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WO2001012727A1 - Détection de composés polyhydroxylés - Google Patents

Détection de composés polyhydroxylés Download PDF

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Publication number
WO2001012727A1
WO2001012727A1 PCT/GB2000/002685 GB0002685W WO0112727A1 WO 2001012727 A1 WO2001012727 A1 WO 2001012727A1 GB 0002685 W GB0002685 W GB 0002685W WO 0112727 A1 WO0112727 A1 WO 0112727A1
Authority
WO
WIPO (PCT)
Prior art keywords
compound
formula
test sample
optionally substituted
polyhydroxyl
Prior art date
Application number
PCT/GB2000/002685
Other languages
English (en)
Inventor
Prakash Patel
Christopher James Ward
Tony David James
Original Assignee
Avecia Limited
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 Avecia Limited filed Critical Avecia Limited
Priority to EP00948109A priority Critical patent/EP1210393A1/fr
Priority to JP2001517617A priority patent/JP2003507390A/ja
Priority to AU61682/00A priority patent/AU6168200A/en
Publication of WO2001012727A1 publication Critical patent/WO2001012727A1/fr

Links

Classifications

    • G—PHYSICS
    • G01—MEASURING; TESTING
    • G01N—INVESTIGATING OR ANALYSING MATERIALS BY DETERMINING THEIR CHEMICAL OR PHYSICAL PROPERTIES
    • G01N33/00—Investigating or analysing materials by specific methods not covered by groups G01N1/00 - G01N31/00
    • G01N33/48—Biological material, e.g. blood, urine; Haemocytometers
    • G01N33/50—Chemical analysis of biological material, e.g. blood, urine; Testing involving biospecific ligand binding methods; Immunological testing
    • G01N33/66—Chemical analysis of biological material, e.g. blood, urine; Testing involving biospecific ligand binding methods; Immunological testing involving blood sugars, e.g. galactose
    • C—CHEMISTRY; METALLURGY
    • C09—DYES; PAINTS; POLISHES; NATURAL RESINS; ADHESIVES; COMPOSITIONS NOT OTHERWISE PROVIDED FOR; APPLICATIONS OF MATERIALS NOT OTHERWISE PROVIDED FOR
    • C09B—ORGANIC DYES OR CLOSELY-RELATED COMPOUNDS FOR PRODUCING DYES, e.g. PIGMENTS; MORDANTS; LAKES
    • C09B23/00—Methine or polymethine dyes, e.g. cyanine dyes
    • C09B23/14—Styryl dyes
    • C09B23/143—Styryl dyes the ethylene chain carrying a COOH or a functionally modified derivative, e.g.-CN, -COR, -COOR, -CON=, C6H5-CH=C-CN
    • C—CHEMISTRY; METALLURGY
    • C09—DYES; PAINTS; POLISHES; NATURAL RESINS; ADHESIVES; COMPOSITIONS NOT OTHERWISE PROVIDED FOR; APPLICATIONS OF MATERIALS NOT OTHERWISE PROVIDED FOR
    • C09B—ORGANIC DYES OR CLOSELY-RELATED COMPOUNDS FOR PRODUCING DYES, e.g. PIGMENTS; MORDANTS; LAKES
    • C09B29/00—Monoazo dyes prepared by diazotising and coupling
    • C09B29/06—Monoazo dyes prepared by diazotising and coupling from coupling components containing amino as the only directing group
    • C09B29/08—Amino benzenes
    • C09B29/0805—Amino benzenes free of acid groups
    • C09B29/0807—Amino benzenes free of acid groups characterised by the amino group
    • C09B29/0809—Amino benzenes free of acid groups characterised by the amino group substituted amino group
    • C09B29/0811—Amino benzenes free of acid groups characterised by the amino group substituted amino group further substituted alkylamino, alkenylamino, alkynylamino, cycloalkylamino aralkylamino or arylamino
    • C—CHEMISTRY; METALLURGY
    • C09—DYES; PAINTS; POLISHES; NATURAL RESINS; ADHESIVES; COMPOSITIONS NOT OTHERWISE PROVIDED FOR; APPLICATIONS OF MATERIALS NOT OTHERWISE PROVIDED FOR
    • C09B—ORGANIC DYES OR CLOSELY-RELATED COMPOUNDS FOR PRODUCING DYES, e.g. PIGMENTS; MORDANTS; LAKES
    • C09B69/00—Dyes not provided for by a single group of this subclass

Definitions

  • the present invention relates to novel compounds, to a method for detecting the presence and/or amount of polyhydroxyl compounds and to a kit for detecting the presence and/or amount of polyhydroxyl compounds. .
  • Glucose-sensors are commonly based on an enzyme such as glucose oxidase.
  • the glucose oxidase method is based on oxidizing glucose with oxygen and measuring the amount of resultant hydrogen peroxide produced by an appropriate means (such as by an electrode). These methods suffer since enzymes can irreversibly change in quality with time and cannot easily be recycled for repeated use. In addition the sample often cannot be provided for other measurement as the glucose or other substrate has been modified in some way during analysis.
  • the present invention provides novel compounds in which a secondary amine is provided in the vicinity of the boronic acid group. These compounds display an unexpectedly large spectral shift on reaction with hydroxyl groups. This makes them particularly suitable for use in the detection of polyhydroxyl compounds.
  • X is a chromophore or a fluorophore
  • W is an optionally substituted aromatic ring; n is 0, 1 or 2; m is 1 or 2; and p is 1 or 2.
  • Preferred chromophores X comprises an optionally substituted azo, disazo, anthraquinone, pyrroline, phthalocyanine, polymethine, aryl-carbonium, triphenodioxazine, diarylmethane, triarylmethane, anthraquinone, phthalocyanine, methine, polymethine, indoaniline, indophenol, stilbene, squarilium, aminoketone, xanthene, fluorone, acridene, quinolene, thiazole, azine, nigrosine, oxazine, thiazine, indigoid quinonioid, quinacridone, lactone, pyrroline, benzodifuranone, or indolene group or a combination of such groups.
  • Preferred fluorophores represented by X are optionally substituted naphthyl, anthryl, pyrenyl, stilbene and phenanthryl or a combination of such groups.
  • X is such that when the compound of Formula (1 ) is dissolved in a solvent the resultant solution is red, orange, yellow, green, blue, indigo or violet. It is preferred that the compound is of Formula (2) or a salt thereof:
  • A-N N-D-(C n H 2n )-NH-(C m H 2m )-E-B(OH) 2
  • A, D and E are each independently an optionally substituted aromatic heterocyclic or homocyclic group and m and n are as hereinbefore defined.
  • n is 0 and m is 1
  • Preferred optionally substituted aromatic heterocyclic groups are diazine, thiazole, benzthiazole, benzdiazine, triazole, isoxazole, benzisoxazole, thiadiazole, oxadiazole, isothiazole, benzisothiazole, pyridiazine, triazine, oxazole, thiophene, benzoxazole, pyrimidine or pyridine.
  • Preferred optionally substituted homocyclic groups are optionally substituted phenyl, naphthyl, pyrenyl, stilbenyl.
  • A is optionally substituted phenyl and D and E are each independently optionally substituted phenylene.
  • the optional substituents which may be present on X, A, D, E and W are each independently selected from alkyl (preferably C ⁇ -alkyl), alkoxy (preferably C, ⁇ - alkoxy), aryl (preferably phenyl), aryloxy (preferably phenoxy), polyalkylene oxide (preferably polyethylene oxide or polypropylene oxide), nitro, cyano, sulpho, halo, ureido, SO 2 F, hydroxy, carboxy, ester; -NR 1 R 2 , -COR 1 , -CONR-R 2 , -NHCOR 1 , phosphato, sulphato, carboxyester, sulphone, and -SO 2 NR 1 R 2 wherein R 1 and R 2 are each independently H or alkyl (especially C 1-4 -alkyl).
  • the optional substituents on X, A, D and E are selected from methyl, ethyl, propyl, butyl, phenyl, methoxy, ethoxy, butoxy, nitro, phenoxy, carboxy, phosphato, sulpho, sulphato, cyano, carboxyester, sulphone, sulphonamide, ureido, SO 2 NR 1 R 2 and NHCOR groups.
  • the compound is of Formula (3) or a salt thereof:
  • R 3 is H, C ⁇ -alkyl, OR 4 , NHCONH 2 , NHCOR 4 , wherein R 4 is H or alkyl.
  • R 4 is H or alkyl.
  • Preferred salts of the compounds described herein are alkali metal salts, especially lithium, sodium and potassium salts, ammonium and substituted ammonium salts and mixtures of the foregoing salts.
  • the compounds in Formula (3) may be prepared by a simple synthetic route in which an optionally substituted 2-formylarylboronic acid is reacted with an aniline (e.g. m-toluidine) to form the respective imine.
  • an aniline e.g. m-toluidine
  • the imine is reduced to the corresponding secondary amine using a sodium borohydride reduction and the resultant compound coupled with the diazonium salt of an optionally substituted aniline to give the desired compound of the invention.
  • a method for detecting the presence and/or amount of a polyhydroxyl compound in a test sample which comprises contacting the test sample with a compound according to the first aspect of the present invention and evaluating any change in the spectral characteristics of the compound.
  • the change in the spectral characteristics of the compound is preferably evaluated visually or, more preferably, using a spectrophotometer, fluorimeter or similar device.
  • the spectral properties of the compounds change, typically in terms of a change in fluorescent properties and/or a bathochromic or hypsochromic shift. There may also be an associated change in the intensity of light absorption.
  • the method is preferably performed at a constant pH preferably in the presence of a pH buffer.
  • Suitable pH buffers are well known in the art, particularly including phosphate, pyrophosphate, acetate, carbonate and citrate and mixed buffers such as citrate/phosphate.
  • the polyhydroxyl compound which may be present in the test sample may be a single compound or a mixture of compounds.
  • Preferred polyhydroxyl compounds contain at least 2 or more hydroxyl groups. Preferably at least two of the hydroxyl groups are vicinal hydroxyl groups.
  • Examples of such polyhydroxyl compounds include saccharides, especially monosaccharides and disaccharides such as glucose, fructose, sucrose, mannose, arabinose, xylose, ribose, galactose, sorbitol.
  • the test sample preferably comprises or is derived from a human or animal body fluid, a plant extract or a microbial fermentation product, more preferably blood or urine.
  • a human or animal body fluid e.g., a plant extract or a microbial fermentation product
  • blood or urine e.g., blood or urine.
  • blood may be centrifuged to remove red blood cells and urine may be decolorised with activated charcoal or some other substance which does not affect the quantity of polyhydroxyl compounds in the test sample.
  • the compound according to the invention used in the method is preferably in solution or attached to a water-insoluble carrier, preferably a cellulosic, glass or polymeric water-insoluble carrier.
  • a water-insoluble carrier preferably a cellulosic, glass or polymeric water-insoluble carrier.
  • the carrier may be in any suitable form, for example as a flat support or beads.
  • the method of this second aspect of the invention preferably comprises the steps of: a) contacting a test sample with a compound according to the first aspect of the present invention, wherein the compound is in solution or attached to a water- insoluble carrier; b) evaluating the spectral characteristics of the compound when in contact with the test sample according to step a); c) evaluating the extent of any change in the spectral characteristics determined in step b) compared to the spectral characteristics of the compound when the compound is not in contact with the test sample.
  • the method may be performed using an automated polyhydroxyl compound detection system, for example where the compound and test sample are automatically introduced into a spectrophotometer or fluorimeter for evaluation of the spectral characteristics of the compound after and optionally before contact with the test sample.
  • the amount of a polyhydroxyl compound may be measured by comparison with a series of predetermined standards or a calibration curve prepared by contacting the compound with a series of known concentrations of the polyhydroxyl compounds.
  • the spectral characteristics of a compound when the compound is not in contact may be evaluated as part of the method normally before step a) or the spectral characteristics may already be known to the user e.g. from a colour chart or the general literature or a manual resulting in no need for this initial spectral characteristic to be measured as part of the method.
  • An advantage of the method is that by using the compounds of the first aspect of the invention the bathochromic or hypsochromic shift observed on reaction with a polyhydroxyl compound is so large that the colour change may be clearly visible to the eye.
  • the reaction of these compounds provides the basis of a rapid colorimetric test for polyhydroxyl compounds by visual comparison with a colour chart, in a similar manner to how pH is determined using litmus paper by comparison with a colour chart.
  • This can be a particular advantage in third world countries and in remote regions where there is no access to spectrometers or fiuorimeters.
  • the visual evaluation is not as valuable or accurate as the use of electronic equipment, but it is better than no evaluation at all and may avoid considerable suffering of patients.
  • kits for the detecting the presence and/or amount of a polyhydroxyl compound which comprises: a) a compound according to the first aspect of the present invention, preferably in one or more pre-weighed portions or attached to a water-insoluble carrier; b) instructions for determining the presence and/or amount of a polyhydroxyl compound using the compound; and c) optionally a pH buffer.
  • the instructions for determining the presence and/or amount of a polyhydroxyl compound using component a) comprise the method according to the second aspect of the present invention.
  • the invention is further illustrated by the following Examples.
  • Step 3 4-nitroaniline (0.10 g, 0.70 mmol) was mixed in water (10 ml), methanol (10 ml) and hydrochloric acid (10 ml, 5.0 M) and then cooled to 0-5°C in an ice-bath. A chilled solution of sodium nitrite (0.12 g, 1.74 mmol, 0.2 M) was added dropwise. Excess nitrite was destroyed by the addition of sulphamic acid.
  • step 2 ie compound (B) (0.16 g, 0.66 mmol) was dissolved in methanol (10 ml) and dilute hydrochloric acid (1 ml, 1 M) then added dropwise to the reaction mixture, which quickly turned a red colour. Sodium acetate was added to raise the pH of the solution to 4 and this was then left to stir at 0-5°C for 3 hours. Sodium hydroxide (2 M) was slowly added to raise the pH to 7.
  • Example 2 The method of Example 1 was repeated except that in Step 3, 3-aminobenzoic acid (0.06 g, 0.41 mmol) was used in place of 4-nitroaniline (all other reagents were used in the same ratios), to give compound (2) (0.10 g, 61 %) as a red solid (Found: C, 64.9; H, 5.2;
  • Example 3 The method of Example 3 was repeated except that p-anisidine (0.61 g, 4.98 mmol, 6 equivalents) was used in place of 4-aminobenzoic acid. Purification was performed by washing the product (in dichloromethane) with 10% sodium hydrogen carbonate solution (w/w) to remove residual acetic acid, to give compound (4) (0.11 g, 35%) as an orange- brown solid: mp 102°C (decomposition); ⁇ max (0.05 M NaCI, 33% MeOH: 67% H 2 O w/w)/nm 400 ( ⁇ /dm 3 mol "1 cm '1 14600, pH 8.42), 453 (13900, pH 11.47).
  • Example 6 The method of Example 1 was repeated with sulphanilic acid (0.08 g, 0.44 mmol) in place of 4-nitroaniline (all other reagents were used in the same ratios) to give compound (5) (0.05 g, 27%) as a dark red solid: mp 219°C (decomposition); ⁇ max (0.05 M NaCI, 33% MeOH: 67% H 2 O w/w)/nm 421 ( ⁇ /dm 3 mol '1 cm "1 19300, pH 8.18), 476 (19800, pH 11.52).
  • sulphanilic acid 0.08 g, 0.44 mmol
  • 4-nitroaniline all other reagents were used in the same ratios
  • Step 3 Compound (B) from step 2 (0.33 g, 1.2 mmol) was mixed in N,N- dimethylformamide (5 ml) and tetracyanoethylene (0.16 g, 1.2 mmol) was added slowly. The reaction mixture was heated to 55 °C for 1 hour. After cooling, the mixture was poured on to ice-water (10 ml) and the precipitate collected by filtration.

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  • Physics & Mathematics (AREA)
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  • General Health & Medical Sciences (AREA)
  • General Physics & Mathematics (AREA)
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  • Investigating Or Analysing Materials By The Use Of Chemical Reactions (AREA)
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Abstract

L'invention concerne un composé représenté par la formule (1) X-[-(CnH2n)n-NH-(CmH2m)-W-B(OH)2]p et ses sels, dans laquelle X est un chromophore ou un fluorophore; W est un noyau aromatique éventuellement substitué; n égale 0, 1 ou 2; m égale 1 ou 2; et p égale 1 ou 2. L'invention concerne aussi l'utilisation des composés représentés par la formule (1) dans un procédé permettant de détecter la présence et/ou la quantité de composés polyhydroxylés, et une trousse pour détecter la présence et/ou la quantité de composés polyhydroxylés.
PCT/GB2000/002685 1999-08-13 2000-07-13 Détection de composés polyhydroxylés WO2001012727A1 (fr)

Priority Applications (3)

Application Number Priority Date Filing Date Title
EP00948109A EP1210393A1 (fr) 1999-08-13 2000-07-13 D tection de compos s polyhydroxyl s
JP2001517617A JP2003507390A (ja) 1999-08-13 2000-07-13 ポリヒドロキシル化合物の検出
AU61682/00A AU6168200A (en) 1999-08-13 2000-07-13 Detection of polyhydroxyl compounds

Applications Claiming Priority (2)

Application Number Priority Date Filing Date Title
GB9919129.8 1999-08-13
GBGB9919129.8A GB9919129D0 (en) 1999-08-13 1999-08-13 Detection of polyhydroxyl compounds

Publications (1)

Publication Number Publication Date
WO2001012727A1 true WO2001012727A1 (fr) 2001-02-22

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PCT/GB2000/002685 WO2001012727A1 (fr) 1999-08-13 2000-07-13 Détection de composés polyhydroxylés

Country Status (5)

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EP (1) EP1210393A1 (fr)
JP (1) JP2003507390A (fr)
AU (1) AU6168200A (fr)
GB (1) GB9919129D0 (fr)
WO (1) WO2001012727A1 (fr)

Cited By (3)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
WO2002025252A1 (fr) * 2000-09-25 2002-03-28 Avecia Limited Procede de detection de fluorure
EP1462484A1 (fr) * 2003-03-05 2004-09-29 Universita Degli Studi di Firenze Colorant contenant un mono- ou disaccharide
CN100379482C (zh) * 2002-12-06 2008-04-09 欧洲过滤股份有限公司 用于过滤器和过滤介质的非织造布层

Citations (4)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
EP0596700A1 (fr) * 1992-11-03 1994-05-11 Hewlett-Packard Company Méthode de détection de cis-diol et supports solides
US5503770A (en) * 1993-11-07 1996-04-02 Research Development Corporation Of Japan Fluorescent compound suitable for use in the detection of saccharides
US5512246A (en) * 1989-09-21 1996-04-30 Anthony P. Russell Method and means for detecting polyhydroxyl compounds
EP0729962A1 (fr) * 1995-03-03 1996-09-04 Research Development Corporation Of Japan Composés de l'acide boronique contenant un groupe binaphthyl

Patent Citations (4)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
US5512246A (en) * 1989-09-21 1996-04-30 Anthony P. Russell Method and means for detecting polyhydroxyl compounds
EP0596700A1 (fr) * 1992-11-03 1994-05-11 Hewlett-Packard Company Méthode de détection de cis-diol et supports solides
US5503770A (en) * 1993-11-07 1996-04-02 Research Development Corporation Of Japan Fluorescent compound suitable for use in the detection of saccharides
EP0729962A1 (fr) * 1995-03-03 1996-09-04 Research Development Corporation Of Japan Composés de l'acide boronique contenant un groupe binaphthyl

Cited By (3)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
WO2002025252A1 (fr) * 2000-09-25 2002-03-28 Avecia Limited Procede de detection de fluorure
CN100379482C (zh) * 2002-12-06 2008-04-09 欧洲过滤股份有限公司 用于过滤器和过滤介质的非织造布层
EP1462484A1 (fr) * 2003-03-05 2004-09-29 Universita Degli Studi di Firenze Colorant contenant un mono- ou disaccharide

Also Published As

Publication number Publication date
JP2003507390A (ja) 2003-02-25
EP1210393A1 (fr) 2002-06-05
GB9919129D0 (en) 1999-10-13
AU6168200A (en) 2001-03-13

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