Chlorotoxin agents and uses thereof
Inventors
MCGONIGLE, Sharon • Majumder, Utpal • Postema, Maarten H. D.
Assignees
Interested in licensing this patent?
MTEC can help explore whether this patent might be available for licensing for your application.
Publication Number
US-11826399-B2
Publication Date
2023-11-28
Expiration Date
Abstract
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).
Core Innovation
The disclosure provides chlorotoxin-based agents for detecting and treating cancers that express Neuropilin 1 (NRP1). The chlorotoxin agent comprises a chlorotoxin polypeptide having an amino acid sequence at least 85% identical to SEQ ID NO: 1, is characterized as binding NRP1, and comprises a carboxylated C-terminal arginine residue rather than an amidated form.
The disclosure further describes chlorotoxin agents comprising payload-conjugated chlorotoxin, including covalent conjugates to detectable or imaging moieties and to therapeutic moieties. The payload categories enumerated include anti-cancer agents, nucleic acids, photosensitizers, radiosensitizers, radioisotopes, superantigens, prodrug-activating enzymes, and anti-angiogenic agents, together with multiple detectable labeling options including radionuclides, paramagnetic ions, and fluorescent dyes.
The chlorotoxin polypeptide may be a reduced-lysine chlorotoxin polypeptide with limited conjugation lysines. The disclosure also includes chlorotoxin payload conjugation strategies and CTX-derived peptides and CTX-Cryptophycin conjugates intended to deliver payload to Neuropilin 1-expressing tumors and exert antitumor activity.
Claims Coverage
The independent claim coverage centers on treating a Neuropilin 1-expressing tumor using a chlorotoxin agent defined by sequence identity, a carboxylated C-terminal arginine residue, and Neuropilin 1 binding. Dependent claims further refine payload selection, conjugation constraints, and characterization of Neuropilin 1 binding, including an in vitro binding assay context.
Neuropilin 1 expression determination in tumor sample
Determining expression of Neuropilin 1 in a tumor sample from a subject.
Chlorotoxin agent with high sequence identity to SEQ ID NO: 1
Administering to the subject having the tumor determined to express Neuropilin 1 a chlorotoxin agent wherein the chlorotoxin agent comprises a chlorotoxin polypeptide having an amino acid sequence at least 85% identical to SEQ ID NO: 1.
Carboxylated C-terminal arginine chlorotoxin binding agent
Wherein the chlorotoxin polypeptide comprises a carboxylated C-terminal arginine residue.
Neuropilin 1 binding characterization
Wherein the chlorotoxin polypeptide is characterized as binding Neuropilin 1.
Limited lysine availability for conjugation
The chlorotoxin polypeptide is not more than one lysine available as a site for conjugation.
In vitro Neuropilin 1 binding assay characterization
The chlorotoxin polypeptide is characterized as binding Neuropilin 1 in an in vitro binding assay.
Therapeutic moiety comprising anti-cancer agent
The method includes a therapeutic moiety comprising an anti-cancer agent selected from a listed group or a combination thereof.
Imaging or detectable moiety selection
The method includes an imaging moiety or detectable moiety selected from a group including fluorescent labels, radioactive or paramagnetic isotopes or ions, ligands, chemiluminescent or bioluminescent agents, photosensitizers, quantum dots, microparticles, metal nanoparticles, nanoclusters, enzymes, colorimetric labels, haptens, molecular beacons, aptamer beacons, biotin, and dioxigenin.
Optional administration of chemotherapeutic agent
The method further comprises administering a chemotherapeutic agent.
Overall, the claim coverage centers on treating a Neuropilin 1-expressing tumor by determining Neuropilin 1 expression and administering a chlorotoxin polypeptide at least 85% identical to SEQ ID NO: 1 that contains a carboxylated C-terminal arginine residue and is characterized as binding Neuropilin 1, optionally further refined by limited lysine availability for conjugation, in vitro binding assay characterization, selectable therapeutic anti-cancer moieties or imaging or detectable moieties, and optional chemotherapeutic agent administration.
Stated Advantages
Greater efficacy is described for carboxylated C-terminal arginine CTX versions than amidated C-terminal arginine counterparts.
Carboxylated CTX(R—COOH) binds Neuropilin 1 and competes with VEGF165 for Neuropilin 1.
Carboxylated CTX-cryptophycin is described as more effective at lower dose in a Neuropilin 1-dependent manner.
Anti-Neuropilin 1 antibody blunts CTX-cryptophycin efficacy, supporting Neuropilin 1 dependency of the therapeutic effect.
Documented Applications
Treating a tumor that expresses Neuropilin 1 by determining Neuropilin 1 expression in a tumor sample and administering a chlorotoxin agent.
Delivering CTX(-derived) peptides and CTX-Cryptophycin conjugates to Neuropilin 1-expressing tumors for antitumor activity, including support from Neuropilin 1 binding and blockade experiments.
Imaging and/or detection using selectable imaging moieties or detectable moieties.
Therapeutic testing of CTX-cryptophycin conjugates in Neuropilin 1 WT versus KO PC-3 xenografts, including comparisons of amidated versus carboxylated chlorotoxin C-termini.
Assessment of CTX bioactivity in crayfish as described for CTX and CTX-derived peptides or fragments with amidated versus carboxylated status in tumor lysates after CTX administration.
Biodistribution or PK assessment describing similar parent conjugate exposure but reduced active cryptophycin metabolite (Metabolite 1) in Neuropilin 1 KO tumors.
Interested in licensing this patent?