Nanoluc suicide substrates
Compounds that may inhibit Oplophorus -derived luciferases are disclosed as well as compositions and kits comprising the compounds and methods of using the compounds.
1 . A method of detecting an Oplophorus -derived luciferase in a sample, the method comprising contacting the sample with a compound of m formula (I):
or a tautomer or a salt thereof, wherein:
R 1 is optionally substituted aryl, optionally substituted heteroaryl, optionally substituted heterocycle, or optionally substituted cycloalkyl, wherein R 1 is substituted with at least one group that is -Q-L-Z;
Q is —O—, —NR Q —, —NR Q —CO—, —CO—NR Q —, —O—CO—NR Q —, or —NR Q —CO—O—;
L is —(CR 1a R 1b ) m —, —(CR 1x R 1y —CR 1x R 1y —O) t1 —(CR 1x R 1y ) t2 —O 1 —, —CR 1x R 1y ) t2 -A-(CR 1x R 1y —CR 1x R 1y —O) t1 -Q 1 -, wherein each Q 1 is independently a bond, —O—, or —NR Q1 —, and A is a bond, —O—, —NR Q —, —NR Q —CO—, —CO—NR Q —, —O—CO—NR Q —, or —NR Q —CO—O—;
Z is —COOR 2 , —SO 2 —OR 3 , —PO(OR 4 )(OR 5 ), halo, azide, C 2 -C 10 alkynyl, a biotin moiety, —NR 6 R 7 , or —NR 8 —CO—R 9 ;
R 2 , R 3 , R 4 , R 5 , R 8 , R 9 , R 10 , R 11 , and R 12 at each occurrence are independently hydrogen, optionally substituted C 1 -C 8 alkyl, optionally substituted aryl, optionally substituted cycloalkyl, optionally substituted heteroaryl, optionally substituted heterocycle, optionally substituted heteroalkyl, or an energy acceptor;
R 6 and R 7 at each occurrence are independently hydrogen, optionally substituted C 1 -C 8 alkyl, optionally substituted aryl, optionally substituted cycloalkyl, optionally substituted heteroaryl, optionally substituted heterocycle, optionally substituted heteroalkyl, or an energy acceptor; or R 6 and R 7 , together with the nitrogen atom to which they are attached, together form an optionally substituted ring;
R 1a , R 1b , R Q , R Q1 , R 1x , and R 1y at each occurrence are independently hydrogen, C 1 -C 4 alkyl, or C 1 -C 4 haloalkyl;
q is 0, 1, or 2;
m at each occurrence is independently 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 11, or 12;
t1 at each occurrence is independently 1, 2, 3, 4, 5, 6, 7, 8, 9, or 10; and
t2 is an integer from 0-5;
wherein, when a group is optionally substituted, it is either unsubstituted or substituted with one or more substituents independently selected from halogen, ═O, ═S, cyano, nitro, alkyl, alkenyl, alkynyl, haloalkyl, haloalkoxy, heteroalkyl, cycloalkyl, cycloalkenyl, aryl, heteroaryl, heterocycle, cycloalkylalkyl, heteroarylalkyl, arylalkyl, hydroxy, hydroxyalkyl, alkoxy, alkoxyalkyl, aryloxy, benzyloxy, amino, alkylamino, acylamino, aminoalkyl, arylamino, sulfonylamino, sulfinylamino, sulfonyl, alkylsulfonyl, arylsulfonyl, aminosulfonyl, sulfinyl, —COOH, amide, carbamate, and acyl; and
wherein the compound is not 4-[4-[[8-benzyl-6-(4-hydroxyphenyl)-3-oxo-7H-imidazo[1,2-a]pyrazin-2-yl]methyl]phenoxy]butanoic acid.
2 . The method of claim 1 , further comprising detecting luminescence in the sample.
3 . A method of detecting luminescence in a sample, the method comprising:
(a) contacting the sample with a compound of formula (I):
or a tautomer or a salt thereof, wherein:
R 1 is optionally substituted aryl, optionally substituted heteroaryl, optionally substituted heterocycle, or optionally substituted cycloalkyl, wherein R 1 is substituted with at least one group that is -Q-L-Z;
Q is —O—, —NR Q —, —NR Q —CO—, —CO—NR Q —, —O—CO—NR Q —, or —NR Q —CO—O—;
L is —(CR 1a R 1b ) m —, —(CR 1x R 1y —CR 1x R 1y —O) t1 —(CR 1x R 1y ) t2 -Q 1 -, —(CR 1x R 1y ) t2 -A-(CR 1x R 1y —CR 1x R 1y —O) t1 -Q 1 -, wherein each Q 1 is independently a bond, —O—, or —NR Q1 —, and A is a bond, —O—, —NR Q —, —NR Q —CO—, —CO—NR Q —, —O—CO—NR Q —, or —NR Q —CO—O—;
Z is —COOR 2 , —SO 2 —OR 3 , —PO(OR 4 )(OR 5 ), halo, azide, C 2 -C 10 alkynyl, a biotin moiety, —NR 6 R 7 , or —NR 8 —CO—R 9 ;
R 2 , R 3 , R 4 , R 5 , R 8 , R 9 , R 10 , R 11 , and R 12 at each occurrence are independently hydrogen, optionally substituted C 1 -C 8 alkyl, optionally substituted aryl, optionally substituted cycloalkyl, optionally substituted heteroaryl, optionally substituted heterocycle, optionally substituted heteroalkyl, or an energy acceptor;
R 6 and R 7 at each occurrence are independently hydrogen, optionally substituted C 1 -C 8 alkyl, optionally substituted aryl, optionally substituted cycloalkyl, optionally substituted heteroaryl, optionally substituted heterocycle, optionally substituted heteroalkyl, or an energy acceptor; or R 6 and R 7 , together with the nitrogen atom to which they are attached, together form an optionally substituted ring;
R 1a , R 1b , R Q , R Q1 , R 1x , and R 1y at each occurrence are independently hydrogen, C 1 -C 4 alkyl, or C 1 -C 4 haloalkyl;
q is 0, 1, or 2;
m at each occurrence is independently 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 11, or 12;
t1 at each occurrence is independently 1, 2, 3, 4, 5, 6, 7, 8, 9, or 10; and
t2 is an integer from 0-5;
wherein, when a group is optionally substituted, it is either unsubstituted or substituted with one or more substituents independently selected from halogen, ═O, ═S, cyano, nitro, alkyl, alkenyl, alkynyl, haloalkyl, haloalkoxy, heteroalkyl, cycloalkyl, cycloalkenyl, aryl, heteroaryl, heterocycle, cycloalkylalkyl, heteroarylalkyl, arylalkyl, hydroxy, hydroxyalkyl, alkoxy, alkoxyalkyl, aryloxy, benzyloxy, amino, alkylamino, acylamino, aminoalkyl, arylamino, sulfonylamino, sulfinylamino, sulfonyl, alkylsulfonyl, arylsulfonyl, aminosulfonyl, sulfinyl, —COOH, amide, carbamate, and acyl; and
wherein the compound is not 4-[4-[[8-benzyl-6-(4-hydroxyphenyl)-3-oxo-7H-imidazo[1,2-a]pyrazin-2-yl]methyl]phenoxy]butanoic acid;
(b) contacting the sample with an Oplophorus -derived luciferase, if it is not present in the sample; and
(c) detecting luminescence in the sample.
4 . The method of claim 1 , wherein the Oplophorus -derived luciferase comprises a polypeptide sequence of SEQ ID NO: 2.
5 . A method of modulating luminescence of an Oplophorus -derived luciferase in a sample, the method comprising
(a) contacting the sample with a coelenterazine substrate and the compound of formula (I):
or a tautomer or a salt thereof, wherein:
R 1 is optionally substituted aryl, optionally substituted heteroaryl, optionally substituted heterocycle, or optionally substituted cycloalkyl, wherein R 1 is substituted with at least one group that is -Q-L-Z:
Q is —O—, —NR Q —, —NR Q —CO—, —CO—NR Q —, —O—CO—NR Q —, or —NR Q —CO—O—;
L is —(CR 1a R 1b ) m —, —(CR 1x R 1y —CR 1x R 1y —O) t1 —CR 1x R 1y ) t2 -Q 1 -, —(CR 1x R 1y ) t2 -A-(CR 1x R 1y —CR 1x R 1y —O) t1 -Q 1 -, wherein each Q 1 is independently a bond, —O—, or —NR Q1 —, and A is a bond, —O—, —NR Q —, —NR Q —CO—, —CO—NR Q —, —O—CO—NR Q —, or —NR Q —CO—O—;
Z is —COOR 2 , —SO 2 —OR 3 , —PO(OR 4 )(OR 5 ), halo, azide, C 2 -C 10 alkynyl, a biotin moiety, —NR 6 R 7 , or —NR 8 —CO—R 9 ;
R 2 , R 3 , R 4 , R 5 , R 8 , R 9 , R 10 , R 11 , and R 12 at each occurrence are independently hydrogen, optionally substituted C 1 -C 8 alkyl, optionally substituted aryl, optionally substituted cycloalkyl, optionally substituted heteroaryl, optionally substituted heterocycle, optionally substituted heteroalkyl, or an energy acceptor;
R 6 and R 7 at each occurrence are independently hydrogen, optionally substituted C 1 -C 8 alkyl, optionally substituted aryl, optionally substituted cycloalkyl, optionally substituted heteroaryl, optionally substituted heterocycle, optionally substituted heteroalkyl, or an energy acceptor; or R 6 and R 7 , together with the nitrogen atom to which they are attached, together form an optionally substituted ring;
R 1a , R 1b , R Q , R Q1 , R 1x , and R 1y at each occurrence are independently hydrogen, C 1 -C 4 alkyl, or C 1 -C 4 haloalkyl;
q is 0, 1, or 2;
m at each occurrence is independently 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 11, or 12;
t1 at each occurrence is independently 1, 2, 3, 4, 5, 6, 7, 8, 9, or 10; and
t2 is an integer from 0-5;
wherein, when a group is optionally substituted, it is either unsubstituted or substituted with one or more substituents independently selected from halogen, ═O, ═S, cyano, nitro, alkyl, alkenyl, alkynyl, haloalkyl, haloalkoxy, heteroalkyl, cycloalkyl, cycloalkenyl, aryl, heteroaryl, heterocycle, cycloalkylalkyl, heteroarylalkyl, arylalkyl, hydroxy, hydroxyalkyl, alkoxy, alkoxyalkyl, aryloxy, benzyloxy, amino, alkylamino, acylamino, aminoalkyl, arylamino, sulfonylamino, sulfinylamino, sulfonyl, alkylsulfonyl, arylsulfonyl, aminosulfonyl, sulfinyl, —COOH, amide, carbamate, and acyl; and
wherein the compound is not 4-[4-[[8-benzyl-6-(4-hydroxyphenyl)-3-oxo-7H-imidazo[1,2-a]pyrazin-2-yl]methyl]phenoxy]butanoic acid; and
(b) detecting luminescence in the sample,
wherein the compound of formula (I) causes a decrease in the luminescence from the Oplophorus -derived luciferase.
6 . The method of claim 5 , comprising contacting the sample with the coelenterazine substrate prior to contacting the sample with the compound of formula (I).
7 . The method of claim 5 , comprising contacting the sample with the compound of formula (I) prior to contacting the sample with the coelenterazine substrate.
8 . The method of claim 1 , wherein the sample comprises a cell.
9 . The method of claim 8 , wherein the cell comprises the Oplophorus -derived luciferase.
10 . The method of claim 9 , wherein the cell expresses the Oplophorus -derived luciferase.
11 . The method of claim 5 , wherein the coelenterazine substrate is selected from a coelenterazine, coelenterazine derivatives, coelenterazine analogs, pro-coelenterazine, and quinone-masked coelenterazine.
12 . The method of claim 3 , wherein the Oplophorus -derived luciferase comprises a polypeptide sequence of SEQ ID NO: 2.
13 . The method of claim 5 , wherein the Oplophorus -derived luciferase comprises a polypeptide sequence of SEQ ID NO: 2.
14 . The method of claim 3 , wherein the sample comprises a cell.
15 . The method of claim 14 , wherein the cell comprises the Oplophorus -derived luciferase.
16 . The method of claim 15 , wherein the cell expresses the Oplophorus -derived luciferase.
17 . The method of claim 5 , wherein the sample comprises a cell.
18 . The method of claim 17 , wherein the cell comprises the Oplophorus -derived luciferase.
19 . The method of claim 18 , wherein the cell expresses the Oplophorus -derived luciferase.
20 . The method of claim 5 , wherein the modulating comprises inhibiting Oplophorus -derived luciferase activity in the sample.
21 . The method of claim 1 , wherein the compound of formula (I) is selected from the group consisting of:
2-(4-((6-aminohexyl)oxy)-3-fluorobenzyl)-8-benzyl-6-(4-hydroxyphenyl)imidazo[1,2-a]pyrazin-3(7H)-one;
N-(6-(4-((8-benzyl-6-(4-hydroxyphenyl)-3-oxo-3,7-dihydroimidazo[1,2-a]pyrazin-2-yl)methyl)-2-fluorophenoxy)hexyl)-3′,6′-bis(dimethylamino)-3-oxo-3H-spiro[isobenzofuran-1,9′-xanthene]-6-carboxamide;
6-(4-((8-benzyl-6-(4-hydroxyphenyl)-3-oxo-3,7-dihydroimidazo[1,2-a]pyrazin-2-yl)methyl)-2-fluorophenoxy)hexane-1-sulfonic acid;
N-(6-(4-((8-benzyl-6-(4-hydroxyphenyl)-3-oxo-3,7-dihydroimidazo[1,2-a]pyrazin-2-yl)methyl)-2-fluorophenoxy)hexyl)-3′,6′-dihydroxy-3-oxo-3H-spiro[isobenzofuran-1,9′-xanthene]-6-carboxamide;
N-(6-(4-((8-benzyl-6-(4-hydroxyphenyl)-3-oxo-3,7-dihydroimidazo[1,2-a]pyrazin-2-yl)methyl)-2-fluorophenoxy)hexyl)-3-(5,5-difluoro-7-(1H-pyrrol-2-yl)-5H-514,614-dipyrrolo[1,2-c:2′,1′-f][1,3,2]diazaborinin-3-yl)propanamide;
N,N-(6-((6-(4-((8-benzyl-6-(4-hydroxyphenyl)-3-oxo-3,7-dihydroimidazo[1,2-a]pyrazin-2-yl)methyl)-2-fluorophenoxy)hexyl)carbamoyl)-3-oxo-3H-spiro[isobenzofuran-1,9′-xanthene]-3′,6′-diyl)bis(2,2,2-trifluoroacetamide);
N-(8-(4-((8-benzyl-6-(4-hydroxyphenyl)-3-oxo-3,7-dihydroimidazo[1,2-a]pyrazin-2-yl)methyl)-2-fluorophenoxy)octyl)-3′,6′-dihydroxy-3-oxo-3H-spiro[isobenzofuran-1,9′-xanthene]-6-carboxamide;
N-(8-(4-((8-benzyl-6-(4-hydroxyphenyl)-3-oxo-3,7-dihydroimidazo[1,2-a]pyrazin-2-yl)methyl)-2-fluorophenoxy)octyl)-3′,6′-bis(dimethylamino)-3-oxo-3H-spiro[isobenzofuran-1,9′-xanthene]-6-carboxamide;
3-(2-(2-aminoethoxy)ethoxy)-N-(8-(4-((1-benzyl-3-(4-hydroxyphenyl)-6-oxo-2,6-dihydropyrrolo[1,2-a]pyrazin-7-yl)methyl)-2-fluorophenoxy)octyl)propanamide;
N-(2-(2-(3-((8-(4-((1-benzyl-3-(4-hydroxyphenyl)-6-oxo-2,6-dihydropyrrolo[1,2-a]pyrazin-7-yl)methyl)-2-fluorophenoxy)octyl)amino)-3-oxopropoxy)ethoxy)ethyl)-3′,6′-dihydroxy-3-oxo-3H-spiro[isobenzofuran-1,9′-xanthene]-6-carboxamide;
N-(2-(2-(3-((8-(4-((1-benzyl-3-(4-hydroxyphenyl)-6-oxo-2,6-dihydropyrrolo[1,2-a]pyrazin-7-yl)methyl)-2-fluorophenoxy)octyl)amino)-3-oxopropoxy)ethoxy)ethyl)-3′,6′-bis(dimethylamino)-3-oxo-3H-spiro[isobenzofuran-1,9′-xanthene]-6-carboxamide;
6-((8-(4-((8-benzyl-6-(4-hydroxyphenyl)-3-oxo-3,7-dihydroimidazo[1,2-a]pyrazin-2-yl)methyl)-2-fluorophenoxy)octyl)carbamoyl)-3-oxo-3H-spiro[isobenzofuran-1,9′-xanthene]-3′,6′-diyl diacetate;
2-(4-((6-azidohexyl)oxy)-3-fluorobenzyl)-8-benzyl-6-(4-hydroxyphenyl)imidazo[1,2-a]pyrazin-3(7H)-one;
5-((8-(4-((8-benzyl-6-(4-hydroxyphenyl)-3-oxo-3,7-dihydroimidazo[1,2-a]pyrazin-2-yl)methyl)-2-fluorophenoxy)octyl)carbamoyl)-2-(2,7-difluoro-3,6-dihydroxyxanthylium-9-yl)benzoate;
5-((8-(4-((8-benzyl-6-(4-hydroxyphenyl)-3-oxo-3,7-dihydroimidazo[1,2-a]pyrazin-2-yl)methyl)-2-fluorophenoxy)octyl)carbamoyl)-2-(3,11-dihydroxydibenzo[c,h]xanthen-14-ium-7-yl)benzoate; and
N1-(8-(4-((8-benzyl-6-(4-hydroxyphenyl)-3-oxo-3,7-dihydroimidazo[1,2-a]pyrazin-2-yl)methyl)-2-fluorophenoxy)octyl)-N4-(2-(5,5-difluoro-1,3-dimethyl-5H-5λ 4 ,6λ 4 -dipyrrolo[1,2-c:2′,1′-f][1,3,2]diazaborinin-10-yl)ethyl)succinamide,
or a tautomer or a salt thereof.
22 . The method of claim 3 , wherein the compound of formula (I) is selected from the group consisting of:
2-(4-((6-aminohexyl)oxy)-3-fluorobenzyl)-8-benzyl-6-(4-hydroxyphenyl)imidazo[1,2-a]pyrazin-3(7H)-one;
N-(6-(4-((8-benzyl-6-(4-hydroxyphenyl)-3-oxo-3,7-dihydroimidazo[1,2-a]pyrazin-2-yl)methyl)-2-fluorophenoxy)hexyl)-3′,6′-bis(dimethylamino)-3-oxo-3H-spiro[isobenzofuran-1,9′-xanthene]-6-carboxamide;
6-(4-((8-benzyl-6-(4-hydroxyphenyl)-3-oxo-3,7-dihydroimidazo[1,2-a]pyrazin-2-yl)methyl)-2-fluorophenoxy)hexane-1-sulfonic acid;
N-(6-(4-((8-benzyl-6-(4-hydroxyphenyl)-3-oxo-3,7-dihydroimidazo[1,2-a]pyrazin-2-yl)methyl)-2-fluorophenoxy)hexyl)-3′,6′-dihydroxy-3-oxo-3H-spiro[isobenzofuran-1,9′-xanthene]-6-carboxamide;
N-(6-(4-((8-benzyl-6-(4-hydroxyphenyl)-3-oxo-3,7-dihydroimidazo[1,2-a]pyrazin-2-yl)methyl)-2-fluorophenoxy)hexyl)-3-(5,5-difluoro-7-(1H-pyrrol-2-yl)-5H-514,614-dipyrrolo[1,2-c:2′,1′-f][1,3,2]diazaborinin-3-yl)propanamide;
N,N-(6-((6-(4-((8-benzyl-6-(4-hydroxyphenyl)-3-oxo-3,7-dihydroimidazo[1,2-a]pyrazin-2-yl)methyl)-2-fluorophenoxy)hexyl)carbamoyl)-3-oxo-3H-spiro[isobenzofuran-1,9′-xanthene]-3′,6′-diyl)bis(2,2,2-trifluoroacetamide);
N-(8-(4-((8-benzyl-6-(4-hydroxyphenyl)-3-oxo-3,7-dihydroimidazo[1,2-a]pyrazin-2-yl)methyl)-2-fluorophenoxy)octyl)-3′,6′-dihydroxy-3-oxo-3H-spiro[isobenzofuran-1,9′-xanthene]-6-carboxamide;
N-(8-(4-((8-benzyl-6-(4-hydroxyphenyl)-3-oxo-3,7-dihydroimidazo[1,2-a]pyrazin-2-yl)methyl)-2-fluorophenoxy)octyl)-3′,6′-bis(dimethylamino)-3-oxo-3H-spiro[isobenzofuran-1,9′-xanthene]-6-carboxamide;
3-(2-(2-aminoethoxy)ethoxy)-N-(8-(4-((1-benzyl-3-(4-hydroxyphenyl)-6-oxo-2,6-dihydropyrrolo[1,2-a]pyrazin-7-yl)methyl)-2-fluorophenoxy)octyl)propanamide;
N-(2-(2-(3-((8-(4-((1-benzyl-3-(4-hydroxyphenyl)-6-oxo-2,6-dihydropyrrolo[1,2-a]pyrazin-7-yl)methyl)-2-fluorophenoxy)octyl)amino)-3-oxopropoxy)ethoxy)ethyl)-3′,6′-dihydroxy-3-oxo-3H-spiro[isobenzofuran-1,9′-xanthene]-6-carboxamide;
N-(2-(2-(3-((8-(4-((1-benzyl-3-(4-hydroxyphenyl)-6-oxo-2,6-dihydropyrrolo[1,2-a]pyrazin-7-yl)methyl)-2-fluorophenoxy)octyl)amino)-3-oxopropoxy)ethoxy)ethyl)-3′,6′-bis(dimethylamino)-3-oxo-3H-spiro[isobenzofuran-1,9′-xanthene]-6-carboxamide;
6-((8-(4-((8-benzyl-6-(4-hydroxyphenyl)-3-oxo-3,7-dihydroimidazo[1,2-a]pyrazin-2-yl)methyl)-2-fluorophenoxy)octyl)carbamoyl)-3-oxo-3H-spiro[isobenzofuran-1,9′-xanthene]-3′,6′-diyl diacetate;
2-(4-((6-azidohexyl)oxy)-3-fluorobenzyl)-8-benzyl-6-(4-hydroxyphenyl)imidazo[1,2-a]pyrazin-3(7H)-one;
5-((8-(4-((8-benzyl-6-(4-hydroxyphenyl)-3-oxo-3,7-dihydroimidazo[1,2-a]pyrazin-2-yl)methyl)-2-fluorophenoxy)octyl)carbamoyl)-2-(2,7-difluoro-3,6-dihydroxyxanthylium-9-yl)benzoate;
5-((8-(4-((8-benzyl-6-(4-hydroxyphenyl)-3-oxo-3,7-dihydroimidazo[1,2-a]pyrazin-2-yl)methyl)-2-fluorophenoxy)octyl)carbamoyl)-2-(3,11-dihydroxydibenzo[c,h]xanthen-14-ium-7-yl)benzoate; and
N1-(8-(4-((8-benzyl-6-(4-hydroxyphenyl)-3-oxo-3,7-dihydroimidazo[1,2-a]pyrazin-2-yl)methyl)-2-fluorophenoxy)octyl)-N4-(2-(5,5-difluoro-1,3-dimethyl-5H-5λ 4 ,6λ 4 -dipyrrolo[1,2-c:2′,1′-f][1,3,2]diazaborinin-10-yl)ethyl)succinamide,
or a tautomer or a salt thereof.
23 . The method of claim 5 , wherein the compound of formula (I) is selected from the group consisting of:
2-(4-((6-aminohexyl)oxy)-3-fluorobenzyl)-8-benzyl-6-(4-hydroxyphenyl)imidazo[1,2-a]pyrazin-3(7H)-one;
N-(6-(4-((8-benzyl-6-(4-hydroxyphenyl)-3-oxo-3,7-dihydroimidazo[1,2-a]pyrazin-2-yl)methyl)-2-fluorophenoxy)hexyl)-3′,6′-bis(dimethylamino)-3-oxo-3H-spiro[isobenzofuran-1,9′-xanthene]-6-carboxamide;
6-(4-((8-benzyl-6-(4-hydroxyphenyl)-3-oxo-3,7-dihydroimidazo[1,2-a]pyrazin-2-yl)methyl)-2-fluorophenoxy)hexane-1-sulfonic acid;
N-(6-(4-((8-benzyl-6-(4-hydroxyphenyl)-3-oxo-3,7-dihydroimidazo[1,2-a]pyrazin-2-yl)methyl)-2-fluorophenoxy)hexyl)-3′,6′-dihydroxy-3-oxo-3H-spiro[isobenzofuran-1,9′-xanthene]-6-carboxamide;
N-(6-(4-((8-benzyl-6-(4-hydroxyphenyl)-3-oxo-3,7-dihydroimidazo[1,2-a]pyrazin-2-yl)methyl)-2-fluorophenoxy)hexyl)-3-(5,5-difluoro-7-(1H-pyrrol-2-yl)-5H-514,614-dipyrrolo[1,2-c:2′,1′-f][1,3,2]diazaborinin-3-yl)propanamide;
N,N-(6-((6-(4-((8-benzyl-6-(4-hydroxyphenyl)-3-oxo-3,7-dihydroimidazo[1,2-a]pyrazin-2-yl)methyl)-2-fluorophenoxy)hexyl)carbamoyl)-3-oxo-3H-spiro[isobenzofuran-1,9′-xanthene]-3′,6′-diyl)bis(2,2,2-trifluoroacetamide);
N-(8-(4-((8-benzyl-6-(4-hydroxyphenyl)-3-oxo-3,7-dihydroimidazo[1,2-a]pyrazin-2-yl)methyl)-2-fluorophenoxy)octyl)-3′,6′-dihydroxy-3-oxo-3H-spiro[isobenzofuran-1,9′-xanthene]-6-carboxamide;
N-(8-(4-((8-benzyl-6-(4-hydroxyphenyl)-3-oxo-3,7-dihydroimidazo[1,2-a]pyrazin-2-yl)methyl)-2-fluorophenoxy)octyl)-3′,6′-bis(dimethylamino)-3-oxo-3H-spiro[isobenzofuran-1,9′-xanthene]-6-carboxamide;
3-(2-(2-aminoethoxy)ethoxy)-N-(8-(4-((1-benzyl-3-(4-hydroxyphenyl)-6-oxo-2,6-dihydropyrrolo[1,2-a]pyrazin-7-yl)methyl)-2-fluorophenoxy)octyl)propanamide;
N-(2-(2-(3-((8-(4-((1-benzyl-3-(4-hydroxyphenyl)-6-oxo-2,6-dihydropyrrolo[1,2-a]pyrazin-7-yl)methyl)-2-fluorophenoxy)octyl)amino)-3-oxopropoxy)ethoxy)ethyl)-3′,6′-dihydroxy-3-oxo-3H-spiro[isobenzofuran-1,9′-xanthene]-6-carboxamide;
N-(2-(2-(3-((8-(4-((1-benzyl-3-(4-hydroxyphenyl)-6-oxo-2,6-dihydropyrrolo[1,2-a]pyrazin-7-yl)methyl)-2-fluorophenoxy)octyl)amino)-3-oxopropoxy)ethoxy)ethyl)-3′,6′-bis(dimethylamino)-3-oxo-3H-spiro[isobenzofuran-1,9′-xanthene]-6-carboxamide;
6-((8-(4-((8-benzyl-6-(4-hydroxyphenyl)-3-oxo-3,7-dihydroimidazo[1,2-a]pyrazin-2-yl)methyl)-2-fluorophenoxy)octyl)carbamoyl)-3-oxo-3H-spiro[isobenzofuran-1,9′-xanthene]-3′,6′-diyl diacetate;
2-(4-((6-azidohexyl)oxy)-3-fluorobenzyl)-8-benzyl-6-(4-hydroxyphenyl)imidazo[1,2-a]pyrazin-3(7H)-one;
5-((8-(4-((8-benzyl-6-(4-hydroxyphenyl)-3-oxo-3,7-dihydroimidazo[1,2-a]pyrazin-2-yl)methyl)-2-fluorophenoxy)octyl)carbamoyl)-2-(2,7-difluoro-3,6-dihydroxyxanthylium-9-yl)benzoate;
5-((8-(4-((8-benzyl-6-(4-hydroxyphenyl)-3-oxo-3,7-dihydroimidazo[1,2-a]pyrazin-2-yl)methyl)-2-fluorophenoxy)octyl)carbamoyl)-2-(3,11-dihydroxydibenzo[c,h]xanthen-14-ium-7-yl)benzoate; and
N1-(8-(4-((8-benzyl-6-(4-hydroxyphenyl)-3-oxo-3,7-dihydroimidazo[1,2-a]pyrazin-2-yl)methyl)-2-fluorophenoxy)octyl)-N4-(2-(5,5-difluoro-1,3-dimethyl-5H-5λ 4 ,6λ 4 -dipyrrolo[1,2-c:2′,1′-f][1,3,2]diazaborinin-10-yl)ethyl)succinamide,
or a tautomer or a salt thereof.