Embodiments of the present disclosure provide for conjugated polymer nanoparticle, method of making conjugated polymer nanoparticles, method of using conjugated polymer nanoparticle, polymers, and the like.

This invention is in the field of fluorescence imaging and relates to a new near infrared (NIR) reactive oxygen species (ROS) sensor designed with controlled fluorescence on-off switching mechanism.

The present invention provides, among other things, compositions and methods relating to detection and/or treatment cancer (e.g., one or more tumors) that expresses Neuropilin 1 (NRP1). The present invention provides methods of treating cancer that include administering a chlorotoxin agent to a subject (e.g., to a subject suffering from or susceptible to the cancer which may, in some embodiments, be a cancer that expresses NRP1). In some embodiments, a chlorotoxin agent for use in accordance with the present invention can be or comprise a chlorotoxin polypeptide and a payload moiety (e.g., as a covalent conjugate).

There is disclosed a composition in the form of a nanoparticle. The nanoparticle composition has a diameter from 5 to 500 nanometers. The nanoparticle composition has i) a central core portion including magnetic Fe.sub.3O.sub.4 nanoparticles adapted to act as a heat source when subjected to a magnetic field and a chemotherapeutic agent configured to treat cancer tissues, ii)a shell portion including a shell member encapsulating said core portion, and iii)antibodies configured to target cancer stem cells and adhered to surface of said shell member. The chemotherapeutic agent is a heat shock protein inhibitor and is releasable on activation of the heat source due to the magnetic field, and the shell member is made of silica or a silica based material. Surface of the nanoparticle is modified with the antibodies capable of binding with a cluster of differentiation molecules on the cell surface of the target cancer stem cells, whereby by way of combination of specificity of the nanoparticle composition due to the antibody, thermo-therapeutic effect of the Fe.sub.3O.sub.4 nanoparticles, and release of the heat shock protein inhibitor on site at the target cancer stem cells, inhibition of the target cancer stem cells is synergistically and additionally enhanced is increased.

A fluorescent compound or a salt thereof represented by General Formulae (I) or (II) shown below:

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In the General Formulae (1) and (II) shown above, R.sup.1 and R.sup.11 represent an alkyl group or an -aminoalkyl group, R.sup.2 and R.sup.12 represent a hydrogen atom or a alkyl group, R.sup.3 and R.sup.13 represent an atomic group represented by a formula (CH.sub.2).sub.m (m is a natural number of 10 or less), R.sup.4 and R.sup.14 represent an atomic group represented by a formula CH.sub.2 or NR.sup.6 (NR.sup.16) (R.sup.6 and R.sup.16 represent an alkyl group), R.sup.5 and R.sup.15 represent an atomic group represented by a formula (CH.sub.2).sub.n (n is a natural number of or less), R represents an atomic group represented by any one of formulae NH.sub.2, NHR.sup.7, NR.sup.7R.sup.8 and NR.sup.7R.sup.8R.sup.9 (NHR.sup.17, NR.sup.17R.sup.18 and N.sup.+R.sup.17R.sup.18R.sup.19) (R.sup.7, R.sup.8, R.sup.9, R.sup.17, R.sup.18 and R.sup.19 independently represents an alkyl group, respectively) and when R.sup.2 and R.sup.12 are alkyl groups and R.sup.4 (R.sup.14) is an atomic group represented by the formula NR.sup.6(NR.sup.16), R.sup.2 and R.sup.6 and R.sup.12 and R.sup.16 may bind with each other to form a ring.

A conjugate containing a fluorescent chromophore, which has any structure selected from C1 to C3. The conjugate containing the fluorescent chromophore provided by the described embodiments includes one fluorescent chromophore and two highly reactive groups R1 and R2 linked to the fluorescent chromophore by a covalent bond. The fluorescent chromophore in the conjugate initially has no or only weak fluorescence emission capability, and only after the two highly reactive groups react together with the corresponding molecule, the fluorescent chromophore has strong fluorescence emission. Therefore, the efficiency of conjugation of drug molecules to targeting molecules can be monitored in situ by the infrared fluorescence emission intensity and applied to the target-mediated drug delivery.

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Methods of optically marking and sorting adherent cells are provided. The methods include providing a plurality of adherent cells attached to a substrate, each adherent cell of the plurality of adherent cells having an optical marker. The methods also include selectively applying light energy to a subset of the plurality of adherent cells, and detaching the plurality of adherent cells from the substrate. These methods also provide the sorting of the plurality of adherent cells.

Embodiments related to methods of use of an image analysis system for identifying residual cancer cells after surgery are disclosed. In some embodiments, the image analysis system collects a surgical site image and indicates on a display one or more locations of the identified cancer cells. In some embodiments, the method for identifying residual cancer cells comprises determining and selecting a portion of the surgical site image responsive to an intensity parameter; modifying the selected portion of the surgical site image to determine one or more groups of residual cancer cells based on size; and identifying at least one of the one or more groups of residual cancer cells from the modified portion of the surgical site image using a local-based threshold.

The present invention provides a fluorescent silica-based nanoparticle that allows for precise detection, characterization, monitoring and treatment of a disease such as cancer. The nanoparticle has a range of diameters including between about 0.1 nm and about 100 nm, between about 0.5 nm and about 50 nm, between about 1 nm and about 25 nm, between about 1 nm and about 15 nm, or between about 1 nm and about 8 nm. The nanoparticle has a fluorescent compound positioned within the nanoparticle, and has greater brightness and fluorescent quantum yield than the free fluorescent compound. The nanoparticle also exhibits high biostability and biocompatibility. To facilitate efficient urinary excretion of the nanoparticle, it may be coated with an organic polymer, such as poly(ethylene glycol) (PEG). The small size of the nanoparticle, the silica base and the organic polymer coating minimizes the toxicity of the nanoparticle when administered in vivo. In order to target a specific cell type, the nanoparticle may further be conjugated to a ligand, which is capable of binding to a cellular component associated with the specific cell type, such as a tumor marker. In one embodiment, a therapeutic agent may be attached to the nanoparticle. To permit the nanoparticle to be detectable by not only optical fluorescence imaging, but also other imaging techniques, such as positron emission tomography (PET), single photon emission computed tomography (SPECT), computerized tomography (CT), bioluminescence imaging, and magnetic resonance imaging (MRI), radionuclides/radiometals or paramagnetic ions may be conjugated to the nanoparticle.

Carbamate and beta-amino acid urea-based scaffolds that have high binding affinity to PSMA are disclosed. These scaffolds can be radiolabeled and used for imaging cells and tumors that express PSMA or for cancer radiotherapy. These compounds also can comprise a fluorescent dye and be used for imaging cells and tumors that express PSMA or for photodynamic therapy.