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1. WO1996041166 - DETECTION OF TRANSMEMBRANE POTENTIALS BY OPTICAL METHODS

Publication Number WO/1996/041166
Publication Date 19.12.1996
International Application No. PCT/US1996/009652
International Filing Date 06.06.1996
Chapter 2 Demand Filed 16.12.1996
IPC
C07D 209/08 2006.01
CCHEMISTRY; METALLURGY
07ORGANIC CHEMISTRY
DHETEROCYCLIC COMPOUNDS
209Heterocyclic compounds containing five-membered rings, condensed with other rings, with one nitrogen atom as the only ring hetero atom
02condensed with one carbocyclic ring
04Indoles; Hydrogenated indoles
08with only hydrogen atoms or radicals containing only hydrogen and carbon atoms, directly attached to carbon atoms of the hetero ring
C07D 239/60 2006.01
CCHEMISTRY; METALLURGY
07ORGANIC CHEMISTRY
DHETEROCYCLIC COMPOUNDS
239Heterocyclic compounds containing 1,3-diazine or hydrogenated 1,3-diazine rings
02not condensed with other rings
24having three or more double bonds between ring members or between ring members and non-ring members
28with hetero atoms or with carbon atoms having three bonds to hetero atoms with at the most one bond to halogen, directly attached to ring carbon atoms
46Two or more oxygen, sulfur or nitrogen atoms
60Three or more oxygen or sulfur atoms
C07D 311/16 2006.01
CCHEMISTRY; METALLURGY
07ORGANIC CHEMISTRY
DHETEROCYCLIC COMPOUNDS
311Heterocyclic compounds containing six-membered rings having one oxygen atom as the only hetero atom, condensed with other rings
02ortho- or peri-condensed with carbocyclic rings or ring systems
04Benzo pyrans, not hydrogenated in the carbocyclic ring
06with oxygen or sulfur atoms directly attached in position 2
08not hydrogenated in the hetero ring
16substituted in position 7
C07F 5/00 2006.01
CCHEMISTRY; METALLURGY
07ORGANIC CHEMISTRY
FACYCLIC, CARBOCYCLIC, OR HETEROCYCLIC COMPOUNDS CONTAINING ELEMENTS OTHER THAN CARBON, HYDROGEN, HALOGEN, OXYGEN, NITROGEN, SULFUR, SELENIUM OR TELLURIUM
5Compounds containing elements of Groups 3 or 13 of the Periodic System
C07F 5/02 2006.01
CCHEMISTRY; METALLURGY
07ORGANIC CHEMISTRY
FACYCLIC, CARBOCYCLIC, OR HETEROCYCLIC COMPOUNDS CONTAINING ELEMENTS OTHER THAN CARBON, HYDROGEN, HALOGEN, OXYGEN, NITROGEN, SULFUR, SELENIUM OR TELLURIUM
5Compounds containing elements of Groups 3 or 13 of the Periodic System
02Boron compounds
C09B 23/02 2006.01
CCHEMISTRY; METALLURGY
09DYES; PAINTS; POLISHES; NATURAL RESINS; ADHESIVES; COMPOSITIONS NOT OTHERWISE PROVIDED FOR; APPLICATIONS OF MATERIALS NOT OTHERWISE PROVIDED FOR
BORGANIC DYES OR CLOSELY-RELATED COMPOUNDS FOR PRODUCING DYES; MORDANTS; LAKES
23Methine or polymethine dyes, e.g. cyanine dyes
01characterised by the methine chain
02containing an odd number of CH groups
CPC
C07D 209/08
CCHEMISTRY; METALLURGY
07ORGANIC CHEMISTRY
DHETEROCYCLIC COMPOUNDS
209Heterocyclic compounds containing five-membered rings, condensed with other rings, with one nitrogen atom as the only ring hetero atom
02condensed with one carbocyclic ring
04Indoles; Hydrogenated indoles
08with only hydrogen atoms or radicals containing only hydrogen and carbon atoms, directly attached to carbon atoms of the hetero ring
C07D 239/60
CCHEMISTRY; METALLURGY
07ORGANIC CHEMISTRY
DHETEROCYCLIC COMPOUNDS
239Heterocyclic compounds containing 1,3-diazine or hydrogenated 1,3-diazine rings
02not condensed with other rings
24having three or more double bonds between ring members or between ring members and non-ring members
28with hetero atoms or with carbon atoms having three bonds to hetero atoms with at the most one bond to halogen, directly attached to ring carbon atoms
46Two or more oxygen, sulfur or nitrogen atoms
60Three or more oxygen or sulfur atoms
C07D 239/62
CCHEMISTRY; METALLURGY
07ORGANIC CHEMISTRY
DHETEROCYCLIC COMPOUNDS
239Heterocyclic compounds containing 1,3-diazine or hydrogenated 1,3-diazine rings
02not condensed with other rings
24having three or more double bonds between ring members or between ring members and non-ring members
28with hetero atoms or with carbon atoms having three bonds to hetero atoms with at the most one bond to halogen, directly attached to ring carbon atoms
46Two or more oxygen, sulfur or nitrogen atoms
60Three or more oxygen or sulfur atoms
62Barbituric acids
C07D 239/66
CCHEMISTRY; METALLURGY
07ORGANIC CHEMISTRY
DHETEROCYCLIC COMPOUNDS
239Heterocyclic compounds containing 1,3-diazine or hydrogenated 1,3-diazine rings
02not condensed with other rings
24having three or more double bonds between ring members or between ring members and non-ring members
28with hetero atoms or with carbon atoms having three bonds to hetero atoms with at the most one bond to halogen, directly attached to ring carbon atoms
46Two or more oxygen, sulfur or nitrogen atoms
60Three or more oxygen or sulfur atoms
66Thiobarbituric acids
C07D 311/16
CCHEMISTRY; METALLURGY
07ORGANIC CHEMISTRY
DHETEROCYCLIC COMPOUNDS
311Heterocyclic compounds containing six-membered rings having one oxygen atom as the only hetero atom, condensed with other rings
02ortho- or peri-condensed with carbocyclic rings or ring systems
04Benzo[b]pyrans, not hydrogenated in the carbocyclic ring
06with oxygen or sulfur atoms directly attached in position 2
08not hydrogenated in the hetero ring
16substituted in position 7
C07F 5/003
CCHEMISTRY; METALLURGY
07ORGANIC CHEMISTRY
FACYCLIC, CARBOCYCLIC OR HETEROCYCLIC COMPOUNDS CONTAINING ELEMENTS OTHER THAN CARBON, HYDROGEN, HALOGEN, OXYGEN, NITROGEN, SULFUR, SELENIUM OR TELLURIUM
5Compounds containing elements of Groups 3 or 13 of the Periodic System
003without C-Metal linkages
Applicants
  • THE REGENTS OF THE UNIVERSITY OF CALIFORNIA [US]/[US] (AllExceptUS)
  • TSIEN, Roger, Y. [US]/[US] (UsOnly)
  • GONZALEZ, Jesus, E., III [US]/[US] (UsOnly)
Inventors
  • TSIEN, Roger, Y.
  • GONZALEZ, Jesus, E., III
Agents
  • FISH & RICHARDSON, P.C.
Priority Data
08/481,97707.06.1995US
Publication Language English (EN)
Filing Language English (EN)
Designated States
Title
(EN) DETECTION OF TRANSMEMBRANE POTENTIALS BY OPTICAL METHODS
(FR) DETECTION PAR PROCEDES OPTIQUES DES POTENTIELS TRANSMEMBRANAIRES
Abstract
(EN)
Methods and compositions are provided for determining the potential of a membrane. In one aspect, the method comprises: (a) introducing a first reagent comprising a hydrophobic fluorescent ion capable of redistributing from a first face of the membrane to a second face of the membrane in response to changes in the potential of the membrane, as described by the Nernst equation, (b) introducing a second reagent which labels the first face or the second face of the membrane, which second reagent comprises a chromophore capable of undergoing energy transfer by either (i) donating excited state energy to the fluorescent ion, or (ii) accepting excited state energy from the fluorescent ion, (c) exposing the membrane to radiation; (d) measuring energy transfer between the fluorescent ion and the second reagent, and (e) relating the energy transfer to the membrane potential. Energy transfer is typically measured by fluorescence resonance energy transfer. In some embodiments the first and second reagents are bound together by a suitable linker. In one aspect the method is used to identify compounds which modulate membrane potentials in biological membranes.
(FR)
La présente invention concerne des procédés et compositions permettant de déterminer les potentiels d'une membrane. Selon un des aspects de l'invention, le procédé consiste: (a) à introduire un premier réactif comprenant un ion fluorescent hydrophobe capable de se redistribuer d'une première face de la membrane vers une seconde face de la membrane en réaction à des modifications des potentiels de la membrane telles qu'elles sont décrites par l'équation de Nernst, (b) à introduire un second réactif qui marque la première face ou la seconde face de la membrane, lequel réactif comprend un chromophore capable de subir un transfert d'énergie, soit (i) en donnant à l'ion fluorescent de l'énergie d'excitation, soit (ii) en acceptant de l'ion fluorescent de l'énergie d'excitation, (c) à exposer la membrane à un rayonnement, (d) à mesurer le transfert d'énergie entre l'ion fluorescent et le second réactif, et (e) à établir la relation entre le transfert d'énergie et le potentiel de la membrane. En l'occurrence, la mesure du transfert d'énergie se fait selon le principe du transfert d'énergie de résonance de fluorescence. Dans certaines réalisations, le premier réactif et le second réactif sont liés ensemble par un liant approprié. Selon un aspect de l'invention, le procédé décrit convient à l'identification de composés capables de moduler les potentiels membranaires des membranes biologiques.
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