WO2001025171A1 - Constructions chimiques - Google Patents

Constructions chimiques Download PDF

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Publication number
WO2001025171A1
WO2001025171A1 PCT/EP2000/009639 EP0009639W WO0125171A1 WO 2001025171 A1 WO2001025171 A1 WO 2001025171A1 EP 0009639 W EP0009639 W EP 0009639W WO 0125171 A1 WO0125171 A1 WO 0125171A1
Authority
WO
WIPO (PCT)
Prior art keywords
substrate
group
chromophore
fragment
cleavage site
Prior art date
Application number
PCT/EP2000/009639
Other languages
English (en)
Inventor
Robin Arthur Ellis Carr
Sylvie Gehanne
Alfredo Paio
Geoffrey Martyn Williams
Alessio Zaramella
Corinne Kay
Original Assignee
Glaxo Group 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 Glaxo Group Limited filed Critical Glaxo Group Limited
Priority to AU76611/00A priority Critical patent/AU7661100A/en
Priority to JP2001528123A priority patent/JP2003511656A/ja
Priority to EP00966100A priority patent/EP1218319A1/fr
Publication of WO2001025171A1 publication Critical patent/WO2001025171A1/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
    • G01N33/53—Immunoassay; Biospecific binding assay; Materials therefor
    • G01N33/543—Immunoassay; Biospecific binding assay; Materials therefor with an insoluble carrier for immobilising immunochemicals
    • G01N33/54353—Immunoassay; Biospecific binding assay; Materials therefor with an insoluble carrier for immobilising immunochemicals with ligand attached to the carrier via a chemical coupling agent
    • C—CHEMISTRY; METALLURGY
    • C07—ORGANIC CHEMISTRY
    • C07B—GENERAL METHODS OF ORGANIC CHEMISTRY; APPARATUS THEREFOR
    • C07B61/00—Other general methods
    • C—CHEMISTRY; METALLURGY
    • C07—ORGANIC CHEMISTRY
    • C07C—ACYCLIC OR CARBOCYCLIC COMPOUNDS
    • C07C311/00—Amides of sulfonic acids, i.e. compounds having singly-bound oxygen atoms of sulfo groups replaced by nitrogen atoms, not being part of nitro or nitroso groups
    • C07C311/15—Sulfonamides having sulfur atoms of sulfonamide groups bound to carbon atoms of six-membered aromatic rings
    • C07C311/16—Sulfonamides having sulfur atoms of sulfonamide groups bound to carbon atoms of six-membered aromatic rings having the nitrogen atom of at least one of the sulfonamide groups bound to hydrogen atoms or to an acyclic carbon atom
    • C07C311/18—Sulfonamides having sulfur atoms of sulfonamide groups bound to carbon atoms of six-membered aromatic rings having the nitrogen atom of at least one of the sulfonamide groups bound to hydrogen atoms or to an acyclic carbon atom to an acyclic carbon atom of a hydrocarbon radical substituted by nitrogen atoms, not being part of nitro or nitroso groups
    • C—CHEMISTRY; METALLURGY
    • C07—ORGANIC CHEMISTRY
    • C07C—ACYCLIC OR CARBOCYCLIC COMPOUNDS
    • C07C311/00—Amides of sulfonic acids, i.e. compounds having singly-bound oxygen atoms of sulfo groups replaced by nitrogen atoms, not being part of nitro or nitroso groups
    • C07C311/30—Sulfonamides, the carbon skeleton of the acid part being further substituted by singly-bound nitrogen atoms, not being part of nitro or nitroso groups
    • C07C311/37—Sulfonamides, the carbon skeleton of the acid part being further substituted by singly-bound nitrogen atoms, not being part of nitro or nitroso groups having the sulfur atom of at least one of the sulfonamide groups bound to a carbon atom of a six-membered aromatic ring
    • C07C311/38—Sulfonamides, the carbon skeleton of the acid part being further substituted by singly-bound nitrogen atoms, not being part of nitro or nitroso groups having the sulfur atom of at least one of the sulfonamide groups bound to a carbon atom of a six-membered aromatic ring having sulfur atoms of sulfonamide groups and amino groups bound to carbon atoms of six-membered rings of the same carbon skeleton
    • C07C311/39—Sulfonamides, the carbon skeleton of the acid part being further substituted by singly-bound nitrogen atoms, not being part of nitro or nitroso groups having the sulfur atom of at least one of the sulfonamide groups bound to a carbon atom of a six-membered aromatic ring having sulfur atoms of sulfonamide groups and amino groups bound to carbon atoms of six-membered rings of the same carbon skeleton having the nitrogen atom of at least one of the sulfonamide groups bound to hydrogen atoms or to an acyclic carbon atom
    • C07C311/42—Sulfonamides, the carbon skeleton of the acid part being further substituted by singly-bound nitrogen atoms, not being part of nitro or nitroso groups having the sulfur atom of at least one of the sulfonamide groups bound to a carbon atom of a six-membered aromatic ring having sulfur atoms of sulfonamide groups and amino groups bound to carbon atoms of six-membered rings of the same carbon skeleton having the nitrogen atom of at least one of the sulfonamide groups bound to hydrogen atoms or to an acyclic carbon atom to an acyclic carbon atom of a hydrocarbon radical substituted by carboxyl groups
    • C—CHEMISTRY; METALLURGY
    • C40—COMBINATORIAL TECHNOLOGY
    • C40B—COMBINATORIAL CHEMISTRY; LIBRARIES, e.g. CHEMICAL LIBRARIES
    • C40B20/00—Methods specially adapted for identifying library members
    • C40B20/08—Direct analysis of the library members per se by physical methods, e.g. spectroscopy
    • C—CHEMISTRY; METALLURGY
    • C40—COMBINATORIAL TECHNOLOGY
    • C40B—COMBINATORIAL CHEMISTRY; LIBRARIES, e.g. CHEMICAL LIBRARIES
    • C40B40/00—Libraries per se, e.g. arrays, mixtures
    • C40B40/04—Libraries containing only organic compounds
    • C—CHEMISTRY; METALLURGY
    • C40—COMBINATORIAL TECHNOLOGY
    • C40B—COMBINATORIAL CHEMISTRY; LIBRARIES, e.g. CHEMICAL LIBRARIES
    • C40B50/00—Methods of creating libraries, e.g. combinatorial synthesis
    • C40B50/14—Solid phase synthesis, i.e. wherein one or more library building blocks are bound to a solid support during library creation; Particular methods of cleavage from the solid support
    • C—CHEMISTRY; METALLURGY
    • C40—COMBINATORIAL TECHNOLOGY
    • C40B—COMBINATORIAL CHEMISTRY; LIBRARIES, e.g. CHEMICAL LIBRARIES
    • C40B70/00—Tags or labels specially adapted for combinatorial chemistry or libraries, e.g. fluorescent tags or bar codes
    • C—CHEMISTRY; METALLURGY
    • C40—COMBINATORIAL TECHNOLOGY
    • C40B—COMBINATORIAL CHEMISTRY; LIBRARIES, e.g. CHEMICAL LIBRARIES
    • C40B80/00—Linkers or spacers specially adapted for combinatorial chemistry or libraries, e.g. traceless linkers or safety-catch linkers
    • C—CHEMISTRY; METALLURGY
    • C07—ORGANIC CHEMISTRY
    • C07B—GENERAL METHODS OF ORGANIC CHEMISTRY; APPARATUS THEREFOR
    • C07B2200/00—Indexing scheme relating to specific properties of organic compounds
    • C07B2200/11—Compounds covalently bound to a solid support
    • G—PHYSICS
    • G01—MEASURING; TESTING
    • G01N—INVESTIGATING OR ANALYSING MATERIALS BY DETERMINING THEIR CHEMICAL OR PHYSICAL PROPERTIES
    • G01N21/00—Investigating or analysing materials by the use of optical means, i.e. using sub-millimetre waves, infrared, visible or ultraviolet light
    • G01N21/62—Systems in which the material investigated is excited whereby it emits light or causes a change in wavelength of the incident light
    • G01N21/63—Systems in which the material investigated is excited whereby it emits light or causes a change in wavelength of the incident light optically excited
    • G01N21/64—Fluorescence; Phosphorescence
    • G01N21/6428—Measuring fluorescence of fluorescent products of reactions or of fluorochrome labelled reactive substances, e.g. measuring quenching effects, using measuring "optrodes"
    • G01N2021/6439—Measuring fluorescence of fluorescent products of reactions or of fluorochrome labelled reactive substances, e.g. measuring quenching effects, using measuring "optrodes" with indicators, stains, dyes, tags, labels, marks

Definitions

  • the resin beads typically used in the synthesis of combinatorial libraries are derived from cross-linked polystyrene resin and are usually of the order of 90 ⁇ m and 250 ⁇ m in diameter (i.e. just visible to the naked eye).
  • the number of beads in a given mass or volume of resin will depend on the average bead size but, for example, with a bead size of 150 ⁇ m diameter, there will be approximately 500 beads per milligram of bulk resin.
  • Typical compound loadings on the beads are of the order of 0.1 to 0.4 mmol g ⁇ l and hence beads of the dimensions given above will tend to have individual compound loadings of about 1 nanomole of compound per bead.
  • coding tags such as isotopic labels can be incorporated into inter alia a linker group between the solid support and the substrate.
  • This approach has the advantage that it does not require the formation of a separate coding tag using orthogonal chemistry and thereby reduces the overall number of synthesis steps involved.
  • the molar abso ⁇ tivity E is constant for a given compound at a given wavelength and is usually expressed as E max - the molar abso ⁇ tivity at an absorption band maximum.
  • a UV chromophore in the construct enables quantitation of the products of a synthesis to be carried out.
  • quantitation can be (i) absolute quantitation or (ii) relative quantitation, or both.
  • absolute quantitation is meant that the absolute concentrations of a substrate molecule or fragment or construct containing the substrate molecule can be determined, whereas the term " relative quantitation "is used herein to mean the determination of the amount of a substrate (or fragment or construct containing the substrate) relative to another component (such as a side product or starting material or another substrate) in a reaction mixture.
  • chromophores are groups containing an aryl group, preferably a fused polycyclic aryl group, e.g. a C - C30 polycyclic aryl group in which one or more (e.g. 1, 2, or 3) ring carbon atoms are optionally replaced by a heteroatom such as nitrogen, sulphur or oxygen.
  • polycyclic aryl groups are polycyclic hydrocarbons such as naphthyl, phenanthrenyl and anthracenyl groups, and polycyclic heteroaryl groups such as acridine, or phenanthroline.
  • alkylthio e.g. C]_20 alkylthio, preferably C ⁇ .
  • chromophores include anthracenyl and dansyl (5-dimethylamino-l- naphthylsulphonyl groups, anthracenyl being particularly preferred.
  • the quantitation of the substrate R can be either absolute quantitation or relative quantitation in which the relative amounts of the substrate R and another component in the synthesis mixture (e.g. another substrate, or starting material, or a side product for example) are determined.
  • Relative quantitation makes use of the fact that the chromophore C u confers on each substrate a common set of characteristic absorbances that can be used as the basis for the spectrophotometric analysis.
  • UV chromophore-containing constructs of the present invention can be used to provide quantitative information on very small amounts of substrate compounds, for example amounts of the order of 1 nanomolar typically available from single resin beads of the size used in split and mix combinatorial methods.
  • substrate compounds for example amounts of the order of 1 nanomolar typically available from single resin beads of the size used in split and mix combinatorial methods.
  • combinatorial libraries formed by the split and mix method each bead typically will contain only a single substrate compound, but a library will contain a plurality of beads bearing different substrates. The analysis of mixtures of beads bearing different substrates is problematical and hence it is generally necessary to carry out analyses on single beads.
  • Resin 2 (0.35 mmol) was swollen in dichloromethane (3 ml) and phenol (66 mg, 0.7 mmol) added, followed by trifluoroacetic acid (4.5 ml). The resin was gently agitated for 5 minutes using a stream of nitrogen then drained and washed with dichloromethane (2x5 ml). The process was repeated and, after draining, the resin washed (dichloromethane (3x8 ml), 10% diisopropylamine in dichloromethane (2x6 ml) and dichloromethane (4x8 ml)) and dried in vacuo. An analytical sample gave a positive Kaiser test.
  • a solid phase construct was formed on hydroxylated polystyrene resin by coupling together a base cleavable ester group, a diaminoethyl spacer group, an acid cleavable Rink linker and a dansyl group as shown in Scheme 4 below. 3 ⁇
  • the resulting resin 15 was filtered, washed with dimethylformamide (2 x 10ml), dichloromethane (5 x 10ml), methanol (2 x 10ml), dichloromethane (5 x 10ml) and dried under reduced pressure.
  • UV analysis to detect the Fmoc group revealed that the cleavage reaction to release the primary amine 15 had been quantitative. The presence of a primary amine group was confirmed by a positive Kaiser test.
  • the ester 3b (0.40 g, 0.80 mmol) in methanol (2 ml) was treated with IM aqueous sodium hydroxide (1.60 ml, 1.60 mmol) and stirred at room temperature for 1 hour. The reaction was then concentrated in vacuo and the residue dissolved in water (10 ml) then acidified with IM hydrochloric acid (1.80 ml, 1.80 mmol) at 0°C. The resulting white precipitate was extracted with dichloromethane (3 x 10 ml).
  • Resin (6) (0.93 g, 0.28 mmol) was treated with 20% Piperidine in DMF solution (2 x 5 ml) for 10 min. Solvents were removed by filtration and the resin washed with DMF (6 x 5 ml), dichloromethane (6 x 5 ml), and ether (2 x 4 ml). The resin was suspended in DMF (5 ml) and dichloromethane (5 ml) and the Rink acid linker (0.92 g, 1.62 mmol) was added, followed by PyBOP (0.84 g, 1.62 mmol) and HOBT (0.22 g, 1.62 mmol). After 4 min. Hunig's base (0.42g, 3.24 mmol) was added.
  • Resin (7) (70 mg, 0.02 mmol) was treated with 20% piperidine in DMF solution (2 x 1 ml) for 10 min. Solvents were removed by filtration and the resin washed with DMF (6 x 1 ml), dichloromethane (6 x 1 ml), ether (2 x 1 ml). The resin was suspended in DMF (0.5 ml) and dichloromethane (0.5 ml) and the appropriate acid (8a-d) (0.15 mmol) was added, followed by PyBOP (0.08 g, 0.15 mmol) and HOBT (0.02 g, 0.15 mmol). After 4 min. Hunig's base (0.04 g, 0.30 mmol) was added.

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  • Chemical & Material Sciences (AREA)
  • Organic Chemistry (AREA)
  • Health & Medical Sciences (AREA)
  • Life Sciences & Earth Sciences (AREA)
  • Molecular Biology (AREA)
  • Biochemistry (AREA)
  • Medicinal Chemistry (AREA)
  • Chemical Kinetics & Catalysis (AREA)
  • General Chemical & Material Sciences (AREA)
  • Engineering & Computer Science (AREA)
  • Immunology (AREA)
  • Biomedical Technology (AREA)
  • Hematology (AREA)
  • Analytical Chemistry (AREA)
  • Physics & Mathematics (AREA)
  • Urology & Nephrology (AREA)
  • Food Science & Technology (AREA)
  • Cell Biology (AREA)
  • Microbiology (AREA)
  • Biotechnology (AREA)
  • Spectroscopy & Molecular Physics (AREA)
  • General Health & Medical Sciences (AREA)
  • General Physics & Mathematics (AREA)
  • Pathology (AREA)
  • Structural Engineering (AREA)
  • Investigating, Analyzing Materials By Fluorescence Or Luminescence (AREA)
  • Investigating Or Analysing Materials By Optical Means (AREA)
  • Organic Low-Molecular-Weight Compounds And Preparation Thereof (AREA)

Abstract

L'invention concerne une construction chimique utilisée dans une synthèse en phase solide, comprenant un support solide Q lié à un substrat R via un groupe de connexion Y. Le groupe de connexion Y possède un premier et un second sites de clivage qui sont orthogonalement et sélectivement clivables, le second site de clivage étant sélectivement clivable de façon à libérer le substrat, et le premier site de clivage étant placé au niveau d'une position située entre le second site de clivage et le support solide, et étant sélectivement clivable de façon à libérer un fragment Fru. Ce fragment comprend le substrat et au mois une partie du groupe de connexion Y, cette partie renfermant un chromophore Cu qui facilite l'analyse dudit fragment Fru par spectroscopie ultra violette, visible ou fluorescente. Ledit chromophore Cu possède une valeur principale log E¿max? d'au moins 2,5, cette valeur principale logEmax étant au moins 1,5 supérieure à la valeur principale logEmax du substrat R; ou présente un pic d'absorption à une longueur d'onde distante des absorptions dues au substrat R. L'invention concerne également des procédés d'analyse de produits de synthèse en phase solide utilisant lesdites constructions.
PCT/EP2000/009639 1999-10-05 2000-10-03 Constructions chimiques WO2001025171A1 (fr)

Priority Applications (3)

Application Number Priority Date Filing Date Title
AU76611/00A AU7661100A (en) 1999-10-05 2000-10-03 Chemical constructs
JP2001528123A JP2003511656A (ja) 1999-10-05 2000-10-03 化学構築物
EP00966100A EP1218319A1 (fr) 1999-10-05 2000-10-03 Constructions chimiques

Applications Claiming Priority (2)

Application Number Priority Date Filing Date Title
GBGB9923577.2A GB9923577D0 (en) 1999-10-05 1999-10-05 Chemical constructs
GB9923577.2 1999-10-05

Publications (1)

Publication Number Publication Date
WO2001025171A1 true WO2001025171A1 (fr) 2001-04-12

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PCT/EP2000/009639 WO2001025171A1 (fr) 1999-10-05 2000-10-03 Constructions chimiques

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EP (1) EP1218319A1 (fr)
JP (1) JP2003511656A (fr)
AU (1) AU7661100A (fr)
GB (1) GB9923577D0 (fr)
WO (1) WO2001025171A1 (fr)

Cited By (3)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
US7488602B2 (en) * 2002-10-18 2009-02-10 Roche Diagnostics Operations, Inc. Method for detecting and compensating an underdosage of test strips
WO2014089680A1 (fr) 2012-12-11 2014-06-19 Nano Safe Coatings Incorporated (A Florida Corporation 3 P14000024914) Revêtements antimicrobiens à base d'ammonium quaternaire à terminaison benzophénone durcis par uv pour surfaces
USRE48510E1 (en) 2013-02-19 2021-04-13 Nano Safe Coatings Incorporated Phosphorus functional antimicrobial coatings for metal surfaces

Families Citing this family (1)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
US6787635B2 (en) * 2001-04-05 2004-09-07 3M Innovative Properties Company Solid phase synthesis supports and methods

Citations (2)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
WO1998033865A1 (fr) * 1997-02-03 1998-08-06 Ciba Specialty Chemicals Holding, Inc. Chromophore fluorescent, lie par covalence a un materiau de support organique
WO2000020357A2 (fr) * 1998-10-05 2000-04-13 Glaxo Group Limited Produits de synthese chimiques

Patent Citations (2)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
WO1998033865A1 (fr) * 1997-02-03 1998-08-06 Ciba Specialty Chemicals Holding, Inc. Chromophore fluorescent, lie par covalence a un materiau de support organique
WO2000020357A2 (fr) * 1998-10-05 2000-04-13 Glaxo Group Limited Produits de synthese chimiques

Non-Patent Citations (2)

* Cited by examiner, † Cited by third party
Title
CARRASCO M R ET AL: "Direct Monitoring of Organic Reactions on Polymeric Supports", TETRAHEDRON LETTERS,NL,ELSEVIER SCIENCE PUBLISHERS, AMSTERDAM, vol. 38, no. 36, 8 September 1997 (1997-09-08), pages 6331 - 6334, XP004087929, ISSN: 0040-4039 *
GEYSEN H M ET AL: "ISOTOPE OR MASS ENCODING OF COMBINATORIAL LIBRARIES", CHEMISTRY AND BIOLOGY,GB,CURRENT BIOLOGY, LONDON, vol. 3, no. 8, 1 August 1996 (1996-08-01), pages 679 - 688, XP002035873, ISSN: 1074-5521 *

Cited By (4)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
US7488602B2 (en) * 2002-10-18 2009-02-10 Roche Diagnostics Operations, Inc. Method for detecting and compensating an underdosage of test strips
WO2014089680A1 (fr) 2012-12-11 2014-06-19 Nano Safe Coatings Incorporated (A Florida Corporation 3 P14000024914) Revêtements antimicrobiens à base d'ammonium quaternaire à terminaison benzophénone durcis par uv pour surfaces
EP2931702A4 (fr) * 2012-12-11 2016-08-10 Nano Safe Coatings Inc Revêtements antimicrobiens à base d'ammonium quaternaire à terminaison benzophénone durcis par uv pour surfaces
USRE48510E1 (en) 2013-02-19 2021-04-13 Nano Safe Coatings Incorporated Phosphorus functional antimicrobial coatings for metal surfaces

Also Published As

Publication number Publication date
JP2003511656A (ja) 2003-03-25
GB9923577D0 (en) 1999-12-08
AU7661100A (en) 2001-05-10
EP1218319A1 (fr) 2002-07-03

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