Conjugates including a detectable moiety
Disclosed herein are detectable moieties and detectable conjugates comprising one or more detectable moieties. In some embodiments, the disclosed detectable moieties have a narrow wavelength and are suitable for multiplexing. Also disclosed are methods of labeling one or more targets within a biological specimen using any of the detectable conjugates and/or detectable moieties described herein.
1 . A compound having Formula (I):
[Z]-[Q] m -[W] (I),
wherein
Z is (i) a tissue reactive moiety selected from a tyramide, a tyramide derivative, or a quinone methide precursor, or (ii) a functional group or a moiety including a functional group capable of participating in a click chemistry reaction, wherein the functional group or the moiety including the functional group capable of participating in the click chemistry reaction is a dibenzocyclooctyne, a trans-cyclooctene, an alkyne, an alkene, an azide, a tetrazine, a maleimide, a N-hydroxysuccinimide, a thiol, a 1,3-nitrone, an aldehyde, a ketone, a hydrazine, a hydroxylamine, an amino group;
Q is a branched or unbranched, linear or cyclic, substituted or unsubstituted group having between 2 and 40 carbon atoms, and optionally having one or more heteroatoms selected from O, N, or S;
m is 0, 1, or 2; and
W has Formula (VIIA):
wherein R x is H or a branched or unbranched C 1 -C 4 alkyl group optionally substituted with one or more halogen atoms;
R m is H, a branched or unbranched C 1 -C 4 alkyl group which is optionally substituted with one or more halogen atoms and or one or more —S(O)(O)(OH) groups, or a branched or unbranched C 1 -C 20 alkyl group optionally including one or more heteroatoms selected from O or N, and optionally including one or more carbonyl groups, provided that the C 1 -C 20 alkyl group terminates in a moiety capable of participating in a click chemistry reaction, wherein the moiety capable of participating in the click chemistry reaction is a dibenzocyclooctyne, a trans-cyclooctene, an alkyne, an alkene, an azide, a tetrazine, a maleimide, a N-hydroxysuccinimide, a thiol, a 1,3-nitrone, an aldehyde, a ketone, a hydrazine, a hydroxylamine, an amino group;
R q and R r are each independently H, a branched or unbranched C 1 -C 4 alkyl group optionally substituted with one or more halogen atoms, or a group R s , where R s is a saturated or unsaturated C 1 -C 20 alkyl group comprising at least one amide group, and which is optionally substituted with one or more heteroatoms, provided that the group R s terminates in a moiety capable of participating in a click chemistry reaction, wherein the moiety capable of participating in the click chemistry reaction is a dibenzocyclooctyne, a trans-cyclooctene, an alkyne, an alkene, an azide, a tetrazine, a maleimide, a N-hydroxysuccinimide, a thiol, a 1,3-nitrone, an aldehyde, a ketone, a hydrazine, a hydroxylamine, an amino group,
provided that at least one of R q or R r comprises a group R s , and further provided that R q and R r are both not R s .
2 . The compound of claim 1 , wherein R m is the branched or unbranched C 1 -C 4 alkyl group which is optionally substituted with the one or more halogen atoms and or the one or more —S(O)(O)(OH) groups.
3 . The compound of claim 1 , wherein R m is the branched or unbranched C 1 -C 20 alkyl group optionally including the one or more heteroatoms selected from O or N, and optionally including the one or more carbonyl groups, provided that the C 1 -C 20 alkyl group terminates in the moiety capable of participating in the click chemistry reaction.
4 . The compound of claim 1 , wherein W has any one of Formulas (VIIB) and (VIIC):
5 . The compound of claim 1 , wherein W is selected from the group consisting of:
6 . The compound of claim 1 , wherein Z is the tissue reactive moiety.
7 . The compound of claim 6 , wherein the tissue reactive moiety is the quinone methide precursor.
8 . The compound of claim 6 , wherein the tissue reactive moiety is the tyramide or a tyramide derivative.
9 . The compound of claim 1 , wherein Z is the functional group or the A moiety including the functional group capable of participating in the click chemistry reaction.
10 . The compound of claim 9 , wherein the functional group or the moiety A including the functional group capable of participating in the click chemistry reaction is selected from the group consisting of azide, diarylcyclooctyne, alkyne, and Trans-cyclooctene.
11 . The compound of claim 2 , wherein R m is an unsubstituted branched or unbranched C 1 -C 4 alkyl group.
12 . The compound of claim 2 , wherein R m is an unbranched C 1 -C 4 alkyl group including one —S(O)(O)(OH) group.
13 . The compound of claim 1 , wherein R q and R m both include a group R s .
14 . The compound of claim 13 , wherein R s includes a C 3 or C 4 alkyl group and includes one amide group.
15 . The compound of claim 1 , wherein R m is the branched or unbranched C 1 -C 20 alkyl group optionally including the one or more heteroatoms selected from O or N, and optionally including the one or more carbonyl groups, provided that the C 1 -C 20 alkyl group terminates in the moiety capable of participating in the click chemistry reaction; and R q is a methyl group.
16 . The compound of claim 15 , wherein the moiety capable of A participating in a click chemistry reaction is selected from the group consisting of azide, diarylcyclooctyne, alkyne, and Trans-cyclooctene.
17 . The compound of claim 1 , wherein R q is H.
18 . The compound of claim 1 , wherein the tyramide or the tyramide derivative has the structure of any one of Formulas (XA) or (XB):
wherein each R group is independently selected from hydrogen or lower alkyl group having between 1 and 4 carbon atoms.
19 . The compound of claim 1 , wherein the quinone methide has the structure of any one of Formulas (IXA) or (IXE):
where R 1 is selected from the group consisting of phosphate, amide, nitro, urea, sulfate, methyl, ester, beta-lactam, and a sugar; R 2 is a halide; R 3 , R 4 , R 5 , and R 6 are independently selected from hydrogen or an aliphatic group having between 1 and 4 carbon atoms; and R 7 is —(CH 2 ) w NH—, —O(CH 2 ) w NH—, —N(H)C(O)(CH 2 ) w NH—, —C(O)N(H)(CH 2 ) w NH—, —(CH 2 ) w O—, —O(CH 2 ) w O—, —O(CH 2 CH 2 O) w —, —N(H)C(O)(CH 2 ) w O—, —C(O)N(H)(CH 2 ) w O—, —C(O)N(H)(CH 2 CH 2 O) w —, —(CH 2 ) w S—, —O(CH 2 ) w S—, —N(H)C(O)(CH 2 ) w S—, —C(O)N(H)(CH 2 ) w S—, —(CH 2 ) w NH—, —C(O)N(H)(CH 2 CH 2 O) w CH 2 CH 2 NH, —C(O)(CH 2 CH 2 O) w CH 2 CH 2 NH—, —C(O)N(H)(CH 2 )NHC(O)CH(CH 3 )(CH 2 ) w NH—, or —N(H)(CH 2 ) w NH—, where w is an integer ranging from 1 to 12.