Diarylamine-based fluorogenic probes for detection of peroxynitrite

Patent metadata

Field	Value
Publication number	US-9651528-B2
Application number	US-201313754499-A
Country	US
Kind code	B2
Filing date	Jan 30, 2013
Priority date	Jan 30, 2012
Publication date	May 16, 2017
Grant date	May 16, 2017

Abstract

Provided herein are improved fluorogenic compounds and probes that can be used as reagents for measuring, detecting and/or screening peroxynitrite. The fluorogenic compounds of the invention can produce fluorescence colors, such as green, yellow, red, or far-red. Also provided herein are fluorogenic compounds for selectively staining peroxynitrite in the mitochondria of living cells. Provided also herein are methods that can be used to measure, directly or indirectly, the presence and/or amount of peroxynitrite in chemical samples and biological samples such as cells and tissues in living organisms. Also provided are high-throughput screening methods for detecting or screening peroxynitrite or compounds that can increase or decrease the level of peroxynitrite in chemical and biological samples.

First claim

We claim:

1. A compound of formula (I) or (II):

or a tautomer thereof;
wherein N is a nitrogen atom, and is linked to Q and R 1 through single covalent bonds;
R 1 is H, alkyl, halogenated alkyl, alkenyl, alkynyl, alkoxyalkyl, heteroalkyl, cycloalkyl, cycloalkenyl, cycloalkynyl, heterocyclyl, aminoalkyl, arylalkyl, alkyloxy, carboxyalkyl, alkylamino, alkoxyamino, alkylamido, alkoxyamido, or acyl;
each of R 2 , R 3 , R 4 , R 5 , R 6 and R 7 is independently H, F, Br, I, CN, alkyl, halogenated alkyl, heteroalkyl, alkenyl, alkynyl, aralkyl, aryl, alkaryl, heterocyclyl, cycloalkyl, cycloalkenyl, cycloalkynyl, hydroxyalkyl, aminoalkyl, amino, alkylamino, arylamino, dialkylamino, alkylarylamino, diarylamino, acylamino, hydroxy, thiol, thioalkyl, alkoxy, alkylthio, alkoxyalkyl, aryloxy, arylalkoxy, acyloxy, nitro, carbamoyl, trifluoromethyl, phenoxy, benzyloxy, phosphonic acid, phosphate ester, sulfonic acid (—SO 3 H), sulfonate ester, sulfonamide, —C(═O)—P 1 or —C(═O)-M-P 2 ;
each of P 1 and P 2 is independently hydrogen, halo, alkoxy, hydroxy, thiol, alkyl, alkenyl, alkynyl, cycloalkyl, cycloalkenyl, cycloalkynyl, aryl, alkaryl, arylalkyl, carbamate, amino, alkylamino, arylamino, dialkylamino, alkylarylamino, diarylamino, alkylthio, heteroalkyl, alkyltriphenylphosphonium, or heterocyclyl having from 3 to 7 ring atoms; M is alkylene, alkenylene, alkynylene, arylene, aralkylene or alkarylene;
A is NR 11 R 12 and B is N + R 11 R 12 with a biologically compatible counterion selected from chloride, bromide, iodide, sulfate, alkanesulfonate, arylsulfonate, phosphate, perchlorate, tetrafluoroborate, tetraphenylboride, nitrate and anions of aromatic or aliphatic carboxylic acids;
wherein: R 11 in combination with R 4 or R 12 in combination with R 3 , but not both, forms a 5- or 6-membered ring that is saturated or unsaturated, or further fused with an aryl or heteroaryl ring, and is optionally substituted by one or more alkyls, carboxylic acids, sulfonic acids (—SO 3 H), or their salts, ester or amide derivatives the other of R 11 or R 12 is H, alkyl, halogenated alkyl, alkenyl, alkynyl, alkoxyalkyl, heteroalkyl, cycloalkyl, cycloalkenyl, cycloalkynyl, heterocyclyl, aminoalkyl, arylalkyl, alkyloxy, acyl, carboxyalkyl, sulfoalkyl, a salt of carboxyalkyl, a salt of sulfoalkyl, or an ester or amide of carboxyalkyl or sulfoalkyl; or R 11 in combination with R 12 forms a piperazine, which is optionally substituted by alkyl, carboxylic acid, a salt of carboxylic acid, or a carboxylic acid ester of an alcohol;
Z is O;
R 8 has the formula

wherein each of R 17 , R 18 , R 19 , R 20 and R 21 is independently H, F, Cl, Br, I, CN, nitro, a carboxylic acid, a salt of carboxylic acid, sulfonate ester (—SO 3 R 15 ), hydroxy, azide, alkyl, alkenyl, alkynyl, heteroalkyl, cycloalkyl, cycloalkenyl, cycloalkynyl, aryl, alkylaryl, arylalkyl, heterocyclyl, alkoxy, alkoxyalkyl, alkoxyalkoxy, acyl, alkylcarbonylalkyl, halogenated alkylcarbonylalkyl, trifluoromethylcarbonylalkyl, aminoalkyl, carboxyalkyl, thiol, alkylthio, amino, alkylamino, dialkylamino, alkoxycarbonyl, alkoxycarbonylalkyl, aminocarbonyl, alkylaminocarbonyl, dialkylaminocarbonyl, or arylcarboxamido, the alkyl or aryl of which is optionally substituted by one or more F, Cl, Br, I, a carboxylic acid, a salt of carboxylic acid, a carboxylic acid ester of an alcohol, amino, alkylamino, dialkylamino, alkoxy, alkyltriphenylphosphonium, sulfonic acid (—SO 3 H), sulfonate ester (—SO 3 R 15 ), or sulfonamide (—SO 2 NR 15 R 16 ) or R 17 and R 18 together, R 18 and R 19 together, R 19 and R 20 together, or R 20 and R 21 together form a 5- or 6-membered cycloalkyl, heterocyclyl, aryl or heteroaryl ring fused with the phenyl ring of formula (III) that is optionally further substituted by one or more F, Cl, Br, I, a carboxylic acid, a salt of carboxylic acid, a carboxylic acid ester of an alcohol, amino, alkylamino, dialkylamino, alkoxy, thiol, alkylthio, alkyltriphenylphosphonium, sulfonic acid (—SO 3 H), sulfonate ester (—SO 3 R 15 ), or sulfonamide (—SO 2 NR 15 R 16 ), and wherein at least one of R 17 , R 18 , R 19 , R 20 and R 21 is not H or alkyl, wherein each of R 15 and R 16 represents a saturated or unsaturated, cyclic or acyclic alkyl that is optionally substituted by one or more F, Cl, Br, I, a carboxylic acid, a salt of carboxylic acid, a carboxylic acid ester of an alcohol, amino, alkylamino, dialkylamino, alkoxy, or alkyltriphenylphosphonium;
R 9 is H, hydroxy, CN or alkoxy; or R 9 in combination with R 17 or R 21 forms a 5- or 6-membered spirolactone, spirosultone, spirolactam or spirosultam ring that is optionally and independently substituted by H, F or CH 3 ; and
Q is a substituted phenyl represented by formula (IV):

wherein each of R 22 , R 23 , R 24 , R 25 , and R 26 is independently H, hydroxy, alkyl, alkenyl, alkynyl, heteroalkyl, cycloalkyl, cycloalkenyl, cycloalkynyl, aryl, alkylaryl, arylalkyl, heterocyclyl, alkoxy, alkoxyalkyl, alkoxyalkoxy, acyl, alkylcarbonylalkyl, halogenated alkylcarbonylalkyl, trifluoromethylcarbonylalkyl, aminoalkyl, carboxyalkyl, thiol, alkylthio, amino, alkylamino, dialkylamino, alkoxycarbonyl, alkoxycarbonylalkyl, aminocarbonyl, alkylaminocarbonyl, dialkylaminocarbonyl, or arylcarboxamido, wherein at least one of R 22 , R 23 , R 24 , R 25 , and R 26 is not H, wherein any of the alkyl or aryl of which is optionally substituted by one or more F, Cl, Br, I, a carboxylic acid, a salt of carboxylic acid, a carboxylic acid ester of an alcohol, amino, alkylamino, dialkylamino, alkoxy, alkyltriphenylphosphonium, sulfonic acid (—SO 3 H), sulfonate ester (—SO 3 R 15 ), or sulfonamide (—SO 2 NR 15 R 16 ); or wherein R 22 and R 23 together, R 23 and R 24 together, R 24 and R 25 together, or R 25 and R 26 together form a 5- or 6-membered cycloalkyl, heterocyclyl, aryl or heteroaryl ring fused with the phenyl ring of formula (IV) that is optionally further substituted by one or more F, Cl, Br, I, a carboxylic acid, a salt of carboxylic acid, a carboxylic acid ester of an alcohol, amino, alkylamino, dialkylamino, alkoxy, thiol, alkylthio, alkyltriphenylphosphonium, sulfonic acid (—SO 3 H), sulfonate ester (—SO 3 R 15 ), or sulfonamide (—SO 2 NR 15 R 16 ) and the remainder of R 22 , R 23 , R 24 , R 25 , and R 26 are as previously defined; or wherein one of R 22 , R 24 , or R 26 is OR 27 , CH 2 CH 2 COR 28 , or NR 29 R 30 ; wherein R 27 is hydrogen or a group selected from alkyl, alkoxyalkyl, alkanoyl, and polyether; R 28 is an electron-withdrawing group selected from halogen-substituted lower alkyl, or (C═O)—O—W 1 , wherein W 1 is a group selected from alkyl, alkenyl, alkynyl, cycloalkyl, cycloalkenyl, cycloalkynyl, aryl, alkaryl or arylalkyl, R 29 and R 30 are independently hydrogen, alkyl, alkenyl, alkynyl, alkoxyalkyl, alkanoyl, alkenoyl, alkynoyl, cycloalkyl, cycloalkenyl, cycloalkynyl, aryl, alkaryl, arylalkyl, aryloyl, or polyether and the remainder of R 22 , R 23 , R 24 , R 25 , and R 26 are as previously defined.

2. The compound of claim 1 , wherein the compound has one of formulae 11-19:

3. A compound of formula (I) or (II):

or a tautomer thereof;
w

Assignees

• Univ Hong Kong

Inventors

• Yang Dan
• Peng Tao
• Shen Jiangang
• Chen Xingmiao

Classifications

• G01N33/84

  involving inorganic compounds or pH · CPC title

• C09B11/24

  Phthaleins containing amino groups {; Phthalanes; Fluoranes; Phthalides; Rhodamine dyes; Phthaleins having heterocyclic aryl rings; Lactone or lactame forms of triarylmethane dyes} · CPC title

• C07D493/10

  Spiro-condensed systems · CPC title

• C07D491/052

  the oxygen-containing ring being six-membered · CPC title

• C09B69/008

  Dyes containing a substituent, which contains a silicium atom · CPC title