Cell-permeable probes for identification and imaging of sialidases

Patent metadata

Field	Value
Publication number	US-2015225766-A1
Application number	US-201314422310-A
Country	US
Kind code	A1
Filing date	Aug 16, 2013
Priority date	Aug 18, 2012
Publication date	Aug 13, 2015
Grant date	—

Abstract

Provided herein are novel irreversible sialidase inhibitors. These compounds can be conjugated with a detectable tagging moiety such as azide-annexed biotin via CuAAC for isolation and identification of sialidases. The provided compounds and the corresponding detectable conjugates are useful for detecting sialidase-containing pathogens and imaging in situ sialidase activities under physiological conditions.

First claim

1 - 79 . (canceled)

80 . A compound of formula (I):

or a salt thereof,
wherein
F atom at the C3-position is axial or equatorial;
R 1 is H or optionally substituted C 1-6 alkyl;
R 2 is OR 2O , N 3 , N(R 2N ) 2 , or guanidine;
each instance of R 2O is independently hydrogen, optionally substituted C 1-6 alkyl, optionally substituted acyl, or an oxygen protecting group;
each instance of R 2N is independently hydrogen, optionally substituted C 1-6 alkyl, optionally substituted acyl, or a nitrogen protecting group;
each instance of R 3a and R 3b is independently hydrogen, —C(═O)—R 3r , or an oxygen protecting group;
each instance of R 3r is optionally substituted C 1-6 alkyl, optionally substituted aryl, optionally substituted heteroaryl, or optionally substituted heterocycle, optionally substituted alkylaryl, optionally substituted alkylheteroaryl, or optionally substituted alkylheterocycle;
X is selected from the group consisting of —O—, —O(C═O)—, —NH—, —NH(C═O)—, —(C═O)NH—, —O(C═O)NH—, —O(C═S)NH—, —NH(C═O)NH—, and —NH(C═S)NH—;
R 4 is H, optionally substituted C 1-6 alkyl, or -L-Z;
Y is optionally substituted C 1-6 alkyl or -L-Z;
each instance of L is independently selected from the group consisting of —(CH 2 ) n —, —(CH 2 ) n C═O—, —(CH 2 ) n NH—, —(C═O)(CH 2 ) n —, —(CH 2 ) n NH(C═O)—, —(C═O)(CH 2 ) n NH(C═O)—, —(CH 2 ) n SCH 2 (C═O)—, and —(CH 2 CH 2 O) n —;
each instance of n is an integer from 1 to 8, inclusive;
each instance of Z is a functional group for further ligation; and
provided that the compound is not of the formula

81 . The compound of claim 80 , wherein
Z is optionally substituted alkyne, optionally substituted alkene, halogen, —N 3 , N(R N ) 2 , OR O , SR S , or CO 2 R O ; each instance of R N is independently hydrogen, optionally substituted C 1-6 alkyl, optionally substituted acyl, or a nitrogen protecting group; each instance of R O is independently hydrogen, optionally substituted C 1-6 alkyl, optionally substituted acyl, or an oxygen protecting group; and each instance of R S is independently hydrogen, optionally substituted C 1-6 alkyl, or a sulfur protecting group.

82 . The compound of claim 80 of formula (II-a):

or a salt thereof,
wherein R 3c is independently hydrogen, optionally substituted C 1-6 alkyl, optionally substituted acyl, or an oxygen protecting group.

83 . The compound of claim 82 of formula (II-b):

or a salt thereof.

84 . The compound of claim 83 , having a formula (II-b1):

or a salt thereof.

85 . The compound of claim 84 , having a formula selected from (II-b2) and (II-b3):

wherein R y1 is hydrogen, halogen, or optionally substituted C 1-6 alkyl, and

or a salt thereof.

86 . The compound of claim 80 of formula (II-c):

or a salt thereof.

87 . The compound of claim 86 , having a formula selected from (II-c1), (II-c2) and (II-c3):

wherein R y2 is hydrogen or optionally substituted C 1-6 alkyl, and

or a salt thereof.

88 . The compound of claim 86 , wherein Y is optionally substituted C 1-6 alkyl.

89 . The compound of claim 88 , wherein Y is methyl or CF 3 .

90 . The compound of claim 80 , wherein R 1 is H or methyl.

91 . The compound of claim 80 , wherein the F atom at the C3-position is axial.

92 . The compound of claim 80 , wherein the F atom at the C3-position is equatorial.

93 . The compound of claim 80 , wherein R 3a is —C(═O)—R 3r , wherein R 3r is optionally substituted alkyl or aryl.

94 . The compound of claim 80 , wherein R 3a is optionally substituted benzoyl.

95 . The compound of claim 80 , wherein R 3a is CH 3 CO—, C 2 H 5 CO—, C 3 H 7 CO—, t-BuCO—, CF 3 CO—, PhCH 2 CO—, or C 6 H 5 CO—.

96 . The compound of claim 80 , wherein the compound is one of the following formulae:

97 . A sialidase protein adduct comprising a compound of claim 80 , wherein the sialidase protein is covalently conjugated to the compound.

98 . The sialidase protein adduct of claim 97 , having a formula:

99 . The sialidase protein adduct of claim 98 , wherein the sialidase is a human, viral or bacterial sialidase.

100 . The sialidase protein adduct of claim 98 , wherein the sialidase is a bacterial sialidase selected from the group consisting of nanA, nanB, nanC, nanJ, nanI, and nanH.

101 . The sialidase protein adduct of claim 98 , wherein the compound is DFSA-5-yne or DFSA-7-yne.

102 . The sialidase protein adduct of claim 98 , wherein the compound is covalently linked to one or more tyrosine residues within any peptide of SEQ ID NOS: 1-6, wherein the peptide is a fragment of nanA, nanB, nanC, nanJ, nanI or nanH.

103 . The sialidase protein adduct of claim 98 , wherein
the sialidase is an influenza virus neuraminidase (NA).

104 . The sialidase protein adduct of claim 98 , wherein the sialidase is a human sialidase selected from the group consisting of Neu1, Neu2, Neu3 and Neu4.

105 . A detectable conjugate comprising a compound of claim 80 , wherein the compound is covalently conjugated to a detectable tagging moiety.

106 . A detectable sialidase protein conjugate comprising a sialidase protein adduct of claim 98 , wherein the sialidase protein adduct is covalently conjugated to a detectable tagging moiety.

107 . The detectable conjugate of claim 105 or detectable sialidase conjugate of claim 106 , wherein the tagging moiety comprises a label.

108 . The detectable conjugate or detectable sialidase conjugate of claim 107 , having a formula selected from:

or a salt thereof,
wherein
F atom at the C3-position is axial or equatorial;
R K is H, optionally substituted C 1-6 alkyl, or a protein;
R 2 is OR 2O , N 3 , N(R 2N ) 2 , or guanidine;
each instance of R 2O is independently hydrogen, optionally substituted

Assignees

• Academia Sinica

Inventors

• Wong Chi-Huey
• Fang Jim-Min
• Cheng Yih-Shyun E
• Tsai Charng-Sheng

Classifications

• C12N9/2402

  hydrolysing O- and S- glycosyl compounds (3.2.1) · CPC title

• C12Q1/40 Primary

  involving amylase · CPC title

• G01N2333/11

  Orthomyxoviridae, e.g. influenza virus · CPC title

• C12Y302/01018

  Exo-alpha-sialidase (3.2.1.18), i.e. trans-sialidase · CPC title

• C12Q1/04

  Determining presence or kind of microorganism; Use of selective media for testing antibiotics or bacteriocides; Compositions containing a chemical indicator therefor {(C12Q1/6897 takes precedence)} · CPC title