BORISOVNA, MOSCOW (RU))." AND ITS RUSSIAN TRANSLITERATION, AND REPLACE "(LISOVENKO, VLADIMIR TROFIMOVICH, MOSCOW (RU))." AND ITS RUSSIAN TRANSLITERATION BY "(LISOVENKO, VASILY TROFIMOVICH, MOSCOW (RU))." AND ITS RUSSIAN TRANSLITERATION

NENP Non-entry into the national phase

Ref country code: CA

NENP Non-entry into the national phase

Ref country code: JP

Ref document number: 1999502192

Format of ref document f/p: F

REG Reference to national code

Ref country code: DE

Ref legal event code: 8642

122 Ep: pct application non-entry in european phase