Multivalent fluorescent probes
Inventors
Singh, Rajendra • Zhang, Ning • Kim, Jae Beom • Diwu, Zhenjun
Assignees
Interested in licensing this patent?
MTEC can help explore whether this patent might be available for licensing for your application.
Publication Number
US-9289515-B2
Publication Date
2016-03-22
Expiration Date
Abstract
Multivalent fluorescent probes and methods of using these multivalent fluorescent probes for in vitro and in vivo imaging are described.
Core Innovation
The invention relates to multivalent fluorescent probes for detecting light from a sample or subject. The probes include a fluorochrome that emits light at 550 to 1200 nm and one or more targeting moieties that specifically target the multivalent fluorescent probe to a specific cell present in the sample or the subject. The multivalent fluorescent probe architecture includes four targeting moieties arranged so that the probe is multivalent, with X and Y selected from O, S, Se or CR15R16, and n selected from 0 to 3.
The targeting moieties are linked to X, Y, N, and N+ via linkers L1, L2, L3 and L4, and the linkers comprise 10 to 50 linear atoms selected from carbon, nitrogen, oxygen, phosphorus and sulfur. The targeting moieties include groups that may be an RGD amino acid residue, zinc (Zn), or a ligand that binds to a cell surface receptor, as specified within the linkers.
A method is disclosed for detecting light by administering the multivalent fluorescent probes to the sample or subject, exposing the sample or subject to light that excites the fluorochrome, and detecting light emitted by the fluorescent probe. The document further describes imaging workflows in which probe localization time can be allowed before excitation and detection, and imaging can be repeated over time to obtain time-dependent labeling signals.
The document describes in vitro and in vivo optical imaging using excitation and fluorescence detection, including applications involving tumor labeling and bacterial infection labeling. Example targets described include glucose transport targeting using 2-deoxy-D-glucose (2-DG), cyclooxygenase targeting using indomethacin-IF750, integrin targeting using RGD constructs, and bacterial infection targeting using bis(zinc(II)-dicolylamine) (DPA) including DPA-tetra.
Claims Coverage
The document includes one independent claim covering a method of detecting light using multivalent fluorescent probes with defined probe structure, multivalent targeting moiety/linker architecture, and fluorescence excitation/detection. The remaining claims are dependent and narrow the independent claim by adding constraints on targeting-moiety composition, timing/localization, imaging modalities, and detection/excitation hardware.
Multivalent fluorescent probe administration with defined probe structure and targeting-moiety/linker architecture
Administering to the sample or subject one or more multivalent fluorescent probes having a specified structure in which X and Y are O, S, Se or CR15R16, n is 0 to 3, and four targeting moieties are linked to X, Y, N and N+ via linkers L1, L2, L3 and L4, wherein the linkers comprise 10 to 50 linear atoms selected from carbon, nitrogen, oxygen, phosphorus and sulfur.
Fluorochrome excitation and fluorescence detection of specifically targeted cells
Exposing the sample or subject to light of a wavelength that excites the fluorochrome of the fluorescent probe and detecting light emitted by the fluorescent probe, wherein the probe emits light having a wavelength of 550 to 1200 nm and the targeting moiety specifically targets the multivalent fluorescent probe to a specific cell present in the sample or the subject.
Localization time between probe administration and excitation/detection
Allowing the fluorescent probe time to localize in the sample or subject after administering the multivalent fluorescent probe and before exposing the sample or subject to excitation light and detecting emitted light.
Targeting moiety composition constraints in the multivalent probe
Confining the multivalent probe targeting-moiety set by requiring that at least two targeting moieties are the same targeting moieties.
Targeting moiety class limitations
Selecting at least one targeting moiety from small molecule, sugar, polypeptide, cytokine, neurotransmitter, or combinations thereof.
Combined imaging modalities including PET, MRI, CT, and X-ray
Selecting additional imaging methods from the group consisting of PET, MRI, CT, and X-ray.
Specified excitation/detection hardware for performing the method
Performing steps (b) and (c) using one of an endoscope, a camera, a catheter, a tomographic system, a hand-held optical imaging system, surgical goggles, or an intraoperative microscope.
The independent claim covers fluorescence-based detection using administered multivalent fluorescent probes with defined probe structure and four targeting moieties linked through specified linkers, followed by excitation and detection of emitted light from specifically targeted cells. Dependent claims further restrict targeting-moiety composition and classes, add localization time, specify combined imaging modalities, and constrain the excitation/detection hardware.
Stated Advantages
Improved specificity/affinity for multivalent fluorescent probes versus single/bivalent probes.
Reduced nonspecific/background signal for multivalent constructs versus controls.
Documented Applications
In vitro and in vivo optical imaging of tumor targets using multivalent fluorescent probes with time-dependent labeling signals.
In vivo labeling of bacterial infection targets, including bacterial labeling with reduced background signal for multivalent constructs versus controls.
Targeted detection of glucose transporter activity using 2-deoxy-D-glucose (2-DG).
Targeted detection of cyclooxygenase COX-1 and COX-2 using indomethacin-IF750.
Targeted detection using RGD integrin targeting, including RGD4 and RGD-HPM2.
Targeted bacterial infection labeling using bis(zinc(II)-dicolylamine) (DPA) and DPA-tetra.
Interested in licensing this patent?