EP0000063B1 - Dipeptides dérivés de la 7-(N-alpha-substitué ou non substitué X-arginyl)-amino-4-méthyl-coumarine - Google Patents

Dipeptides dérivés de la 7-(N-alpha-substitué ou non substitué X-arginyl)-amino-4-méthyl-coumarine Download PDF

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Publication number
EP0000063B1
EP0000063B1 EP78100111A EP78100111A EP0000063B1 EP 0000063 B1 EP0000063 B1 EP 0000063B1 EP 78100111 A EP78100111 A EP 78100111A EP 78100111 A EP78100111 A EP 78100111A EP 0000063 B1 EP0000063 B1 EP 0000063B1
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EP
European Patent Office
Prior art keywords
amino
methylcoumarin
arginyl
group
mixture
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.)
Expired
Application number
EP78100111A
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German (de)
English (en)
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EP0000063A1 (fr
Inventor
Shumpei Sakakibara
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Ajinomoto Co Inc
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Ajinomoto Co Inc
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Priority claimed from JP52066517A external-priority patent/JPS5842862B2/ja
Priority claimed from JP52067718A external-priority patent/JPS5811424B2/ja
Priority claimed from JP6771977A external-priority patent/JPS543074A/ja
Application filed by Ajinomoto Co Inc filed Critical Ajinomoto Co Inc
Publication of EP0000063A1 publication Critical patent/EP0000063A1/fr
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    • C—CHEMISTRY; METALLURGY
    • C07—ORGANIC CHEMISTRY
    • C07K—PEPTIDES
    • C07K5/00—Peptides containing up to four amino acids in a fully defined sequence; Derivatives thereof
    • C07K5/04—Peptides containing up to four amino acids in a fully defined sequence; Derivatives thereof containing only normal peptide links
    • C07K5/10—Tetrapeptides
    • C07K5/1002—Tetrapeptides with the first amino acid being neutral
    • C07K5/1005—Tetrapeptides with the first amino acid being neutral and aliphatic
    • C07K5/101—Tetrapeptides with the first amino acid being neutral and aliphatic the side chain containing 2 to 4 carbon atoms, e.g. Val, Ile, Leu
    • C—CHEMISTRY; METALLURGY
    • C07—ORGANIC CHEMISTRY
    • C07D—HETEROCYCLIC COMPOUNDS
    • C07D311/00—Heterocyclic compounds containing six-membered rings having one oxygen atom as the only hetero atom, condensed with other rings
    • C07D311/02—Heterocyclic compounds containing six-membered rings having one oxygen atom as the only hetero atom, condensed with other rings ortho- or peri-condensed with carbocyclic rings or ring systems
    • C07D311/04—Benzo[b]pyrans, not hydrogenated in the carbocyclic ring
    • C07D311/06—Benzo[b]pyrans, not hydrogenated in the carbocyclic ring with oxygen or sulfur atoms directly attached in position 2
    • C07D311/08—Benzo[b]pyrans, not hydrogenated in the carbocyclic ring with oxygen or sulfur atoms directly attached in position 2 not hydrogenated in the hetero ring
    • C07D311/16—Benzo[b]pyrans, not hydrogenated in the carbocyclic ring with oxygen or sulfur atoms directly attached in position 2 not hydrogenated in the hetero ring substituted in position 7
    • C—CHEMISTRY; METALLURGY
    • C07—ORGANIC CHEMISTRY
    • C07K—PEPTIDES
    • C07K5/00—Peptides containing up to four amino acids in a fully defined sequence; Derivatives thereof
    • C07K5/04—Peptides containing up to four amino acids in a fully defined sequence; Derivatives thereof containing only normal peptide links
    • C07K5/06—Dipeptides
    • C07K5/06008—Dipeptides with the first amino acid being neutral
    • C07K5/06017—Dipeptides with the first amino acid being neutral and aliphatic
    • C07K5/06026—Dipeptides with the first amino acid being neutral and aliphatic the side chain containing 0 or 1 carbon atom, i.e. Gly or Ala
    • C—CHEMISTRY; METALLURGY
    • C07—ORGANIC CHEMISTRY
    • C07K—PEPTIDES
    • C07K5/00—Peptides containing up to four amino acids in a fully defined sequence; Derivatives thereof
    • C07K5/04—Peptides containing up to four amino acids in a fully defined sequence; Derivatives thereof containing only normal peptide links
    • C07K5/06—Dipeptides
    • C07K5/06008—Dipeptides with the first amino acid being neutral
    • C07K5/06078—Dipeptides with the first amino acid being neutral and aromatic or cycloaliphatic
    • C—CHEMISTRY; METALLURGY
    • C07—ORGANIC CHEMISTRY
    • C07K—PEPTIDES
    • C07K5/00—Peptides containing up to four amino acids in a fully defined sequence; Derivatives thereof
    • C07K5/04—Peptides containing up to four amino acids in a fully defined sequence; Derivatives thereof containing only normal peptide links
    • C07K5/06—Dipeptides
    • C07K5/06139—Dipeptides with the first amino acid being heterocyclic
    • C07K5/06165—Dipeptides with the first amino acid being heterocyclic and Pro-amino acid; Derivatives thereof
    • C—CHEMISTRY; METALLURGY
    • C07—ORGANIC CHEMISTRY
    • C07K—PEPTIDES
    • C07K5/00—Peptides containing up to four amino acids in a fully defined sequence; Derivatives thereof
    • C07K5/04—Peptides containing up to four amino acids in a fully defined sequence; Derivatives thereof containing only normal peptide links
    • C07K5/08—Tripeptides
    • C07K5/0802—Tripeptides with the first amino acid being neutral
    • C07K5/0804—Tripeptides with the first amino acid being neutral and aliphatic
    • C07K5/0808—Tripeptides with the first amino acid being neutral and aliphatic the side chain containing 2 to 4 carbon atoms, e.g. Val, Ile, Leu
    • C—CHEMISTRY; METALLURGY
    • C07—ORGANIC CHEMISTRY
    • C07K—PEPTIDES
    • C07K5/00—Peptides containing up to four amino acids in a fully defined sequence; Derivatives thereof
    • C07K5/04—Peptides containing up to four amino acids in a fully defined sequence; Derivatives thereof containing only normal peptide links
    • C07K5/08—Tripeptides
    • C07K5/0819—Tripeptides with the first amino acid being acidic
    • C—CHEMISTRY; METALLURGY
    • C07—ORGANIC CHEMISTRY
    • C07K—PEPTIDES
    • C07K5/00—Peptides containing up to four amino acids in a fully defined sequence; Derivatives thereof
    • C07K5/04—Peptides containing up to four amino acids in a fully defined sequence; Derivatives thereof containing only normal peptide links
    • C07K5/08—Tripeptides
    • C07K5/0821—Tripeptides with the first amino acid being heterocyclic, e.g. His, Pro, Trp
    • C07K5/0823—Tripeptides with the first amino acid being heterocyclic, e.g. His, Pro, Trp and Pro-amino acid; Derivatives thereof
    • 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
    • Y02P—CLIMATE CHANGE MITIGATION TECHNOLOGIES IN THE PRODUCTION OR PROCESSING OF GOODS
    • Y02P20/00—Technologies relating to chemical industry
    • Y02P20/50—Improvements relating to the production of bulk chemicals
    • Y02P20/55—Design of synthesis routes, e.g. reducing the use of auxiliary or protecting groups

Definitions

  • This invention relates to peptide derivatives, more particularly 7-(N"-substituted or non- substituted-X-arginyl)-amino-4-methyl-coumarin wherein X is a residue of an amino acid selected from the group of glycine, phenylalanine and proline, and to acid salts thereof, which are useful as fluorogenic substrates for determining enzymatic activities.
  • a method which comprises reacting a material which is specifically acted upon by an enzyme with the enzyme and comparing the state of the material before reaction with that after reaction.
  • This invention responds to such need and provides synthetic substrates which show specificity to enzymes such as Trypsin and Thrombin.
  • Compounds of the present invention are novel peptide derivatives, which are not described in any reference, more particularly 7-(N *- substituted or non-substituted-X-arginyl)-amino-4-methyl coumarin wherein X is a residue of an amino acid selected from the group of glycine, phenylalanine and proline, and acid salts thereof.
  • R is a hydrogen atom
  • the compound is 7-(glycylarginyl)-amino-4-methylcoumarin.
  • R is a prolyl group
  • the compound is 7-(prolylglycylarginyl)-amino-4-methylcoumarin.
  • R 1 is a glutamyl group
  • the compound is 7-(glutamylglycylarginyl)-amino-4-methylcoumarin.
  • R, is a seryl group the compound is 7-(serylglycylarginyl)-amino-4-methylcoumarin.
  • R is a leucyl group
  • the compound is 7-(leucylglycylarginyl)-amino-4-methyl-coumarin.
  • R is a glutamyl group carrying an isoleucyl group in the N ⁇ -position thereof, namely an isoleucylglutamyl group
  • the compound is 7-(isoleucylglutamylglycylarginyl)-amino-4.-methylcoumarin.
  • R is a seryl group having a valyl group in the N"-position thereof, namely a valylseryl group
  • the compound is 7-(valylserylglycyl- arginyl)-amino-4-methylcoumarin.
  • R is a leucyl group having a valyl group in the N ⁇ -position thereof, namely valylleucyl group
  • the compound is 7-(valylleucylglycylarginyl)-amino-4-methylcoumarin.
  • the compounds of the present invention are represented by the following formula: and salts thereof, wherein R 2 is a radical selected from the group consisting of a hydrogen atom and a prolyl group.
  • R 2 is a radical selected from the group consisting of a hydrogen atom and a prolyl group.
  • R 2 is a hydrogen atom
  • the compound is 7-(phenylalanylarginyl)-amino-4-methylcoumarin.
  • R 2 is a prolyl group
  • the compound is 7-(prolylphenylalanylarginyl)-amino-4-methylcoumarin.
  • R 3 When R 3 is a hydrogen atom, the compound is 7-(prolylarginyl)-amino-4-methylcoumarin. When R 3 is a valyl group, the compound is 7-(valylprolylarginyl)-amino-4-methylcoumarin.
  • N"-amino and imino groups in such compounds may be protected by conventional protecting grqups for peptides, such as an acyl group (for example, acetyl group and benzoyl group), carbobenzoxy group, tert-alkyloxycarbonyl group, tosyl group and glutaryl group.
  • an acyl group for example, acetyl group and benzoyl group
  • carbobenzoxy group for example, acetyl group and benzoyl group
  • carbobenzoxy group for example, tert-alkyloxycarbonyl group, tosyl group and glutaryl group.
  • the hydroxy group thereof may be protected by protecting groups which are used in the conventional method for synthesizing peptides, such as alkyl groups and benzyl groups.
  • the y-carboxyl group thereof may be protected by an ester group with an alcohol such as benzyl alcohol.
  • the guanidino group of arginine which constitutes such compounds may be protected by N-guanidino-protecting groups which are used in the conventional method for synthesizing peptides, such as nitro groups, tosyl groups and p-methoxy-benzenesulfonyl groups, and by adding a proton, as in the form of an acid.
  • the compounds of the present invention can be produced by conventional methods for synthesizing peptides using 7-arginyl-amino-4-methylcoumarin.
  • 7-(glycylarginyl)-amino-4-methylcoumarin can be produced by reacting glycine whose amino group has been protected, with 7-arginylamino-4-methylcoumarin in the presence of condensing agents such as dicyclohexyl-' carbodiimide (DCCI), or by reacting an active ester of glycine whose amino group has been protected; with 7-arginylamino-4-methylcoumarin, and then removing the protecting-group of the amino group therein.
  • DCCI dicyclohexyl-' carbodiimide
  • 7-(glutamylglycylarginyl)-amino-4-methylcoumarin By reacting this compound with glutamic acid whose amino group and y-carboxyl group are protected and then removing protecting-groups from both the amino group and y-carboxyl group of thus obtained compound, 7-(glutamylglycylarginyl)-amino-4-methylcoumarin can be obtained.
  • 7-(isoleucyl-glutamylglycylarginyl)-amino-4-methylcoumarin can be produced by reacting 7-(glutamyl- glycylarginyl)-amino-4-methylcoumarin as produced above, with an active ester of isoleucine whose amino group has been protected, an then treating such reacted material in the same manner as above.
  • 7-(prolylglycylarginyl)-amino-4-methylcoumarin can be produced by reacting 7-(glycylarginyl)-amino-4-methylcoumarin obtained above with proline whose imino group has been protected and then removing the protecting group from the thus obtained compound.
  • the component having the carboxyl group which reacts with the component having the amino group is in the active ester form.
  • Suitable examples of the active esters are N-hydroxysuccinimide ester and p-nitrophenyl ester. Reactions using active ester can be operated sufficiently at room temperature, but can be accelerated by heating, if necessary.
  • the reaction mixture is concentrated to yield a solid material and the material is purified by column chromatography and then lyophilized.
  • the compounds thus obtained are usually white powders and are decomposed slowly by heating at more than 140°C.
  • such protecting-groups can be removed by conventional methods or synthesizing peptides.
  • the carbobenzoxy group can be removed by catalytic hydrogenation in a solvent such as alcohol, and the tert-butoxycarbonyl group can be removed by reacting the compound in hydrogen fluoride for 30 minutes or by reacting the compound with p-toluene sulfonic acid for 90 minutes.
  • the compounds of the present invention can be produced in free form or acid salt form according to the method for production.
  • the free form can be easily converted to the acid salt thereof and an acid salt can be easily converted to the free form thereof.
  • Examples of the acid salts are salts of inorganic acids such as hydrochloric acid, sulfuric acid, nitric acid and phosphoric acid, and salts of organic acids such as acetic acid, oxalic acid, tartaric acid, succinic acid, citric acid and toluene sulfonic acid.
  • inorganic acids such as hydrochloric acid, sulfuric acid, nitric acid and phosphoric acid
  • organic acids such as acetic acid, oxalic acid, tartaric acid, succinic acid, citric acid and toluene sulfonic acid.
  • the chemical structure of the compounds of the present invention was identified by elemental analysis, amino acid analysis, ultraviolet spectrum, and comparison of ultraviolet spectrum of material hydrolyzed with trypsin with 7-amino-4-methylcoumarin.
  • the compounds of the present invention are all hydrolyzed specifically by one or more enzyme(s) such as Thrombin, Horseshoe Crab Blood Coagulating Enzyme, urokinase and/or Blood Coagulating Factor XII . , and therefore are suitable as synthetic substrates of these compounds.
  • enzyme(s) such as Thrombin, Horseshoe Crab Blood Coagulating Enzyme, urokinase and/or Blood Coagulating Factor XII .
  • Amino acids which compose the compounds of the present invention may have any stereochemical configuration, L, D and DL, if the compounds may be enzymatically hydrolyzed.
  • 7-Arginylamino-4-methylcoumarin and the sals thereof are also new compounds.
  • 7-Arginylamino-4-methylcoumarin in the free form can be easily produced by reacting the acid salts thereof with an alkaline material.
  • N"-carbobenzoxy-L-arginine hydrochloric acid salt (69.0 g, 0.2 M) and 7-amino-4-methyl- coumarin (17.5g, 0.1 M) were dissolved in dimethylformamide (DMF) (300 ml).
  • DMF dimethylformamide
  • DCCD Dicyclohexylcarbodiimide
  • 21 g, 0.1 M was added to the mixture with stirring at room temperature. This mixture was stirred overslav at room temperature, and the produced dicyclohexyl urea was removed by filtration. The obtained filtrate was concentrated under reduced pressure and to the obtained residue methyl alcohol (50 ml) and ethyl acetate (500 ml) were added.
  • N°-carbobenzoxy-N G- tosyl-L-arginine (See J. A.C.S. 82, 4576) (6.0 g, 13 mM) was dissolved in DMF (20 ml), and 4-methyl-7-aminocoumarin (1.75 g, 10 mM) was added thereto.
  • DCCD (2.47 g, 12 mM) was added with ice-cooling. The mixture was stirred overnight at room temperature and the precipitated dicyclohexyl urea was removed by filtration. DMF was distilled off under reduced pressure, and to the residue chloroform (200 ml) was added for extraction.
  • the chloroform solution was washed with 1 N-HCI, 5% NaHC0 3 and water successively, and then was dried over anhydrous sodium sulfate.
  • chloroform was distilled off under reduced pressure, and the residue was purified by column chromatography using silicagel and a binary mixture of chloroform and ethyl acetate in 2:1 volume ratio. The main fractions were combined and concentrated.
  • the residue was dissolved in chloroform (10 ml) and ether was added thereto to precipitate desired material. This material was separated to give 7-(N ⁇ -carbobenioxy-N G -tosyl-L-arginyl)amino-4-methylcoumarin (2.31 g, yield: 37%).
  • the precipitate was separated by filtration and dried.
  • the precipitate was dissolved in DMF (8 ml), and N-hydroxysuccinimide ester of tert-butyloxycarbonyl- ⁇ -benzy)-L-glutamic acid 1478 mg. 1.1 mM).
  • This mixture was stirred for 2 days at room temperature, and excess ether was added to yield a precipitate.
  • This precipitate was separated by filtration and purified by column chromatography (2 x 10 cm) using silica gel as adsorbent and a ternary eluent of chloroform, ethyl acetate and ethyl alcohol in 5:5:2 volume ratio. The main fractions were combined and concentrated to yield a solid material.
  • the material was purified by column chromatography (2 x 15 cm) using silica gel as adsorbent and ternary mixtures of chloroform, methylalcohol and acetic acid in 95:5:3 (adsorption) and then 85:20:5 (elution) in volume ratio.
  • the main fraction was concentrated to yield a solid material and dissolved in acetic acid (20 ml). This solution was lyophilized to give 7-(N"- tert-butyloxycarbonyl-O-benzyl-L-serylglycyl-L-arginyl)-amino-4-methyl-coumarin hydrochloric acid salt [Xi] (180 mg).
  • the material was purified by column chromatography (2 x 15 cm) using silica gel as adsorbent and ternary mixtures of chloroform, methylalcohol and acetic acid in 95:5:3 (adsorption) and then 85:20:5 (elution) in volume ratio.
  • the main fraction was concentrated to yield a solid material, dissolved in acetic acid (20 ml) and lyophilized to give 7-(N°-tert-butyloxycarbonyl-L-valyl-L-leucyl-glycyl-L-arginyl)amino-4-methylcoumarin hydrochloric acid salt [XII] (350 mg).
  • the carbobenzoxy group of the compound thus obtained can be removed by applying a hydrogenation reaction in alcohol thereto.
  • the compound produced above (0.1-0.2 mM) was dissolved in a mixture of dimethylsulfoxide (5 ml) and water (5 ml) and this solution was diluted with 0.05 M Tris-HCI buffer (pH 8.0) containing both 0.1 M NaCl and 10 mM CaCI 2 to adjust the total volume thereof to 500 ml and thereby a substrate solution was prepared.
  • Tris-HCI buffer pH 8.0
  • each substrate solution (2 ml) was added in a test tube and the obtained test tube was kept at 37°C for 5 minutes. Then, each enzyme solution (20 ⁇ l) was added thereto and incubated at 37°C for 20 minutes with stirring. The incubation was stopped by adding 100% acetic acid (0.5 ml) thereto.
  • the hydrolysis degree was determined by measuring the fluorescence intensity at 460 nm with excitation at 380 mn in the fluorescence spectra. Results are listed in the following table.
  • Each value is the fluorescence intensity ratio when the fluorescence intensity of 7-amino-4-methylcoumarin solution (40 ⁇ M, 40 ⁇ mol/l) being 100.
  • a value having the mark -- *-- is one with the fluorescence intensity of 7-amino-4-methylcoumarin solution (200 ⁇ M, 200 ⁇ mol/l) being 100.

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  • Chemical & Material Sciences (AREA)
  • Organic Chemistry (AREA)
  • Genetics & Genomics (AREA)
  • Biochemistry (AREA)
  • Biophysics (AREA)
  • General Health & Medical Sciences (AREA)
  • Health & Medical Sciences (AREA)
  • Medicinal Chemistry (AREA)
  • Molecular Biology (AREA)
  • Proteomics, Peptides & Aminoacids (AREA)
  • Life Sciences & Earth Sciences (AREA)
  • Peptides Or Proteins (AREA)
  • Medicines That Contain Protein Lipid Enzymes And Other Medicines (AREA)

Claims (1)

1. 7-(N"-R-X-arginyl)-amino-4-méthylcoumarines de formule générale:
Figure imgb0033
et leurs sels d'acides,
où X est un résidu d'un aminoacide choisi dans le groupe constitué par la glycine, le phénylalanine et la proline, et,
a) lorsque X est le résidu de la glycine, R est choisi parmi un atome d'hydrogène et les groupes prolyle, glytamyle, séryle, leucyle, isoleucylglutamyle, varylséryle et valylleucyle,
b) lorsque X est le résidu de la phénylalanine, R est choisie parmi un atome d'hydrogène et un groupe prolyle, et
c) lorsqûe X est le résidu de la proline, R est choisi parmi un atome d'hydrogène et un group valyle,

et où les groupes N"-amino et imino, les groupes hydroxy, les groupes carboxy et les groupes guanidino des composés peuvent être protégés par des groupements protecteurs classiques pour les peptides.
EP78100111A 1977-06-06 1978-06-06 Dipeptides dérivés de la 7-(N-alpha-substitué ou non substitué X-arginyl)-amino-4-méthyl-coumarine Expired EP0000063B1 (fr)

Applications Claiming Priority (6)

Application Number Priority Date Filing Date Title
JP52066517A JPS5842862B2 (ja) 1977-06-06 1977-06-06 ペプチド誘導体
JP66517/77 1977-06-06
JP52067718A JPS5811424B2 (ja) 1977-06-08 1977-06-08 新規ポリペプチド
JP67719/77 1977-06-08
JP67718/77 1977-06-08
JP6771977A JPS543074A (en) 1977-06-08 1977-06-08 Novel peptide derivative

Publications (2)

Publication Number Publication Date
EP0000063A1 EP0000063A1 (fr) 1978-12-20
EP0000063B1 true EP0000063B1 (fr) 1981-06-10

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EP78100111A Expired EP0000063B1 (fr) 1977-06-06 1978-06-06 Dipeptides dérivés de la 7-(N-alpha-substitué ou non substitué X-arginyl)-amino-4-méthyl-coumarine

Country Status (3)

Country Link
US (1) US4215047A (fr)
EP (1) EP0000063B1 (fr)
DE (1) DE2860749D1 (fr)

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SE7801373L (sv) * 1978-02-07 1979-08-08 Kabi Ab Lett spjelkbara substrat for kvantifiering av proteaser
US4275153A (en) * 1978-08-03 1981-06-23 American Hospital Supply Corporation Analytical fluorogenic substrates for proteolytic enzymes
US4294923A (en) * 1979-04-23 1981-10-13 Smith Robert E Substrates and method for determining enzymes
DE2936543A1 (de) * 1979-09-10 1981-04-09 Behringwerke Ag, 3550 Marburg Chromogene verbindungen
FR2497798A1 (fr) * 1981-01-09 1982-07-16 Pharmindustrie Nouveaux peptides portant un fluorophore, procede pour leur preparation et leur application au dosage fluorimetrique des endotoxines
US4388233A (en) * 1981-05-15 1983-06-14 The Regents Of The University Of California Synthetic substrates for enzyme analysis
US4448715A (en) * 1981-11-02 1984-05-15 University Of Miami Tagged pyroglu-L-Phe-L-Arg derivatives, substrates and assays for kallikrein
JPH0611237B2 (ja) * 1982-04-14 1994-02-16 ラジオメーター コーポレイト ディベロップメント リミテッド 微生物学的試験方法
US5079144A (en) * 1982-04-14 1992-01-07 Radiometer Corporate Development Ltd. Microorganism testing with a hydrolyzable fluorogenic substrate
JPS59192099A (ja) * 1983-04-12 1984-10-31 Ajinomoto Co Inc 微生物菌数の測定法
US5252484A (en) * 1988-11-29 1993-10-12 Minnesota Mining And Manufacturing Company Rapid read-out biological indicator
US5223401A (en) * 1988-11-29 1993-06-29 Minnesota Mining And Manufacturing Company Rapid read-out sterility indicator
US5073488A (en) * 1988-11-29 1991-12-17 Minnesota Mining And Manufacturing Company Rapid method for determining efficacy of a sterilization cycle and rapid read-out biological indicator
US5055594A (en) * 1990-07-19 1991-10-08 Becton, Dickinson And Company Fluorogenic trypotophanase substrates
FR2677023B1 (fr) * 1991-05-30 1994-02-25 Eurobio Laboratoires Derives de coumarine hydrosolubles, leur preparation et leur utilisation comme substrat d'enzyme.
DE19834374B4 (de) * 1998-07-30 2004-03-04 Preh-Werke Gmbh & Co. Kg Drehknopf eines Steuergerätes
US8895239B2 (en) 2006-09-20 2014-11-25 American Sterilizer Company Genetically engineered biological indicator
US8043845B2 (en) 2006-09-20 2011-10-25 American Sterilizer Company Sterilization indicator
US8173388B2 (en) 2008-09-30 2012-05-08 American Sterilizer Company Self-contained biological indicator
JP5934231B2 (ja) 2010-11-01 2016-06-15 スリーエム イノベイティブ プロパティズ カンパニー 生物学的滅菌インジケーター
CN103189524B (zh) 2010-11-01 2015-01-28 3M创新有限公司 生物灭菌指示器及其使用方法
EP3269822B1 (fr) 2010-11-24 2019-11-27 3M Innovative Properties Company Procédés pour détecter une activité biologique
US20180237821A1 (en) 2017-02-23 2018-08-23 Ethicon, Inc. Apparatus and method to read biological indicator
CN109265373A (zh) * 2018-11-03 2019-01-25 郑州安图生物工程股份有限公司 一种氨基酸类底物及其制备方法和用途

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US2929822A (en) * 1955-10-31 1960-03-22 Geigy Ag J R 3-substituted 7-carbalkoxyamino-coumarins
DE1293160B (de) * 1964-03-07 1969-04-24 Bayer Ag Verfahren zur Herstellung von Cumarinverbindungen
CA1007555A (en) * 1971-08-16 1977-03-29 Robert E. Smith Carbobenzoxydiglycyl-1-arginyl-4-methoxy-2-naphthylamide and process for determining enzyme concentrations

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US4215047A (en) 1980-07-29
EP0000063A1 (fr) 1978-12-20
DE2860749D1 (en) 1981-09-17

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