[go: up one dir, main page]

WO2009002209A1 - Substance destinée aux produits médicamenteux dermatologiques à base de collagénase d'origine végétale - Google Patents

Substance destinée aux produits médicamenteux dermatologiques à base de collagénase d'origine végétale Download PDF

Info

Publication number
WO2009002209A1
WO2009002209A1 PCT/RU2007/000581 RU2007000581W WO2009002209A1 WO 2009002209 A1 WO2009002209 A1 WO 2009002209A1 RU 2007000581 W RU2007000581 W RU 2007000581W WO 2009002209 A1 WO2009002209 A1 WO 2009002209A1
Authority
WO
WIPO (PCT)
Prior art keywords
collagenase
substance
activity
microbiotic
ultrafiltrate
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/RU2007/000581
Other languages
English (en)
Russian (ru)
Inventor
Nina Sergeevna Demina
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.)
Otkrytoe Akzionernoe Obschestvo 'moscow Committee Of Science And Technologies'
Original Assignee
Otkrytoe Akzionernoe Obschestvo 'moscow Committee Of Science And Technologies'
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 Otkrytoe Akzionernoe Obschestvo 'moscow Committee Of Science And Technologies' filed Critical Otkrytoe Akzionernoe Obschestvo 'moscow Committee Of Science And Technologies'
Publication of WO2009002209A1 publication Critical patent/WO2009002209A1/fr
Anticipated expiration legal-status Critical
Ceased legal-status Critical Current

Links

Classifications

    • 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
    • C12P1/00—Preparation of compounds or compositions, not provided for in groups C12P3/00 - C12P39/00, by using microorganisms or enzymes
    • C12P1/04—Preparation of compounds or compositions, not provided for in groups C12P3/00 - C12P39/00, by using microorganisms or enzymes by using bacteria
    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61P—SPECIFIC THERAPEUTIC ACTIVITY OF CHEMICAL COMPOUNDS OR MEDICINAL PREPARATIONS
    • A61P17/00—Drugs for dermatological disorders
    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61P—SPECIFIC THERAPEUTIC ACTIVITY OF CHEMICAL COMPOUNDS OR MEDICINAL PREPARATIONS
    • A61P43/00—Drugs for specific purposes, not provided for in groups A61P1/00-A61P41/00

Definitions

  • the invention relates to biotechnology, in particular to the creation of a substance for medicines on the basis of collagenase of microbial origin.
  • a group of microbial collagenases is known, which is widely used in biotechnology and for laboratory purposes, for example, collagenases synthesized by the bacterium Clostridium histoluticum .. Collagenase producers are also insects Nuroderma lipeatum and Kamchatka crabs [1].
  • Known analogues of the invention may be preparations obtained from bacteria Clostridium histoluticum and crab collagenase [1].
  • Clostridium histoluticum is a causative agent of gas gangrene, a strict anaerobic, requiring special growing conditions.
  • the technology for producing the drug from this bacterium is complex and expensive.
  • Crab collagenase is inferior in bacterial properties, as it does not completely neutralize collagen.
  • the inherent fibrenolytic and thrombolytic properties of the drug can cause unwanted effects when applied to the skin.
  • the closest prototype of the proposed biologically active ultrafiltrate is collagenase isolated from the culture fluid of the strain Stromatosis strain VKPM S-910 [2].
  • the disadvantage of this invention is the low collagenolytic activity, a more complex and expensive way to obtain the named enzyme, as well as a limited area of use in medical biotechnology.
  • SUBSTITUTE SHEET (RULE 26)
  • the technical task of this invention is to obtain a highly effective biologically active ultrafiltrate containing collagenase and protease of microbial origin, intended for use as a substance for drugs in various wound healing preparations, preparations for the lysis of postoperative or post-traumatic hemothorax, for the lysis of postoperative adhesions, in cosmetology.
  • the claimed biologically active substance contains collagenase and protease from Stromatosis lavepdulae VKPM S-910 with the following enzymatic activities:
  • the strain strainomes lavepdulae VKPM S-910 is used, which is grown on solid nutrient medium - on potato agar, and then on a fermentation nutrient medium of the following composition (g / l): corn flour - 20; peptone - 0.5; dry fodder yeast - 0.05; MgSOdLNgO - 0.4; gelatin - 0.5; tap water pH 6.4 - 6.8.
  • Cultivation is carried out in Erlenmeyer flasks with a volume of 250 ml on a rocking chair (200 rpm) at a temperature of 28 degrees Celsius for 72 - 96 hours. The mycelium is separated by centrifugation.
  • the procedure for obtaining biologically active ultrafiltrate from the filtrate of the culture fluid begins with its concentration while separating low molecular weight substances and protein fractions at a temperature of 15 - 20 degrees Celsius and a pressure of 760 - 800 mm Hg.
  • the filtrate of the culture fluid is placed in an ultrafiltration laboratory unit with a working surface of 2 square decimeters.
  • the receiving tank is filled with filtrate.
  • Ultrafiltration of the native solution is carried out under a pressure of 0.4 + 0.02 MPa.
  • SUBSTITUTE SHEET (RULE 26) ultrafiltrate in the work using a membrane with a low permeability of the protease.
  • the obtained biologically active ultrafiltrate is stored at a temperature of +1 - +20 degrees Celsius in the presence of a stabilizer.
  • Ultralizin a substance for dermatological medicines, meets the requirements of the developed technical specifications TU 9336-001-11684510-03, effective from 07.07.2003, in compliance with sanitary norms and rules in force at the enterprises of the microbiological industry of the Russian Federation.
  • Ultralizin corresponds to the following indicators:
  • the pH of the aqueous solution is 5.0 - 7.0;
  • - collagenolytic activity is from 1800 to 2500 collagenolytic units of activity per 1 ml;
  • total proteolytic activity is from 150 to 200 proteolytic units per 1 ml
  • Collagenolytic activity is determined by the method of Rosen [4]. Proteolytic activity was determined according to Kunitz [3].
  • a unit of collagenolytic activity is the amount of the enzyme that exposes the hydrolysis products equivalent to 1 ⁇ g of L-lycine under standard experimental conditions to the collagen.
  • test tubes with a capacity of 20 ml each, add a pipette of 2 ml of a suspension of collagen in a phosphate buffer pH 7.4.
  • test tubes In 1, 2 and 3 test tubes (experimental) add 2 ml of the test solution of ultralizine (before analysis, carefully shake the contents of the ampoule or vial).
  • tubes 4 and 5 (control) add 2 ml of water. All tubes are closed with stoppers and placed in a thermostat at 37 + 0.5 degrees Celsius for 18 hours. Cool.
  • the absorbance values are determined on a spectrophotometer at a wavelength of 600 nm in cuvettes with a layer thickness of 10 mm. As a comparison solution, water is used. According to the calibration graph, the L-lycine concentration values are found in the experimental and control tubes, and arithmetic mean values are calculated. From the value of the experimental concentration, the value of the control is subtracted. This value is used to calculate the activity of ultralysin.
  • Reagent B 0.2 M Na 2 HPO 4 "7 H 2 O - 53.65 g / l. Take 81 ml of reagent B.
  • Experimental and control tubes containing 2 ml of two percent casein are thermostated for 10 minutes at a temperature of 30 degrees Celsius.
  • 2.5 ml of trichloroacetic acid (TCA) are added to the control.
  • 0.5 ml of ultrafiltrate is added to the experimental and control tubes.
  • 2.5 ml of TCA are added to the experiment, after 30 seconds, into each tube.
  • the optical density (OD) of the ultrafiltrate is determined at 280 nm.
  • the resulting indicator is multiplied by dilution.
  • the enzymatic activity of the ultraconcentrate varied in the following ranges:
  • Example 1 The ultraconcentrate had the following characteristics: Collagenolytic activity of 1800 KEA / ml
  • the proteolytic activity is 120 PE / ml.
  • Example 2 collagenolytic activity - 2000 KEA / ml; proteolytic activity - 150 PE / ml.
  • mice males and females with a body weight of 19-21 g.
  • Ultralizin was administered to mice intravenously, once, in a volume of 0.2-0.5 ml, at a rate of OD ml / s.
  • mice The duration of observation of mice was 10-15 days. During the experiment, we monitored the behavior, appearance, motor activity, and the reaction of animals to external stimuli. In cases of death, the clinical picture was recorded and an autopsy was performed of dead mice.
  • mice With the intravenous administration of the drug to mice in doses up to 3500-4000 U / kg, no signs of intoxication were observed. Animals remained motile, reacted normally to the environment. With a further increase in the dose of the administered substance, the death of individual animals was noted.
  • mice administration to mice was and 4500 U / kg. In these cases, when observing animals for 2 weeks, death was not observed.
  • SUBSTITUTE SHEET (RULE 26) Studies have been conducted for the presence of pyrogenic impurities in the drug. The test was carried out on chinchilla rabbits, kept on a standard diet of vivarium.
  • a biologically active ultrafiltrate based on collagenase isolated from the strain of Stertomas lavepdulae VKPM S-910 with collagenolytic activity of 1800 - 2500 KEA / ml and proteolytic activity of 120 - 200 PE / ml is a highly effective, non-toxic, non-allergenic drug. Having in its composition collagenase and a protease enhancing its action, the proposed biologically active ultrafiltrate can be used in various wound healing preparations and dermatology.

Landscapes

  • Health & Medical Sciences (AREA)
  • Life Sciences & Earth Sciences (AREA)
  • Chemical & Material Sciences (AREA)
  • Organic Chemistry (AREA)
  • Engineering & Computer Science (AREA)
  • Bioinformatics & Cheminformatics (AREA)
  • Chemical Kinetics & Catalysis (AREA)
  • General Chemical & Material Sciences (AREA)
  • General Health & Medical Sciences (AREA)
  • Medicinal Chemistry (AREA)
  • Wood Science & Technology (AREA)
  • Animal Behavior & Ethology (AREA)
  • Nuclear Medicine, Radiotherapy & Molecular Imaging (AREA)
  • Public Health (AREA)
  • Veterinary Medicine (AREA)
  • Zoology (AREA)
  • Pharmacology & Pharmacy (AREA)
  • Biotechnology (AREA)
  • Microbiology (AREA)
  • Mycology (AREA)
  • Dermatology (AREA)
  • Biochemistry (AREA)
  • General Engineering & Computer Science (AREA)
  • Genetics & Genomics (AREA)
  • Medicines That Contain Protein Lipid Enzymes And Other Medicines (AREA)

Abstract

Selon l'invention, une substance pour produits dermatologique à base d'une collagénase d'origine végétale se présente comme un ultrafiltrat obtenu à partir d'un bouillon de culture de la souche Streptomyces lavandulae VPKM S-910 à activité collagénolythique 1800 - 2500 KEA/ml et à activité protéolytique 120 - 200 PE/ml.
PCT/RU2007/000581 2007-06-19 2007-10-22 Substance destinée aux produits médicamenteux dermatologiques à base de collagénase d'origine végétale Ceased WO2009002209A1 (fr)

Applications Claiming Priority (2)

Application Number Priority Date Filing Date Title
RU2007122707/13A RU2340371C1 (ru) 2007-06-19 2007-06-19 Субстанция для дерматологических лекарственных средств на основе коллагеназы микробного происхождения "ультрализин"
RU2007122707 2007-06-19

Publications (1)

Publication Number Publication Date
WO2009002209A1 true WO2009002209A1 (fr) 2008-12-31

Family

ID=40185854

Family Applications (1)

Application Number Title Priority Date Filing Date
PCT/RU2007/000581 Ceased WO2009002209A1 (fr) 2007-06-19 2007-10-22 Substance destinée aux produits médicamenteux dermatologiques à base de collagénase d'origine végétale

Country Status (2)

Country Link
RU (1) RU2340371C1 (fr)
WO (1) WO2009002209A1 (fr)

Cited By (2)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
US8849416B2 (en) 2004-05-04 2014-09-30 University Of Rochester Implantable bio-electro-physiologic interface matrix
US8938300B2 (en) 2004-05-04 2015-01-20 University Of Rochester Leadless implantable intravascular electrophysiologic device for neurologic/cardiovascular sensing and stimulation

Families Citing this family (1)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
RU2484811C2 (ru) * 2011-08-15 2013-06-20 Общество с ограниченной ответственностью "Межотраслевая компания по науке и технологиям ООО "МКНТ" Ферментный ранозаживляющий препарат

Citations (5)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
RU2075219C1 (ru) * 1995-03-23 1997-03-10 Нина Сергеевна Демина Коллагеназа
US5989888A (en) * 1996-01-24 1999-11-23 Roche Diagnostics Corporation Purified mixture of collagenase I, collagenase II and two other proteases
RU2166950C1 (ru) * 2000-06-30 2001-05-20 Демина Нина Сергеевна Фармацевтическая композиция на основе коллагеназы микробного происхождения
RU2180002C2 (ru) * 1999-08-16 2002-02-27 Пермское научно-производственное объединение "Биомед" Способ получения коллагеназы
RU2005128329A (ru) * 2005-09-13 2007-03-27 Открытое акционерное общество "Система-Венчур" (RU) Ферментная композиция "ультрализин"

Family Cites Families (1)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
SU1735364A1 (ru) * 1989-12-22 1992-05-23 Институт микробиологии АН СССР Штамм актиномицета SтRертомUсеS LaVeNDULae - продуцент протеолитических ферментов

Patent Citations (5)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
RU2075219C1 (ru) * 1995-03-23 1997-03-10 Нина Сергеевна Демина Коллагеназа
US5989888A (en) * 1996-01-24 1999-11-23 Roche Diagnostics Corporation Purified mixture of collagenase I, collagenase II and two other proteases
RU2180002C2 (ru) * 1999-08-16 2002-02-27 Пермское научно-производственное объединение "Биомед" Способ получения коллагеназы
RU2166950C1 (ru) * 2000-06-30 2001-05-20 Демина Нина Сергеевна Фармацевтическая композиция на основе коллагеназы микробного происхождения
RU2005128329A (ru) * 2005-09-13 2007-03-27 Открытое акционерное общество "Система-Венчур" (RU) Ферментная композиция "ультрализин"

Cited By (2)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
US8849416B2 (en) 2004-05-04 2014-09-30 University Of Rochester Implantable bio-electro-physiologic interface matrix
US8938300B2 (en) 2004-05-04 2015-01-20 University Of Rochester Leadless implantable intravascular electrophysiologic device for neurologic/cardiovascular sensing and stimulation

Also Published As

Publication number Publication date
RU2340371C1 (ru) 2008-12-10

Similar Documents

Publication Publication Date Title
Dobell et al. On the cultivation of Entamoeba histolytica and some other entozoic amoebae
Abraham et al. Further observations on penicillin
Wadström et al. Studies on extracellular proteins from Staphylococcus aureus: VII. Studies on β-haemolysin
RU2663136C2 (ru) Способ получения высокочистого нейротоксического компонента ботулотоксина и его применения
Correa et al. Antarctic microorganisms as sources of biotechnological products
RU2340371C1 (ru) Субстанция для дерматологических лекарственных средств на основе коллагеназы микробного происхождения "ультрализин"
US10499655B2 (en) Reagents and methods for inhibiting or disrupting biofilm
DE69324918T2 (de) Arznei gegen cystische fibrosen
Jennison Bacterial collagenase
Surendra et al. Antimicrobial activity of crude venom extracts in honeybees (Apis cerana, Apis dorsata, Apis florea) tested against selected pathogens
Bonventre et al. The biologic activities of Bacillus anthracis and Bacillus cereus culture filtrates
RU2129375C1 (ru) Биопрепарат фитоспорин жидкий для защиты растений от болезней
RU2193063C2 (ru) Бактериолитический комплекс, способ его получения и штамм для осуществления способа
CN100469391C (zh) 人溶菌酶在制备治疗痤疮的药物中的应用
RU2403283C2 (ru) Штамм staphylococcus epidermidis bvm-1987 - продуцент стафилолитического ферментного комплекса
Brogden et al. Changes in ovine erythrocyte morphology due to sphingomyelin degradation by Corynebacterium pseudotuberculosis phospholipase D
Pitamber et al. Production of L-Asparaginase by marine Actinobacteria isolated from the rhizosphere of mangrove plants
RU2285038C2 (ru) Способ получения протеолитического ферментного препарата и протеолитический ферментный препарат
US2601350A (en) Hydrolysis of beta lactam linkage by penicillinase
CN118755644B (zh) 一株头状葡萄球菌、一种菌剂及应用和一种头皮护理产品
Bunyan et al. Phenotypic detection and biofilm formation among Pseudomonas aeruginosa isolated from different sites of infection
SU1735364A1 (ru) Штамм актиномицета SтRертомUсеS LaVeNDULae - продуцент протеолитических ферментов
Smith Bacteria with their coats off: spheroplasts, protoplasts and L-forms
RU2313577C2 (ru) Рекомбинантная плазмида, экспрессирующая клонированный ген cef vibrio cholerae (варианты), и штамм escherichia coli - продуцент cef vibrio cholerae (варианты)
Ryazanova et al. Effect of the proteolytic enzymes of Bacillus licheniformis and the lysoamidase of Lysobacter sp. XL1 on Proteus vulgaris and Proteus mirabilis cells

Legal Events

Date Code Title Description
121 Ep: the epo has been informed by wipo that ep was designated in this application

Ref document number: 07861037

Country of ref document: EP

Kind code of ref document: A1

NENP Non-entry into the national phase

Ref country code: DE

122 Ep: pct application non-entry in european phase

Ref document number: 07861037

Country of ref document: EP

Kind code of ref document: A1