US20030199017A1 - Method for releasing a product comprising chemical oxidation, method for detecting said product and uses thereof - Google Patents

Method for releasing a product comprising chemical oxidation, method for detecting said product and uses thereof Download PDF

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US20030199017A1
US20030199017A1 US10/307,490 US30749002A US2003199017A1 US 20030199017 A1 US20030199017 A1 US 20030199017A1 US 30749002 A US30749002 A US 30749002A US 2003199017 A1 US2003199017 A1 US 2003199017A1
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formula
group
compound
groups
substrate
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Jean-Louis Reymond
Denis Wahler
Frabrizzio Badalassi
Hong-Khanh Nguyen
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Universitaet Bern
Proteus SA
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Proteus SA
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Priority claimed from FR0006952A external-priority patent/FR2809819A1/fr
Application filed by Proteus SA filed Critical Proteus SA
Assigned to PROTEUS, UNIVERSITE DE BERNE reassignment PROTEUS ASSIGNMENT OF ASSIGNORS INTEREST (SEE DOCUMENT FOR DETAILS). Assignors: BADALASSI, FABRIZZIO, NGUYEN, HONG-KHANH, REYMOND, JEAN-LOUIS, WAHLER, DENIS
Publication of US20030199017A1 publication Critical patent/US20030199017A1/en
Priority to US11/601,649 priority Critical patent/US8741593B2/en
Abandoned legal-status Critical Current

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    • C—CHEMISTRY; METALLURGY
    • C07—ORGANIC CHEMISTRY
    • C07D—HETEROCYCLIC COMPOUNDS
    • C07D407/00—Heterocyclic compounds containing two or more hetero rings, at least one ring having oxygen atoms as the only ring hetero atoms, not provided for by group C07D405/00
    • C07D407/02—Heterocyclic compounds containing two or more hetero rings, at least one ring having oxygen atoms as the only ring hetero atoms, not provided for by group C07D405/00 containing two hetero rings
    • C07D407/12—Heterocyclic compounds containing two or more hetero rings, at least one ring having oxygen atoms as the only ring hetero atoms, not provided for by group C07D405/00 containing two hetero rings linked by a chain containing hetero atoms as chain links
    • 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
    • C12—BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
    • C12Q—MEASURING OR TESTING PROCESSES INVOLVING ENZYMES, NUCLEIC ACIDS OR MICROORGANISMS; COMPOSITIONS OR TEST PAPERS THEREFOR; PROCESSES OF PREPARING SUCH COMPOSITIONS; CONDITION-RESPONSIVE CONTROL IN MICROBIOLOGICAL OR ENZYMOLOGICAL PROCESSES
    • C12Q1/00—Measuring or testing processes involving enzymes, nucleic acids or microorganisms; Compositions therefor; Processes of preparing such compositions
    • C12Q1/26—Measuring or testing processes involving enzymes, nucleic acids or microorganisms; Compositions therefor; Processes of preparing such compositions involving oxidoreductase
    • C—CHEMISTRY; METALLURGY
    • C12—BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
    • C12Q—MEASURING OR TESTING PROCESSES INVOLVING ENZYMES, NUCLEIC ACIDS OR MICROORGANISMS; COMPOSITIONS OR TEST PAPERS THEREFOR; PROCESSES OF PREPARING SUCH COMPOSITIONS; CONDITION-RESPONSIVE CONTROL IN MICROBIOLOGICAL OR ENZYMOLOGICAL PROCESSES
    • C12Q1/00—Measuring or testing processes involving enzymes, nucleic acids or microorganisms; Compositions therefor; Processes of preparing such compositions
    • C12Q1/34—Measuring or testing processes involving enzymes, nucleic acids or microorganisms; Compositions therefor; Processes of preparing such compositions involving hydrolase
    • C—CHEMISTRY; METALLURGY
    • C12—BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
    • C12Q—MEASURING OR TESTING PROCESSES INVOLVING ENZYMES, NUCLEIC ACIDS OR MICROORGANISMS; COMPOSITIONS OR TEST PAPERS THEREFOR; PROCESSES OF PREPARING SUCH COMPOSITIONS; CONDITION-RESPONSIVE CONTROL IN MICROBIOLOGICAL OR ENZYMOLOGICAL PROCESSES
    • C12Q1/00—Measuring or testing processes involving enzymes, nucleic acids or microorganisms; Compositions therefor; Processes of preparing such compositions
    • C12Q1/34—Measuring or testing processes involving enzymes, nucleic acids or microorganisms; Compositions therefor; Processes of preparing such compositions involving hydrolase
    • C12Q1/37—Measuring or testing processes involving enzymes, nucleic acids or microorganisms; Compositions therefor; Processes of preparing such compositions involving hydrolase involving peptidase or proteinase
    • C—CHEMISTRY; METALLURGY
    • C12—BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
    • C12Q—MEASURING OR TESTING PROCESSES INVOLVING ENZYMES, NUCLEIC ACIDS OR MICROORGANISMS; COMPOSITIONS OR TEST PAPERS THEREFOR; PROCESSES OF PREPARING SUCH COMPOSITIONS; CONDITION-RESPONSIVE CONTROL IN MICROBIOLOGICAL OR ENZYMOLOGICAL PROCESSES
    • C12Q2326/00—Chromogens for determinations of oxidoreductase enzymes
    • C—CHEMISTRY; METALLURGY
    • C12—BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
    • C12Q—MEASURING OR TESTING PROCESSES INVOLVING ENZYMES, NUCLEIC ACIDS OR MICROORGANISMS; COMPOSITIONS OR TEST PAPERS THEREFOR; PROCESSES OF PREPARING SUCH COMPOSITIONS; CONDITION-RESPONSIVE CONTROL IN MICROBIOLOGICAL OR ENZYMOLOGICAL PROCESSES
    • C12Q2334/00—O-linked chromogens for determinations of hydrolase enzymes, e.g. glycosidases, phosphatases, esterases
    • C—CHEMISTRY; METALLURGY
    • C12—BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
    • C12Q—MEASURING OR TESTING PROCESSES INVOLVING ENZYMES, NUCLEIC ACIDS OR MICROORGANISMS; COMPOSITIONS OR TEST PAPERS THEREFOR; PROCESSES OF PREPARING SUCH COMPOSITIONS; CONDITION-RESPONSIVE CONTROL IN MICROBIOLOGICAL OR ENZYMOLOGICAL PROCESSES
    • C12Q2334/00—O-linked chromogens for determinations of hydrolase enzymes, e.g. glycosidases, phosphatases, esterases
    • C12Q2334/20—Coumarin derivatives
    • G—PHYSICS
    • G01—MEASURING; TESTING
    • G01N—INVESTIGATING OR ANALYSING MATERIALS BY DETERMINING THEIR CHEMICAL OR PHYSICAL PROPERTIES
    • G01N2333/00—Assays involving biological materials from specific organisms or of a specific nature
    • G01N2333/90—Enzymes; Proenzymes
    • G01N2333/914—Hydrolases (3)
    • G01N2333/916—Hydrolases (3) acting on ester bonds (3.1), e.g. phosphatases (3.1.3), phospholipases C or phospholipases D (3.1.4)
    • G01N2333/918—Carboxylic ester hydrolases (3.1.1)

Definitions

  • This present invention relates to a method for detecting a chemical transformation of a substrate such that said chemical transformation of this substrate releases a product, whereby said released product is then a detectable product.
  • the invention also relates to substrates that can be used in this method.
  • the invention also relates to the use of this method in a process for detection and optionally quantification of a chemical transformation in a sample.
  • a second class of substrate leads to the revealing of a product that is obtained after enzymatic secondary reaction and/or spontaneous secondary reaction.
  • This second class of substrate is more stable. This second class of substrate, however, is limited to particular uses, because the fact of using an enzyme for the secondary reaction represents a drawback in terms of cost and flexibility.
  • This invention has as its object a method for releasing at least one product that can be detected, characterized in that a compound of Formula (II′) below:
  • R′ 7 to R′ 10 which are identical or different, correspond to a hydrogen atom, a substituted or unsubstituted alkyl group, or a substituted or unsubstituted functional group,
  • X and Y which are identical or different, are selected from among an oxygen atom, a sulfur atom, an amine of Formula -NR 11 R 12 , and R 11 is selected from among: a hydrogen atom, an alkyl group, and s substituted or unsubstituted aryl group, and R 12 is not a hydrogen atom.
  • This chemical oxidation reaction is carried out with an oxidizing chemical agent that can correspond advantageously and in a non-limiting way to one or more of the following reagents: H 5 IO 6 , RuO 2 , OSO 4 , (CH 3 CH 2 CH 2 ) 4 N(RuO 4 ), NaClO 4 , NaIO 4 , Na 3 H 2 IO 6 , NaMnO 4 , K 2 O s O 4 , KIO 4 , KMnO 4 , KRuO 4 , K 2 RuO 4 , LiOCl, lead acetate, tetrapropylammonium periodate, chromic acid or salts of the latter, NaBiO3,Ph3BiCO3, Ca(OCl) 2 , the reagents Ce(IV), Cr(VI), the salts of Co (II), IOAc, I(OAc)3, N-iodosuccinimide, VO(OAc), Pb(OAc) 4 , Mn
  • the oxidizing chemical agent is a periodate salt.
  • Said product released directly or indirectly, can be a volatile molecule or an active substance that can modulate a function such as in particular a pharmacological substance or else a detectable compound.
  • Volatile molecule one or more molecules by themselves or in combination that have the property of being volatile, in particular any aldehyde-type or volatile ketone-type molecule, such as, for example, numerous perfumes or numerous bioactive molecules such as the pheromones.
  • Active substance one or more molecules by themselves or in combination that can modulate a function such as in particular a pharmaceutical substance that can modulate, for example, a hormonal action, a physiological action (blood pressure, mood, waking state), or a malignant development (tumor).
  • the active substances can also correspond to molecules that have properties that can cure or prevent a pathology.
  • the active substances can also have an antibiotic or insecticide effect or exhibit specific properties of odor or taste.
  • Specific compound one or more compounds that can exhibit by themselves or in combination a variation of their biophysical, biological or chemical properties before and after the chemical oxidation stage, such as in particular a spectral-type variation or a variation of the solubility.
  • Chemical oxidation any chemical reaction that is carried out in the presence of a chemical reagent that can oxidize the product of Formula (II′) and that induces a cleavage of the bond between carbons C1 and C2.
  • Functional group any chemical group that belongs to a class of organic compounds that is characterized by chemical properties.
  • a functional group it is possible to cite: amides, acyls, alkoxy, nitrites, aryls, heteroaryls, alkenyls, carbonyls, thiocarbonyls, carboxyls, thiocarboxyls, carbamyls, thiocarbamyls, thiocarbamides, alcohols, thiols, or amines that may or may not be substituted.
  • Direct release the release of product that is obtained immediately after said chemical oxidation reaction.
  • Indirect release the release of the product that is obtained after one or more secondary reactions, in particular a chemical reaction, after said chemical oxidation reaction.
  • the secondary reaction can correspond, by way of example, to a beta-elimination reaction.
  • Indirect releases are also those that result from the exposure of a product that is obtained immediately after said chemical oxidation reaction to irradiation or to the action of enzymes that are more or less specific. These secondary reactions can allow in particular the monitoring of the release of said product.
  • the released product corresponds to the Formula
  • the released product corresponds to the product of at least one secondary reaction that is carried out on at least one compound of Formula
  • R′ 7 to R′ 10 , X, and Y have the same meanings as above.
  • detectable product that corresponds to a detectable volatile molecule, to a detectable active substance or else to a detectable specific compound.
  • the compound of Formula (II′) makes it possible to release, after the chemical oxidation, respectively at least one volatile molecule, at least one active substance or at least one specific compound.
  • the pinacol groups or the vicinal diol groups are not sensitive to the oxidation of air and are not volatile. They can therefore be used as stable derivatives of the volatile molecule, which will be released in particular in a controlled manner by exposure of said derivatives to an oxidizing agent, such as periodate.
  • the method of the invention can be used to implement an atomizer of a volatile molecule, and said volatile molecule will be released gradually from a substrate by a chemical oxidation of the latter.
  • the method of the invention can be used to release a galenical form of a medication.
  • the invention therefore also has as its object a composition that comprises a compound of Formula (II′) from which it is possible to release a product directly or indirectly by the method of the invention.
  • Such a composition can be useful, by way of example, for the preparation of an atomizer of a volatile molecule, comprising a mixture of at least one compound of Formula (II′) and an oxidizing agent such as periodate.
  • the composition of the invention comprises an inert solid matrix or a galenical form of a medication.
  • the volatile molecule can be a perfume or a bioactive molecule such as a pheromone. Respectively a perfume atomizer or a mosquito-repellant sprayer is then considered.
  • the volatile molecule is selected from among the group that comprises: benzaldehyde (artificial almond), butanal, citronellal, anisaldehyde, menthone, cuminaldehyde, salicylaldehyde (used in perfumery), vanillin, phenylacetaldehyde (hyacinth) or isovaleraldehyde (lemon, mint, eucalyptus).
  • the active substance as defined in the invention is selected from among the group that comprises: testosterone, estrone or nicotine.
  • the method for releasing the product according to the invention can also comprise a preliminary stage for preparing the compound of Formula (II′) by any technique that is known to one skilled in the art.
  • the invention has as its object a method for releasing at least one product that comprises the following stages:
  • Preparation stage (a) can comprise at least one chemical transformation and/or at least one enzymatic transformation.
  • the preparation of the compound of Formula (II′) can be done, for example, by obtaining the reduced form of the ketone or aldehyde function of at least one of said products of Formula
  • stage (a) can be, for example, a reaction for dimerization of at least one volatile molecule, in particular an aldehyde or a ketone.
  • reaction for dimerization of stage (a) of the method for releasing a product of the invention can be carried out by biosynthesis of vicinal diols.
  • Such reactions for dimerization of a product to obtain a stable derivative are, for example, those described in FIG. 7, attached, in which:
  • a first dimerization reaction according to stage (a) is a reducing dimerization of aldehyde 1 or ketone 2 of the released product in the presence of zinc, so as to obtain a pinacol group.
  • a second dimerization reaction according to stage (a) is carried out in two stages, by olefination of carbonyl in the presence of a phosphorated ylide, followed by a dihydroxylation by osmium tetroxide.
  • This second approach leads to a terminal diol that, after oxidation by the sodium periodate, releases 1 equivalent of formaldehyde.
  • the first two examples correspond to direct releases of the active substance and the last example to an indirect release in which the oxidation reaction is followed by a beta elimination.
  • the released product is a detectable product; it can then be used to detect a chemical transformation.
  • the compound of Formula (II′) is a compound of Formula (II)
  • X and Y which are identical or different, are selected from among an oxygen atom, a sulfur atom, an amine of Formula -NR 11 R 12 , and R 11 is selected from among: a hydrogen atom, an alkyl group, an aryl group that may or may not be substituted, and R 12 is not a hydrogen atom,
  • R 7 to R 10 which are identical or different, are selected from among a hydrogen atom, an alkyl group that may or may not be substituted, or a functional group that may or may not be substituted, and at least one of groups R 7 to R 10 is a group Di, which is insensitive to the stage (b) chemical oxidation reaction and which exhibits detectable properties, directly or indirectly, alone or in combination, after the cleavage of the bond C 1 -C 2 .
  • these properties can be biophysical, biological or chemical, such as in particular a spectral-type variation or a solubility variation.
  • the compound of Formula (II) corresponds to a particular implementation of the compound of Formula (II′), thus the statements made regarding the compound of Formula (II′) are valid for the compound of Formula (II).
  • the detectable product can then be released after having carried out a first chemical transformation stage of a substrate to obtain a compound of Formula (II) then a second chemical oxidation stage of said compound of Formula (II).
  • the detection of this transformation is obtained by using the method for releasing a detectable product or by carrying out a chemical transformation of a substrate that produces the compound of Formula (II), then by chemically oxidizing the compound of Formula (II) to release the product that can be detected.
  • the release of the detectable product comprises the following stages:
  • At least one of groups R 1 to R 6 is a group Di as defined above,
  • Groups R 1 to R 6 which are identical or different, are selected from among: a hydrogen atom, an alkyl group that may or may not be substituted, a functional group that may or may not be substituted, and in the case of a functional group of Formulas OR 12 , —SR 12 , and -NR 11 R 12 , R 11 is selected from among: a hydrogen atom, an alkyl group, an aryl group that may or may not be substituted, and R 12 is not a hydrogen atom,
  • Group or groups R 1 to R 6 that form after stage (a) the groups of Formulas -XH and -YH are insensitive to stage (b) oxidation or can be sensitive to this oxidation if the corresponding compound of Formula (I) does not produce the compound of Formula (II) during an oxidation of the type of that of stage (b),
  • X and Y which are identical or different, are selected from among an oxygen atom, a sulfur atom, or an amine of Formula -NR 11 R 12 ,
  • R 7 to R 10 which are identical or different, are either identical to at most four of the groups R 1 to R 6 , or, because of the transformation of one or more of groups R 1 to R 6 , during the reaction of stage (a), selected from among a hydrogen atom, an alkyl group that may or may not be substituted, or a functional group that may or may not be substituted, and at least one of groups R 7 to R 10 is a group Di as defined above.
  • stage (b) the method of the invention obviously comprises a stage for direct or indirect detection of the detectable product to carry out, for example, the detection of the chemical transformation of stage (a).
  • the method of the invention accepts several preferred meanings of groups R 1 to R 6 , among which the following cases are considered more particularly:
  • R 1 to R 6 are selected so that the bond C 1 -C 2 is part of at least one cycle
  • At least one pair of groups R 1 to R 6 together form an oxygen atom, a sulfur atom, or a group of Formula -NR 11 R 12 ,
  • At most one pair of groups R 1 to R 6 one of which is selected from among R 1 , R 2 , and R 6 , and the other is selected from among R 3 , R 4 and R 5 form a double bond between carbons C 1 and C 2 ,
  • At most two pairs of groups R 1 to R 6 one of which is selected from among R 1 , R 2 and R 6 , and the other is selected from among R 3 , R 4 and R 5 , form a triple bond.
  • FIG. 1 A diagram example for detecting a chemical transformation of a substrate according to the invention is shown in FIG. 1, attached, whereby the compounds of Formulas (III) and (IV) are directly or indirectly detectable.
  • An advantageous implementation of the method for releasing a detectable product comprises the following stages:
  • Groups R′ 1 , to R′ 6 which are identical or different, are selected from among: a hydrogen atom, an alkyl group that may or may not be substituted, a functional group that may or may not be substituted, and in the case of a functional group of Formulas —OR 12 , —SR 12 , and -NR 11 R 12 , R 11 is selected from among: a hydrogen atom, an alkyl group, an aryl group, that may or may not be substituted, and R 12 is not a hydrogen atom,
  • Group or groups R′ 1 to R′ 6 that form after stage (a) the groups of Formulas -XH and -YH are insensitive to stage (b) oxidation or can be sensitive to this oxidation if the corresponding compound of Formula (I′) does not produce the compound of Formula (II′) during an oxidation of the type of that of stage (b),
  • X and Y which are identical or different, are selected from among an oxygen atom, a sulfur atom, an amine of Formula -NR 11 R 12 , and R 11 is selected from among: a hydrogen atom, an alkyl group, an aryl group, that may or may not be substituted, and R 12 is not a hydrogen atom,
  • R′ 7 to R′ 10 which are identical or different, are either identical to at most four of groups R′ 1 to R′ 6 , or, because of the transformation of one or more of groups R 1 to R 6 , during the reaction of stage (a), selected from among a hydrogen atom, an alkyl group that may or may not be substituted, or a functional group that may or may not be substituted, and in the compound of Formula (II):
  • R 7 to R 10 which are identical or different, correspond to a hydrogen atom, an alkyl group that may or may not be substituted, a functional group that may or may not be substituted, and at least one of groups R 7 to R 10 is a group Di as defined above,
  • X and Y which are identical or different, are selected from among an oxygen atom, a sulfur atom, an amine of Formula -NR 11 R 12 , and R 11 is selected from among: a hydrogen atom, an alkyl group, an aryl group, that may or may not be substituted, and R 12 is not a hydrogen atom.
  • This advantageous implementation puts the compound of Formula (II′), obtained from the chemical transformation of the compound of Formula (I′), into competition with the compound of Formula (II) during oxidation stage (b).
  • the oxidation speed of the compound of Formula (IT) will therefore decrease proportionately to the amount of compound of Formula II′ that is formed.
  • the signal that is associated with the transformation of the compound of Formula (II), detectable thanks to the presence of a group Di that is a precursor of a detectable product, will therefore also decrease proportionately to the amount of compound of Formula (II′) that is produced.
  • the substrate of Formula (I′) can correspond to the non-modified specific substrate of the chemical transformation such as, for example, the non-modified specific substrate of an enzyme.
  • the method of the invention also accepts a modification of one of groups R 7 to R 10 except for Di, by the chemical oxidizing agent of stage (b), provided that this disturbs the cleavage of the bond C 1 -C 2 neither during stage (b) nor during the subsequent detection.
  • stage (a) and (b) are simultaneous, group or groups R 1 to R 6 are insensitive to stage (b) oxidation.
  • stages (a) and (b) are not simultaneous.
  • Groups R 1 to R 6 then may or may not be sensitive to the oxidation of stage (b).
  • Transformation reaction (a) can take place under experimental conditions that may or may not be similar to the oxidation reaction (b).
  • stages (a) and (b) are not simultaneous, a reaction speed that is proportional to the amount of accumulated product will be obtained.
  • the method of the invention can be used with a substrate that comprises several chemical groups that can be transformed in stage (a) and several chemical groups that are precursors of detectable products.
  • the method of the invention can be carried out in a suitable reaction medium that is selected from among an aqueous, organic, two-phase or solid medium.
  • chemical transformation or chemical reaction means any transformation of a substrate that may or may not be spontaneous and that can call for particular experimental conditions such as heat, UV, etc.
  • the reaction can call for a chemical reagent or a catalyst such as an enzyme.
  • This chemical transformation of stage (a) can be carried out with several sequential reactions that may or may not be simultaneous.
  • Groups R 1 to R 10 can consist of or comprise an isotope, such as, for example, deuterium.
  • groups R 1 to R 10 are stable in a reaction medium, in particular an aqueous, organic, two-phase, or solid medium, etc.
  • the substrate that is used in the process of the invention comprises one or more chiral centers.
  • the substrate that is used in the process of the invention can therefore consist of a mixture of enantiomers, or diastereomers or enantiomers that are pure.
  • a first class of substrate (I) corresponds to Formulas (V) to (IX) below:
  • Di is a group as defined above
  • R 1 to R 3 and X and Y have the same meanings as above,
  • P 1 and P 2 are a hydrogen atom.
  • P 1 and P 2 which are identical or different, are selected from among an acyl group that is substituted by an aryl or alkyl or peptidyl group, a phosphate group, a phosphate ester group, a phosphonate group, a carbamyl group that is substituted by an aryl or acyl or peptidyl group, a glycosyl group and a sulfate group,
  • P 3 is selected from among a carbonyl group, a group —PO 2 R 11 or a group R 11 PO—, where R 11 has the same meaning as above, a group —SO 2 , a group —CHOR 13 where R 13 represents an aryl, alkyl or glycosyl group, a group SiR 14 R 15 where R 14 and R 15 , which are identical or different, represent an aryl, alkyl, aryloxy or alkoxy group, and a group AsO 2 H—,
  • G is selected from among an oxygen atom, a sulfur atom, or an amine group of Formula NR 11 R 12 where R 11 has the same meaning as above.
  • groups X and Y of the compound of Formula (XV) are hydroxyl groups.
  • the detection of the chemical transformation according to the invention is used either by a direct revelation or by an indirect revelation of the compound that is obtained after the chemical oxidation of the compound of Formulas (II) or (X) carried out with an oxidizing chemical agent according to stage (b).
  • Tetrapropyl ammonium periodate soluble in organic phase, is advantageously used when the method of the invention is carried out in an organic medium.
  • direct revelation according to which the compound that corresponds to Formula (XI) obtained after the chemical oxidation reaction that is carried out on the derivative that corresponds to Formulas (II) or (X) after stage (b) is directly detectable according to the diagram below.
  • An embodiment of the invention with a direct revealing uses the direct detection of a property of the compound of Formula (XI), not present in the compounds of Formulas (I) or (II) or (X).
  • Di in the compound of Formula (X) is a precursor of a detectable product
  • compound (XI) a physical property, such as solubility; a physicochemical property, such as a spectral property; or a biological property, such as the activation of an enzyme, an odor, or the action of a pheromone.
  • a physical property such as solubility
  • a physicochemical property such as a spectral property
  • a biological property such as the activation of an enzyme, an odor, or the action of a pheromone.
  • the molecule that corresponds to Formula (XI) can correspond to aromatic ketones, for example a beta-aromatic ketone that is detected by a spectral variation, an aldehyde such as benzaldehyde or citronellal that is detected by odor, or to a pheromone that is detected by the attraction of insects.
  • aromatic ketones for example a beta-aromatic ketone that is detected by a spectral variation, an aldehyde such as benzaldehyde or citronellal that is detected by odor, or to a pheromone that is detected by the attraction of insects.
  • R 16 , R 17 and R 18 which are identical or different, represent a hydrogen atom, an alkyl group that may or may not be substituted or a functional group that may or may not be substituted, and
  • Z is a precursor of detectable product ZH.
  • Di in the compounds of Formulas (X) and (XII) is the precursor of said detectable product that will be released after a secondary beta-elimination reaction.
  • the detection of a property of product ZH that is obtained by a beta-elimination reaction that is carried out on the compound of Formula (XII) that is obtained after chemical oxidation stage (b) is carried out.
  • This beta-elimination reaction advantageously spontaneous, is preferably carried out in the presence of a base called B that can correspond to bovine serum albumin (BSA).
  • B bovine serum albumin
  • Compound ZH is selected from among an aromatic alcohol, a heteroaromatic alcohol, a heteroaromatic amine, a halogen atom, or a phosphoric ester.
  • an aromatic alcohol a heteroaromatic alcohol, a heteroaromatic amine, a halogen atom, or a phosphoric ester.
  • fluorescein phenolphthalein, phenol red, p-nitrophenol, o-nitrophenol, 2,4-dinitrophenol, 6-hydroxynaphthoic acid, 8-hydroxy-pyrene 1,3,6-trisulfonic acid, tyrosine, luciferin, indolyl, 5-bromo-4-chloro-indolyl, quinolinium, nitro-anilinium or pyridoxamine.
  • a non-limiting example of interaction between the two groups Di is a FRET-type energy transfer.
  • FIG. 2 A diagram that shows this revelation using a FRET-type energy transfer is shown in FIG. 2, attached.
  • Examples of substrates that allow the implementation of the invention with a direct revelation are compounds of Formula (V) in which groups X, Y, R 1 , R 2 , R 3 and Di are selected from among those that are described in the following table.
  • Examples of substrates that allow the implementation of the invention with an indirect detection are compounds of Formula (V) in which groups X, Y, R 11 , R 12 , R 13 as well as groups R 1 , R 2 , R 3 and ZH are selected from among those that are described in Table II below. TABLE II Di No.
  • the diamines can be mono- or disubstituted, whereby groups P 1 and P 2 cannot both correspond to H.
  • P 1 and/or P 2 can correspond to:
  • the diols can be mono- or disubstituted.
  • hydroxyl radicals can be monofunctionalized as above, and in addition as follows:
  • R′′′′′ 5′ end of a DNA or RNA chain
  • stage (b) the revelation of stage (b) is carried out sequentially by using substrates that are at least disubstituted such as the disubstituted substrate of Formula XVII below:
  • a preferred embodiment of the method of the invention that uses a substrate that corresponds to Formula (V) above comprises the transformation of mono- or diester derivatives of diols (1 a/b) by a lipase or an esterase to produce compound (2) according to the diagram below.
  • Another preferred embodiment of the method of the invention that uses a substrate that belongs to the second class above comprises the transformation of mono- or diester derivatives of diols (3 a/b) by a lipase or an esterase to produce compound (4) according to the diagram below.
  • Coum and NP have the same meanings as above.
  • a preferred embodiment of the method of the invention that uses a substrate that corresponds to Formula (VII) above comprises the transformation of epoxide (7) by an epoxide esterase to produce compound (2) according to the diagram below.
  • Another preferred embodiment of the method of the invention that uses a substrate that corresponds to Formula (VII) above comprises the transformation of epoxide (8) by an epoxide hydrolase to produce compound (4) according to the diagram below.
  • a preferred embodiment of stage (a) of the method of the invention that uses a substrate according to Formula (VIII) above comprises the transformation of olefin (9) by a dihydroxylase or a chemical catalyst such as an alkaloid or an amino alcohol in the presence of OsO4 to produce compound (2) according to the diagram below:
  • stage (a) of the method of the invention that uses a substrate according to Formula (VIII) above comprises the transformation of olefin (10) by a dihydroxylase to produce compound (4) according to the diagram below.
  • the invention also relates to the substrates that correspond to Formulas (V), (VI), (VII), (VIII) or (IX).
  • these substrates are stable in the reaction medium, in particular in the very wide ranges of pH and temperature, but also in very diverse solvents.
  • substrates that can be used in the method of the invention are shown in Table XI, attached.
  • the invention also has as its object a composition that comprises at least one compound of Formula (II′).
  • This compound of Formula (II′) can correspond to a form that is stabilized by a volatile molecule, an active substance or a specific compound.
  • composition according to the invention comprises an oxidizing agent.
  • the method of the invention is noteworthy in that it makes it possible to detect a chemical transformation that is optionally present in a sample by selecting a substrate that is the most suitable for the transformation that it is desired to analyze.
  • these substrates have various degrees of specificity. These degrees of specificity are provided by the structure of one or more groups R 1 to R 6 of Formula (I) or R 1 to R 3 , or G, or P 1 to P 3 of Formulas (V), (VI), (VII), (VIII) or (IX).
  • group or groups R 1 to R 6 are selected so that the substrates that correspond to Formula (I) are close to the structure of the specific substrate. In particular, they can correspond to a fatty acid chain.
  • group or groups R 1 to R 6 are selected so that the substrates that correspond to Formula (I) are close to the structure of the specific substrate. In particular, they can correspond to a fatty acid chain.
  • One skilled in the art will know, based on the type of chemical transformation to be detected, to adapt the selection of group or groups R 1 to R 6 .
  • the substrate of Formula (I) can correspond to the non-modified specific substrate of the enzyme, which represents an advantage in the case of use of the method of the invention for the identification of new catalysts or of the new activity of known catalysts that can transform a substrate of Formula (I) or (I′) respectively into a compound of Formula (II) or (II′).
  • the method of the invention makes it possible to identify an enantioselective or stereoselective chemical transformation in a sample.
  • the chemical transformation is carried out with a catalyst and in particular an enzyme.
  • an enzyme that is selected from among: lipase, esterase, protease, glycosidase, glycosyl transferase, phosphatase, kinase, mono- or dioxygenase, haloperoxidase, lignin peroxidase, diarylpropane peroxidase, epoxide hydrolase, nitrile hydratase, nitrilase, transaminase, amidase, acylase, dihydroxylase, phytase, xylanase, nuclease, and reductase.
  • the invention therefore finally relates to a process for detecting and/or quantifying a known chemical transformation in a sample that consists of using the method for detecting a chemical transformation described above in the presence of said sample and a substrate that is suitable for the desired activity.
  • the invention also makes it possible to detect the substrate of Formula (I) or (I′) in the presence of a chemical or biochemical catalyst reagent.
  • the method of the invention can be used for screening, particularly at a high flow rate, a catalyst from an in vivo or in vitro expression library.
  • These libraries can be prepared, for example, from microorganisms or microalgae that preferably have extremophile properties.
  • the method of the invention can also be used to identify catalysts that have a different activity relative to a starting activity. These catalysts will be, for example, products of directed mutagenesis or directed evolution.
  • a particular embodiment of the method of the invention relates to the case where the substrate of Formula (I) or (I′) is itself the product of a first unknown chemical transformation, whose activity is desired to be identified.
  • the method of the invention is noteworthy in that it also makes possible the identification and the isolation of new chemical or biochemical catalysts that can transform a substrate of Formula (I) or (I′) respectively into a compound of Formula (II) or (II′).
  • the method of the invention makes it possible to disclose the presence of a catalyst that, thanks to chemical oxidation, generates a signal that is directly or indirectly detectable.
  • the invention also has as its object the use of the method for detecting a chemical transformation, as described above, for the identification of new catalysts, or the new activity of a known catalyst.
  • the invention also relates to a catalyst that can be identified by the method of the invention.
  • FIG. 3 represents the enzymatic activity of the Candida antarctica lipase based on the temperature.
  • the enzymatic reactions were carried out in the PIPES buffer (0.1 M, pH 7) at 50, 60, 70, 80 and 95° C. for 40 minutes by using 2-hydroxy-4-p-nitrophenyl-butyl-decanoate.
  • FIG. 4 illustrates the enzymatic activity of the Candida antarctica lipase based on the temperature.
  • the enzymatic reactions were carried out in the PIPES buffer (0.1 M, pH 7) at 50, 60, 70, 80 and 95° C. for 40 minutes by using para-nitrophenylbutyrate.
  • FIG. 5 illustrates the activity of the Candida antarctica lipase based on the pH of the solution. The tests were carried out by using 2-hydroxy-4-p-nitrophenyl-butyl-decanoate.
  • FIG. 6 illustrates the activity of the Candida antarctica lipase based on the pH of the solution. The tests were carried out by using para-nitrophenolbutyrate.
  • a suspension of 7-hydroxycoumarin (3 g, 19.6 mmol) in DMF (20 ml) is treated with NaH (60% suspension in oil, 1.03 g, 25.9 mmol), then with 4-bromo-1-butene (2.65 g, 19.6 mmol). It is heated to 60° C. for 24 hours, then it is diluted with ethyl acetate (400 ml), and it is washed with water (3 ⁇ 300 mL) then with 1 M NaOH (3 ⁇ 200 ml). The residue is chromatographed with the mixture (ethyl acetate-hexane) to obtain 4-(7-coumarinyloxy)-1-butene (2.7 g, 67%).
  • AD-alpha-mix suspension (4.2 g) is stirred in 30 ml of t-BuOH/water 1:1 until a single phase is obtained (5 minutes). It is then cooled to 0° C., and olefin is added (0.648 g, 3 mmol). After 16 hours at 0° C., it is quenched with Na 2 S 2 O 5 (4.5 g). After one hour at ambient temperature, it is extracted with CH 2 Cl 2 and chromatographed (CH 2 Cl 2 /acetone elution 7:3) to obtain 0.6 g (80%) of pure diol (S).
  • Acetyl chloride (0.36 mmol, 0.024 ml) is added to a solution of (S)-4-(7-coumarinyloxy)-1,2-butanediol (0.090 g, 0.36 mmol) and triethylamine (0.1 ml, 0.72 mmol) in 8 ml of dry CH 2 Cl 2 . After 40 minutes at 0° C., it is diluted with CH 2 Cl 2 , and it is washed with an aqueous solution of NaHCO 3 . The chromatography of the residue after evaporation provides monoacetate S (0.058 g, 0.22 mmol, 60%).
  • 6-Methoxy-2-naphthaldehyde is prepared by successive treatment of 2-bromo-6-methoxynaphthalene in solution in dry ether with n-butyllithium and then dimethylformamide. (Literature: J. Med. Chem. 1998, 1308-1312). 6-Dimethylamino-2-naphthaldehyde is prepared according to the described procedure (Barbas et al., Proc. Natl. Acad. Sci. USA 1998, 95, 15351) as follows: gaseous dimethylamine is introduced into a mixture of 2.4 ml of dry benzene and 2.4 ml of hexamethylphosphoramide (HMPA) until 750 mg (16.7 mmol) is dissolved.
  • HMPA hexamethylphosphoramide
  • n-butyllithium (1.6 M in hexane, 16.7 mmol) is added, then after 15 minutes, 6-methoxy-naphthaldehyde (390 mg, 2.09 mmol) is added. Stirring is continued for 14 hours at 20° C., then it is poured into aqueous phosphate buffer, pH 7.4, and it is extracted with ether. Purification by chromatography provides 6-dimethylamino-2-naphthaldehyde (350 mg, 84%).
  • 6-Methoxy-naphthaldehyde (432 mg, 2.35 mmol) is added to a solution of instantaneous ylide (Ph 3 P + Br ⁇ +NaNH 2 , 1.48 g, 2.4 mmol) in anhydrous THF (5 ml). After one hour of stirring at ambient temperature, it is diluted with ether, it is filtered on Celite, and it is chromatographed (hexane/AcOEt 5:1) to obtain 6-methoxy-2-vinyl-naphthalene (390 mg, 90%).
  • Diesters (R)- (S) as well as monoesters (R)- and (S)-represented by Formula (I) constitute excellent substrates for lipases and esterases given their structural similarity with glycerides.
  • the amides that are shown by Formula I (compound 5) react with the amidases and peptidases.
  • Epoxides (R)- and (S)- that are represented by Formula (II) (compounds 7 and 8) react with the epoxy-hydrolase enzymes. The applicant thus found that the transformation undergone by these fluorogenic substrates transformed by sodium periodate, in the presence of hydrolytic enzymes, provides a new method for detecting said enzymes.
  • the substrates acetates 1, amide 5 and epoxide 6 are stable in the presence of periodate and BSA without the aqueous media. Under the same conditions, diol 2 and amino alcohol 6 release umbelliferone. Under optimized reaction conditions using 100 ⁇ m of substrate, 1 mmol of sodium periodate, 2 mg/ml of BSA in a borate buffer with a pH of 8.8, diol 2 is transformed into umbelliferone with a yield of 72% without observable secondary reactions. The oxidation yield is independent of the presence of BSA, which indicates that these reagents do not interfere with one another in a notorious manner. The oxidation of amino alcohol 6 takes place with a yield of 85% under the same conditions.
  • Acetates 1, amide 5 and epoxides (R)-7 and (S)-7 were tested in the presence of corresponding hydrolytic enzymes.
  • the acetates were tested compared to 25 different esterases and lipases.
  • the epoxides were tested compared to the epoxy-hydrolase extracted from Aspergillus Niger (X-J. Chen, A. Archelas, R. Furstoss, J. Org. Chem. 1993, 58, 5528) and Rhodotorula glutinis (C. A. G. M. Weijers, A. L. Botes, M. S. van Dyk, J. A. M. de Bont, Tetrahedron Asymm. 1998, 9, 467).
  • Phenylacetamide 5 was tested against penicillin G acylase, chymotrypsin and papain. An increase of fluorescence that is clearly dependent on time is observed when the enzymatic activity is present. The analyses that are carried out by high-pressure liquid chromatography (HPLC) confirm that the increase of fluorescence is caused by the release of umbelliferone from substrates.
  • the hydrolytic activity of the Candida antarctica lipase was tested on 2 substrates: 2-hydroxy-4-p-nitrophenyl-butyl-decanoate and p-nitro-phenylbutyrate.
  • the lipase activity is detected after hydrolysis of ester functions of substrates that produce the release (direct or indirect) of yellow-colored para-nitrophenol that is measured by colorimetry at 414 nm. The tests were carried out at different temperatures and pH.
  • V.1.1 Measurement of Lipase Activity with 2-Hydroxy-4-p-nitrophenyl-butyl-decanote.
  • the activity of the lipase based on the incubation temperature was measured by using 2-hydroxy-4-p-nitrophenyl-butyl-decanoate and the para-nitrophenylbutyrate commercial substrate.
  • Table VIII shows the measurements of the activity of the Candida antarctica lipase that are made by using 2-hydroxy-4-p-nitrophenyl-butyl-decanoate and para-nitrophenylbutyrate. The activities are expressed in %.
  • the enzymatic reactions were carried out in the PIPES buffer (0.1 M, pH 7) at 50, 60, 70, 80 and 95° C. for 40 minutes. A negative control without enzyme was also carried out by using the same experimental conditions.

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US10238767B2 (en) * 2012-07-25 2019-03-26 Dsm Ip Assets, B.V. Mono-layer thin film adhesive compounds and methods of synthesis and use

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FR2809820B1 (fr) 2000-05-30 2002-10-25 Proteus Methode de detection d'une transformation d'un substrat et ses applications
FR2832729B1 (fr) 2001-11-28 2004-01-16 Proteus Methode de detection d'une activite catalytique d'un echantillon mettant en oeuvre la detection de la transformation d'un substrat
FR2847590B1 (fr) * 2002-11-27 2007-05-18 Proteus Methode et kit pour le dosage specifique de ribonucleosides
WO2013044118A2 (en) * 2011-09-23 2013-03-28 Merial Limited Indirect modeling of new repellent molecules active against insects, acarids, and other arthropods

Family Cites Families (15)

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FR2341865A1 (fr) * 1976-02-19 1977-09-16 Api Labor Support pour le dosage d'activites enzymatiques et procede de dosage a l'aide de ce support
EP0018825B1 (en) * 1979-05-02 1985-02-13 National Research Development Corporation A process for the identification of bacteria and a kit of reagents for use in this process
US4778757A (en) * 1982-04-08 1988-10-18 Toyo Boseki Kabushiki Kaisha Method for the determination of substrates or enzyme activities
JPH07114709B2 (ja) * 1987-11-13 1995-12-13 協和メデックス株式会社 酵素活性の定量法
US5122602A (en) * 1989-02-13 1992-06-16 Miles Inc. Chromogenic merocyanine enzyme substrates
US5162203A (en) * 1989-09-07 1992-11-10 President And Fellows Of Harvard College Methods of measuring isozymes and isozyme classes of alcohol dehydrogenase
CA2038406C (en) 1990-04-12 1995-11-21 Mark L. Sussman Identifying microorganisms by measuring enzymatic activity in the presence of enzyme activity affecting agents
US5338843A (en) * 1992-01-30 1994-08-16 Becton, Dickinson And Company Fluorogenic and chromogenic β-lactamase substrates
JP3885900B2 (ja) * 1996-03-22 2007-02-28 大日本インキ化学工業株式会社 ベンゾピラン誘導体及びそれを有効成分とする心臓疾患治療剤
US5846754A (en) 1996-05-28 1998-12-08 Bayer Corporation Enzyme determination with protease inactivation
WO2001036662A2 (en) 1999-11-05 2001-05-25 Iconix Pharmaceuticals, Inc. Hydrolytic enzyme substrates and assay method
AU2001238407A1 (en) 2000-02-16 2001-08-27 Thermogen, Inc. Esterase enzymes having selective activity
US20020058329A1 (en) 2000-02-18 2002-05-16 Sharat Singh Multiple-site reaction device and method
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