Compounds as stimuli-responsive probes, methods and applications thereof
Inventors
Thimmaiah, Govindaraju • Narayanaswamy, Nagarjun
Assignees
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Publication Number
US-10544167-B2
Publication Date
2020-01-28
Expiration Date
Abstract
The present disclosure relates to the field of synthetic pharmaceutical chemistry and biology. The disclosure provides a compound of Formula I and a process of preparation thereof. The disclosure further relates to methods and use of Formula I compounds as stimuli-responsive probes. Said Formula I compounds are employed for detecting and quantifying reactive oxygen species (ROS), and have related applications including but not limited fluorescence spectroscopy, diagnostics, treatment, imaging and biomedical applications.
Core Innovation
The invention relates to a compound of Formula I that is a stimuli-responsive fluorescent/cleavable ROS probe for detecting and/or quantifying reactive oxygen species in biological samples and in vivo subjects. The compound includes defined structural substituent selections (X, R1-R10) that include boronic ester or phenyl boronic acid functionality as the ROS-cleavable moiety. In particular embodiments, aryl boronate masked DNA-binding quinone-cyanine chemistry is described, including QCy-BA and the cleaved fluorescent species QCy-DT.
The invention addresses the problem of detecting and quantifying reactive oxygen species, including hydrogen peroxide (H2O2), with fluorescence turn-on and NIR signal selectivity through ROS-driven cleavage. The ROS cleavage triggers release of QCy-DT, which binds AT-rich DNA at the Drew-AT DNA minor groove, thereby providing a measurable fluorescent response correlated to the presence and level of ROS.
The invention further describes use for in vivo ROS imaging and quantification, including diagnosing disease conditions associated with abnormal ROS. The described detection modalities include near infrared fluorescence and fluorescence microscopy, spectroscopy, confocal laser scanning microscopy, and total internal reflection fluorescence microscopy.
Claims Coverage
The independent claims include a compound defined by Formula I and methods directed to detecting and/or quantifying ROS, diagnosing ROS-associated diseases, treating such diseases, and providing a kit. Across the independent claim set, the coverage centers on a stimuli-responsive fluorescent/cleavable ROS probe that cleaves in the presence of ROS to release a DNA-binding fluorescent species (QCy-DT) and produces a fluorometric NIR fluorescence response.
Compound of formula I
A compound of Formula I wherein X is selected from oxygen, sulphur, and selenium; R1 is selected from specified groups including methyl phenyl boronic acid, boronic ester, and related groups; R2 is selected from H, OH, halogen, alkyl and substituted alkyl; R3 or R4 is selected from H, alkyl, aryl, nitrile, acid, and halogen; R5-R8 are selected from H, OH, alkyl, aryl, halogen, nitro, sulfonates (SO3−), and nitrile; R9 is selected from H and —(CH2)n— where n is 1-6; and R10 is selected from hydrogen, —OH, methyl, amine, terminal alkyne, alkene, alkyl acid, amine acid, and sulfonates (SO3−).
Ros detection using a combination probe with dna-binding and fluorescence turn-on
A method for detecting and/or quantifying reactive oxygen species using a combination probe comprising the compound of Formula I and DNA sequences, wherein ROS cleaves the compound to release QCy-DT, and QCy-DT binds AT-rich DNA (Drew-AT DNA minor groove) to provide fluorometric ROS detection and quantification, including fluorescence microscopy and fluorescence spectroscopy modalities.
Disease diagnosis by detecting ros in a sample
A method for diagnosing a disease in a mammal or plant by detecting and optionally quantifying reactive oxygen species (ROS) in a sample, with the diagnosis based on the detected ROS.
Therapeutic treatment of a ros-abnormality disease
A method of treating a disease characterized by abnormal levels of ROS in a subject by administering the compound of Formula I.
Kit for detecting ros
A kit for detecting reactive oxygen species (ROS) in a sample, wherein the compound of Formula I is present in an amount effective to detect the presence of ROS.
Overall, the claims cover a Formula I compound and related methods that use ROS-driven cleavage to generate a DNA-binding fluorescent response for detecting and/or quantifying ROS, including applications to diagnosing and treating ROS-associated diseases and providing a kit formulation.
Stated Advantages
Provides H2O2/ROS selectivity.
Enables a switch-on near-infrared fluorescence response with AT-rich DNA (Drew-AT).
Supports ratiometric fluorescence behavior (500/650 nm) for ROS sensing.
Enables fluorometric detection and quantification of ROS using DNA-binding fluorescence (QCy-DT with AT-rich DNA).
Enables in vivo ROS imaging and quantification.
Documented Applications
Detection and/or quantification of reactive oxygen species (ROS) using cells, tissue, and biological fluids, including hydrogen peroxide and superoxide.
In vivo ROS imaging and quantification in a subject.
Diagnosis of diseases in a mammal or plant sample by detecting and optionally quantifying ROS, including cancer, cardiovascular dysfunction, neurodegenerative disease, gastroduodenal pathogenesis, inflammatory disorder, metabolic dysfunction of organ, premature aging, and any disorder related to ROS.
Therapy of a disease characterized by abnormal levels of ROS in a subject by administering the Formula I compound.
Use of a kit to detect reactive oxygen species (ROS) in a sample.
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