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US 12,576,164 B2Granted
Imidazopyrazine derivatives, process for preparation thereof, and their uses as luciferins
AssigneesInstitut Pasteur, Centre National De La Recherche Scientifique
InventorsYves-Louis Janin, Eloi Paul Coutant, Vincent Hervin, Glwadys Gagnot, Yves Jacob, Sophie Goyard, Thierry Rose
Life of this patent
Estimating the term…
Abstract
The present invention is in the field of bioluminescence in biology and/or medicine. In particular, the invention provides imidazopyrazine derivatives, processes for preparation thereof, and their uses as luciferins.
Claims11
A method to detect and/or measure an enzymatic activity of a luminogenic protein, comprising:
- (a) contacting a compound of following formula (I) with a solution comprising a strong acid to obtain a deprotected form of the compound of formula (I);
- wherein:
- R1 represents H or a group selected from C1-C6 alkyl, C3-C7 cycloalkyl, C6-C10 aryl, aralkyl and C5-C10-membered heteroaryl groups, said C1-C6 alkyl, C3-C7 cycloalkyl, C6-C10 aryl, aralkyl and C5-C10-membered heteroaryl groups optionally substituted by at least one Y1 group;
- R2 represents a group selected from C6-C10 aryl and C5-C10-membered heteroaryl groups, said C6-C10 aryl and C5-C10-membered heteroaryl groups optionally substituted by at least one Y2 group;
- or R1 and R2 together form with the two carbon atoms to which they are respectively attached a C5-C7 cycloalkene group, a C4-C7 heterocycloalkene group, or a C6-C10 arene, said C5-C7 cycloalkene group and C4-C7 heterocycloalkene groups fused with a C6-C10 arene, said C5-C7 cycloalkene group, C4-C7 heterocycloalkene, C6-C10 arene, C5-C7 cycloalkene group and C4-C7 heterocycloalkene groups optionally substituted by at least one Y12 group;
- R3 represents a C1-C6 alkyl, an aralkyl group, a hetaralkyl group or a heterocycloalkyl-CH2— group, said C1-C6 alkyl, aralkyl group, hetaralkyl group and heterocycloalkyl-CH2— group being optionally substituted by at least one Y3 group;
- R4 represents H or a group selected from C1-C6 alkyl and C3-C7 cycloalkyl groups, said C1-C6 alkyl and C3-C7 cycloalkyl groups optionally substituted by at least one Y4 group;
- R5 represents a —C(═O)Ra group or a —C(═O)ORa group;
- wherein when R5 is the —C(═O)Ra group:
- Ra represents H, a C1-C6 alkyl group, a C3-C7 cycloalkyl group, a C6-C10 aryl, or an aralkyl group, said C1-C6 alkyl group, C3-C7 cycloalkyl group, C6-C10 aryl, and aralkyl group optionally substituted by at least one Ya group, selected from a C1-C6 alkyl, a C3-C7 cycloalkyl, a C6-C10 aryl, a C5-C10-membered heteroaryl group, a halogen, a —CF3 group, a —CN group, a —ORi group, a —OSO3H group, a —NRiRii group, and a guanidinyl group;
- and
- wherein when R5 is the —C(═O)ORa group:
- (i) Ra represents H, a C1-C6 alkyl group, a C3-C7 cycloalkyl group optionally substituted by at least one Ya group selected from a C1-C6 alkyl;
- a C3-C7 cycloalkyl;
- a C6-C10 aryl;
- a C5-C10-membered heteroaryl group;
- a halogen;
- a —CF3 group;
- a —CN group;
- a —ORi group;
- a —OSO3H group;
- a —NRiRii group;
- and a guanidinyl group;
- or
- (ii) Ra represents a C6-C10 aryl group optionally substituted by at least one Ya group selected from a C1-C6 alkyl;
- a C3-C7 cycloalkyl;
- a C6-C10 aryl;
- a C5-C10-membered heteroaryl group;
- a —CF3 group;
- a —CN group;
- a —ORi group;
- a —OSO3H group;
- a —NRiRii group;
- and a guanidinyl group;
- Z represents a C6-C10 aryl, a C5-C10-membered heteroaryl groups, a C1-C6 alkyl, a C3-C7 cycloalkyl, a C4-C7 heterocycloalkyl, said C6-C10 aryl, C5-C10-membered heteroaryl groups, C1-C6 alkyl, C3-C7 cycloalkyl, and C4-C7 heterocycloalkyl optionally substituted by at least one Yz group;
- said Y1, Y2, Y12, Y3, Y4, Y5, and Yz, groups each independently selected from:
- a C1-C6 alkyl;
- a C3-C7 cycloalkyl;
- a C6-C10 aryl;
- a C5-C10-membered heteroaryl group;
- a halogen;
- a —CF3 group;
- a —CN group;
- a —ORi group;
- a —OSO3H group;
- a —NRiRii group;
- and
- a guanidinyl group;
- Ri and Rii each independently represent H, a C1-C6 alkyl group, a C3-C10 cycloalkyl group or an aralkyl group;
- or together form with the nitrogen atom to which they are attached a C4-C7 heterocycloalkyl group;
- (b) contacting the deprotected form of the compound of formula (I) with a buffered media to obtain a solution comprising the deprotected form of the compound of formula (I);
- (c) contacting the solution of the deprotected form of the compound of formula (I) with a luminogenic protein;
- and
- (d) detecting a luminescence signal, so as to detect and/or measure the enzymatic activity of the luminogenic protein.
51 more elements
A method to detect and/or measure an enzymatic activity of a luminogenic protein, comprising:
- (a) contacting a compound of following formula (I) with a solution comprising a strong acid to obtain a deprotected form of the compound of formula (I);
- wherein the deprotected form of the compound of formula (I) is a compound according to formula (III);
- wherein:
- R1 represents H or a group selected from C1-C6 alkyl, C3-C7 cycloalkyl, C6-C10 aryl, aralkyl and C5-C10-membered heteroaryl groups, said C1-C6 alkyl, C3-C7 cycloalkyl, C6-C10 aryl, aralkyl and C5-C10-membered heteroaryl groups optionally substituted by at least one Y1 group;
- R2 represents a group selected from C6-C10 aryl and C5-C10-membered heteroaryl groups, said C6-C10 aryl and C5-C10-membered heteroaryl groups optionally substituted by at least one Y2 group;
- R3 represents a C1-C6 alkyl, an aralkyl group, a hetaralkyl group or a heterocycloalkyl-CH2— group substituted by at least one Y3 group;
- R4 represents H or a group selected from C1-C6 alkyl and C3-C7 cycloalkyl groups, said C1-C6 alkyl and C3-C7 cycloalkyl groups optionally substituted by at least one Y4 group;
- R5 represents a —C(═O)Ra group or a —C(═O)ORa group;
- wherein when R5 is the —C(═O)Ra group:
- Ra represents H, a C1-C6 alkyl group, a C3-C7 cycloalkyl group, a C6-C10 aryl, or an aralkyl group, said C1-C6 alkyl group, C3-C7 cycloalkyl group, C6-C10 aryl, and aralkyl group optionally substituted by at least one Ya group, selected from a C1-C6 alkyl, a C3-C7 cycloalkyl, a C6-C10 aryl, a C5-C10-membered heteroaryl group, a halogen, a —CF3 group, a —CN group, a —ORi group, a —OSO3H group, a —NRiRii group, and a guanidinyl group;
- and
- wherein when R5 is the —C(═O)ORa group:
- (i) Ra represents H, a C1-C6 alkyl group, a C3-C7 cycloalkyl group optionally substituted by at least one Ya group selected from a C1-C6 alkyl;
- a C3-C7 cycloalkyl;
- a C6-C10 aryl;
- a C5-C10-membered heteroaryl group;
- a halogen;
- a —CF3 group;
- a —CN group;
- a —ORi group;
- a —OSO3H group;
- a —NRiRii group;
- and a guanidinyl group;
- or
- (ii) Ra represents a C6-C10 aryl group optionally substituted by at least one Ya group selected from a C1-C6 alkyl;
- a C3-C7 cycloalkyl;
- a C6-C10 aryl;
- a C5-C10-membered heteroaryl group;
- a —CF3 group;
- a —CN group;
- a —ORi group;
- a —OSO3H group;
- a —NRiRii group;
- and a guanidinyl group;
- Z represents a group of following formula:
- an oxazolyl;
- or an oxadiazolyl;
- R, R′, R″ and R′″ each independently represent H or a group selected from:
- a C1-C6 alkyl group;
- a C3-C7 cycloalkyl group;
- a halogen;
- a —ORi group;
- and
- a —CF3 group;
- and
- X represents O or S;
- said Y1, Y2, Y3 and Y4, groups each independently selected from:
- a C1-C6 alkyl group;
- a C3-C7 cycloalkyl group;
- a halogen;
- and
- a —ORii group;
- and
- Ri and Rii each independently represent H, a C1-C6 alkyl group, a C3-C10 cycloalkyl group or an aralkyl group;
- provided that:
- when Z═Z1:
- at least one of R, R′ and R″ does not represent H;
- or
- when R, R′ and R″ represent H, then R2 represents a group selected from C6-C10 aryl and C5-C10-membered heteroaryl groups substituted by at least one halogen group;
- or R3 represents a group selected from C6-C10 aralkyl groups substituted by at least one halogen group;
- when Z═Z2:
- at least one of R, R′, R″ and R″ does not represent H;
- when R, R′, R″ and R′″ represent H;
- then R2 represents a group selected from C6-C10 aryl and C5-C10-membered heteroaryl groups substituted by at least one halogen group;
- or R3 represents a group selected from C6-C10 aralkyl groups substituted by at least one halogen group;
- when R, R′ and R′″ represent H and R″ represents a C1-C6 alkyl group or a —ORi group;
- then R2 does not represent a 4-HO-Ph- group;
- or R3 represents a group selected from C6-C10 aralkyl groups substituted by at least one halogen group;
- (b) contacting the deprotected form of the compound of formula (I) with a buffered media to obtain a solution comprising the deprotected form of the compound of formula (I);
- (c) contacting the solution of the deprotected form of the compound of formula (I) with a luminogenic protein;
- and
- (d) detecting a luminescence signal, so as to detect and/or measure the enzymatic activity of the luminogenic protein.
69 more elements
A method to detect and/or measure an enzymatic activity of a luminogenic protein, the method comprising:
- (a) contacting a compound of formula (I) with a solution comprising a strong acid to produce a deprotected form of compound of formula (I);
- or providing the deprotected form of the compound of formula (I);
- wherein:
- R1 represents H, a C1-C6 alkyl group, a C3-C7 cycloalkyl group, a C6-C10 aryl group, a aralkyl group or a C5-C10-membered heteroaryl group;
- R2 represents one of the following groups, optionally, substituted by at least one Y2 group:
- a C6-C10 aryl or a C5-C10-membered heteroaryl group;
- R3 represents H;
- or one of the following groups substituted by at least one Y3 group:
- a C1-C6 alkyl group, an aralkyl group, a hetaralkyl group or a heterocycloalkyl-CH2— group;
- R4 represents H, a C1-C6 alkyl group, or a C3-C7 cycloalkyl group;
- R5 represents a —C(═O)Ra group or a —C(═O)ORa group;
- Ra represents H, a C1-C6 alkyl group, a C3-C7 cycloalkyl group, or an aralkyl group;
- Z represents a C6-C10 aryl group, a C5-C10-membered heteroaryl group, a C1-C6 alkyl group, a C3-C7 cycloalkyl group, or a C4-C7 heterocycloalkyl group;
- Y2 and Y3 are each independently selected from:
- a C1-C6 alkyl;
- a C3-C7 cycloalkyl;
- a C6-C10 aryl;
- a C5-C10-membered heteroaryl group;
- a halogen;
- a —CF3 group;
- a —CN group;
- a —ORi group;
- a —OSO3H group;
- a —NRiRii group;
- and
- a guanidinyl group;
- and
- Ri and Rii each independently represent H, a C1-C6 alkyl group, a C3-C10 cycloalkyl group or an aralkyl group;
- or together with the nitrogen atom to which they are attached form a C4-C7 heterocycloalkyl group;
- (b) contacting the deprotected form of the compound of formula (I) with a buffered media to obtain a solution comprising the deprotected form of the compound of formula (I);
- (c) contacting the solution of the deprotected form of the compound of formula (I) with a luminogenic protein;
- and
- (d) detecting a luminescence signal, wherein the luminescence signal correlates to the presence of enzymatic activity of the luminogenic protein.
30 more elements
Dependencies are inferred from the claim text.