Use of fluorescein dicarbonate derivatives as a cell marker
OwnerPhilippe GAIN, Gilles THURET +6inventors
InventorsPhilippe GAIN, Gilles THURET, Emilie COURRIER, Zhiguo HE, Gilles ULRICH, Antoinette DE NICOLA, Corantin MAURIN, Corentin MARET
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Abstract
The invention relates to fluorescein derivative compounds and the use thereof as fluorescent markers. Said markers can be used in particular in biology and in medicine, particularly in the field of ophthalmology. The non-fluorescent fluorescein derivatives are activated in the cell, thereby allowing the detection of living cells as well as the quantification of cell viability by fluorescence, without being toxic to cells.
Claims20
A method of cellular marking of cells comprising the steps of contracting the cells with a compound of general formula (I),
- wherein:
- R1 is selected from
- a C3-C20 alkyl,
- a polyethylene glycol or a polypropylene glycol,
- malic acid derivatives and corresponding esters,
- sugars, and
- natural polysaccharides;
- X1 is selected from H, Cl, F, Br and NO2;
- X2 is selected from H, Cl, Br and N(CH2CO2H)2;
- R2 is selected from H, NCS and CO2R3;
- and
- R3 is selected from H or a C1-C3 alkyl,
- and its pharmaceutically acceptable salts,
- assessing the presence of stain or fluorescence in said cells.
A method for preparing a compound of formula (I)
- wherein
- R1 is selected from
- a C3-C20 alkyl,
- a polyethylene glycol or a polypropylene glycol,
- malic acid derivatives and corresponding esters,
- sugars, and
- natural polysaccharides;
- X1 is selected from H, Cl, F, Br and NO2;
- X2 is selected from H, Cl, Br and N(CH2CO2H)2;
- R2 is selected from H, NCS and CO2R3;
- and
- R3 is selected from H or a C1-C3 alkyl
- and its pharmaceutically acceptable salts,
- wherein the method comprises the steps of:
- a) Reaction between a diphenol (II),
- with phosgene or a derivative thereof in the presence of base leading to the formation of the synthesis intermediate of formula (III)
- Reaction of the intermediate (III),
- with an alcohol of formula (IV), R1OH, in the presence of base to obtain the compound of formula (I).
A method for assessing the cellular viability of cells which comprises the steps of contacting the cells with a compound of general formula (I)
- wherein
- R1 is selected from
- a C3-C20 alkyl,
- a polyethylene glycol or a polypropylene glycol,
- malic acid derivatives and corresponding esters,
- sugars, and
- natural polysaccharides;
- X1 is selected from H, Cl, F, Br and NO2;
- X2 is selected from H, Cl, Br and N(CH2CO2H)2;
- R2 is selected from H, NCS and CO2R3;
- and
- R3 is selected from H or a C1-C3 alkyl
- and its pharmaceutically acceptable salts,
- assessing the presence of staining or fluorescence of the cells, the staining or fluorescence being indicative of the presence of living cells.
Dependencies are inferred from the claim text.